THE CHINESE UNIVERSITY OF HONG KONG

Li Dak Sum Yip Yio ChinR & D Centre

for Chinese Medicine

Shiu-Ying Hu HerbariumSchool of Life Sciences

Organized by: Supporting Organization:

Chung Chi CollegeCUHK

Institute for Control of Chinese Traditional Medicine and Ethnic Medicine, National Institutes for Food and Drug

Control (NIFDC), China

Department of Health, Hong Kong Special

Administrative Region

International Conference on DNA technology for authentication, quality control

and conservation of herbal material

LI DAK SUM YIP YIO CHIN R & D CENTRE FOR CHINESE MEDICINE

12-14 December, 2016

香港中文大學 李達三葉耀珍中醫藥研究發展中心LI DAK SUM YIP YIO CHIN R & D CENTRE FOR CHINESE MEDICINE

THE CHINESE UNIVERSITY OF HONG KONG

FOREWORD

While organoleptic and chemical techniques are conventionally used, DNA technologyprovides an independent and alternative approach for authentication and quality control ofherbal material. Nowadays, molecular techniques have been recognized by regulatory bodiesaround the world for the identification of selected herbal material. Besides authenticationpurpose, DNA technology is also useful in investigating the subtle genetic differences ofherbs grown in different localities, which often results in the differences of pharmacologicalproperties. DNA technology may also be used to select, breed and conserve the variety thathas the best pharmacological properties and to investigate the genetic diversity of apopulation.

In the view of the rapid development in genome sequencing and analyses, and the increaseapplication of DNA technologies in herbal materials in many countries, our newly formed R &

D Centre takes the initiative to organize this international conference. This conference is inline with our missions in promoting the quality control and assurance of Chinese medicinalmaterial and products, and in developing personnel and expertise in herbal industry onquality control and assurance. In this conference, key researchers will present on the mostupdated molecular technologies and their applications for identification, analysis and

conservation of herbal material. We hope this conference will be the beginning of similarconference series and participants will have a fruitful time in this conference, as well as inHong Kong.

Pang Chui Shaw

ChairmanOrganizing Committee

香港新界沙田香港中文大學 TEL 電 話: (852) 2609 6122 WEBSITE網址: http://rdccm.cuhk.edu.hk/CUHK, Shatin, NT, HK FAX 圖文傅真: (852) 2603 5646 E-MAIL 電郵: pcshaw@cuhk.edu.hk

Organizing Committee

Chairperson:Prof. Pang Chui Shaw

Li Dak Sum Yip Yio Chin R & D Centre for Chinese Medicine, School of Life Sciences, Institute of Chinese Medicine,

The Chinese University of Hong Kong

Co-chair:Prof. Karl Wah Keung Tsim

Centre for Chinese Medicine R & D and Division of Life Science, Hong Kong University of Science and Technology

Members:Dr. David Tai Wai Lau

Shiu-Ying Hu Herbarium, School of Life Sciences, The Chinese University of Hong Kong

Dr. Stanley Chun Kai CheungLi Dak Sum Yip Yio Chin R & D Centre for Chinese Medicine,

The Chinese University of Hong Kong

Prof. Clara Bik San Lau Institute of Chinese Medicine,

The Chinese University of Hong Kong

Mr. Kenneth Chi Fai Leung School of Life Sciences,

The Chinese University of Hong Kong

Venue: Yasumoto International Academic Park - YIA LT2, CUHK8:15- 9:00 Registration& set up poster

9:00 - 9:45

Opening Ceremony

Dr. Edwin Lok Kin TsuiAssistant Director (Traditional Chinese Medicine), Department of Health, HKSAR

Prof. Fanny Mui Ching CheungPro-Vice-Chancellor, The Chinese University of Hong Kong

Prof. Pang Chui Shaw Director, Li Dak Sum Yip Yio Chin R&D Centre for Chinese Medicine, The Chinese University of Hong Kong

Signing Ceremony

Memorandum of CollaborationRoyal Botanic Gardens, Kew, UK

Group Photo

Chairperson: Prof. Pang Chui Shaw

9:45 - 10:20Prof. Kelvin Kam Chuen ChanDepartment of Health, HKSAR, ChinaTitle: Future Development in the Testing Centre for Chinese Medicines

10:20 – 11:00 Tea Break & set up of poster

Chairperson: Prof. Clara Bik San Lau

11:00 – 11:45

Prof. Monique S.J. Simmonds Royal Botanic Gardens, Kew, UKTitle: Authentication of Plants Used in Traditional Medicines: Identification and Quality Control

11:45 - 12:30

Prof. Pang Chui Shaw Li Dak Sum Yip Yio Chin R&D Centre for Chinese Medicine, The ChineseUniversity of Hong KongTitle: An Introduction to Li Dak Sum R & D Centre for Chinese Medicine and Research on DNA Authentication of Herbs for Cool Beverage

12:30 - 14:00Lunch

Venue : Chung Chi College Chung Chi Tang Student Canteen (G/F)

12 December, 2016 (Monday)

Theme: Molecular Authentication and DNA Databases.

Chairperson: Prof. Christopher Hon Ki Cheng

14:00 - 14:20

Prof. Jerome Ho Lam HuiSchool of Life Sciences, The Chinese University of Hong Kong, Hong Kong SAR,ChinaTitle: A Molecular and Morphometric Database of Heung trees Aquilariasinensis (Lour.) Spreng in Hong Kong

14:20 - 14:55

Prof. Tae Jin YangPlant Sciences, Seoul National University, Seoul, KoreaTitle: Authentication of Plant Materials Based on Genomics and DNA Barcoding To Prohibit Economically Motivated Adulteration

14:55 - 15:30

Dr. Natalia IvanovaCanadian Centre for DNA Barcoding, Biodiversity Institute of Ontario, University of Guelph, Ontario, CanadaTitle: A Biocomplexity Perspective on Authentication of Herbal Supplements

15:30 - 16:10 Tea Break & Poster Presentation (odd number posters)

Chairperson: Prof. Kwok Pui Fung

16:10 - 16:45

Dr. David Tai Wai LauShiu-Ying Hu Herbarium, The Chinese University of Hong Kong, Hong Kong SAR, China Title: Taxonomic Archive System (TAS) – A Database of New Coding Methods for Botanical and Herbal Authentication

16:45 - 17:05

Prof. Kang NingBioinformatics, School of Life Science and Technology, Huazhong University of Science and Technology, Hubei Sheng, ChinaTitle: Consolidated ITS2 Database and Search Engines for Plant and Animal Biomarker Analyses

17:05 - 17:25

Mr. Tin Hang WongShiu-Ying Hu Herbarium, School of Life Sciences & Li Dak Sum Yip Yio Chin R&D Centre for Chinese Medicine, The Chinese University of Hong Kong, Hong Kong SAR, ChinaTitle: Medicinal Materials DNA Barcode Database (MMDBD) v1.5 - One Stop Solution of Storage, BLAST, Alignment & Primer Design

17:25 - 17:45

Miss Jin Kyung Kim Department of Plant Sciences, Seoul National University, Seoul, KoreaTitle: Comparative Analysis of The Complete Chloroplast Genome Sequences of Seven Taraxacum Species for Phylogenetic and Taxonomic Study

Venue: Yasumoto International Academic Park - YIA LT2, CUHK8:30 - 9:00 Registration

Chairperson: Dr. David Tai Wai Lau

9:00 - 9:35

Prof. David Chi Cheong WanSchool of Biomedical Sciences, The Chinese University of Hong Kong, Hong Kong SAR, China Title: Herbalog: Computational Herbal Screening for Evidence-based Study of Traditional Chinese Medicine

9:35 - 10:10

Prof. Shi Lin ChenInstitute of Chinese Materia Medica, Chinese Academy of Chinese Medical Sciences, Beijing, ChinaTitle: Traditional Medicine Identification Using DNA Barcoding

10:10 - 10:40 Tea Break & Poster Presentation (even number posters)

Chairperson: Dr. Natalia Ivanova

10:40 - 11:15Prof. Pete HollingsworthRoyal Botanic Garden, Edinburgh, UKTitle: DNA Barcodes, Genomics and Plant Identification

11:15 - 11:35

Ms. Hana LeeDepartment of plant sciences, Seoul national university, Seoul, KoreaTitle: The Complete Chloroplast Genome Sequences of Six Hydrangea SerrataAccessions and Their Comparative Analysis for Developing DNA Barcoding Marker

11:35 - 11:55

Dr. Lee Hong TnahForestry Biotechnology, Forest Research Institute Malaysia(FRIM), MalaysiaTitle: DNA Barcoding for Species Authentication of Common Herbal Plants in Malaysia

11:55 - 12:15

Dr. Yat Tung Lo Li Dak Sum Yip Yio Chin R & D Centre for Chinese Medicine, Institute of Chinese Medicine & School of Life Sciences, The Chinese University of Hong Kong, Hong Kong SAR, ChinaTitle: Molecular Authentication on Processed Herbal Products: Case Study on Animal-derived Concentrated Chinese Medicine Granules

12:15 - 14:00Lunch

Venue : Chung Chi College Chung Chi Tang Student Canteen (G/F)

13 December, 2016 (Tuesday)

Theme: DNA Barcodes --- Development and Application.

Chairperson: Prof. Tae Jin Yang

14:00 - 14:35Prof. De Zhu LiKunming Institute of Botany, Chinese Academy of Sciences, Yunnan, ChinaTitle: Beyond Flora of China: DNA Barcoding and iFlora

14:35 - 14:55

Dr. Aekkhaluck IntharuksaPharmaceutical Sciences, Faculty of Pharmacy, Chiang Mai University, Chiang Mai, ThailandTitle: A Comparison of Different DNA Barcoding Markers for Identification of Terminalia Plants and Their Crude Drugs Collected from Thailand.

14:55 - 15:15

Prof. Sathishkumar RamalingamBiotechnology, Bharathiar University, Tamil Nadu , IndiaTitle: Application of Universal DNA Barcode System- Experience with Indian Herbal Industries

15:15 - 15:45 Tea Break & Poster Session

Chairperson: Prof. Chun Kwok Wong

15:45 - 16:20

Prof. Madasamy ParaniGenetic Engineering, Center for DNA Barcoding, SRM University, Tamil Nadu, IndiaTitle: Authentication of Commercial saffron Samples by HPLC and DNA Barcoding

16:20 - 16:40

Mr. Masashi Kitamura Graduate School of Medical Sciences, Kanazawa University, Ishikawa, JapanTitle: Development of Loop-mediated Isothermal Amplification (LAMP) Assay for Rapid Identification of Cannabis sativa

16:40 - 17:00Mr. Saravanan MohanasundaramBiotechnology, Bharathiar University, Tamil Nadu , IndiaTitle: DNA Barcoding for Authentication of the Commercial Indian Herbal Tea

17:00 - 17:20

Dr. Meng Hua WuJinan University, Guangdong Sheng, ChinaNew Book Announcement - Molecular Identification of Chinese Medicinal Materials: Technology And Application

17:20 - 17:40

Ms. Mahima KarthikeyanDepartment of Biotechnology, Bharathiar University, Tamil Nadu , IndiaTitle: Application Of DNA Barcoding To Unravel The Taxonomical Complexity In Ficus L. (Moraceae) Of Weatern Ghats, India

18:00 - 21:00Banquet (Invited & registered guests)Venue: Chung Chi College Staff Club

Venue: Yasumoto International Academic Park - YIA LT2, CUHK8:30 - 9:00 Registration

Chairperson: Prof. Jerome Ho Lam Hui

09:00 - 09:35

Prof. David L. EricksonGenetics, DNA4 Technologies LLC, Maryland, USATitle: Application of Metagenomic Methods to Forensic Identification of Commercial Herbal Medicines

09:35 - 10:10

Prof. Karl Wah Keung TsimCenter for Chinese Medicine R & D, The Hong Kong University of Science and Technology, Hong Kong SAR, ChinaTitle: The Osteogenic Properties of Danggui Buxue Tang, a Chinese Herbal Decoction Containing Astragali Radix and Angelicae Sinensis Radix: Genomics Analysis of Specific Chemical Knock-out Herbal Decoction

10:10 - 10:40 Tea Break & Dismount of Posters

Chairperson: Prof. Karl Wah Keung Tsim

10:40 - 11:15

Prof. Xi Wen Li Institute of Chinese Materia Medica, Chinese Academy of Chinese Medical Sciences, Beijing, ChinaTitle: Complete Chloroplast Genomes of An Endangered Species Magnolia Officinalis Subsp. Biloba: Organization and Implications for Selecting Specific

Barcodes

11:15 - 11:35

Dr. Dhivya SelvarajDepartment of Biotechnology, Bharathiar University, Tamil Nadu , IndiaTitle: Development of DNA Barcode Based CHIP for Visual Detection of Adulterants in Herbal Products

11:35 - 11:55

Dr. Chun Xia Zeng Kunming Institute of Botany, Chinese Academy of Sciences, Yunnan, ChinaTitle: Unveiling Genome Information of Herbarium Specimens in Next Generation Sequencing Era

11:55 - 12:15

Mr. Yang Liu Institute of Medicinal Plant Development, Beijing, ChinaTitle: Nucleotide Signature Method for the Identification of Chinese Patent Medicines

12:15 - 12:20

Closing CeremonyProf. Pang Chui ShawDirector, Li Dak Sum Yip Yio Chin R & D Centre for Chinese Medicine, School of Life Sciences, Institute of Chinese Medicine, CUHK

12:20 - 14:00Lunch

Venue : Chung Chi College Chung Chi Tang Student Canteen (G/F)

14 December, 2016 (Wednesday)

Theme: New Authentication Technology.

Opening Ceremony - Invited Guest

Edwin Lok Kin TsuiAssistant Director (Traditional Chinese Medicine), Department of Health, The Government of the Hong Kong Special Administrative Region, ChinaEmail: edwintsui@dh.gov.hk

Biography

Dr. Edwin Lok Kin Tsui is the Assistant Director (Traditional Chinese Medicine) of the Department of Health. He oversees the implementation of the Chinese Medicine Ordinance and its subsidiary legislations, the promotion of safe and efficacious use of Chinese medicine and practice, the conducting of Chinese medicine related public health and health promotion activities and the planning and establishment of the Testing Centre for Chinese Medicines which is expected to operate in phases from 2017.

He is the Director of the World Health Organization (WHO) Collaborating Centre for Traditional Medicine, and is also serving as focal point of Hong Kong SAR of the International Regulatory Cooperation for Herbal Medicines under the WHO.

Prior to this, he had been responsible for and contributed in various fields in public health protection and promotion including port health measures, food safety, contingency planning, surveillance, investigations and control of public health crisis. He had assisted the WHO in 2011 and 2014 to develop WHO strategic documents on International Health Regulations and border quarantine measures. In 2016, he had been invited to be WHO Temporary Advisor and provided professional advice in development of traditional medicines in the Western Pacific regions of the WHO.

Abstracts of Invited Speakers

Future Development in the Testing Centre for Chinese

Medicines

Kelvin Kam Chuen ChanDepartment of Health, The Government of the Hong Kong Special Administrative RegionEmail: kelvin_kc_chan@dh.gov.hk

Biography

Prof. Kelvin Kam Chuen Chan (DSc PhD FRSM) is currently Senior Advisor in Chinese Medicines at the Department of Health, Hong Kong SAR, PR China; concurrently Visiting Professor at Liverpool John Moores University, UK and Adjunct Professor of Traditional Chinese Medicine (TCM) at University of Technology Sydney and Western Sydney University. Initially trained in Industrial Pharmacy, then specialized in Clinical Pharmacology in the UK, he started R&D of TCM at the Chinese University of Hong Kong (1981-1992) while teaching medical pharmacology in orthodox medicine; subsequently he has taught pharmacy, and biomedical sciences in TCM at universities in Hong Kong, UK, UAE and Australia, while focusing his research on the role TCM in integrative healthcare.

Research publications include 250 full SCI papers, 3 reference/specialist books in toxic Chinese Medicinal Materials (CMM), interactions between pharmaceuticals and TCM products and several monographs of pharmaceutical drugs and CMM, and over 300 international keynotes/presentations at conferences.

Over 30 PhD and post-graduates were trained by his research teams on drug disposition/interaction/metabolism/pharmacokinetics and several projects in TCM research, some involved the following themes:• QC and pharmacopoeia methodology of CMM and related products;• Chemometrics evaluation linked with synergy-mechanistic investigation for R&D of

TCM products with industrial partners;• Integrative research linking TCM principles with biomedical research methods

involving systems biology as proof of principles according conventional science;• Establishing TCM-appropriate Quality of Life instrument in assessing patient reported

outcomes as evidence-based approaches for TCM practice and research; • Development of training curriculum for future human resources for integrative

healthcare.

Abstract

Kelvin Kam Chuen Chan

Chinese Medicine Division, Department of Health, The Government of the Hong Kong

Special Administrative Region

Corresponding Author E-mail : kelvin_kc_chan@dh.gov.hk

The Government had accepted the recommendation of the Chinese Medicine Development Committee and announced in the 2015 Policy Address that it would plan and develop a Testing Centre for Chinese Medicines (Testing Centre) to be managed by the Department of Health.

The Testing Centre will specialize in the testing of, and scientific research on Chinese medicines with a view to setting authoritative reference quality standards for the safety, quality and testing methods of Chinese medicines, thereby facilitating the internationalisation of Hong Kong’s Chinese medicine industry and enhancing Hong Kong as an international hub of Chinese medicines testing.

The planning work of setting up the Testing Centre has already commenced. Since it may take a few years to establish the Testing Centre, a temporary Testing Centre in reduced scale for interim operation is planned to commence operation in phases in 2017.

The Testing Centre will continue promoting the Hong Kong Chinese Materia MedicaStandards (HKCMMS) as authoritative international benchmark for reference testing standards and methods in order to facilitate the internationalization of Hong Kong’s Chinese medicines industry. Since the HKCMMS project first launched in 2002, reference quality standards for 236 Chinese Materia Medica (CMM) has been developed. It is envisaged that the setting of applicable and adoptable reference standards for CMM will facilitate the business of Chinese medicines.

The temporary Testing Centre will consist of several laboratories. The laboratories are planned to build up its capacity to promote scientific research on TCM as well as strengthening the capability for quality control and identification of CMM. Analytical work, including both chemistry and DNA, will be explored to develop standards for application in the quality control of Chinese medicines by employing new-emerging technologies, such as DNA barcoding. The methods and standards developed by the Testing Centre will be fully validated and the Centre will aim to provide technology transfer to the Chinese medicines trade and industry in order to promote Hong Kong as the international hub for Chinese medicines testing.

Authentication of Plants Used in Traditional Medicines:

Identification and Quality Control

Monique S.J. SimmondsRoyal Botanic Gardens, Kew, UKEmail: M.Simmonds@kew.org

Biography

Prof. Monique S.J. Simmonds is Deputy Director of Science at the Royal Botanic Gardens, Kew. She has 30 years’ experience of research into different aspects of plant and fungal chemistry with an emphasis on the economic value of natural products. This includes using different methods such as DNA for the authentication and quality control of medicinal plants. She has co-authored patent for compounds in the area of malaria, cancer, pest control, cosmetics and have been involved in getting food, medicinal and cosmetic products to market. For example, the Botanics range with Walgreen/Boots and Active Botanical with Procter & Gamble. These projects involve ensuring a good supply chain and the work was awarded the Guardian Natural Capital prize in 2014. Her research illustrates how Kew’s diverse and validated unique range of collections as well as the expertise of its scientists and horticultural staff can help address economical important issues associated with plant/fungal use. She has worked in 81 different countries and supervised 37 PhD students.

She has experience in working on authentication with different business sectors including retail, pharmaceutical and agricultural companies. Her academic work is well cited with an H-Index 55. Recent paper (Michl, J. et al. (2016) LC-MS- and H-1 NMR-Based MetabolomicAnalysis and in Vitro Toxicological Assessment of 43 Aristolochia Species. Journal of Natural Products, 79, 30-37.) was awarded the American Society of Pharmacognosy 2016 Arthur E. Schwarting Award for the best paper published in the Journal of Natural Products.

Abstract

Monique S.J. Simmonds, Felix Forest, Christine J. Leon, Melanie-Jayne Howes

Royal Botanic Gardens, Kew, Richmond, Surrey TW 9 3AB, UK

Corresponding Author E-mail : M.Simmonds@kew.org

Advances in DNA technology, decrease in costs and increased availability of equipment, have meant that DNA methods are now starting to be considered as viable tests to be included in national Pharmacopoeias, along with the traditional morphology and chemical tests. The use of “DNA barcodes” (pre-designated DNA regions) has been widely advocated in the past decade as the golden standard in DNA-based authentication, but it has not been constantly successful at identifying a particular sample at the species level. Furthermore, the approach is not always suitable for degraded material, often the template requiring identification in the trade of traditional medicines. The advent of high-throughput sequencing technologies offer new tools in DNA-based authentication that promise to resolved some of the issues related to the use of only a few DNA barcodes. Going forward DNA methods, particularly those using high-throughput sequencing technologies, along with conventional methods provide approaches that can discriminate among species of plants as well as provide an indication of quality. They can also identify contaminates, harmful substitutes and adulterants. This review provides an overview of some of the recent DNA barcoding methods, their advantages and limitations, as well as some of the new molecular methods including gene capture, genome skimming, or a combination of both.

The talk will be illustrated by research at Kew as well as with others working on different aspects of DNA barcoding. It will cover the challenges of finding authenticated plant material especially substitutes and problems dealing with processed and unprocessed material entering the trade.

Challenges in Assessing the Authenticity and Quality

of Herbal Material

Ikhlas A. KhanThe University of Mississippi, USAEmail: ikhan@olemiss.edu

Biography

Prof. Ikhlas Khan is the Associate Director of the National Center for Natural Products Research, Director of the FDA Center of Excellence, Research Professor, as well as Professor in the Department of Pharmacognosy at the National Center for Natural Products Research at the University of Mississippi. Additionally, he is the Director for Sino-US TCM Research Center; Director, Center for Research of Indian Systems of Medicine

(CRISM); Research Professor and Coordinator for Natural Products Research Center for Water and Wetlands Research; a visiting Professor at Hunan University of Chinese Medicine, China; an Adjunct Professor, Chinese University of Hong Kong; visiting professor for King Saud University, Saudi Arabia since 2010; Soochow University, since 2010 and Heilongjiang University of Chinese Medicine, The People’s Republic of China since 2010. He received a D. Litt (Honoris Causa) from University of Hamdard, Delhi, India 2012; a B.S. in Chemistry in 1980 and a M.S. in Organic Chemistry in 1982 from the Aligarh Muslim University in Aligarh, India. He then received a Ph.D. in Pharmacy from the Institute fuerPharmaceutische Biology in Munich, West Germany in 1987 and postdoctoral studies at Swiss Federal Institute of Technology (ETH) Zurich. After completing his education, He began his career with the University of Mississippi in 1992 as a research scientist. He became a Research Assistant Professor in 1995, and was promoted to Research Associate Professor and Director of the FDA program in 2001, Assistant Director of NCNPR in Oct. 2002, Professor in July 2005 and to Associate Director in 2015.

His primary research interests include analytical fingerprinting for standardization of herbal products, and bio-analytical approaches to improvement of product quality and safety.

Prof. Ikhlas Khan has authored or co-authored over 700 original research articles, publications, or reviews. He has been an invited speaker at numerous events. He serves as reviewer for several journals such as foreign editor for J. Traditional Chinese Medicine. He was previously Co-editor of Planta Medica.

He is the recipient of numerous awards, including: most recently he was awarded “Cumberland Pharmaceuticals, Inc. Research Award, 2016”, “UM Distinguished Researcher Award, 2016”, “IASTAM Zandu International Award for Excellence in Field of Ayurvedic and/or Natural Products”, 2015, “Outstanding Contribution in Natural Product Research”, Water’s Corporation, given at the ICSB 2013, “The Center for Food Safety and Applied Nutrition” (CFSAN) Director’s Special Citation Award, 2012; the Varro E. Tyler Prize, the American Society of Pharmacognosy, 2011; the Nutrition Business Journal Education Award, 2011; the 2nd Hakim Ahmed Ashraf Global Award, December 2010; the 5th Annual AHPA Herbal Insight Award, April 2010; Food and Drug Administration’s Commissioner’s Special Citation, June, 2009; the Norman R. Farnsworth Excellence in Botanical Research Award (2009); the Cumberland Pharmaceuticals Inc Faculty Research Award, 2007-2008, School of Pharmacy; ISHS Award for Meritorious Service, 2005; Researcher of the Year in the National Center for Natural Products Research, 2001; Faculty Research Award, School of Pharmacy, 2001-2002.

He serves on several committees of National Center for Complementary and Alternative Medicines, Washington, D.C. which include serving as Invited Reviewer and as a Committee Member of the Product Quality Working Group. Additional appointments to

national and/or international committees include membership of the First Editorial Board of World Journal of Traditional Chinese Medicine (WJTCM); membership of expert panel of United States of Pharmacopoeia (USP) Committee; Advisory Board of Women’s Health & Asian Traditional Medicine (WHAT); Advisory Board of American Herbal Product Association, Washington, D.C.; Committee of AOAC Dietary Supplement Task Force; Board of Missouri Botanical Garden; Advisory Board of the American Botanical Council; Expert Advisory Committee, Natural Health Directorate, Health Canada.

Memberships to scientific organizations include fellowship of the American Institute of Chemists (FAIC); fellowship of the Royal Society of Chemists (FRSC); member of the International Society for Ethnopharmacology; member of the American Chemical Society; the Society of Medicinal Plant Research; The American Association for the Advancement of Sciences, and the New York Academy of Sciences, American Association of Pharmaceutical Scientists, International Society for Horticultural Science, National Advisory Council for Complementary and Alternative Medicine, American Society for Clinical Pharmacology and Therapeutics, and the American Association for the Advancement of Science. He is a member of the American Society of Pharmacognosy, where he served as Chair of the Constitution of By-laws Committee in 1998. He also served as a WHO consultant in Vietnam in 2002. He is an honorary memberships of Sigma Xi, the Scientific Research Society; Rho Chi, the Pharmaceutical Honor Society.

Abstract

Ikhlas A. Khan

National Center for Natural Products Research, School of Pharmacy, The University of Mississippi, University, MS, 38677.

Corresponding Author E-mail: ikhan@olemiss.edu

With the ever-increasing necessity for new and improved chromatographic techniques for the authentication and detection of adulterants of botanicals, the need for the development of effectual chemical fingerprinting methodologies has become a valued alternative to being dependent on the use of single entity relevant biomarker(s). This is particularly true for botanicals where no unique relevant biomarker is present. Too often, many of the established analytical methods rely solely on the use of a few common plant metabolites for identification purposes. The utility of building a full analytical fingerprint profile for identification purposes is that it provides a solid scientific basis for authentication purposes as well as the ability to identify contaminants, adulterants as well as “spiked” samples. Through the use of a detailed evaluation of a broader spectrum of metabolites it is also possible to identify species/varietal, seasonal, and regio-specific differences between various populations of a given plant. However, this detailed approach comes at a significant cost in time, effort and expense but the end result provides researchers a firmer scientific basis for making informed decisions for authentication purposes.

New Book Announcement - Molecular Identification of Chinese Medicinal Materials: Technology And

Application

Dr. Meng Hua WuJinan University, Guangdong Sheng, China

Biography

Dr. Meng Hua Wu comes from School of Pharmacy, Jinan University in Guangzhou. She majors in Pharmacognosy and was graduated from Hong Kong Baptist University in 2013. She has participated in the compilation of some books such as Encyclopedia on Contemporary Medicinal Plants and Illustrated Chinese Materia Medica. Now she will introduce a new book to us, Molecular identification of Chinese Medicinal Materials:

Technology and Application.

Authentication of Plant Materials Based on Genomics

and DNA Barcoding To Prohibit Economically Motivated Adulteration

Tae Jin YangSeoul National University, KoreaEmail: tjyang@snu.ac.kr

Biography

Prof. Tae Jin Yang is in the department of plant science at Seoul National University since 2006. Before, he worked as researcher in rural development administration, Korea, for 10 years. He studied and got master and Ph.D. degree at Seoul National University. He did research as postdoctoral researcher at Clemson University and Arizona University Genomics institute for 4 years.

His research interests are on the plant genomics, evolution, molecular breeding and NGS-based high-throuput plastid genome sequencing and phylogenomics. He involved in genome sequencing of rice, Brassica crops and now focusing on genome sequencing and evolution of Panax ginseng. He developed novel NGS-based pipeline for high throughput complete sequencing of plastid and nuclear ribosomal DNA, simultaneously, which is useful for phylogenomics.

Recently, he is developing unique DNA markers using the complete sequences of CP genomes and ribosomal DNA and chemical markers to authenticate various medicinal plants utilized for functional foods or natural medicines which is supported from Ministry of Food and Drug Safety, Korea.

Abstract

Tae Jin Yang

Department of Plant Science and Plant Genomics and Breeding Institute, College of

Agriculture and Life Sciences, Seoul National University, Seoul, 151-921, Korea

Corresponding Author E-mail: tjyang@snu.ac.kr

Cytoplasmic chloroplast (cp) genome and 45S nuclear ribosomal DNA gene cluster (nrDNA) regions are main targets for elucidation of plant genetics diversity, barcoding and evolution. We developed a high throughput method for de novo sequence assembly and finishing to obtain complete whole cp genome and a major unit for 45S nrDNA tandem array as well as characterization of major repeats simultaneously using only small scale whole genome shotgun sequence (dnaLCW method). The dnaLCW method can be applied for any plant species and will open a new era for elucidation of genetic diversity and fundamental understanding of tree of life for plants as well as for development of practical barcoding markers not only at species level but also intra species cultivar level. The genome information produced by dnaLCW method can be applied to improve breeding of the target species and also to develop the practical barcode markers which can be applied to authenticate specific species or specific cultivars among plants using raw materials as well as the processed functional foods. The species -unique or cultivar-specific markers can be applied to prohibit the illegal trade of wrong plant materials instead of the target plants for economically motivated adulteration in the production of functional foods or traditional oriental medicinal drugs.

Acknowledgement :This work was supported by the grant funded by Next-Generation BioGreen21 Program (No. PJ01103001, PJ01100801), Rural Development Administration, the Bio & Medical Technology Development Program (NRF-2015M3A9A5030733) of the NRF funded by the Korean government, and “16172MFDS229” from Ministry of Food and Drug Safety in 2016, Republic of Korea.

A Biocomplexity Perspective on Authentication of

Herbal Supplements

Natalia IvanovaUniversity of Guelph, Guelph, Ontario, CanadaEmail: nivanova@uoguelph.ca

Biography

Dr. Natalia Ivanova received her Ph.D. in Molecular Biology from Lomonosov Moscow State University in 1998. Most of her Ph.D. data on the molecular systematics of lichens was gathered at the NMNH, Smithsonian Institution. In 1998-2004 she worked as a researcher at the Engelhardt Institute of Molecular Biology (Russian Academy of Sciences,

Moscow). She has joined Prof. Hebert’s group at the University of Guelph in 2004, soon after inception of DNA Barcoding concept, and contributed to transition from an academic lab to a largest DNA Barcoding facility in the world processing 1M specimens per year. She considers herself an application specialist with a very simple strategy: design –implement – maintain. Her recent track record in methods development includes

integration of Beckman Coulter Biomek liquid handling stations, DNA extraction protocols for high-throughput robotic environment, room temperature DNA storage and shipment, express DNA barcoding protocols, and development of QA/QC procedures utilizing control panels. She is currently involved in the following research projects: 384-well automated pipeline for HTS sample processing, SPRI-based methods for cycle-sequencing products cleanup and DNA extraction, minimizing PCR bias in NGS workflows, detection and monitoring of toxigenic algal blooms in Great Lakes, biomonitoring of vertebrates in water and soil using eDNA, metabarcoding of insects in bulk samples, and authentication of food and Natural Health Products using NGS.

Abstract

Natalia V. Ivanova1*, Maria L. Kuzmina1, Thomas W. A. Braukmann1, Alex V. Borisenko1 and Evgeny V. Zakharov1

1 – Centre for Biodiversity Genomics, Biodiversity Institute of Ontario, University of Guelph, Guelph, Ontario, Canada

Corresponding Author E-mail : nivanova@uoguelph.ca

Recent advances in DNA-based authentication have enabled fast and sensitive detection of DNA sources in herbal supplements. This is now utilized by some manufacturers for quality assurance of raw plant materials, final products, and contamination control during production. One of the common approaches, DNA barcoding, heavily relies on Sanger sequencing. However, stochastic amplification of multiple DNA sources that are often present even in single-source supplements renders Sanger results non-interpretable or non-reproducible, hence indicating strong need for NGS-based methods. While advances in NGS technology enable rapid and sensitive detection of DNA in complex mixtures, such results should be interpreted from a biocomplexity perspective. Aside from intended or non-intended substitution, possible cross-contamination with trace plant or fungal DNA can occur at any stage during growing, harvesting, manufacturing, handling or laboratory analysis of plant material. Detection of such non-target DNA is not necessarily indicative of technological flaws or deliberate adulteration and such results should be interpreted with caution. Diversity of fungi in herbal supplements is determined by a combination of pathogenic, saprophytic, endophytic and mycorrhizal fungi naturally associated with live plant material, and strains involved in the fermentation during manufacturing of bioactive

components. Although this entire spectrum can be easily detected by NGS, interpretation of test results should focus on potential mycotoxin-producing fungi and human pathogens. We propose NGS-facilitated DNA barcoding as a standardized tool for authentication of herbal supplements. Because manufacturing of extracts leads to DNA degradation or loss, quality control should utilize synergetic approach targeting both bioactive components (HPLC-MS) and DNA.

Traditional Medicine Identification Using DNA

Barcoding

Shi Lin ChenInstitute of Chinese Materia Medica, Chinese Academy of Chinese Medical Sciences, ChinaEmail: slchen@implad.ac.cn

Biography

Prof. Shi Lin Chen, the Director of the Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, now also works as the Director of the World Health Organization Collaborating Center for Traditional Medicine, Academician of International Academy of Sciences for Europe and Asia, Vice President of the Consortium for Globalization of Chinese Medicine. Prof. Chen obtained his Ph.D from Chengdu University of Traditional Chinese Medicine and was trained in Royal Botanic Gardens, Kew, UK. And he has been the Visiting Professor in Hong Kong Polytechnic University, Visiting Professor of Tokyo University of Pharmacy and Life Sciences, Japan. He has established the DNA Barcoding identification system for Chinese herbal medicine species, and he has written some related works, including Standard DNA barcodes of Chinese Materia Medica in Chinese Pharmacopoeia. He has so far published totally more than 400 academic works, including 200 SCI papers on famous academic journals, such as Nature Commun, Biotech Adv and PNAS. His academic works have been cited more than 10,000 times, H-index: 48(Google Scholar). He has been honored the Wujieping Award for Medical Innovation, the Star of Nobel Prize and Agilent Thought Leadership Award etc.

Abstract

CHEN Xiao-chen1, Xiang Li2, SONG Jing-yuan1*, YAO Hui1, HAN Jian-ping,SHI Lin-chun1, Pang Xiao-hui,CHEN Shi-lin1,2*

1.Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100193, China;2.Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China

Corresponding Author E-mail : slchen@implad.ac.cn

A standard barcode database based on the ITS2 region and the psbA-trnHregion for Kampo crude materials was developed. This tool provides users with the basic information on each crude drug recorded in the Japanese pharmacopoeia, herbal material DNA barcoding identification, and the standard operating procedure (SOP) from sampling to data analysis. A total of 576 samples were collected to establish the database. An additional 100 samples were identified using the database. 71% of test samples were identified at species level as same as the names listed on their label. The remaining 29% were correctly identified at genus level. We report the first database that includes the standard barcode sequences from 97.3% of the crude drugs recorded in the JP (16thedition), which has abeneficial demonstration effect throughout the other pharmacopoeias.

DNA Barcodes, Genomics and Plant Identification

Pete M. HollingsworthRoyal Botanic Garden Edinburgh, UKEmail: p.hollingsworth@rbge.org.uk

Biography

Prof. Pete Hollingsworth is Director of Science at the Royal Botanic Garden Edinburgh, and a Visiting Professor at the University of Edinburgh and an Honorary Professor of the Chinese Academy of Sciences’ Kunming Institute of Botany. His research focuses on understanding and conserving plant biodiversity. In recent years he has contributed to the international efforts of building a unified DNA based-index of life on earth, including various roles within the International Barcode of Life project and the Consortium for the Barcode of Life. He has a strong interest in linking scientific research to practical conservation outcomes, with particular interests in reintroductions/translocations and the integration of genetic data into conservation planning.

http://www.rbge.org.uk/science/genetics-and-conservation/pete-hollingsworth-home-pagehttps://scholar.google.co.uk/citations?user=9_dz4HwAAAAJ&hl=en

Abstract

Pete M. Hollingsworth

Royal Botanic Garden Edinburgh, 20A Inverleith Row, Edinburgh, EH3 5LR, UK

Corresponding Author E-mail : p.hollingsworth@rbge.org.uk

DNA barcoding involves the standardised use of DNA sequences to tell species apart. A key strength of the approach is scalability – harnessing efforts of the global community to build a shared DNA-based identification resource. In plants, the barcode databases have been built with a focus on 2-3 regions of plastid DNA and the internal transcribed spacers of nuclear ribosomal DNA. For many applications these regions provide adequate resolution, but in other cases they do not. Rapid advances in sequencing technologies now provide the opportunity to increase the efficiency of screening standard barcodes, and provide additional characters to improve species resolution. In this presentation, I will review progress to date in using standard plant barcodes, and explore opportunities for future improved protocols using new sequencing platforms.

Beyond Flora of China: DNA Barcoding and iFlora

De Zhu LiKunming Institue of Botany, Chinese Academy of Sciences, ChinaEmail: dzl@mail.kib.ac.cn

Biography

Prof. De Zhu Li received his PhD from the Chinese Academy of Sciences (CAS)’ Kunming Institute of Botany (KIB) in 1990. He worked at the Royal Botanic Garden Edinburgh from 1993 to 1994 as a Ferguson Fellow and at the Cambridge University Botanic Garden as the Taxonomist from 1994 to 1996. He is now a Professor of KIB/CAS, the President of CAS Kunming Branch, as well as Vice President of the Botanical Society of China, and the Representative of China for the International Barcode of Life project (iBOL). Prof. Li is an associate editor for Proceedings of the Royal Society B - Biological Sciences, BMC Evolutionary Biology and PhytoKeys, and on the editorial boards of Journal of Systematics and Evolution and Kew Bulletin. He was awarded to an OBE in September 2010 for his contributions to China-UK botanic exchange and the promotion of biodiversity conservation. His research interests focus on plant systematics, biogeography, plant DNA barcoding and germplasm conservation. He has authored some 300 papers, including 210 in ISI-index journals, including PNAS, Trends in Plant Science, Systematic Biology, New Phytologist and Molecular Ecology. He has authored or contributed to 21 monographs and books, including the bamboo account of the Flora of China, and is the editor of the Chinese translation of Plant Systematics, A Phylogenetic Approach (3rd Edition).

Abstract

De-Zhu Li1,2, Jun-Bo Yang1, Hong-Tao Li1, Chun-Xia Zeng1, Ting-Shuang Yi1, Lian-Ming Gao2, Yu-Hua Wang1, Ting Zhang1, Jie Cai1, & Hong Wang2

1. Germplasm Bank of Wild Species, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, Yunnan 650201, China;2. Key Laboratory for Plant Diversity and Biogeography in East Asia, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, Yunnan 650201, China

Corresponding Author E-mail : dzl@mail.kib.ac.cn

There are 10-100 million estimated species on earth. However, Since the Species Plantarum of Linnaeus, we described 1.7-2.3 m species in 263 years, with about 39,100 species of vascular plants (Kew, 2016: The State of the World’s Plants). The bulk of biodiversity are still not described. The Flora of China described 3,1000 vascular plants or 8% of world’s total by 2013. We illustrate progresses in advancing an iFlora of China. By 2015, 67,869 accessions of seeds of 9129 species of seed plants or 30% of Chinese flora, plus some 10,000 plant DNA samples were preserved in the Germplasm Bank of Wild Species in Kunming. Meanwhile, we obtained a total of 110,000 barcodes of 28,519 individuals representing 9280 species of 2,345 genera of 279 families of vascular plants of China, with the support of the China Plant DNA Barcoding Project, and 50% of those data were analyzed. It is projected that a third of the species (ca. 10,000) and 80% of the genera (ca. 2700) of the Chinese Flora will be barcoded by 2016. And finally, we discuss options for enhancing ‘standard’ plant barcodes, focusing on increasing discriminatory power by next-generation sequencing, particularly genome skimming.

Identification of Orchid Species in The TCM and Food

Trade Using DNA Barcodes.

Gunter Fischer The Flora Conservation Department at Kadoorie Farm and Botanic Garden (KFBG), Hong KongEmail: gfischer@kfbg.org

Biography

Dr. Gunter Fischer is the head of The Flora Conservation Department at Kadoorie Farm and Botanic Garden (KFBG), Hong Kong, where he is responsible for the daily operation and management of the Estate (Botanical Garden), the Orchid Conservation, the Restoration Ecology, the Genetic Laboratory and the Ecological Statistics (GIS) sections.

Born and raised in Salzburg, Austria, he graduated from the University of Vienna with a PhD in molecular ecology in 2007 and subsequently hold research positions at the Universities of Salzburg and Vienna. His research and conservation interests have taken him into many areas in China, Indochina, South-East Asia, Africa and Madagascar. He joined KFBG in 2010 and has since than established a number of conservation programmes and projects in the fields of in-situ/ex-situ species conservation, ecosystem conservation and restoration, conservation genetics, environmental statistics and modeling/GIS, climate change monitoring, urban greening and awareness raising for conservation and sustainability in Hong Kong, South China and the greater Indo-Burma biodiversity hotspot region. In 2016 he has been appointed as Honorary Associate Professor at the University of Hong Kong (HKU).His research findings have been published in scientific reports and papers.

Abstract

Gunter Fischer

The Flora Conservation Department at Kadoorie Farm and Botanic Garden (KFBG),

Lam Kam Road, Tai Po, New Territories, Hong Kong

Corresponding Author E-mail : gfischer@kfbg.org

Traditional Chinese Medicine (TCM) uses thousands of different plant species and many of them are traded in high quantities throughout Asia. One of the most commonly used ones are orchid species from the genus Dendrobium. Out of the 1200 taxa described globally, only ~ 15 species originating from China and Indochina, are used as medicine or food supplement. Despite the fact that most of the 30 species can easily be farmed in large quantities, many are still collected from wilderness areas because of traditional beliefs, that wild collected plants have “better” health benefits. This leads to an uncontrolled overexploitation of these species and many of them are already at the brink of extinction in the wild. Therefore many of these species are protected by law in countries of origin and the trade is regulated by CITES.During the process of preparing the orchids as medicine, the stems are dried, cut into small pieces and often boiled and coiled into small balls, which makes a morphological identification virtually impossible. Most of the time the orchids are traded just as medicinal herbs or as “ Herba Dendrobii", or "Dendrobii Caulis” making it difficult for law enforcement agents to distinguish between protected and non protected species.In this study we developed new DNA barcodes specific to Dendrobium to identify the traded species and tested the discriminatory power of these barcodes with randomly purchased “ Herba Dendrobii", or "Dendrobii Caulis” samples from different traditional pharmacies in Hong Kong.Our survey revealed that more than 20 orchid species are traded as “ Herba Dendrobii", or "Dendrobii Caulis” belonging to three different orchid genera, whereby Dendrobiumwas the most commonly found genus, accounting for 89% of the trade. The rest are similar looking species from other genera suggesting that these species are used as surrogates. Most of the traders claim that their products belong to D. huoshanense or D. officinale, the most “potent” medicinal species, but our data suggests that the majority of the traded species belong either to D. devonianum or D. aphyllum, both more common and easy to grow species.We hope that our newly developed DNA barcodes can help to improve law enforcement and help to set standards for quality control to prevent the sale of surrogates.

Authentication of Commercial Saffron Samples by

HPLC and DNA Barcoding

Madasamy ParaniSRM University, IndiaEmail: parani.m@ktr.srmuniv.ac.iin

Biography

Prof. Madasamy Parani has completed his undergraduate degree in Agriculture and Postgraduate degree in Plant Breeding and Genetics from Tamil Nadu Agricultural University, India. He joined as Research Assistant in M. S. Swaminathan Research Foundation, Chennai, India in 1993 and left the institute as Scientist in 2002. During this

period, he completed his PhD from Anna University, Chennai. Then he worked as Post Doctoral Fellow in University of Florida, Florida, USA and Research Assistant Professor in University of Toledo, Ohio, USA between the year 2002 and 2006. He joined SRM University, Chennai, India as Assistant Professor in 2006, and currently he is heading the Department of Genetic Engineering. He is the General Secretary of the Indian Youth

Science Congress Association since 2010. He has been a visiting scientist to University of Heidelberg (Germany), University of Bologna (Italy), and International Rice Research Institute (Philippines). He has been a visiting professor to University of Guelph (Canada) and Texas A&M University (USA). He has received Prof. Uma Kanth Sinha Memorial Award for Young Scientist from Indian Science Congress (2002) and India DBT-Cutting Edge Research Enhancement and Scientific Training Award (2012). He has 50 publications and 2 patents to his credit. His areas of research include plant made pharmaceuticals, plant genomics and DNA barcoding of plants.

Abstract

Varadharajan Bhooma, Sophie Vassou, Mariappan Vairamani, Madasamy Parani*

Center for DNA Barcoding, Department of Genetic Engineering, SRM University, Kattankulathur, Kanchipuram District, Tamil Nadu, India 603 203

Corresponding Author E-mail : parani.m@ktr.srmuniv.ac.iin

Saffron (Crocus sativus L.) is most expensive spice, which is used for its flavor, color and medicinal properties. Owing to its high economic value and restricted production, it is often subjected to adulteration. We have used HPLC and DNA barcoding for the authentication of the commercial saffron samples. Safranal was extracted in acetonitrile for 2 hours at 70°C, and run in HPLC along with safranal standard. All the authentic saffron samples contained safranal peak and a specific HPLC fingerprint. Among the 94 samples collected from 9 countries, safranal and saffron HPLC fingerprint were detected in 51 samples, and all of them were confirmed to be pure saffron by DNA barcoding. Neither the safranal peak nor the saffron HPLC fingerprint was detected in 21 samples, and these samples were also found to be saffron negative in DNA barcoding. Sixteen samples showed safranal peak but not the saffron HPLC fingerprint. DNA barcoding revealed that these saffron samples were subjected to fraudulent substitution with plant material derived from other species. Safranal in these samples seems to originate from externally added synthetic safranal, which could be detected by HPLC after 5 minutes extraction at room temperature. Six samples showed safranal peak and saffron HPLC fingerprint but DNA barcoding showed mixed peaks characteristic of species admixture. These results show that DNA barcoding is superior to HPLC in reliable authentication of commercial

saffron samples.

Application of Metagenomic Methods to Forensic

Identification of Commercial Herbal Medicines

David L. EricksonDNA4 Technologies LLC, USAEmail: david@dna4tech.com

Biography

Dr. David L. Erickson is the founder and CEO of DNA4 Technologies LLC, a biotechnology company in Baltimore, Maryland (USA), that focuses on developing new methods for the identification of natural products. In particular, he directs the development of computational tools that leverage Next Generation Sequencing to diagnose the species content of natural materials including supplements, food, timber and environmental samples. Before founding DNA4 Technologies, he was a biologist at the Smithsonian Institution’s National Museum of Natural History where he developed laboratory and statistical analysis methods applied to DNA barcoding. While at the Smithsonian Institution he contributed to over 50 peer-reviewed publications, many of which have been seminal in the development of DNA barcodes and their application in a range of scientific endeavors. Prior to his work at the Smithsonian, he held postdoctoral fellowships in Quantitative Genetics at the University of Maryland, and in ancient DNA analysis of agricultural species at the Smithsonian Institution. He received his Ph.D. in

Botany from the University of Georgia where he received an NSF Fellowship in Mechanisms of Plant Molecular Evolution, and where his love of plants and genetics merged.

Abstract

David L. Erickson

DNA4 Technologies LLC, bwtech@UMBC Research & Technology Park, Baltimore, MD 21227, USA

Corresponding Author E-mail : david@dna4tech.com

The forensic identification of commercial herbal medicines, or nutraceuticals, is essentially the same process as that used to deduce the species content ofenvironmental mixtures. This field of deducing environmental samples is evolving rapidly in light of ever cheaper sequencing and improved analytics. This presentation will review current methods that extend beyond PCR amplicon approaches, and which utilize a wider spectrum of genomic data. This will include the use of RNA baits and Whole-Genome-Shotgun methods for use in forensic identifications. The relative merits of different approaches will be contrasted and an example data using Echinacea supplements will be provided.

The Osteogenic Properties of Danggui Buxue Tang, A

Chinese Herbal Decoction Containing Astragali Radix and Angelicae Sinensis Radix: Genomics Analysis of

Specific Chemical Knock-out Herbal Decoction

Karl Wah Keung TsimThe Hong Kong University of Science and Technology, Hong Kong

Email: botsim@ust.hk

Biography

Prof. Karl Wah Keung Tsim received his BSc and MPhil from The Chinese University of Hong Kong, and PhD in Molecular Neurobiology from the University of Cambridge, UK. After his post-doctoral training at Stanford University, USA, he joined The Hong Kong University of Science and Technology (HKUST) in 1992. Currently, he is a Chair Professor of Division of Life Science, and the Director of Center for Chinese Medicine R & D at the university. Prof. Tsim has developed molecular technique to determine the genetic and chemical properties of Chinese herbs. He has published over 300 scientific papers and serves as editors for many scientific journals internationally. His works on Chinese herbal medicine have been awarded twice for Research Excellence in Natural Sciences from Ministry of Education of China. He also serves as an advisor/consultant/member to various organizations, both nationally and internationally, in the standardization of Chinese herbs, which include WHO and HKSAR Government in Testing and Certification of Chinese herbs. He is an active entrepreneur and is a founding director of few companies. He also serves as Independent Non-executive Director of Lee’s Pharmaceutical Holding Ltd.

(HK stock # 950).

Abstract

Amy G.W. Gong, Tina T.X. Dong, Karl W.K. Tsim

Division of Life Science and Center for Chinese Medicine, The Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong; HKUST Shenzhen Research Institute, Hi-Tech Park, Shenzhen 518000, China

Corresponding Author E-mail : botsim@ust.hk

Danggui Buxue Tang (DBT), a Chinese herbal decoction, contains 2 herbs of Astragali Radix (AR) and Angelicae Sinensis Radix (ASR) at the ratio of 5:1. Clinically, DBT was utilized to mitigate menopausal osteoporosis; however, the signaling mechanism and the active compounds contributed in bone formation remained unclear. Calycosin and ferulic acid, being the most abundant distinct chemicals in AR and ASR, respectively, were depleted specifically from DBT by HPLC separation. Authentic DBT, a calycosin-depleted DBT (DBT∆cal) and a ferulic acid-depleted DBT (DBT∆fa) were applied onto cultured rat osteoblasts. Mineralization assay illustrated that both DBT and DBT∆fa significantly induced osteoblast differentiation, while DBT∆cal exhibited a reduced activation in mineralization. The total RNA from herbal decoction-treated osteoblasts was isolated for transcriptome analysis by RNA-seq. The gene expressions of osteogenic markers during differentiation, including Alpl, Runx2, Sparc, Sp7 and Spp1, were significantly increased under the treatments of DBT and DBT∆fa. Signaling pathway analysis by KEGG and GO-Elite revealed that 11 pathways were activated by DBT, including MAPK/ERK and Wnt/β-catenin signalings, as compared to the untreated control; these signaling pathways were the well-known cascades for osteoblast formation. In contrast, DBT∆cal was not able to trigger Wnt/β-catenin signaling. Thus, our findings indicated that calycosin could be an indispensable chemical in DBT to orchestrate multi-components of DBT as to achieve maximal osteogenic properties.

Acknowledgement: Supported by Hong Kong Research Grants Council GRF (662713, M-HKUST604/13), TUYF15SC01, The Hong Kong Jockey Club Charities Trust (HKJCCT12SC01), Foundation of The Awareness of Nature (TAON12SC01), JCYJ20160229205726699 to Karl Tsim. JCYJ20160229205812004 to Tina Dong.

Abstracts of Oral Presentations

Taxonomic Archive System (TAS) – A Database of New

Coding Methods for Botanical and Herbal Authentication

DTW Lau*#, TH Wong*, SK Wong*, YM Chan*

Shiu-Ying Hu Herbarium, School of Life Sciences, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong, China# Corresponding author

Presenting Author Email : lautaiwai@cuhk.edu.hkCorresponding Author E-mail : lautaiwai@cuhk.edu.hk

Abstract

Authentication is an important first step of herbal quality control, mainly because of the principle in “Origin governs product genuineness”. Authenticating the source plant of a traditional Chinese medicine (TCM) could be multi-directional as many methods have been

developed in recent decades along with new technology and research capacity. Nonetheless, organoleptic characteristics have never been abandoned because of its primitiveness, stability and genuineness in an authentic voucher specimen. Hence, the diagnostic terms of plant description including the major organoleptic characteristics of TCM, can be used in both TCM and their source plants authentication.

Identification skills in plant taxonomy can be applied in TCM authentication under a number of significant situations such as 1.) Authentication of voucher plant specimens, 2.) Species that are not documented in TCM references, 3.) Novel uses of a specific plant organ, 4.) New records of a medicinal plant and 5.) Rare adulterants and substitutes.

This research and educational project is to build up a Taxonomic Archive System (TAS) database with new coding methods from the concepts and capacity of traditional plant taxonomy. This new system would consist of documentation and computational manipulation of thousands of plant characteristics in a web-based database. The major deliverable of TAS is given in form of factsheets and interactive keys for plant authentication.

Consolidated ITS2 Database and Search Engines for Plant and Animal Biomarker Analyses

Hongjun Li1┼, Hong Bai1┼, Maozhen Han1, Kang Ning 1, 2,*

1 Key Laboratory of Molecular Biophysics of the Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, Hubei 430074, China┼ These authors contributed equally to this work.

Presenting Author Email: ningkang@hust.edu.cnCorresponding Author E-mail: ningkang@hust.edu.cn

Abstract

With the development of sequencing technologies, several representative molecular sequences become the standard digital biomarkers for various species on earth. ITS2 is one of such biomarkers for both plant and animals. Though several studies have shown that ITS2 has shown high sensitivity and specificity for differentiating plant and animal

species, there is currently a lack of well-curated ITS2 database and search engine for species identification.

In this study, we have collected more than 200,000 ITS2 sequences representing more than 100,000 species, and manually curated these sequences to ensure that all ITS2 sequences that we have used are of high quality and representative enough for the plant and animal species. The current database contains 149,730 sequences representing 91,014 species and 34,919 genus. We have also designed the dual-engine search scheme for using this database, including the local alignment based search mode (BLAST and UCLUSTER) and the global alignment based search mode (KRAKEN). Our test on several thousands of ITS2 sequences as queries have shown that the search accuracies could reach above 85% and 95%, based on the local and global alignment based search mode, respectively.

Therefore, the ITS2 database represents the most comprehensive ITS2 database, and it also represents the most advanced search engine for biomarker matches, based on which unprecedented accuracies could be achieved for plant and animal species identification based on biomarkers.

Medicinal Materials DNA Barcode Database (MMDBD)

v1.5 - One Stop Solution of Storage, BLAST, Alignment & Primer Design

TH Wong1,3, Grace WC But2, David TW Lau3 and, PC Shaw1,2

1. LDS YYC R & D Centre for Chinese Medicine, 2. Institute of Chinese Medicine3. SY Hu Herbarium, School of Life Sciences, The Chinese University of Hong Kong

Presenting Author Email: wongtinhang@cuhk.edu.hkCorresponding Author E-mail : pcshaw@cuhk.edu.hk

Abstract

Authentication of medicinal materials by DNA technology is increasingly popular in the herbal industry and regulatory bodies. MMDBD v1.0 established by our team in 2010 is the first of its kind to provide a reference database of documenting DNA sequences for

medicinal materials barcoding purpose. This database contains DNA sequences of medicinal materials listed in the Chinese Pharmacopoeia and the US Pharmacopoeia and the common adulterants. The data archive is updated from time to time and now has over 40,000 DNA sequences representing 1,300 medicinal materials. Our team has recently improved the interfaces and incorporated some essential bioinformatics tools to facilitate the authentication work.

DNA sequences of MMDBD v1.5 can be retrieved by Latin names and Chinese character stroke search, etc. It also includes a BLAST based engine for searching DNA sequences. Since primer design is a key step in DNA sequencing, we have integrated the Clustal Omega alignment tool and Primer3 in form of web interface into the original MMDBD. With the detail information such as material name, medical part and pharmacopeia information being shown in the search results, users may now directly check an unknown DNA sequence whether it is included in the Pharmacopoeias and then design appropriate primers for the amplification of a target sequence.

Comparative analysis of The Complete Chloroplast

Genome sequences of Seven Taraxacum Species for Phylogenetic and Taxonomic Study

Jin-Kyung Kim1, Jee Young Park1, Yun Sun Lee1, Hyun Oh Lee2, Hyun-Seung Park1, Sang-ChoonLee1, Jung Hwa Kang3, Taek Joo Lee3, Sang Hyun Sung4 and Tae-Jin Yang1

1. Department of Plant Science, Plant Genomics and Breeding Institute, and Research Institute of Agriculture and Life Sciences, College of Agriculture and Life Sciences, Seoul National University, Seoul, Korea,

2. Phyzen Genome Institute, Seongnam, Korea3. Hantaek Botanical Garden, Yongin, Gyeonggi-Do, Korea, 4College of Pharmacy and

Research Institute of Pharmaceutical Science, Seoul National University, Seoul, Korea

Presenting Author Email : luvjkkim@gmail.comCorresponding Author Email : tjyang@snu.ac.kr

Abstract

The genus Taraxacum, known as dandelion, belongs to the family Asteraceae and widely distributed in the temperate areas of the Northern Hemisphere. More than 2500 species are known in the genus Taraxacum. Taraxacum species have several curative properties including choleretic, diuretic and anti-inflammatory activities and so have long been used for traditional medicinal herbs. The genus Taraxacum is an evolutionary and taxonomical complex taxa, because of complex hybridity and coexistence of agamospermy, which causes great difficulties in the phylogenetic study of this genus. Accordingly, the complete chloroplast genome sequences of seven Taraxacum species were generated by de novo assembly with whole genome sequencing data. The cp genomes of the seven Taraxacumspecies were circular quadripartite DNA with 151,173 bp to 151,451 bp in length, which was similar in range with those from other angiosperms. A total of 112 genes including 79 protein-coding genes, 29 tRNA genes and four rRNA genes were identified in these chloroplast genomes. Genome-wide diversity analysis revealed that around 900 variations including SNPs and InDels and a number of polymorphic sites such as repetitive sequences

and SSRs were observed. The characterized chloroplast genome sequences will be basic genetic resource for phylogenetic analysis, species identification and further molecular study of Taraxacum species with other species in the family Asteraceae. This work was supported by “15172MFDS246” from Ministry of Food and Drug Safety in 2015, Republic of Korea.

The Complete Chloroplast Genome Sequences of Six

Hydrangea Serrata Acessions and Their comparative Analysis for Developing DNA Barcoding Marker

Hana Lee1, Jee Young Park1, Jin-Kyung Kim1, Ho Jun Joh1, Sang-Choon Lee1, Jonghoon Lee2, Jung Hwa Kang3, Taek Joo Lee3, and Tae-Jin Yang1

1. Department of Plant Science, Plant Genomics and Breeding Institute, and Research Institute of Agriculture and Life Sciences, College of Agriculture and Life Sciences, Seoul National University, Seoul, Korea

2. Joeun seeds Co., Ltd, Goesan-gun, Chungbuk, Korea3. Hantaek Botanical Garden, Yongin, Gyeonggi-Do, Korea

Presenting Author Email : hanamingo@hanmail.net Corresponding Author Email : tjyang@snu.ac.kr

Abstract

The Hydrangea genus consists of about 75 species belonging to the Hydrangeaceae family, most of which are cultivated as popular flowering shrubs worldwide. Among those, H. serrata is naturally distributed in mountain regions in Korea, Japan, China and Americas, and furthermore commercially valuable due to its flowers with various colors such as white, blue, and pink. Traditionally, classification of Hydrangea species have mainly been based on morphological characters and thus problematic. Meanwhile, DNA barcoding markers have been considered as a rapid and accurate tool for identification of plant species. On this account, we assembled chloroplast (cp) genomes of six H. serratavarieties and developed DNA barcoding markers based on cp genome sequences. The genomes are the first complete cp genomes in the Hydrangeaceae family and even in the Cornales order. The complete six H. serrata cp genomes ranged from 157,617 bp to 157,730 bp in size and had overall GC contents of 38.5%. The cp genomes were a circular molecules consisting of four distinct regions: inverted repeat (IR) regions of 26,115 bp -26,124 bp, large single copy (LSC) regions of 86,672 bp – 86,789 bp and small single copy (SSC) regions of 18,697 bp – 18,712 bp. Through comparison of cp genomes, polymorphic sites such as SNPs and InDels were identified and used to develop DNA barcoding markers for identifying six H. serrata accessions. This research will provide genomic information for understanding the genetic diversity of Hydrangea species and contributing to natural germplasm conservation strategies. This research was supported by the Bio & Medical Technology Development Program of the NRF funded by the Korean government, MSIP (NRF-2015M3A9A5030733).

DNA barcoding for Species Authentication of Common

Herbal Plants in Malaysia

Tnah L.H., Lee S.L, Tan A.L., Lee, C.T, Ng K.K.S., Ng C.H., Ling S.K., Nurul Farhanah Z.

Forest Research Institute Malaysia, 52109 Kepong, Selangor Darul Ehsan, Malaysia

Presenting Author Email : leehong@frim.gov.myCorresponding Author Email : leehong@frim.gov.my

Abstract

Global market for herbal plants and their related products had become increasingly important over the past two decades. However, along with the growing herbal market, the adulteration of herbal products is rising as a global concern. Adulteration can occur due to misidentification of herbs or intentional substitution with in-expensive herbs or synthetic drugs, which may lead to public health hazard and poor quality product formulations. As the fundamental step to confirm the quality of herbal product is through species authentication, DNA barcoding technique offers a good alternative for detecting plant based adulterants in traded herbal medicinal materials. In the present study, we established a reference DNA barcoding database for 100 common herbal plants in Malaysia for species identification and authentication. DNA barcodes were generated using rbcL and trnH-psbA plastid markers. The performance of the plastid markers in recovering species groups were evaluated using similarity BLAST, phylogenetic tree and barcoding gap. Our analyses showed that rbcL provided apparent resolution at the species level, and was able to separate the majority of the species. To improve the species concepts, trnH-psbA appeared promisingly as a second tier locus. The analysis indicates the potential of using multigene tiered approach for DNA barcoding. Additionally, a blind test of the authenticity for herbal products sampled from local market was conducted. As a whole, authentication of herbal products is challenging, but with the establishment of new DNA barcoding authentication system in Malaysia, this would provide a measure of quality assurance in the herbal industry.

A Molecular and Morphometric Database of Heung Trees Aquilaria sinensis (Lour.) Spreng in Hong Kong

David Tai Wai LAU1,2, Lee Man CHU1, Jerome Ho Lam HUI1,3

1. School of Life Sciences, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong, China

2. Shiu-Ying Hu Herbarium, School of Life Sciences, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong, China

3. State Key Laboratory of Agrobiotechnology,The Chinese University of Hong Kong, Shatin, N.T., Hong Kong, China

Presenting Author Email : jeromehui@cuhk.edu.hk Corresponding Author Email : jeromehui@cuhk.edu.hk

Abstract

The Heung or Incense tree, Aquilaria sinensis, is endemic to the south China and Hong Kong. Indeed, it is also commonly believed that the name of Hong Kong derives its name from this evergreen tree (i.e. Incense Harbour). Under pathological conditions such as natural fungal infection and external wounding, resin is produced which becomes the valuable agarwood (Chen Xiang) that are used as incense and traditional Chinese medicine. Owing to the decline in its distribution due to habitat loss in China, the Heung tree is now categorised under the “List of Wild Plant Under State Protection (Category II)” in mainland China and is defined as vulnerable species under the IUCN Red List of Threatened Species. Nevertheless, Aquilaria sinensis can be commonly found in the countryside of Hong Kong without any major natural threats, which in turn have attracted illegal harvesting and created an increasing threat to local wildlife and citizens. We are now carrying out a large-scale analysis and constructing a morphometric and DNA barcoding database of Heung trees in light to aid its conservation in Hong Kong. Molecular authentication of agarwood (Chen Xiang) for different purposes including its medicinal use will also be discussed in this talk.

A Comparison of Different DNA Barcoding Markers for

Identification of Terminalia Plants and Their Crude Drugs Collected from Thailand

Aekkhaluck Intharuksa1, Hirokazu Ando2, Wannaree Charoensup1, RatchupornSuksathan3, Kittipong Kertsawang3, Panee Sirisa-ard1, Masayuki Mikage4, Yohei Sasaki2

1. Faculty of Pharmacy, Chiang Mai University, Thailand2. Graduate School of MedicalSciences, Kanazawa University, Japan3. Queen Sirikit Botanic Garden, Thailand, 4Department of Agriculture, Tokyo University of

Agriculture, Japan

Presenting Author Email : identity.int@gmail.comCorresponding Author Email : sasaki@p.kanazawa-u.ac.jp

Abstract

[Purpose] Dried fruits of Terminalia chebula and T. bellirica (Combretaceae)are used as the main ingredients in Triphala, a famous polyherbal formulation in Ayurvedicmedicine and Thai folk medicine. The present study aimed to discriminate species in genus Terminalia by DNA barcoding technique and compare chemical profile of medicinal Terminalia fruits.

[Material and Methods] A total of 221 closely related sequences were obtainedfrom nine Terminalia species collected from Thailand and DDBJ database. Aneffective DNA marker for authentication of Terminalia species was selected from six DNA

regions (matK, rbcL, psbA-trnH, ITS, ITS1, and ITS2) and two-locuscombination. All sequences were appraised in three major methods: (1) BLAST (2) genetic method using Kimura 2-parameter (K2P) distance matrices (3) tree topology based on Neighbor-Joining (NJ).

[Results and Discussion] The result of comparison of six candidate DNA locirevealed that ITS identified Terminalia with 100% accuracy at the species and genus level in BLAST1 method. ITS2 ranked first in comparable K2P variabilityconsequences by distance method. The data from single markers and two-locus combination were analyzed and the phylogenetic tree was obtained using the NJ method. No individual markers exhibited clear resolution among nine Terminalia species. But the two-locus combination, ITS + psbA-trnH and rbcL + psbA-trnH clearly discriminated all species. From DNA sequence analysis and the three methods, ITS2 is recommended for the identification of Terminalia species, and psbA-trnH may supplement it. Terminalia crude drugs will be authenticated by ITS2 and evaluated by chemical markers using HPLC in future plan.

Application of Universal DNA Barcode System-Experience with Indian Herbal Industries

Ramalingam Sathishkumar

Plant Genetic Engineering Laboratory, Department of Biotechnology, Bharathiar University,Coimbatore, Tamil Nadu, India.

Presenting Author Email : rsathish@buc.edu.inCorresponding Author Email : rsathish@buc.edu.in

Abstract

DNA based identification methods are routinely used for plant authentication. Recent studies have proved the sensitivity and specificity of DNA barcodes in accurate identification of the botanical species, unknown adulterants and contaminants. But none of these studies have attempted to provide the extent of practicality of this method as quality control standards for herbal industries. Currently, herbal industries have set of standards to assess the quality of bulk plant materials, which is determined by identity, purity and the content of bioactive constituents. Hence, our study involved testing of industrial samples to determine the feasibility of DNA barcodes to authenticate the raw bulk materials and processed products. We tested around 104 commercial samples, which covered 76 species (45 families) and 23 different types of sample materials in the form of fresh, dried, extract and powdered substances. A standard experimental protocol (EP) was used to test all the samples. Among the tested samples, nearly 53% of thesamples from 35 families that covered 17 different types of sample materials were validated with accurate identification. Approximately, 28% of the samples could not be authenticated due to many reasons, 1) degraded DNA from the processed samples; 2) adulteration/contamination ofsamples with more than one species; 3) lack of EP for extracting DNA, PCR amplification and sequencing of certain herbal species and their products etc., From this study, we confirmed that DNA barcode based method can be used to test the specific product. So, it can be concluded that DNA-based tests should be complemented by conventional techniques like phytochemical and botanical morphological (microscopic/macroscopic) for herbal industries.

Development of Loop-mediated Isothermal

Amplification (LAMP) Assay for Rapid Identification of Cannabis Sativa

Masashi Kitamura1, Masako Aragane2, Kou Nakamura2, Tatsushi Adachi3,Kazuhito Watanabe4, Yohei Sasaki1*

1. Laboratory of Molecular Pharmacognosy, Division of Pharmaceutical Sciences, Graduate School of Medical Sciences, Kanazawa University

2. Medicinal Plant Garden, Tokyo Metropolitan Institute of Public HealthFaculty of Pharmaceutical Sciences

3. KanekaCorporation4. Daiichi Universityof Pharmacy.

Presenting Author Email : dna@stu.kanazawa-u.acCorresponding Author Email : sasaki@p.kanazawa-u.ac.jp

Abstract

Tetrahydrocannabinol (THC) is the characteristic psychoactive compound inCannabissativaL.In many countries including Japan, the C. sativaplant is illegal and its cultivation and possession are prohibited by law. C. sativa can bediscriminated from other species by morphological analysis using cystolithic hairs on the leaves or chemical analyses detecting cannabinoids.As thoseanalyses cannot identify the plant in some cases, asimple yet accurate DNA-based method for the identification of C. sativais desired.We have developed a loop-mediated isothermal amplification (LAMP)assay for the rapid identification of C. sativa.1)By optimizing the conditions for the LAMP reaction that targets a highly conserved region of tetrahydrocannabinolic acid(THCA)synthase gene,C. sativawas identified within 40min.The sensitivityof the assay was the same as or higher than that of conventional PCR. The LAMP assay detected all 21specimens of C. sativa, showing high specificity. Using oursimpleprotocol, the identification of C. sativacould be accomplished within 60min from sample treatment to detectionwith aportable equipment.Our rapid, sensitive, specific and simple methodisexpected to be applicableto not only laboratory detection but also on-site detectioninforensic investigationand industrial quality control.1) M. Kitamura, et al., Biol. Pharm. Bull., 2016 (in press)

DNA Barcoding for Authentication of The Commercial

Indian Herbal Tea

Saravanan Mohanasundaram1, Krishnaveni Citheswaran2 and Sathishkumar Ramalingam3*

1,2. Plant Genetic Engineering Laboratory, Department of Biotechnology,Bharathiar University, Tamil Nadu, India

Presenting Author Email : saravananmohan.bu@gmail.comCorresponding Author Email : rsathish@buc.edu.in

Abstract

Herbal infusions of dried plants, commonly known as ‘Herbal Tea’, are among the most widely consumed hot beverage. The popularity of herbal tea is mainly due to its desirable aroma, taste and potential health benefits. Herbal teas are formulated from the flowers, barriers, peels, seeds, leaves and roots of many different plants. Herbal teas are available in

market as a tea bags and ground or fragmented loose products. It is difficult to identify the botanical origin of the blended or processed product by its appearance. To determine the presence of biological contaminants and toxic elements, the herbal tea must be analyzed for the listed ingredients on its label. In this study, DNA barcoding approach was employed for the authentication of herbal tea using ITS2 and rbcLa. Towards this, genomic DNA was isolated and sequenced for all the 11 Biological Reference Materials (BRM) and 15 commercial herbal tea samples. Our study clearly revealed that the contaminants like Trigonella foenum-graecum and Chukrasia tabularis, which is of least economic importance was present in the tea samples, which has not been listed in the product label. Hence, we suggest that DNA barcoding can be incorporated in a QC pipeline for authentication by the herbal tea industries. This would only amount to a minor cost to the herbal tea industries that would result in high quality, authentic product that will boost the consumer’s confidence. This study involved the blind samples purchased over the counter in Indian market.

Molecular Authentication on Processed Herbal Products:

Case Study on Animal-derived Concentrated Chinese Medicine Granules

Yat-Tung Lo1, Li-Li Jiang2, Pang-Chui Shaw1,2,3*

1. Li Dak Sum Yip Yio Chin R & D Centre for Chinese MedicineThe Chinese University of Hong Kong, Shatin, N.T., Hong Kong, China

2. State Key Laboratory of Phytochemistry and Plant Resources in West China, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong, China

3. School of Life Sciences, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong, China

Presenting Author Email : ytlo@cuhk.edu.hkCorresponding Author Email : pcshaw@cuhk.edu.hk

Abstract

With rapid development of the modern Chinese medicine industry, herbal materials are now manufactured in the form of concentrated Chinese medicine granules (CCMG) which offer patients a convenient way of consumption and reduce the risk of damaging of the raw herbs due to insect infestation, moisture or temperature. However, the morphological and microscopic characteristics of the herbs are lost and this poses difficulties in authentication. People usually believe that molecular authentication is not applicable for herbal products due to DNA degradation. This study is thus aimed to evaluate the feasibility of using DNA techniques to authenticate animal-derived CCMG, which has no definite mean for authentication so far. Species-specific primers for two types of CCMG, namely Zaocys and Scorpio, were designed to differentiate the genuine drug from their adulterants. Results showed that DNA fragments with sizes generally less than 200 bp could be isolated and amplified in CCMG. Species-specific primers could be used for diagnosis if there are differences of DNA sequences between the genuine and claimed adulterants. This study thus demonstrated the presence of short-fragment DNA which is available for species identification in CCMG. This work has extended the application of molecular authentication in herbal products and may be further developed into diagnostic kits for quality control and regulatory compliance.

Application of DNA Barcoding To Unravel The

Taxonomical Complexity in Ficus L. (Moraceae) of Western Ghats, India

Mahima Karthikeyan1, Sathishkumar Ramalingam2*, J.V Sudhakar3

1, 2. Plant Genetic Engineering Laboratory, Department of Biotechnology, Bharathiar University

3. Botanical Survey of India, Southern Regional Centre, TNAU Campus, Coimbatore, Tamil Nadu, India

Presenting Author Email : mahimakarthikeyan863@gmail.com Corresponding Author Email : rsathish@buc.edu.in

Abstract

The genus Ficus L. is one of the largest and utmost diverse genera of the flowering plants, with 735 species is distributed in most tropical and subtropical forests of all over the world and in India it is represented by 91 species and 24 infraspecific taxa. The species under the genus Ficus commonly called as ‘Figs’. They are the keystone species in many tropical ecosystems and intimate association between figs and pollinating wasps perhaps represent the reproductively based mutualism, considered as an example for the study of co-evolution. The figs are important as both food and traditional medicine and contain laxative substances, flavonoids, sugars, vitamins A and C, acids and enzymes.

Ficus L. was included in Taxonomically Complex Groups (TCGs), because of large variation

between the habitats. Generally taxonomical parameters like leaves and fruits are frequently used which are very sensitive and dependent upon environmental conditions. Therefore, a simple and accurate molecular identification of Ficus species is indispensable. In this study, DNA barcoding approach was employed for the identification of 8 selected species of Ficusgenus collected from Western Ghats using ITS2 and trnH- psbA. Our study clearly revealed

that ITS2 and psbA-trnH showed 100% amplification and sequencing success. ITS2 showed 100% discrimination between the species F.virens and F.middletonni (complex II), F.amplocarpa and F.guttata (complex III) and F.beghalensis and F.krishnae (complex IV) but not for F.anamalayana and F.dalhousiae (complex I). Hence, F.anamalayana and F.dalhousiae(complex I) were further analyzed with trnH-psbA marker for better understanding. Results showed that plastid gene trnH-psbA performed best in terms of discriminatory power as it contains high variable informative sites (3.34%) in F.anamalayana and F.dalhousiae (complex I), where as ITS2 shows only 0.3%. This study has clearly proved that DNA barcoding is a potential tool that can be used to unravel the taxonomical complex groups. These universal molecular markers will be very useful in conservation and in resolving the numerous operational, logistical and biological questions that are faced by gene bank managers.

Nucleotide Signature Method for The Identification of Chinese Patent Medicines

Yang Liu, Xiaoyue Wang, Zitong Gao, Jianping Han*

Institute of Medicinal Plant Development, Chinese Academy of Medicinal Science &Peking Union Medicinal College, Beijing 100193, P.R. China

Presenting Author Email ：Fkyoung@163.comCorresponding Author Email ： jphan@implad.ac.cn

Abstract

Chinese patent medicines are produced using herbals as raw materials in different dosage forms, including pills, powders and ointment, with different manufacturing processes. Chinese patent medicines have played a big role on the global pharmaceuticals market, however the identification system for these products is not enough, which leads to a serious adulteration problem and put a threat to the consumers’ health. It’s very difficult to identify the ingredients within the complicated compound prescription of Chinese patent medicines with morphological or chemical methods. And because of the serious DNA degradation occurred during the manufacturing processes, traditional DNA-barcoding method is not suitable for dealing with DNA-degraded products. Therefore, a new technique with one extremely short molecular marker for the identification of processed materials is urgently needed. In this study, we developed nucleotide signatures based on SNP sites for American ginseng, Ophiocordyceps Sinensisand Angelicae Sinensis Radix and designed specific primer pairs to amplify those signature sequences from processed materials or products. In order to detect whether the Chinese patent medicines were adulterated or substituted, we also developed nucleotide signatures for some adulterants ,including Eucommia ulmoides (the adulterants of Lonicerae JaponicaeFlos extracts ), Cynomorium songaricum, Cistanche sinensis, Boschniakia rossica, and Orobanche coerulescens (the adulterants of Cistanches Herba). And both the aimed species and adulterated species are successfully identified within different kinds of Chinese patent medicines using nucleotide signature. Thus, the nucleotide signature method, with only 30~37-bp, will broaden the application of DNA barcoding to identify the components in extract powders, Chinese patent medicines and other products with degraded DNA.

Development of DNA Barcode Based CHIP for Visual Detection of Adulterants in Herbal Products

Dhivya Selvaraj and Sathishkumar Ramalingam

Plant Genetic Engineering Laboratory, Department of Biotechnology, Bharathiar University, Coimbatore, India.

Presenting Author Email : bioinfodhivya@gmail.comCorresponding Author Email : rsathish@buc.edu.in

Abstract

Herbal medicine is being revived by day-to-day practice for its long-lasting curative effect, easy availability, natural way of healing, and less side-effects. Today herbal medicines are gaining importance and expanding throughout the world. The adulteration and substitution of the herbal drug is the burning problem in herbal industry. By intentionally or accidentally the crude drugs are substituted with the inferior commercial varieties, which may or may not have any thereupatic potential as that of original drug. Numerous reports of toxic effects contradict the popular view that herbals are natural and therefore harmless. Several herbals have been identified as a cause of acute and chronic hepatitis, cholestasis, drug-induced autoimmunity, vascular lesions, and even hepatic failure due to the presence of pyrrolizidinealkaloids, germander, greater celandine, kava, atractylis gummifera and senna alkaloids. Hepatotoxicity can also result from misidentification or mislabelling of a plant as contaminants. Towards this, we have developed a chip based colorimetric detection approach to distinguish herbal products from its common adulterants. This was achieved by

digesting the PCR products with lambda exonuclease by specific probe for each species in the herbal samples, which are printed on the chip, creating G-quadruplexes. These, G-quadruplexes bind to hemin and catalyze the oxidization of ABT to give a visual green product in the presence of hydrogen peroxide. Depending upon the intensity of the green color, the correct species from the herbal products and the adulterant were detected. DNA barcode chip is an rationalised approach to authenticate the herbal products that can become a useful tool for the quality assurance of drug and health commodities.

Complete Chloroplast Genomes of An Endangered

Species Magnolia Officinalis Subsp. Biloba: Organization and Implications for Selecting Specific Barcodes

Xiwen Li*§, Huanhuan Gao*, Yingjie Zhu, Shilin Chen§

Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China*The authors contributed equally to this work

Presenting Author Email : xwli@icmm.ac.cnCorresponding Author Email : Xiwen Li. (xwli@icmm.ac.cn); Chen, S (slchen@icmm.ac.cn)

Abstract

Despite many investigations of the Magnoliidae, the relationship of Magnoliaceae at low taxonomic level is still not clear. Here we present the complete chloropast (cp) genome sequence of Magnolia officinalis subsp. biloba using 454 high-throughput sequencing technology. Genomic comparisons between M. officinalis susup. biloba and six Magnoliaceae cp genomes reveal that gene content and gene order are similar. They all encode 84 protein genes, 8 rRNA, 37 tRNA, giving a total of 129 genes and 17 of which are duplicated in the IRs. The mVISTA alignment was conducted to screen low intra- species genetic divergence regions and scanned the entire chloroplast genomes of 9 species in Magnoliaceae to search for high inter-species variable regions. A total of 14 highly variable regions were identified including genes and intergenic regions, which were amplified and sequenced to select the suitable candidate barcodes for species identification or classification. With BLAST and the nearest distance methods, data analysis showed that rps15-ycf1was a potential barcode for Magnolia species identification. We also identified SSR loci are rich in Magnolia cp genomes ranging from 218 to 239, which have the potential to be used for further phylogenetic studies in Magnoliaceae. The cp genomes of Magnoliaceae evolved slowly, and phylogenomics based on high variable cp locus in this study might be used to resolve major relationships within one of the major clade of angiosperm.

Unveiling Genome Information of Herbarium Specimens in Next Generation Sequencing Era

Chun-Xia Zeng1, Jun-Bo Yang1, Jing Yang1, Hong-Tao Li1, Zheng-Shan He, Zhi-Rong Zhang, De-Zhu Li1,2*

1. Germplasm Bank of Wild Species, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, Yunnan 650201, China

2. Key Laboratory for Plant Diversity and Biogeography of East Asia, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, Yunnan 650201, China

Presenting Author Email : zengcx@mail.kib.ac.cnCorresponding Author Email : dzl@mail.kib.ac.cn

Abstract

Unlocking the vast genomic diversity stored in natural history collections would create unprecedented opportunities for genome-scale evolutionary, phylogenetic, domestication

and population genomic studies. In today’s next-generation sequencing world, opportunities and prospects for historical DNA have changed dramatically, as most NGS methods are actually designed for taking short fragmented DNA molecules as templates. A “genome skimming” approach, focusing on highly repetitive genome regions such as rDNA or organelle genomes, can yield highly useful sequence data at relatively low sequence depth. Here we used 25 specimens with ages up to 80 years old from 16 different angiosperm families. The minimal (500 pg) input DNA was tested for library construction. We also explored the relationship between the numbers of PCR cycles of library construction and assembled chloroplast coverage. The result show that using a standard multiplex and paired-end Illumina sequencing approach, complete chloroplast sequences and rDNA were assembled for all specimens. We find no significant correlation between chloroplast coverage and PCR cycle times in our samples. Finally, we conclude that chloroplast sequencing from herbarium specimens is feasible and cost-effective, and can be performed with limited sample destruction.

List of Poster Titles

Number Title Institute CountryCorresponding

Author

IC16-A4Endangered Uyghur Medicinal Plant

Ferula Identification Through the Second Internal Transcribed

Institute of Chinese Materia Medica and

Ethnical MateriaChina Xiao Jin Li

IC16-A11DNA Barcoding for Species

Authentication of Common Herbal Plants in Malaysia

Forest Research Institute Malaysia(FRIM)

Malaysia Lee Hong Tnah

IC16-A14

Isolation and Functional Characterization of A Lonicera

Japonica HydroxycinnamoylTransferase Involved in Chlorogenic

Acid Synthesis

Shandong University of Traditional Chinese

MedicineChina Gao Bin Pu

IC16-A15Consolidated ITS2 Database and

Search Engines for Plant and Animal Biomarker Analyses

Huazhong University of Science and Technology

China Kang Ning

IC16-A15

Assessing the Quality of TCM Preparations by Combination of

Chemical Ingredient and Biological Ingredient Analysis: Story for Liuwei

Dihuang Wan

Huazhong University of Science and Technology

China Kang Ning

IC16-A17Application of Universal DNA

Barcode System- Experience with Indian Herbal Industries

Bharathiar University IndiaRamalingam Sathishkumar

IC16-A21

Development of Loop-mediated Isothermal Amplification (LAMP) Assay for Rapid Identification of

Cannabis Sativa

Kanazawa University Japan Yohei Sasaki

IC16-A23

A Comparison of Different DNA Barcoding Markers for Identification

ofTerminalia Plants and Their Crude

Drugs Collected from Thailand.

Chiang Mai University Thailand Yohei Sasaki

IC16-A25

Comparative Analysis of The Complete Chloroplast Genome Sequences of Seven Taraxacum

Species for Phylogenetic and Taxonomic Study

Seoul National University

Korea Tae Jin Yang

IC16-A27

Genetic Diversity of Panax Species Revealed by Chloroplast Genome

Sequences and The Practical Marker Application for The Authentication of

Panax Species

Seoul National University

Korea Tae Jin Yang

Number Title Institute CountryCorresponding

Author

IC16-A28

The Complete Chloroplast Genome Sequences of Six Hydrangea Serrata

Acessions and Their Comparative Analysis for Developing DNA

Barcoding Marker

Seoul National University

Korea Tae Jin Yang

IC16-A31

Identification of Traditional She Medicine Shi-Liang Tea Species and

Closely Related Species Based on the ITS2 Barcode

Institute of Medicinal Plant Development (IMPLAD), Chinese

Academy of Medical Sciences

China Ke Jun Cheng

IC16-A32DNA Barcoding the Commercial

Bombyx Batryticatus

Institute of Medicinal Plant Development (IMPLAD), Chinese

Academy of Medical Sciences

ChinaJing Yuan Song,

Shi Lin Chen

IC16-A33

The Complete Chloroplast Genome Sequence of 4 Species Belong To The Family Asteraceae and Development

of DNA Barcoding Markers for Its Authentication

Seoul National University

Korea Tae Jin Yang

IC16-A35

Potential Chloroplast DNA Barcodes in Highly Complex Plant System for

Herbal Authentication: A Case Study From “Parpadagam” (Genus:

Mollugo; Family Molluginaceae)

SRM University IndiaParani

Madasamy

IC16-A36Development of DNA Barcode Based

CHIP for Visual Detection of Adulterants in Herbal Products

Bharathiar University IndiaSathishkumar Ramalingam

IC16-A37

Authentication of Medicinal Plant Bacopa Monieri Using PCR-based DNA Marker Based on psbA-trnH

Region

Chulalongkorn University

ThailandSuchadaSukrong

IC16-A39

PCR-RFLP Analysis of Derris Reticulata As An Adulterant of

Albizia Myriophylla Purchased in Crude Drug Markets

Chulaongkorn University ThailandSuchadaSukrong

IC16-A41DNA Barcoding for Authentication of The Commercial Indian Herbal Tea

Bharathiar University IndiaSathishkumarRamalingam

Number Title Institute CountryCorresponding

Author

IC16-A48

Complete Chloroplast Genomes of An Endangered Species Magnolia

Officinalis Subsp. Biloba: Organization and Implications for

Selecting Specific Barcodes

Institute of Chinese Materia Medica, Chinese

Academy of Chinese Medical Sciences, China.

ChinaXi Wen Li,

Shi Lin Chen

IC16-A53

Application of DNA Barcoding to Unravel The Taxonomical Complexity

In Ficus L. (Moraceae) of WeaternGhats, India

Bharathiar University IndiaSathishkumarRamalingam

IC16-A54

DNA Identification Reveals Substitution and Mislabeling of Confusing Medicinal Plants and Crude Drugs Purchased in Thai

Markets

Chulalongkorn University ThailandSupawet

Rattanasiriwongwut

IC16-A58

Development of Loop-Mediated Isothermal Amplification (LAMP) for

the Authentication of Transgenic Papaya

The Chinese University of Hong Kong

Hong Kong,China

Pang Chui Shaw

IC16-A59

Authentication of Plant Materials Based on Genomics and DNA

Barcoding To Prohibit Economically Motivated Adulteration

Seoul National University

Korea Tae Jin Yang

IC16-A61Is it possible to identify constituent herbs in decoctions and granules by

molecular approach?

The Chinese University of Hong Kong

Hong Kong,China

Pang Chui Shaw

IC16-A62Using DNA Barcodes To Identify

Plants or Biological Resources: Some Case Studies in Yews (Taxus)

Kunming Institute of Botany, Chinese

Academy of SciencesChina Lian Ming Gao

IC16-A63Influence of Grafting on The

Morphological Variation of Spina Gleditsia Revealed by DNA Barcoding

Shenzhen Institute for Drug Control

China Tie Jie Wang

IC16-A66Nucleotide Signature Method for the

Identification of Chinese Patent Medicines

The Institute of Medicinal Plant Development

China Jian Ping Han

Number Title Institute CountryCorresponding

Author

IC16-A68Unveiling Genome Information of

Herbarium Specimens in Next Generation Sequencing Era

Kunming Institute of Botany, Chinese

Academy of SciencesChina De Zhu Li

IC16-A71Genetical Identification of

Aristolochia Materials by ITS2 DNA Barcoding

Eastern Asia University ThailandPiroonrat

Dechbumroong

IC16-A72

The Application of HRM Combined with DNA Barcoding To Analysis of The Armeniacae Semen Amarum

Mixed in Persicae Semen

Institute of Chinese Materia Medica

China Shi Lin Chen

IC16-A74DNA Barcode Evaluation for The

Genus Ilex in China

South China Botanical Garden, Chinese

Academy of SciencesChina Xue Jun Ge

IC16-A83Celastri Orbiculati Caulis and

Tripterygii Wilfordii CaulisHubei University of Chinese Medicine

ChinaZhi Gang Hu, Shi Lin Chen

IC16-A86DNA Authenticity for Brand

Protection: How Industry Can B0enefit from Product Testing

University of Guelph CanadaAmanda Naaum

IC16-A89

A Nucleotide Signature for The Identification of Angelicae Sinensis

Radix (Danggui)and Its Products

The Institute of Medicinal Plant

Development (IMPLAD)China

Jian Ping Han, Shi Lin Chen

IC16-A90Nucleotide Signature Method for the

Identification of Cistanches Herba (Rou Cong Rong) Products

The Institute of Medicinal Plant

Development (IMPLAD)China Jian Ping Han

IC16-A92An Authenticity Survey of

Commercial Wuweizi UsingITS2 Barcode

Peking union medical college

China Xiaohui Pang

IC16-A93Identification of Herbal Medicines in

Cynanchum Using ITS2 BarcodePeking union medical

collegeChina Xiaohui Pang

Number Title Institute CountryCorresponding

Author

IC16-A103

Application of Sequence Characterized Amplified Region (SCAR) Analysis to Authenticate

Lycium barbarum and Its Adulterants

The University of Hong Kong

Hong Kong,China

Kalin Yan Bo Zhang

IC16-A103

A DNA Microarray for Differentiation of The Chinese Medicinal Herb

Dendrobium Officinale (FengdouShihu) by its 5 S Ribosomal DNA

Intergenic Spacer Region

The University of Hong Kong

Hong Kong,China

Kalin Yan-Bo Zhang

IC16-A125

Taxonomic Archive System (TAS) – A Database of New Coding Methods

for Botanical and Herbal Authentication

The Chinese University of Hong Kong

Hong Kong,China

Tai Wai Lau

IC16-A126

Medicinal Materials DNA Barcode Database (MMDBD) v1.5 - One stop

solution of BLAST, Alignment & Primer Design for TCM

The Chinese University of Hong Kong

Hong Kong,China

Pang Chui Shaw

IC16-A127A Molecular and Morphometric

Database of Heung Trees AquilariaSinensis (Lour.) Spreng in Hong Kong

The Chinese University of Hong Kong

Hong Kong,China

Ho Lam Hui

IC16-A128The limitations of DNA approach in Quality Control of Herbal Materials

The Chinese University of Hong Kong

Hong Kong,China

Pang Chui Shaw

IC16-A129Rapid Authentication of Medicinal

Cordyceps by Loop-mediated Isothermal Amplification

The Chinese University of Hong Kong

Hong Kong,China

Pang Chui Shaw

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