part i: general information -...

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PROFORMA – I

PROFORMA FOR SUBMISSION OF PROJECT PROPOSALS ON RESEARCH AND DEVELOPMENT, PROGRAMME SUPPORT

(To be filled by the applicant)

PART I: GENERAL INFORMATION 1. Name of the Institute/University/ Organization submitting the Project Proposal:

Division of Genetics, Indian Agricultural Research Institute, New Delhi 110 012

2. State: Delhi 3. Status of the Institute: ICAR Institute (Please see Annexure-I) 4. Name and designation of the Executive Authority of the Institute/University forwarding the application:

Dr. H.S. Gupta Director Indian Agricultural Research Institute, New Delhi 110 012

5. Project Title:

MOLECULAR TAGGING OF FERTILITY RESTORATION GENE(S) AND MARKER FACILITATED DIVERSIFICATION OF RESTORER LINES IN INDIAN MUSTARD (Brassica juncea)

6. Category of the Project (Please tick) :.R&D/ Programme Support 7. Specific Area (Please see Annexure-II): Plant Molecular Biology

(Agriculture & Allied area) 8. Duration: 3 Years 9. Total Cost: (Rs.): 190.90 lakhs 10. Is the project Single Institutional or Multiple-Institutional (S/M)? : M 11. If the project is multi-institutional, please furnish the following:

Name of Project Coordinator: Dr. Naveen Singh, Senior Scientist

Affiliation: Division of Genetics, IARI

Address: Dr. Naveen Singh, Senior Scientist, Division of Genetics, Indian Agricultural Research Institute, New Delhi-110012

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12. Scope of application indicating anticipated product and processes Identification of a large set of polymorphic DNA markers is the first step towards their

utilization in marker assisted breeding. A good number of DNA markers evenly distributed across the genome are required to distinguish genotypes available within the working germplasm. Despite the discovery of a large number of markers in B juncea very few markers are useful in breeding. These markers are suitable for differentiating diverse germplasm of different origin. However, their application in routine breeding is limited due to low levels of polymorphism among Indian mustard accessions. Therefore, identification of additional polymorphic DNA markers in Brassica juncea is need of hour for successful utilization in marker assisted selection (MAS).

Fertility-restoring nuclear gene(s) for alloplasmic Cytoplasmic Male Sterility (CMS) systems are generally not available in natural populations and have to be introgressed from the cytoplasm-donor species. At IARI, CMS lines of B. juncea carrying alien cytoplasms of Moricandia arvensis (mori), Diplotaxis erucoides (eru), Diplotaxis berthautii (ber) and Erucastrum canariense (can) are being developed in the nuclear background of elite good combining lines and these derivatives are in BC3-BC4 generation. The fertility restoring (Rf) genes for the above cytoplasms have been introgressed into B. juncea background but are at present not available in improved backgrounds. The transfer of Rf gene using conventional approach needs 6-7 cycles of backcrossing which is both time consuming and highly labour intensive. Availability of workable molecular markers linked to the Rf gene(s) can help track the gene in individual plants of backcross (BC) generations, saving the time and labour for gene(s) transfer to any genetic background. MAS would cut down the time required for restorer line diversification, in addition to ensuring the presence of Rf gene(s) in BC generations. Linkage drag is minimal through marker assisted background selection. Once tightly linked DNA markers are identified and mapped, these can be further used for cloning of Rf gene(s). Availability of a series of fertility restorer lines, at the end of project, would help in identification of highly heterotic combinations and development of a reliable commercial hybrid seed production system.

At NRCPB, new CMS lines have been isolated from advanced backcross progenies of interspecific hybrids. Diversity of new CMS sources is desirable to avoid uniformity of the CMS background. These new CMS lines will be characterized to establish their mitochondrial and plastid genome constitution and to identify the mt-genes associated with CMS. The anticipated products and processes are as follows: • Polymorphic DNA markers which can differentiate among Indian B. juncea genotypes. • Availability of workable closely linked DNA markers for tracking Rf genes. • Well characterized new CMS lines.

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13. Project Summary (Not to exceed one page. Please use separate sheet)

Hybrid breeding involves identification of heterotic combinations and then converting the parents into CMS and restorer lines. This takes about 10 years. The progress achieved in the interim in pureline breeding overshadows the gains offered by hybrids which are based on combinations identified 10 years earlier. Any breeding option which can reduce the time for conversion of parental lines and floating the fertility restorer gene(s) into large number of diverse germplams would help in overcoming this limitation. A large number of molecular markers reported in B. juncea are suitable for diversity analysis and fingerprinting of germplasm but are unsuitable for routine breeding as they show low levels of polymorphism among Indian mustard accessions. Therefore, background selection cannot be effectively integrated with Marker Assisted Backcross Breeding (MABB). Availability of a large number of polymorphic markers would help in achieving this goal.

Tagging of Rf genes with molecular marker(s) would help in more efficient transfer of these gene(s) to suitable genetic backgrounds. Once DNA markers tightly linked to the desired Rf gene(s) are identified and mapped, they can also be used for cloning of these gene(s). MABB also provides us the opportunity for pyramiding Rf genes in the improved genetic backgrounds.

At IARI in collaboration with NRCPB, nuclear genes from fifteen improved B. juncea genotypes are being transferred to the four sterile cytoplasms viz., mori, eru, ber and can for development of CMS lines in elite parental backgrounds giving highly heterotic hybrids. These lines are in BC2 and BC5 generations and needs to be characterized once they become stable. The list of these CMS lines are as follows:

Donor Cytoplasm

Maintainers under conversion Generation

Moricandia arvensis

Pusa Agrani, Pusa Kisan, SEJ-8, NPJ-93, Pusa Jagannath, NPJ-112, NPJ-139, RGN-48, LES 39, LES-1-27, NRCDR-2, Pusa Bahar, Pusa Tarak, EC-308575, EC 598325 and BioYSR

BC2-BC5

Erucastrum canariense


BC2-BC5

Diplotaxis erucoides


BC2-BC5

Diplotaxis berthautii


BC2-BC5

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Mori restorer also restores fertility in CMS lines derived from eru and berth CMS sources; however, the restorer for can is different. The restorer lines developed in B. juncea background possesses normal phenotypes and exhibit regular meiosis with normal fertility, indicating stable introgression. Fertility restoration in these cytosterility sources is monogenic and gametophytic (mori, eru, and ber) or sporophytic (can) (Prakash et al 2001; Bhat et. al. 2008). The restorer genes available in B. juncea background, however, have yet not been transferred in improved backgrounds as yet, for restoring fertility in the above cytoplasms.

The transfer of Fertility Restorer (Rf) gene using conventional approach, needs making backcrosses which is time consuming and labour intensive job. Although the Rf gene, which restores fertility in CMS developed from mori has been tagged using a SCAR marker (Ashutosh et. al. 2007), but the marker is not polymorphic among elite genetic backgrounds and its position on the chromosome is not yet known. Moreover, no report on their utilization through MAS is available. The fertility restorer gene(s) for the cytoplasms taken in this study have not been cloned or mapped as yet.

More than 3000 chromosome specific microsatellite markers are available in Brassica which need to be validated in active B. juncea genetic stocks. Availability of workable molecular markers, like SSRs, would help in practicing background selection and can be associated with the trait like fertility restoration. Any marker linked with the Rf gene can track it in individual plants of backcross generations, and thus will help in precise transfer of this gene(s) to any genetic background. Moreover, the time required for restorer and CMS line development can be cut down using of MAS. Information and material emanating from this study would help in fast development and characterization of a series of fertility restorer and CMS lines. This would further help in diversification of male sterile and restorer lines in B. juncea. Therefore, the present study is proposed with the following objectives: 1. Identification of polymorphic SSR markers for development of genome-wide covered

framework in AABB genomes for use as polymorphic background selection markers (IARI & NRCPB, New Delhi)

2. Tagging of Rf gene(s) functional to restore fertility in hybrids from four alien cytoplasms with DNA markers (IARI and NRCPB, New Delhi)

3. Convert productive heterotic male parental lines identified earlier with Rf genes using MABB (IARI, New Delhi)

4. Characterization of male-sterility related genomic regions in the cytoplasm of CMS lines (NRCPB, New Delhi)

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PART II: PARTICULARS OF INVESTIGATORS (One or more co-investigators are preferred in every project. Inclusion of co-investigator(s) is mandatory for investigators retiring before completion of the project) Principal Investigator: 14. Name: Dr. Naveen Singh

Date of Birth:16.03.1974 Sex (M/F): Male Designation: Senior Scientist Department: Genetics Institute/University: Indian Agricultural Research Institute Address: Division of Genetics Indian Agricultural Research Institute, New Delhi PIN: 110 012. Telephone: 011 25841481 Fax: 011 25846420 E-mail: [email protected] Number of research projects being handled at present: Two (Institutional)

Co-Principal Investigator 15a. Name: Dr. D.K. Yadava

Date of Birth: 10.04.1966 Sex (M/F): Male Designation: Senior Scientist Department: Genetics Institute/University: Indian Agricultural Research Institute Address: Division of Genetics Indian Agricultural Research Institute, New Delhi PIN: 110 012. Telephone: 011 25841481 Fax: 011 25846420 E-mail: [email protected] Number of research projects being handled at present: Two

15b. Name : Dr. (Mrs.) Sujata

Date of Birth : 07.10.1962 Sex(M/F) : Female Designation : Principal Scientist Department : Genetics Institute/University: Indian Agricultural Research Institute Address: Division of Genetics, Indian Agricultural Research Institute, New Delhi PIN: 110 012. Telephone : 011 25841481 Fax 011 25846420 E-mail : [email protected] Number of Research projects being handled at present: Two

15c. Name : Dr. S.R. Bhat

Date of Birth : 22.02.1954 Sex(M/F) : Male Designation : Principal Scientist Department : NRC on Plant Biotechnology Institute/University: NRC on Plant Biotechnology Address: NRC on Plant Biotechnology, Indian Agricultural Research Institute, New Delhi PIN: 110 012. Telephone : 011 25841787 Fax 011 25843984 E-mail : [email protected] Number of Research projects being handled at present: Two

mailto:[email protected]




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15d. Name : Dr. Naveen Chander Gupta

Date of Birth : Sex(M/F) : Male Designation : Scientist Department : NRC on Plant Biotechnology Institute/University: NRC on Plant Biotechnology Address: NRC on Plant Biotechnology, Indian Agricultural Research Institute, New Delhi PIN: 110 012. Telephone : 011 25841787 Fax 011 25843984 E-mail : [email protected] Number of Research projects being handled at present: One

15e. Name: Dr. K.V. Prabhu

Date of Birth : 20-05-1958 Sex (M/F) :Male Designation : Head Department : Genetics Institute/University: Indian Agricultural Research Institute Address : Division of Genetics Indian Agricultural Research Institute, New Delhi PIN: 110 012. Telephone : 011 25841481 Fax: 25846420 E-mail : [email protected] Number of Research projects being handled at present: Six

Note : Use separate page, if more investigators are involved


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PART III : TECHNICAL DETAILS OF PROJECT (Under the following heads on separate sheets)

16. Introduction (not to exceed 2 pages or 1000 words) 16.1 Origin of the proposal

Production and productivity of rapeseed-mustard in India is almost static since last decade, despite availability of improved high yielding varieties. Productivity is hovering around 1.0 t/ha, which is much below the world average of 1.98 t/ha (FAOSTAT 2009). Heterosis breeding is regarded as one of the most effective means to break the yield barrier. Heterosis in oilseed Brassica can be exploited commercially with the availability of highly effective hybrid seed production. A number of hybrids of rapeseed-mustard are being cultivated in Europe, Canada and Australia (Prakash et al. 2008). Hybrid mustard is most relevant in India for a) enlarging yield potential b) developing short duration hybrids suitable for multiple crop rotations and c) development of canola quality hybrids. These factors shall enable cultivation of mustard in poorly irrigated/rainfed areas of central and peninsular India and hence reduce water requirement of the non-traditional mustard regions. With the availability of suitable hybrids, this crop is having potential to increase area under its cultivation, and the total national production can comfortably be targeted from 8 million tons to 12million tons.

Recent progress in heterosis breeding led to development of a few commercial hybrids in

India. However, the yield increment is marginal and these mustard hybrids have not gained popularity. A major reason for slow progress in heterosis breeding is that the fertility-restoring nuclear gene(s) for alloplasmic CMS systems are generally not available in active germplasm of Brassica spp. and need to be transferred from the initial introgressed lines, which is time consuming, labour intensive and tedious job.

Hybrid programme in Indian mustard generally focuses on identification of heterotic

combinations and then converting the parents of these crosses into CMS and restorer lines which takes about 10 years. In the mean time, the progress achieved by the newly developed improved pure lines overshadows the gain achieved by such commercial hybrids of the same period. Any breeding option which can reduce the time for conversion of parental lines and floating the fertility restorer gene(s) in large number of diverse germplasm lines would help in reducing the time span with assured result. Tagging of fertility restorer genes and their transfer through MAS into any genetic background would save time and efforts of the breeders and also ensure the presence of Rf gene(s) in BC generations. The linkage drag can also be minimized through background selection. Furthermore, CMS lines need to be characterized for their identification and purity testing of the hybrid seed. Identification and mapping of DNA markers tightly linked to Rf gene(s) would help in map-based cloning of the Rf gene(s).

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16.2 (a) Rationale of the study supported by cited literature (b) Hypothesis (c) Key questions:

(a) Rationale: Although a large number of SSR and Intron spanning STMS markers have been reported in B. juncea, their utility in routine breeding is hampered by the low levels of polymorphism among local accessions. Identification of molecular markers which can differentiate the working set of Indian B. juncea genotypes and those linked to male fertility restorer genes in different CMS sources would help in their marker facilitated transfer to good combiner parental lines, which will save time and efforts of the breeders. This ensures the presence of Rf gene(s) in BC generations. Furthermore, once the DNA markers tightly linked to the Rf gene(s) are identified, they can be effectively used for cloning. Therefore, identification of highly polymorphic SSR markers is a priority for MAS. This study thus, aims at addressing the research gaps hindering the hybrid breeding programmes in rapeseed-mustard in India.

(b) Hypothesis: The fertility restoration is governed by single dominant gene acting in gametophytic manner for restoring fertility in CMS lines carrying mitochondrial genomes of M. arvensis, D. erucoides, D. berthautii whereas in E. canariense CMS system, fertility restoration is sporophytic, monogenic and dominant. AFLP markers linked to the fertility restorer gene introgressed from M. arvensis was identified and converted to SCAR marker for its utilization in MAS. However, this SCAR marker is dominant and does not show polymorphism with most of the varieties. Markers for canariense restorer gene have not been identified so far. Identification SSR markers showing high polymorphism among accessions will not only help in tagging of Rf genes but also will aid MAS.

The hypotheses of the proposed study, which will be put to test is: Molecular markers giving high degree of polymorphism among indigenous accessions of B. juncea if identified could accelerate diversification of CMS and fertility restorer lines in this species. Further, identification of marker tightly linked to the Rf gene would help in cloning of Rf genes. Thus in this study:

• DNA marker(s) close to the Rf genes will be identified

• Attempts will be made to convert previously reported dominant SCAR marker into co-dominant marker

• Markers (SSR) that show high polymorphism among B. juncea accessions will be identified for use in MAS

• Molecular characterization of new CMS lines for mitochondrial and plastid genome constitution and expression.

(c) Key Questions:

The transfer of Fertility Restorer (Rf) gene(s), using conventional approach, needs making back crosses which is time consuming and labour intensive job. The Rf gene, which restores fertility in CMS developed from M. arvensis has been tagged with a SCAR marker (Ashutosh et al. 2007). However, this marker is not polymorphic over genetic backgrounds and its position on

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the chromosome is not known. The fertility restorer gene(s) for other cytoplasms taken in this study has/have not been mapped yet. As no tightly linked marker to the Rf gene has been reported in B. juncea, hence, cloning of Rf gene has not been achieved in this species. Chromosome specific SSR primers are available now in B. juncea (www.brassica.info/resource/markers/ssr-exchange.php), therefore, availability of workable molecular markers like SSRs would help in efficient transfer of Rf gene(s), through background and foreground selection, thus saving the time and labour for the transfer of this trait to any genetic background. Availability of these co-dominant markers can differentiate the homozygous and heterozygous genotypes and reduce the time requirement for restorer development through Marker Assisted Selection (MAS). Information and material emanating from this study would help in fast development of fertility restorers and characterization of a series of CMS lines. This would further help in diversification of male sterile and restorer lines in B. juncea.

The key questions to be addressed in the proposed project are: • Are there adequate SSR markers that show high polymorphism among accessions of B.

juncea? If so, what are those? If not, what other markers should be explored? • Whether the already reported dominant SCAR marker can be converted into co-dominant

marker that will work in different backgrounds? • Is MAS approach efficient and cost effective for CMS and restorer line diversification? 16.5 Current status of research and development in the subject (both international and national status)

Genetic variability is of prime importance for the improvement of many crop species including Brassica. Molecular markers are the best tools for differentiating genotypes. A variety of molecular markers have been used to study the extent of genetic variation among the diverse group in the genus Brassica. These include restriction fragment length polymorphism (RFLP), simple sequence repeats (SSR), random amplification of polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP). These techniques differ in their principles and generate varying amounts of data. RFLP analysis is labour intensive, time consuming and expensive. There is increasing number of reports where Random Amplified Polymorphic DNAs (RAPDs) have been successfully used to estimate genetic variability in Brassica. Genic microsatellite markers, also known as functional markers, are preferred over anonymous markers as they reveal the variation in transcribed genes among individuals. Microsatellite or simple sequence repeat (SSR) markers are routinely used for tagging genes and assessing genetic diversity. In spite of their importance, there are limited numbers of polymorphic SSR markers available for Brassica crops.

Fertility-restoring nuclear gene(s) for alloplasmic CMS systems are generally not

available in natural populations and thus must be introgressed from the cytoplasm-donor species. Homoeologous chromosome pairing occurs between the chromosomes of alien genome carrying the fertility restoration gene(s) and crop species resulting into the introgression of Rf gene(s). Raphanus/Ogu was the first CMS to be exploited for commercial purposes. No restorer was found

http://www.brassica.info/resource/markers/ssr-exchange.php


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for this system in any Brassica spp. (Rouselle, 1982). However, restorer genes were identified in European radish cultivars (Bonnet, 1975) and were introgressed through synthetic Raphanobrassica (Raphanus sativus × B. napus) and (R. sativus × B. oleracea) by Heyn (1976) and Rouselle and Dosba (1985), respectively. Restorers were successfully obtained for (Raphanus) B. napus (Paulman and Röbbelen, 1988), (Tournefortii) B. napus (Stiewe and Röbbelen, 1994), (Moricandia) B. juncea (Prakash et al., 1998), (Trachystoma) B. juncea (Kirti et al., 1997), (Canariense) B. juncea (Prakash et al., 2001), and (Enarthrocarpus) B. rapa, B. juncea, and B. napus (Banga et al., 2003; Deol et. al., 2003). These restorer lines possess normal phenotypes and exhibit regular meiosis with normal pollen and seed fertility, indicating that introgressed genetic material is fully integrated with the recipient species chromosomes. Rf genes for (Diplotaxis muralis) B. rapa were frequently found in B. rapa (Hinata and Konno, 1979).

In almost all cases, isolation of Rf genes has been accomplished using linked molecular

markers. For tagging of traits, molecular markers such as random amplified polymorphic DNA (RAPD, Williams et al. 1990) and amplified fragment length polymorphism (AFLP, Vos et al. 1995) have been effectively used in combination with near isogenic lines (NILs; Muehlbauer et al. 1988) or bulk segregant analysis (BSA; Michelmore et al. 1991). Delourme et al. (1994) employed BSA approach and identified four RAPD markers linked to the fertility restorer locus (Rfo) of (ogu) .B napus. These markers were cloned and sequenced to develop co-dominant SCAR markers. The region around the Rfo gene was further saturated (Delourme et al. 1998) and a marker RAPD-13, was identified that co-segregated with the Rfo locus. Similarly, Hanson et al. (1997) have also reported a RAPD marker tightly linked to the Rfo locus. Imai et al. (2003) identified four RAPD-STS and six AFLP-STS markers linked to the fertility restorer Rfk1 of CMS Kosena radish. AFLP technology combined with BSA enabled identification and positional cloning of the Rfk1 gene (Koizuka et al. 2003). Similarly, Jean et al. (1997) identified 11 DNA markers (10 RFLP and 1 RAPD marker) linked to the Rfp1 locus of pol CMS B. napus by BSA. For (tour) B. napus restorer gene, Trendelkamp et al. (1999) identified 11 AFLP markers linked to the restorer gene through BSA method, whereas Janeja et al. (2003b) found 2 AFLP markers using NILs. Likewise, two RAPD markers flanking the Rf1 locus of (Enarthrocarpus lyratus) B. napus were identified (Janeja et al. 2003a). Although the Rf gene, which restores fertility in CMS developed from Moricandia arvensis is tagged using SCAR marker (Ashutosh et. al. 2007), however, the marker is not polymorphic among elite genetic backgrounds. Moreover, no report on utilization of this marker through MAS is available. On the other hand primers sequence of SSRs, a co-dominant marker, is available in public domain (www.brassica.info/resource/markers/ssr-exchange.php).

Moricandia arvensis restorer is unique. It restores fertility in four CMS systems:

(Moricandia) B. juncea, (Catholica) B. juncea, (Erucoides) B. juncea, and (Berthautii) B. juncea (Pathania et al., 2003; Bhat et al., 2005; 2006; 2008). In (Catholica) B. juncea, derived from sexual hybridization, it is sporophytic (Bhat et al., 2005) while in other three systems, the restorer function is required at the gametophytic stage. Moricandia restorer is an introgression from M. arvensis into B. juncea, it may carry more than one restorer gene at this locus.



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The restorer genes in Raphanus/Ogu and Kosena are identical. In Raphanus/Ogu, genetic studies indicate the involvement of two genes; however, one dominant allele of the restorer gene Rfo can fully restore the cybrids (Pellan-Delourme, 1986; Pelletier et al., 1987). Delourme et al. (1994b) subsequently introgressed this gene to cybrid lines of B. rapa and B. juncea. Two nuclear genes Rfk1 and Rfk2 are required to restore Kosena CMS in radish and were introgressed into B. napus following protoplast fusion (Sakai et al., 1996). However, in B. napus cybrids, only one gene (Rfk2) could restore fertility (Koizuka et al., 2000). Listed below are some of the salient achievements during recent years:

• Fairly saturated linkage map in Brassica juncea is available in public domain which will be used in this study (www.brassica.info/resource /markers/ssr-exchange.php)

• Genetics of fertility restoration has been worked out and monogenic and gametophytic control of Rf genes responsible for restoration of fertility in M. arvensis, D. erucoides and D. berthautii cytoplasms was observed, however, in E. canariense CMS system fertility restoration is sporophytic, monogenic and dominant.

• Rf genes introgressed from the cytoplasm-donor species to B. juncea background. • AFLP markers linked to the male fertility restorer gene for M. arvensis cyto-sterility source

and converted to SCAR marker.

16.6 The relevance and expected outcome of the proposed study

A number of hybrid varieties of rapeseed-mustard are being cultivated in Europe, Canada and Australia (Prakash et al. 2008). A good level of economic heterosis is reported in B. juncea and B. napus in India too. Pradhan et al. (1993) reported 29.4 to 91.8% heterosis over better parent for two crosses over two seasons. Heterosis to the extent of 31.2 -71.3 % was also reported by Katiyar et al. (2000). At IARI, we have identified a large number of heterotic combinations in Indian mustard that recorded more than 25% yield superiority over the best checks (Yadava et al. communicated). Recent progress in heterosis breeding led to development of a few commercial hybrids of B. juncea in India viz., NRCHB 506, DMH 1, Coral 432 and Coral 437. However, the level of yield gain from these hybrids is marginal hence they are not getting popularity over the pure lines. Since fertility-restoring nuclear gene(s) for alloplasmic CMS systems are generally not available in Brassica spp. germplasm, hence it needs to be introgressed from the initial introgressed lines which is time consuming, labour intensive and tedious job. As and when heterotic combinations are identified their conversion into A and R lines is initiated and in the mean time progress achieved by the heterotic gains through the hybrids is nullified by the newly developed pure lines released during that period. MAS facilitated transfer of Rf genes into the parental lines would help reduce the number of back crosses by allowing background and foreground selections, thus saving time and efforts of the breeders. Although, a large number of markers have been reported in B. juncea, however, there are very few workable markers available. These markers work well only for differentiating diverse germplasm from different origin, however, their application in routine breeding is limited due to low levels of polymorphism

http://www.brassica.info/resource /markers/ssr-exchange.php

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among Indian mustard accessions. Therefore, The objective of proposed study is to identify additional polymorphic DNA markers and to tag the Rf gene(s). The information emanating from this study can further be used for cloning of these genes. Molecular characterization of CMS lines would help in their easy identification and maintenance of genetic purity. This study thus, aims at addressing the research gaps hindering the hybrid breeding programmes in rapeseed-mustard in India will accelerate the use of CMS systems in developing heterotic hybrid varieties. Expected outcome: • Additional polymorphic DNA markers. • Availability of workable DNA markers for MAS and tagging of Rf gene(s) for restoring

fertility in Moricandia arvensis, Diplotaxis erucoides, Diplotaxis berthautii and Erucastrum canariense cyto-sterility sources.

• Molecular characterized newly developed CMS lines.

16.7 Preliminary work done so far

At IARI, in collaboration with NRCPB, nuclear genes from fifteen improved B. juncea genotypes are being transferred to the four male sterile cytoplasmic backgrounds viz., mori, eru, bar and can for development of CMS lines in elite backgrounds and these lines are in BC2 and BC5 generations. The restorer genes, for restoring fertility in the above cytoplasms, are available in B. juncea background and for their transfer to improved backgrounds, the backcrossing programme has been initiated. During Rabi 2010-11, eight recipient parents viz., Pusa Vijay, Pusa Jagannath, Pusa Agarni, Pusa Mustard 25, SEJ 8, Pusa Mustard 21, Pusa Mustard 22 and RH 30 were crossed with the four fertility restorer donors for the above mentioned cyto-sterility sources and the F1 seeds were harvested. The BC1 F1 crosses will be made during rabi 2011-12 for the development of mapping populations. In addition, new CMS and fertility restorer lines have been recently isolated from advance backcross progenies of somatic hybrids.

17. Specific objectives (should be written in bulleted form, a short paragraph indicating the methods to be followed for achieving the objective and verifiable indicators of progress should follow each specific objective)

1. Identification of polymorphic SSR markers for development of genome-wide covered framework in AABB genomes for use as polymorphic background selection markers (IARI & NRCPB, New Delhi)

2. Tagging of Rf gene(s) functional to restore fertility in hybrids from four alien cytoplasms with DNA markers (IARI and NRCPB, New Delhi)

3. Convert productive heterotic male parental lines identified earlier with Rf genes using MABB (IARI, New Delhi)

4. Characterization of male-sterility related genomic regions in the cytoplasm of CMS lines (NRCPB, New Delhi)

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1. Identification of polymorphic SSR markers for development of genome-wide covered framework in AABB genomes for use as polymorphic background selection markers (IARI & NRCPB, New Delhi) Method: Readily available DNA markers, especially microsatellite markers, will be used to study the parental polymorphism in 100 genotypes of Indian mustard. SSR primers are available in public domain (www.brassica.info/resource/markers/ssr-exchange.php) will be tested to identify the informative markers. Verifiable indicators: Informative/ workable markers

2. Tagging of Rf gene(s) functional to restore fertility in hybrids from four alien cytoplasms with DNA markers (IARI and NRCPB, New Delhi) Method: Readily available DNA markers, especially microsatellite markers, will be used to tag the Rf genes. Parental polymorphism will be studied using the SSR primers available in public domain (www.brassica.info/resource/markers/ssr-exchange.php) to identify the informative primers. Pollen from individual plants will be examined under microscope for phenotyping for fertility/sterility by staining with 2% acetocarmine. For mapping the Rf genes plants from BC1F1 or F2 will be used to construct the sterile bulks and the plants carrying fertile pollens will be used as the fertile bulks. Fertile and sterile bulks, thus, prepared for BSA by pooling equal quantity of DNA from ten fertile and ten sterile individuals of each population, respectively will be genotyped using informative primers. The SSR primers differentiating the bulks will be used for genotyping individual plant and data will be analyzed using suitable software. SCAR marker linked to Rf locus responsible for fertility restoration in M. arvensis (Ashutosh et al. 2007), will be tested for validation in the BC1F1 populations thus generated. Verifiable indicators: New DNA marker(s) linked to Rf gene(s) which restorers fertility in mori, eru, ber and can cyto-sterility sources will be available.

3. Convert productive heterotic male parental lines identified earlier with Rf genes using MABB (IARI, New Delhi) Method: Foreground and background selection will be attempted in BC3 and BC4 for transferring fertility restoration through marker assisted backcross breeding in eight heterotic parental lines viz., Pusa Vijay, Pusa Jagannath, Pusa Agarni, Pusa Mustard 25, SEJ 8, Pusa Mustard 21, Pusa Mustard 22 and RH 30.

Verifiable indicators: Fertility restorer gene(s) in elite genetic backgrounds.

4. Characterization of male-sterility related genomic regions in the cytoplasm of CMS

lines (NRCPB, New Delhi)

Method: Newly developed CMS lines will be characterized through Southern hybridization to identify their mt- and plastid genome constitution and the genes associated with CMS will be identified through northern hybridization with mt- gene probes.

Verifiable indicators: Molecular fingerprints of CMS lines.



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18. Work Plan: should not exceed 3-4 pages (the section can be divided according to the specific aims

and under each specific aim, the following should be stated clearly as sub headings)

18.1 Work plan (methodology/experimental design to accomplish the stated aim)

First year: a) Main season (IARI, New Delhi): i) Raising of of 100 B. juncea genotypes in the field ii) DNA isolation of about 100 genotypes. Genotypic polymorphism will be studied using the

SSR primers available in public domain (www.brassica.info/resource/markers/ssr-exchange.php).

iii) DNA of BC1F1 / F2 plants representing each of the mapping populations, generated for different cyto-sterility sources, will be isolated. DNA from these plants will be used to construct the sterile and fertile bulks.

iv) Phenotyping of the mapping population using microscope for fertility/sterility by staining with 2% acetocarmine.

v) Genetic analysis of fertility restoration in newly isolated CMS lines. vi) Southern analysis of new CMS lines. Off season: (IARI New Delhi and IARI,RS, Wellington)

i) Genotypic polymorphism will be studied using the SSR primers Second year: Main season (IARI, New Delhi and NRCPB, New Delhi): i) Genotypic polymorphism will be studied using the SSR primers ii) Using informative SSR primers BC1F1 plants will be genotyped. Bulk segregant analysis

method (Michelmore et al., 1991) will be used to identify the markers and tag the genes. ii) Marker assisted back crossing programme (BC2) will be undertaken to transfer Rf genes

into eight good combining parental lines viz., Pusa Vijay, Pusa Jagannath, Pusa Agarni, Pusa Mustard 25, SEJ 8, Pusa Mustard 21, Pusa Mustard 22 and RH 30.

iii) Northern analysis of new CMS lines for identification of genes associated with CMS. Off season: (IARI and NRCPB, New Delhi) i) Genotypic polymorphism will be studied using the SSR primers ii) Identification of additional closely linked DNA markers for Rf gene(s) iv) Attempting BC3 with the positive plants to transfer Rf gene(s) into eight good combining

parental lines viz., Pusa Vijay, Pusa Jagannath, Pusa Agarni, Pusa Mustard 25, SEJ 8, Pusa Mustard 21, Pusa Mustard 22 and RH 30.

iii) Molecular characterization of newly developed CMS lines using informative markers. Third year: Main season (IARI and NRCPB, New Delhi):

i) Genotypic polymorphism will be studied using the SSR primers ii) Marker assisted back crossing programme (BC4) will be undertaken to transfer Rf genes

into ten good combining parental lines viz., Pusa Vijay, Pusa Jagannath, NRCDR 02, RGN 145, Pusa Agarni, Pusa Mustard 25, SEJ 8, RH 30, Pusa Bahar and Pusa Mustard 28.



15

iii) Foreground and background selection will be undertaken in BC3F1 plants for identification of restorers close to the recurrent parent

iv) Molecular characterization of newly developed CMS lines using informative markers. Off season: (IARI and NRCPB, New Delhi)

i) Foreground and background selection will be undertaken in BC4F1 plants for identification of restorers close to the recurrent parent

ii) Analysis and report writing

Activities Years I II III

Procurement of equipment and recruitment of RA & SRF

Southern analysis of newly isolated CMS lines

Northern analysis of newly isolated CMS and restorer lines

Genetic study of fertility restoration in new CMS lines

Genotypic polymorphism studies using SSR primers

Polymorphic studies in parental lines for fertility restoration

Tagging of Rf gene(s)

Hybridization programme for restorer transference: backcrossing

Foreground and background selection for transferring Rf genes

Molecular characterization of CMS lines

Report writing

18.2 Connectivity of the participating institutions and investigators (in case of multi- institutional

projects only): IARI and NRCPB New Delhi are already working jointly for the development of A, B and R lines in different genetic backgrounds. The joint efforts are directed towards development of commercial hybrids.

18.3 Alternate strategies (if the proposed experimental design or method does not work what is the alternate strategy): Molecular based mapping and tagging of gene (s) is a well-established method so there are very less chance of failure of the programme. From this study good number of informative markers are expected to be identified which can be used extensively in Indian mustard hybrid breeding programme. Alternatively, new marker systems like SNPs, IP etc. can be explored.

16

19. Timelines: (Please provide quantifiable outputs)

Period of study

Achievable targets

6 Months

• Development of lab infrastructure and recruitment of contractual staff. • DNA isolation of 100 B. juncea genotypes and about 500 plants of each of

BC1F1/F2 populations. • Phenotyping of individual BC1F1 plants for sterility/fertility • Custom synthesis of available primers and procurement of other consumables. • Studying parental polymorphism using SSR primers.

12 Months • Development of lab infrastructure • Studying genotypic polymorphism using SSR primers. • Southern analysis of newly developed CMS lines

18 Months • Identification of polymorphic markers which can differentiate genotypes • Attempting BC2 crosses • Genotyping of mapping populations using informative markers for Rf genes and tagging of Rf gene(s). • Attempting BC2 crosses and harvesting of BC2F1 seeds for transferring Rf gene

through MAS. • Genetics of fertility restoration in new CMS lines

24 Months • Studying genotypic polymorphism using SSR markers • Exploring identification of additional closely link DNA marker to Rf gene(s). • Raising BC2F1 and attempted BC3 crosses with the positive plants of ten elite genotypes

for transferring Rf gene through MAS. • Molecular characterization of new CMS lines using informative markers • Northern analysis of CMS and fertility restorer lines

30 Months

• Studying genotypic polymorphism using SSR markers • Testing of more SSR primers to identify still closer markers to the Rf gene(s). • Foreground and background selection will be done in BC3F1 plants using

informative markers. • Attempting BC4 crosses • Molecular characterization of new CMS lines using informative markers

36 Months • Foreground and background selection will be undertaken in BC4F1 plants for identification of restorers close recurrent parent

• Analysis and report writing

17

20. Name and address of 5 experts in the field

Sr. No. Name Designation Address

1 Dr. S. S. Banga National Professor Department of Plant Breeding and Genetics, PAU, Ludhiana-141004

2 Dr. N. Sarla Principal Scientist Directorate of Rice Research, Rajendranagar, Hyderabad- 500030

3 Dr. S. Rakshit Principal Scientist Directorate of Sorghum Research, Rajendranagar, Hyderabad- 500030

4 Dr. A. Pradhan Professor Division of Genetics, Delhi University (South Campus) New Delhi

5 Dr. P.B. Kirti Professor School of Life Sciences, University of Hyderabad

18

PART IV: BUDGET PARTICULARS

Budget (In lakh rupees)

a) IARI, New Delhi

A. Non-Recurring (e.g. equipments, accessories, etc.)

S. No. Items Year 1 Year 2 Year 3 Total

1. Gel Doc system with accessories 7.00 -- -- 7.00

2. Thermal cycler 5.00 -- -- 5.00

3. Ice Flaking Machine 1.50 -- -- 1.50

4. Hot Water Bath 0.25 -- -- 0.25

5. Horizontal electrophoretic unit with power pack (two)

3.00 -- -- 3.00

6. Table top centrifuge (COLD) 5.50 -- -- 5.50

7. Multichannel pippettes 1.00 -- -- 1.00

8. Air Conditioners 2.0 tones with stabilizers (Two)

0.80 -- -- 0.80

9. Deep freezer (-20) 2.00 -- -- 2.00

10. Minor equipments and lab furniture (Less than 15000)

3.00 -- -- 3.00

15. Online UPS (5 KVA) 1.50 -- -- 1.50

13. Microscope with mounted camera and accessories

6.00 6.00

14. Microscope 1.00 1.00

15 Computer with scanner, coloured laserjet multipurpose printer

1.50 -- -- 1.50

16 Photocopier 1.00 -- -- 1.00

17. Nets for maintenance of CMS/ restorers

-- 4.00 -- 4.00

18 Renovation of laboratory (Two) 8.00 -- -- 8.00

19 Nano drop 4.00 -- -- 4.00

Total 52.05 4.00 -- 56.05

Institute charges @ 15% of total 7.81 0.60 -- 8.41

Grand total 59.86 4.60 -- 64.46 Justification: For strengthening the laboratory and for use in the present programme to fulfill the

objectives.

19

B. Recurring

B.1 Manpower (See guidelines at Annexure-III)

S. No.

Position No.

Consolidated Emolument

Year 1 Year 2 Year 3 Total (Lakhs)

1. Senior Research Fellow 2 (IARI)

Rs. 18000/ plus HRA @ 30% per month

5.62 5.62 5.62 16.86

2. Skilled worker 3 (IARI)

Rs. 8000/- per month (Consolidated)

2.88 2.88 2.88 8.64

Sub-Total (B.1) 25.50 Justification: For carrying out the field observations and laboratory work. B.2 Consumables S. No.

Item Quantity Year 1 Year 2 Year 3 Total (Lakhs)

1. Consumables like molecular biology reagents, kits, primers, glassware, plastic wares, other chemicals, micropipettes etc.

For carrying out the various field activities and lab work at IARI

10.00 5.00

5.00

20.00

Sub-Total (B.2) 20.00

Other items Consolidated Emolument


B.3 Travel

For attending meeting etc.

2.00 2.00 2.00 6.00

B.4 Contingency

a) To meet the day to day requirements other than chemicals

1.00 1.00 1.00 3.00

B.5 Overhead@ 15 % (If applicable)

3.23 2.48 2.48 8.19

Sub-total of B (B.1+B.2+B.3+B.4+B.5)

24.73 18.98 18.98 62.69

Grand Total (A+B) 84.59 23.58 18.98 127.15 *This is essential to effect raising, managing, DNA sampling and making crosses at offseason nursery at IARI Regional Station Wellington (Tamilnadu). There will be at least 2 visits each by three staff with one stay of prolonged nature while effecting crosses and sampling with phenotype (TA + DA will have to be borne to meet the objectives)

20

b.) NRCPB, New Delhi

A. Non-Recurring (e.g. equipments, accessories, etc.)

S. No.

Items Year 1 Year 2 Year 3 Total

1. Thermal cycler 5.00 -- -- 5.00

2. Gel electrophoresis system 1.50 -- -- 1.50

3. Deep freezer (-20) 2.00 -- -- 2.00

4. Multichannel pippettes 1.00 1.00

5 Miscellaneous small equipments and lab furniture (microwave oven etc.) less than Rs.15000

1.50 -- -- 1.50

6. Air Conditioners 2.0 tones with stabilizers (Two) 0.80 -- -- 0.80

7. Total 11.80 -- -- 11.80

Institute charges @ 15% of total 1.77 1.77

Grand total 13.57 13.57 Justification: For strengthening the laboratory and for use in the present programme to fulfill the

objectives.

B. Recurring

B.1 Manpower (See guidelines at Annexure-III)

S. No.

Position/ No. Consolidated Emolument


1. Senior Research Fellow -One

Rs. 18000/ plus HRA @ 30% per month

2.81

2.81 2.81 8.43

2. Skilled worker - One

Rs. 8000/- per month (Consolidated)

0.96 0.96 0.96 2.88


Justification: For carrying out the field observations and laboratory work.

21

B.2 Consumables S. No.

Item

Quantity Year 1 Year 2 Year 3 Total

1. Consumables like molecular biology reagents, kits, primers, glassware, plastic wares, other chemicals, micropipettes etc.

For carrying out the various field activities and lab work at NRCPB

8.00 4.00

4.00

16.00


Other items Consolidated Emolument Year 1 Year 2 Year 3 Total

B.3 Travel

For attending meeting etc. 1.00 1.00 1.00 3.00

B.4 Contingency

a) To meet the day to day requirements other than chemicals

0.50 0.50 0.50 1.50

B.5 Overhead@ 15 % (If applicable)

2.00 1.40 1.40 4.80

Sub-total of B (B.1+B.2+B.3+B.4+B.5)

15.27 10.67 10.67 36.61

Grand Total (A+B) 28.84 10.67 10.67 50.18

Total budget outlay for IARI and NRCPB, New Delhi

Item IARI NRCPB Total

A. Non Recurring (including over head @15%)

64.46 13.57 78.03

B. Recurring

(B.1+B.2+B.3+B.4+B.5)- including over head@15%

62.69 50.18 112.87

Total 127.15 63.75 190.90

22

PART V : EXISTING FACILITIES (IARI, NEW DELHI and NRCPB, New Delhi) Resources and additional information

1. Laboratory: A full fledged well equipped Brassica quality laboratory is available with the following equipments:

Sl. No.

Name of the Equipment Make Funded by Year of Installation

Remarks

IARI, New Delhi 1. Deep fridge (-12) Widson Scientific DBT 1993 Non functional 2. Cold room Blue Star DBT 1993 Non functional 3. Vertical electrophoresis Biorad DBT 1999 Working 4. Horizontal electrophoresis Biorad DBT 1999 Working 5. Circulating water bath Pharmacia DBT 1993 Working 6. pH meter Tarson DBT 2007 Working 7. UPS 5 KVA Vintec DBT 1999 Non functional 8. Centrifuge HettichRotina 35R DBT 1999 Non functional 9 Digital balance Afcoset IARI 1995 Non functional 10 Gas Chromatograph Perkin Elmer DBT 2010 working 11 UV spectriphotometer Spectramax DBT 2010 working 12 -80 deep freezer DBT 2010 working 13 -20 deep freezer DBT 2008 working 14 Digital Balance DBT 2010 working 15 Gel Doc* Biorad DBT 2000 Non functional 16 Thermal Cycler G storm DBT working NRCPB, New Delhi 17 Complete tissue culture

facility 1995 working

18 Refrigerated Centrifuge Thermo ICAR 2007 working 19 Spectrophotometer Jena ICAR 2008 working 20 Confocal and fluorescence

microscopes Leica ICAR 2008 Working

21 Gel documentation system Alfa Innotech ICAR 2007 working 22 Thermal cycler Biometra ICAR 2008 working 23 Microfuge Eppendorf ICAR 2005 working 24 Hybridization oven DBT 2000 working

*: The earlier one has lived its life and is not accessible for documentation analysis now required.

a. Manpower: One Technical Officer each will be assisting in raising the crop and for various field activities. For hybridization, Phenotyping for fertility/ sterility and molecular work, the contractual manpower has been proposed.

2. Other resources such as clinical material, animal house facility, glass house. Experimental garden, pilot plant facility etc.: Fields, Phytotron Facility and off-season nursery (IARI, RS, Wellington, Tamilnadu) growing facilities are available.

23

PART VI: DECLARATION/CERTIFICATION

It is certified that a) the research work proposed in the scheme/project does not in any way duplicate the work already done

or being carried out elsewhere on the subject. b) the same project proposal has not been submitted to any other agency for financial support. c) the emoluments for the manpower proposed are those admissible to persons of corresponding status

employed in the institute/university or as per the Ministry of Science & Technology guidelines (Annexure-III)

d) necessary provision for the scheme/project will be made in the Institute/University/State budget in anticipation of the sanction of the scheme/project.

e) if the project involves the utilization of genetically engineered organisms, we agree to submit an application through our Institutional Bio-safety Committee. We also declare that while conducting experiments, the Bio-safety Guidelines of the Department of Biotechnology would be followed in toto.

f) if the project involves field trials/experiments/exchange of specimens, etc. we will ensure that ethical clearances would be taken from concerned ethical Committees/Competent authorities and the same would be conveyed to the Department of Biotechnology before implementing the project.

g) it is agreed that any research outcome or intellectual property right(s) on the invention(s) arising out of the project shall be taken in accordance with the instructions issued with the approval of the Ministry of Finance, Department of Expenditure, as contained in Annexure-V.

h) we agree to accept the terms and conditions as enclosed in Annexure-IV. The same is signed and enclosed.

i) the institute/university agrees that the equipment, other basic facilities and such other administrative facilities as per terms and conditions of the grant will be extended to investigator(s) throughout the duration of the project.

j) the Institute assumes to undertake the financial and other management responsibilities

Signature of the Executive authority of the Institute with Seal

Dr. Naveen Singh Principal Investigator & Senior Scientist Div. of Genetics, IARI, New Delhi

Dr. D.K. Yadava Co- PI & Senior Scientist Div. of Genetics, IARI, New Delhi

Dr. Sujata V. Co- PI & Principal Scientist Div. of Genetics, IARI, New Delhi

Dr. S.R. Bhat Co-PI & Principal Scientist NRCPB, New Delhi

Dr. Naveen Chander Gupta Co-PI Scientist & NRCPB, New Delhi

Dr. K.V. Prabhu Co-PI & Head Div. of Genetics, IARI, New Delhi

24

PART VII: PROFORMA FOR BIOGRAPHICAL SKETCH OF INVESTIGATORS Provide the following information for the key personnel in the order listed on PART II. Follow this format for each person. DO NOT EXCEED THREE PAGES

Name : Dr. Naveen Singh

Designation : Senior Scientist

Department/Institute/University: Division of Genetics, Indian Agricultural Research Institute, New Delhi - 110012

Date of Birth : 16.03.1974 Sex (M/F) Male SC/ST : x

Education (Post-Graduation onwards & Professional Career)

Sr. No.

Institution / Place Degree Awarded

Year Field of Study

1. CCS Haryana Agricultural University, Hisar

M.Sc. 1996 Plant Breeding & Genetics

2. CCS Haryana Agricultural University, Hisar

Ph. D. 1999 Plant Breeding & Biotechnology

Position and Honors

Position and Employment (Starting with the most recent employment) Sr. No.

Institution Place

Position From (Date) To (date)

1. IARI, New Delhi Senior Scientist 21.01.2010 Contd…

2. Punjab Agricultural University, Ludhiana

Assistant Plant Breeder (SS)

10.07.2005 20.01.2010

3. Punjab Agricultural University, Ludhiana

Assistant Plant Breeder

09.07.2001 09.07.2005

4. CCS HAU, Hisar Research Associate

17.01.2000 08.07.2001

5. CCS HAU, Hisar Senior Research Fellow

17.07.1999 31.12.1999

Honors/Awards

i. University Merit Scholarship holder in M.Sc. degree. ii. University Merit Scholarship holder in Ph.D. degree.

iii. Awarded commendation certificate for outstanding performance in ICAR sponsored winter school on “Advances in hybrid rice technology” held at Directorate of rice research, Hyderabad , India in the year 2003

25

Professional Experience and Training relevant to the Project: I have been working on development of CMS and restorer lines in rice from 2001 to 2009. From the last two years I have undertaken the work on development of heterotic parental lines in B. juncea.

Professional experience: • Co-PI in DBT Programme support for research and development in Agricultural

Biotechnology Theme: Biotechnological intervention to enhance water use efficiency in rice Sub Project: Enhancing water use efficiency and productivity in rice and basmati through molecular marker assisted selection (associated from 2006 to 2009).

• Co-PI in DBT Project on “Development of biotic stress resistant rice through Marker Assisted Selection” (associated from Dec. 2008 to Jan. 2010).

• Associated scientist in the NATP project on “Development of Hybrid (crops)- Rice” (2001-2004)

• Associated scientist in the NATP project on “ Evaluation of basmati rice and wheat genotypes for their response to tillage option and thermal response in system perspective (PSR59-1.1)” (2001-2005)

• Associated in the NATP project on “Breeding high sugared multi-cut sorghum hybrids and varieties for enhanced nutritional quality” (2000-2001)

Professional trainings:

Sr. No.

Title Duration Institution Year

1 Breeding for Natural Resource Management & Abiotic Stress Tolerance in Crop Plants

21 days CCS Haryana Agricultural University, Hisar

2009

2 GGE Biplot Analysis 6 months (Semester I)

Punjab Agricultural University, Ludhiana

2008-09

3 Heterosis Breeding in Crop Improvement

21 days CCS Haryana Agricultural University, Hisar

2007

4 Planning Rice Breeding for Impact 12 days IRRI, Los Banios, Philippines

2005

5 Advances in marker assisted selection 4 days IRRI, Los Banios, Philippines

2005

6 Advances in Hybrid Rice Technology 21 days DRR, Rajendranagar, Hyderabad

2003

7 Research Management 30 days CCS HAU, Hisar 2002

8 Marker Assisted Selection in Pearl Millet Breeding

23 days ICRISAT, Patancheru, Hyderabad

1998

9 Molecular Biology & Crop Biotechnology

13 days Sponsored by DST, at CCS University, Meerut

1996

26

Publications (Numbers only):

1. Research papers 22 2. Symposia/Conference papers 19 3. Book chapters 2 4. Chapters in manuals 10 5. Popular articles 26 6. Bulletin 1 7. Patents/ Varieties: 11 (Varieties Released= Brassica-1, Rice-6, CMS lines-8)

TOTAL 97 Selected peer-reviewed publications (Ten best publications in chronological order)

1. Bhatia,Dharminder Sharma Rajiv, Vikal Yogesh, Mangat G.S., Mahajan Ritu, Sharma Neerja, Singh Jagjeet, Singh Naveen, Bharaj TS and Singh Kuldeep (2011). Marker-assisted Development of Bacterial Blight Resistant, Dwarf and High Yielding Versions of Two Traditional Basmati Rice Cultivars. Crop Science 51: 759-770.

2. Yadava D. K., Giri S. C., Vignesh M., Vasudev S., Yadav A. K., Dass B., Singh Rajendra, Singh Naveen, Mohpatra T. and Prabhu K. V. (2011) Genetic variability and trait association studies in Indian mustard (Brassica juncea) Indian Journal of Agricultural Sciences 81 (8): 712–716.

3. Mahajan, G., Sekhon, N.K., Singh, Naveen, Kaur Rupinder and Sidhu, A.S. (2010). Yield and Nitrogen Use Efficiency in Response to Nitrogen Fertilizer. Journal of New Seeds 11: 356-368. (Taylor & Fransis)

4. Kaur, J.J., Rang Allah, Singh Naveen and Kaur Rupinder. (2010). Genetics of resistance to bacterial blight in rice cultivar PAU 201. Plant Disease Research 25(2): 174-175.

5. Kaur, J.J., Rang Allah, Singh Naveen and Kaur Rupinder. (2010). Genetics of Resistance to Bacterial Blight in Cultivars PR 111, PR 113 and PR 115 of Rice (Oryza sativa L.) Crop Improvement 37 (1): 9-12.

6. Sharma Neerja, Singh Naveen, Bhullar M S and Bharaj T S (2008) Identification of Aromatic Fine Grained Rice Genotypes for Utilization in Improving Basmati. Indian Journal of Agricultural Sciences 78 (1): 44-49.

7. Singh Naveen, Kaur Rupinder, Singh Mandeep, Rang Allah and Bharaj T S (2008) Effect of Age of Nursery and GA3 on Hybrid Rice Seed Yield. Crop Improvement 35 (2): 122-124.

8. Sharma Neerja, Mangat G S, Singh Naveen and Bharaj T S (2005). Identification of donors for quality traits in fine-grained aromatic rice (Oryza sativa). Indian Journal of Agricultural Sciences 75 (12): 831-833.

9. Singh Naveen, Behl R K and Punia M S (2005). Effect of genotypic background on haploid production through embryo rescue in wheat x maize crosses. Plant, Soil and Environment 51(4): 193-196.

10. Singh Naveen, Behl R K and Punia M S (2001). Production of double haploids via maize pollination in wheat. Cereal Research Communication 29 (3-4): 289-296.

27

List maximum of five recent publications relevant to the proposed area of work.

1. Bhatia, Dharminder Sharma Rajiv, Vikal Yogesh, Mangat G.S., Mahajan Ritu, Sharma Neerja, Singh Jagjeet, Singh Naveen, Bharaj TS and Singh Kuldeep (2011). Marker-assisted Development of Bacterial Blight Resistant, Dwarf and High Yielding Versions of Two Traditional Basmati Rice Cultivars. Crop Science 51: 759-770.

2. Yadava D. K., Giri S. C., Vignesh M., Vasudev S., Yadav A. K., Dass B., Singh Rajendra, Singh Naveen, Mohpatra T. and Prabhu K. V. (2011) Genetic variability and trait association studies in Indian mustard (Brassica juncea) Indian Journal of Agricultural Sciences 81 (8): 712–716.

3. Sharma Neerja, Singh Naveen, Bhullar M S and Bharaj T S (2008) Identification of Aromatic Fine Grained Rice Genotypes for Utilization in Improving Basmati. Indian Journal of Agricultural Sciences 78 (1): 44-49.

4. Singh Naveen, Behl R K and Punia M S (2001). Production of double haploids via maize pollination in wheat. Cereal Research Communication 29 (3-4): 289-296.

5. Singh Naveen, Behl R K, Punia M S and Singh Vikram (2002). In vitro screening and production of karnal bunt resistant double haploids in wheat. Wheat Information Service 94: 5-8.

Research Support Ongoing Research Projects Sl No.

Title of Project Funding Agency

Amount (lakhs)

Date of sanction Duration

1. Genetic Enhancement of Rapeseed Mustard for Development of Varieties and Hybrids for Different Agronomical Situations

IARI (ICAR)

616.65 01.04.2009 for 5 years

i. Development of rapeseed mustard varieties and hybrids tolerant to biotic and abiotic stresses

ii. Oil quality enhancement in Indian mustard iii. Development of Indian mustard varieties suitable

for late sown conditions 2. Challenge Programme of the Institute: Hybrid

development in crops (Wheat, pigeonpea and mustard)

IARI (ICAR)

288.88 01.04.2010 for 5 years

3. Development of double low (canola type) Indian mustard (Brassica juncea) through marker assisted selection

DBT 85.14 Feb. 2009 for 5 years

Completed Research Projects (State only major projects of last 3 years): No

Place : New Delhi Signature of Principal Investigator

Date:

28

Name : Dr. D.K. Yadava

Designation : Senior Scientist

Institute : Division of Genetics, Indian Agricultural Research Institute, New Delhi - 12

Date of Birth : 10.04.1966 Sex (M/F): Male SC/ST : x Education (Post-Graduation onwards & Professional Career)

Sl No. Institution Place

Degree Awarded

Year Field of Study

1. Haryana Agricultural University, Hisar

M.Sc. 1990 Studies on hybrid barley

2. Haryana Agricultural University, Hisar

Ph. D. 1993 Seed production technology in sunflower

Position and Honors Position and Employment (Starting with the most recent employment)


Position From (Date) To (date)

1. IARI, New Delhi Division of Genetics

Senior Scientist (PB) 01.04.2003 Contd…

2. RAU, Bikaner Agricultural Research Station, Sriganganagar

Assistant Professor (PBG) & Sr. Scale

15.05.1996 31.03.2003

3. CCS HAU, Hisar Forage Section

Senior Technical Assistant

17.02.1994 14.05.1996

4. CCS HAU, Hisar Rice Research Station, Kaul

Research Associate 21.07.1993 16.02.1994

Honors/Awards i. Recipient of Junior Research Fellowship (ICAR) during M.Sc. ii. Recipient of Senior Research Fellowship (UNDP) during Ph.D. iii. University Merit Scholarship holder in B.Sc. degree.

iv. Awarded Scholarship by Education Department of Haryana from 5th standard to matriculation. v. Fellow, Indian Society of Pulses Research and Development, IIPR, Kanpur

vi. Fellow, Indian Society of Plant Breeding and Genetics, IARI, New Delhi vii. Fellow, Indian Society of Forage Research, HAU, Hisar

viii. Fellow, Indian Society of Oilseed Research, DOR, Hyderabad

Professional Experience and Training relevant to the Project • Associated in the ICAR funded network project entitled “Functional Genomics for draught tolerance

and Alternaria blight resistance in Brassica” • Principal Investigator of concluded project entitled, “Mapping and marker assisted selection for new

genes for resistance to white rust (Albugo candida) in Indian mustard (Brassica juncea (L.) Czern. & Coss)” (DBT).

• Principal Investigator of ongoing project entitled, “Development of double low (Canola type) Indian mustard through marker assisted selection” (DBT).

• Professional trainings

29

Qualifications Date awarded 1. Long term training on “Plant Genetic Engineering and Molecular Breeding” conducted by National research Centre on Plant Biotechnology, IARI, New Delhi Title: Evaluation of microsatellite markers for analysis of genetic diversity in Brassica species & related genera

August 2 - November 1, 2004

2. “Unconventional approaches in Plant Breeding” conducted by Punjab Agricultural University, Ludhiana.

January 23 – February 12, 2001

3. “Computer training for ARIS Cell personal” conducted by Indian Agril. Statistical Research Institute, New Delhi.

August 28 - September 5, 1997

Short courses attended Subject Conducted by Dates Capacity Building Program for Indian Agricultural Research, Extension and Development (RED) Organizations in Globalized Agricultural Economy.

CITA - ICAR, IARI, New Delhi

September 15-16, 2005

Use of computers in Agriculture NIIT - RAU, Bikaner July 17-19, 2000 Orientation training programme for Assistant Professors

Rajasthan Agricultural University, Bikaner

August 20 - September 02, 1998

Plant Genetic Resources: Principles, procedures and approaches in collection, evaluation and conservation

NBPGR, New Delhi July 27 -August 06, 1998

Networking Basic concepts and services NIC, New Delhi July 7-10, 1998 Publications (Numbers only) 1. Varieties Released 16 (Chickpea-1, Mungbean-2, Brassica-13) 2. Research papers 50 3. Symposia/Conference papers 25 4. Book chapters 10 5. Books 4 6. Popular articles 40 7. Bulletins/ Pamphlets 16

TOTAL 161

Selected peer-reviewed publications (Ten best publications in chronological order) Ram Bakshi, Chaudhary, B.S. and Yadava, D.K. (1997). General and specific selection indices

for single stool stage of selection in sugarcane. Euphytica 95 : 39-44. Kalita, M.C., Mohapatra, T., Dhandapani, A., Yadava, D.K., Srinivasan, K., Mukherjee, A.K.,

and Sharma, R. P. (2007). Comparative evaluation of RAPD, ISSR and Anchored-SSR markers in the assessment of genetic diversity and fingerprinting of oilseed Brassica genotypes. J. Plant Biochemistry & Biotechnology 16(1): 41-48.

Koundal Vikas, Parida S.K., Yadava D.K., Ali Arif, Koundal K.R. and Mohapatra T. (2008). Evaluation of microsattelite markers for genome mapping in Indian mustard (Brassica juncea L.). J. Plant Biochemistry & Biotechnology 17(1): 69-72.

30

Yadava, D.K., Parida, S. K., Dwivedi, V.K., Varshney, A., Ghazi, Irfan A., Sujata V. and Mohapatra, T. 2009. Cross-transferability and polymorphic potential of genomic STMS markers of Brassica species. J. Plant Biochemistry & Biotechnology Vol. 18(1): 29-36.

Kemparaju, K.B., Yadava D.K., Sujata Vasudev, Anil Kumar Yadav, Bhagwan Dass, S.C.Giri and K.V.Prabhu. 2009. Genetic Analysis of yield and yield attributing traits in Indian mustard (Brassica juncea). Ind. J. Agric Sciences 79 (7): 552-554.

Yadava, D.K., R L Sapra, V Sujata, B Dass, and K. V. Prabhu 2009. Selection of high diversity with a minimal set of accessions from Indian Mustard {Brassica juncea (L.) Czern & Coss.} germplasm collection Ind. J. Agric Sciences 79 (7): 555-558.

Vignesh, M., Yadava, D.K. Sujata. V., Mohapatra T., Jain Neelu, Yadav Anil Kumar, Malik Divya, Yadav M. S. and Prabhu K. V. 2009. Genetics of white rust resistance in Brassica juncea (L.) Czern. & Coss. and allelic relationship between interspecific sources of resistance. Indian J. of Genet. 69(3): 205-208.

Parida S., Yadava, D.K. and Mohapatra T. 2010. Microsatellites in Brassica unigenes: Relative abundance, marker design and use in comparative physical mapping and genome analysis. Genome 53: 55-67.

Yadava D K, Giri S.C., Sujata V., Yadav Anil Kumar, Dass B, Raje R.S., Vignesh M. Singh Rajendra, Mohapatra T. and K. V. Prabhu 2010. Stability analysis in Indian mustard (B. juncea) varieties. Ind. J. Agric Sciences 80 (9) 761-765.

Yadava D.K., Giri S.C., Vignesh M., Sujata V., Yadav A.K., Dass B., Singh Rajendra, Singh N., Mohapatra T. and Prabhu K.V. 2011. Genetic variability and trait association studies in Indian mustard (Brassica juncea). Indian Journal of Agricultural Sciences 81(8): 712-716.

List maximum of five recent publications relevant to the proposed area of work. Kalita, M.C., Mohapatra, T., Dhandapani, A., Yadava, D.K., Srinivasan, K., Mukherjee, A.K.,

and Sharma, R. P. (2007). Comparative evaluation of RAPD, ISSR and Anchored-SSR markers in the assessment of genetic diversity and fingerprinting of oilseed Brassica genotypes. J. Plant Biochemistry & Biotechnology 16(1): 41-48.

Koundal Vikas, Parida S.K., Yadava D.K., Ali Arif, Koundal K.R. and Mohapatra T. (2008). Evaluation of microsattelite markers for genome mapping in Indian mustard (Brassica juncea L.). J. Plant Biochemistry & Biotechnology 17(1): 69-72.

Yadava, D.K., Parida, S. K., Dwivedi, V.K., Varshney, A., Ghazi, Irfan A., Sujata V. and Mohapatra, T. 2009. Cross-transferability and polymorphic potential of genomic STMS markers of Brassica species. J. Plant Biochemistry & Biotechnology Vol. 18(1): 29-36.

Yadava, D.K., R L Sapra, V Sujata, B Dass, and K. V. Prabhu 2009. Selection of high diversity with a minimal set of accessions from Indian Mustard {Brassica juncea (L.) Czern & Coss.} germplasm collection Ind. J. Agric Sciences 79 (7): 555-558.

Parida S., Yadava, D.K. and Mohapatra T. 2010. Microsatellites in Brassica unigenes: Relative abundance, marker design and use in comparative physical mapping and genome analysis. Genome 53: 55-67.

Yadava D K, Giri S.C., Sujata V., Yadav Anil Kumar, Dass B, Raje R.S., Vignesh M. Singh Rajendra, Mohapatra T. and K. V. Prabhu 2010. Stability analysis in Indian mustard (B. juncea) varieties. Ind. J. Agric Sciences 80 (9) 761-765.

Yadava D.K., Giri S.C., Vignesh M., Sujata V., Yadav A.K., Dass B., Singh Rajendra, Singh N., Mohapatra T. and Prabhu K.V. 2011. Genetic variability and trait association studies in Indian mustard (Brassica juncea). Indian Journal of Agricultural Sciences 81(8): 712-716.

31

Research Support

Ongoing Research Projects Sl No.


Amount (lakhs)



IARI 616.65 01.04.2009 for 5 years


IARI 01.04.2009 for 5 years

ii. Oil quality enhancement in Indian mustard IARI 01.04.2009 for 5 years

iii. Development of Indian mustard varieties suitable for late sown conditions


2. Challenge Programme of the Institute: Hybrid development in crops (Wheat, pigeonpea and mustard)

288.88 01.04.2010 for 5 years



4. Functional Genomics of Alternaria blight resistance and draught tolerance in Brassica

ICAR 93.24 05.08.2005 for 7 years

5. Genomics and Molecular breeding of linseed/ flax

ICAR- Saskatchewan

60.98 XI Plan

Completed Research Projects (State only major projects of last 3 years): Sl No.


Amount (lakhs)


1. Breeding rapeseed mustard for biotic and abiotic stress resistance

IARI In house project

31.03.2009

2. Breeding for quality in mustard (B. juncea) IARI In house project

31.03.2009

3. Molecular Breeding for crop improvement (Brassica) IARI In house project

31.03.2009

4. Mapping and marker assisted selection for new genes for resistance to white rust (Albugo candida) in Indian mustard (Brassica juncea (L.) Czern. & Coss)

DBT 17.02 24.09.2009

Place : New Delhi Signature of Co-Principal Investigator Date: 12.12.2011

32

Name : Dr. Sujata Vasudev

Designation : Principal Scientist Institute : Division of Genetics, IARI, New Delhi 110012 Date of Birth : 07.10.1962 Sex (M/F): Female SC/ST: × Education (Post-Graduation onwards & Professional Career)


Degree Awarded

Year Field of Study

1. Delhi University Delhi 110007

M. Sc. (Botany)

1986 Botany

2. Indian Agricultural Research Institute, New Delhi 110012

Ph.D 1989 Biochemistry

Position and Honors

Position and Employment (Starting with the most recent employment)

Sl No. Institution/Place Position From (Date) To (date) 1. Division of Genetics

(IARI) New Delhi Principal Scientist (Genetics)

Dec 2006 Till date

2. Division of Genetics (IARI) New Delhi

Senior Scientist (Genetics)

Dec 1998 Dec 2006


Scientist Sr. Scale (Genetics)

Dec 1994 Dec 1998

4 Division of Genetics (IARI) New Delhi

Scientist S-1 (Genetics)

Feb 1993 Dec 1994

5. ICRISAT Patancheru, Andhra Pradesh

ICAR-OCRISAT Post Doc Fellow

Feb 1991 Feb 1993

6. Water technology Centre (IARI) New Delhi

Scientist S-1 (Genetics)

May 1990 Feb 1991

7. NAARM Scientist S-1 (Genetics)

Dec 1989 May 1990

Honors/Awards ♦ IARI Gold medal for outstanding academic performance in 1990

♦ The Jawaharlal Nehru award for outstanding Post-graduate research in Agriculture the field of Microbiology (1990)

♦ ICAR-ICRISAT Fellowship from 1991-1993

Professional Experience and Training relevant to the Project Publications 1. Varieties 8 (Indian mustard) 2. Research papers 25 3. Symposia/Conference 10 4. Popular articles 17 5. Book chapters 4 6. Bulletins/ Pumphlets 10 TOTAL 74

33

Selected peer-reviewed publications (Ten best publications in chronological order)

1. Sujata Vasudev, Madan L. Lodha & K.R. Sreekumar. 1990. Transfer and expression of

Bradyrhiobium Japonicum uptake genes in Cicer-rhizobia. Indian journal of Experimental Biology. 28: 1040-1045

2. Sujata Vasudev, Madan L. Lodha & K.R. Sreekumar. 1991. Imparting hydrogen-recycling capability to Cicer-rhizobial strains by plasmid pIJ1008 transfer. Current Science. 60: 600-603.

3. Sujata Vasudev, S. Sivaramakrishnan, K.N.Rai and Seetha Kannan 1994: A new source of cytoplasmic male-sterility in pearl millet: RFLP analysis of mitochondrial DNA. Genome, 37: 482-486.

4. Renu Khanna Chopra, Moinuddin, Sujata Vasudev, M. Maheshwari, Amita Srivastav and D Bahukhandi. 1994. K+, Osmoregulation and drought Tolerance- An Overview. Proc. Indian Natn. Sci Acad. 1: 51-56

5. Arunachalam V, Prabhu K.V. and Sujata V. 1996. Isozyme parameters in genetic evaluation in Brassica – A tribute of Haldane’s vision on the synergy between biochemistry and genetics. Current Science. 70: 1062-1065.

6. Arunachalam V, Shefali Verma, Sujata V, Prabhu K.V 2005. Marker-aided genetic divergence analysis in Brassica. Journal of Genetics 84 (2): pp 123-130

7. Yadava, D.K., Parida, S. K., Dwivedi, V.K., Varshney, A., Ghazi, Irfan A., Sujata V. and Mohapatra, T. 2009. Cross-transferability and polymorphic potential of genomic STMS markers of Brassica species. J. Plant Biochemistry & Biotechnology Vol. 18(1): 29-36.

8. Sujata, V., Yadava, D.K., Malik Divya, Tanwar, R.S. and Prabhu, K.V. 2008. A simplified method for preparation of fatty acid methyl esters of Brassica oil. Indian J. of Genet. 68(4): 456457.

9. Yadava, D.K., R L Sapra, V Sujata, B Dass, and K. V. Prabhu 2009. Selection of high diversity with a minimal set of accessions from Indian Mustard {Brassica juncea (L.) Czern & Coss.} germplasm collection Ind. J. Agric Sciences 79 (7): 555-558.

10. Yadava D K, Giri S.C., Sujata V., Yadav Anil Kumar, Dass B, Raje R.S., Vignesh M. Singh Rajendra, Mohapatra T. and K. V. Prabhu 2010. Stability analysis in Indian mustard (B. juncea) varieties. Ind. J. Agric Sciences 80 (9) 761-765.

List maximum of five recent publications relevant to the proposed area of work. 1. Sujata Vasudev, Madan L. Lodha & K.R. Sreekumar. 1991. Imparting hydrogen-recycling

capability to Cicer-rhizobial strains by plasmid pIJ1008 transfer. Current Science. 60: 600-603.

2. Sujata Vasudev, S. Sivaramakrishnan, K.N.Rai and Seetha Kannan 1994: A new source of cytoplasmic male-sterility in pearl millet: RFLP analysis of mitochondrial DNA. Genome, 37: 482-486.

3. Arunachalam V, Shefali Verma, Sujata V, Prabhu K.V 2005. Marker-aided genetic divergence analysis in Brassica. Journal of Genetics 84 (2): pp 123-130

4. Yadava, D.K., Parida, S. K., Dwivedi, V.K., Varshney, A., Ghazi, Irfan A., Sujata V. and Mohapatra, T. 2009. Cross-transferability and polymorphic potential of genomic STMS markers of Brassica species. J. Plant Biochemistry & Biotechnology Vol. 18(1): 29-36.

5. Yadava, D.K., R L Sapra, V Sujata, B Dass, and K. V. Prabhu 2009. Selection of high diversity with a minimal set of accessions from Indian Mustard {Brassica juncea (L.) Czern & Coss.} germplasm collection Ind. J. Agric Sciences 79 (7): 555-558.

34

Research Support

Ongoing Research Projects

Ongoing Research Projects Sl No.


Amount (lakhs)



IARI 616.65 01.04.2009 for 5 years






2. Challenge Programme of the Institute: Hybrid development in crops (Wheat, pigeonpea and mustard)

288.88 01.04.2010 for 5 years



4. Genomics and Molecular breeding of linseed/ flax ICAR- Saskatchewan

60.98 XI Plan

Completed Research Projects (State only major projects of last 3 years): Sl No.


Amount (lakhs)




31.03.2009


31.03.2009

3. Molecular Breeding for crop improvement (Brassica) IARI In house project

31.03.2009


DBT 17.02 24.09.2009

5. Network project on marker assisted selection for leaf rust resistance in wheat

ICAR 20.00 31.03.2008

Place : New Delhi Signature of Co-Principal Investigator Date: 12.12.2011

35

Name: Dr. S.R. Bhat Date of birth 22.02.1954 Male M SC/ST NA Designation Principal Scientist (Biotechnology) Department National Research Centre on Plant Biotechnology Institute National Research Centre on Plant Biotechnology Address NRC on Plant Biotechnology, LBS Centre, IARI, New Delhi 110 012 Telephone 25841787 FAX 25843984 E-mail: [email protected]

Education (Post graduation onwards) & Professional Career Sr. No.

University Degree awarded Year Award/Prize/Certificate

1. UAS Dharwad M Sc (Agri) 1977 Gold Medal 2. University of

Birmingham, UK Ph. D. 1986

Position and Employment Name of employer/Institute Post held Period ICAR (Indian Inst of Sugarcane Research) Scientist S1 1978-1985 ICAR (NBPGR, New Delhi and IARI, New Delhi)

Scientist S2/Senior Scientist

1985-1998

ICAR (IARI, New Delhi/NRCPB, New Delhi) Principal Scientist 1998 onwards

Awards and Honours Sl. No

Recognition/ Award

Year Awarding Agency/ Institution

Nature of Award

Purpose

1 Commonwealth Fellowship, UK

1983-86 UK For Ph D

2. DBT overseas Associateship

1999 DBT Post-doc

Professional Experience and Training Relevant to the Project

I have been working on development of CMS lines and their molecular/genetic characterization for the past 17 years. I have all the relevant experience for undertaking the work proposed in this programme.

Publications (Numbers only)

Research papers- 60 Book Chapters/Proceedings- 5 Scientific reviews- 6


36

Ten Best Publications 1. Narasimhulu, S. B., Kirti, P. B., Bhat, S. R., Prakash, S. and Chopra, V. L. 1994. Intergeneric

protoplast fusion between Brassica carinata and Camelina sativa. Plant Cell Reports, 13:657-660.

2. Prakash, S., Kirti, P.B., Bhat, S.R., Gaikwad, K., Dinesh Kumar, V., Chopra, V.L. 1998. A Moricandia arvensis based cytoplasmic male sterility and fertility restoration system in Brassica juncea. Theor. Appl. Genet. 97:488-492.

3. Kirti, P.B., Prakash, S., Gaikwad, K., Kumar, D.V., Bhat, S.R., and Chopra, V.L. 1998. Chloroplast substitution overcomes leaf chlorosis in Moricandia arvensis based cytoplasmic male sterile Brassica juncea. Theor. Appl. Genet. 97:1179-1182.

4. Pathania A. Bhat S. R., Dinesh Kumar V., Ashutosh, Kirti P. B. Prakash, S. and Chopra, V. L. 2003. Cytoplasmic male sterility in alloplasmic Brassica juncea carrying Diplotaxis catholica cytoplasm: molecular characterization and genetics of fertility restoration. Theor. Appl. Genet. 107: 455-161

5. Bhat, S. R. Prakash, S., Kirti, P. B., Dinesh Kumar, V. and Chopra, V. L. 2005. A unique introgression from Moricandia arvensis confers male fertility upon two different cytoplasmic male-sterile lines of Brassica juncea. Plant Breeding 124: 117- 120.

6. Prasad A. M., Sivanandan, C., Resminath, R., Thakare, D. R., Bhat, S. R. and Srinivasan, R. 2005. Cloning and characterization of a pentatricopeptide protein encoding gene (LOJ) that is specifically expressed in lateral organ junctions in Arabidopsis thaliana. Gene 353: 67-79.

7. Bhat, S. R., Vijayan, P., Ashutosh, Dwivedi, K. K. and Prakash, S. 2006. Diplotaxis erucoides-induced cytoplasmic male sterility in Brassica juncea is rescued by the Moricandia arvensis restorer: genetic and molecular analyses. Plant Breeding, 125:150-155.

8. Sivanandan, C., Sujatha, T. P., Prasad, A. M., Resminath, R., Thakare, D. R., Bhat, S. R. and Srinivasan. 2005. T-DNA tagging and characterization of a cryptic root-specific promoter in Arabidopsis. Biochimica et Biophysica Acta, 1731: 202-208.

9. Ashutosh, Sharma, P. C., Prakash, S. and Bhat, S. R. 2006. Identification of AFLP markers linked to the male fertility restorer gene of CMS (Moricandia arvensis) Brassica juncea and conversion to SCAR marker. Theor. Appl. Genet. 114: 385-392.

10. Ashutosh, Kumar, P., Kumar, D. V., Sharma, P. C., Prakash, S. and Bhat, S. R. 2008. A novel orf108 co-transcribed with the atpA gene is associated with cytoplasmic male sterility in Brassica juncea carrying Moricandia arvensis cytoplasm. Plant and Cell Physiology. 49: 284-289.

Five Best Publications Relevant to the Area of Work 1. Pathania A. Bhat S. R., Dinesh Kumar V., Ashutosh, Kirti P. B. Prakash, S. and Chopra, V.

L. 2003. Cytoplasmic male sterility in alloplasmic Brassica juncea carrying Diplotaxis catholica cytoplasm: molecular characterization and genetics of fertility restoration. Theor. Appl. Genet. 107: 455-161

2. Bhat, S. R. Prakash, S., Kirti, P. B., Dinesh Kumar, V. and Chopra, V. L. 2005. A unique introgression from Moricandia arvensis confers male fertility upon two different cytoplasmic male-sterile lines of Brassica juncea. Plant Breeding 124: 117- 120.

37

3. Bhat, S. R., Vijayan, P., Ashutosh, Dwivedi, K. K. and Prakash, S. 2006. Diplotaxis erucoides-induced cytoplasmic male sterility in Brassica juncea is rescued by the Moricandia arvensis restorer: genetic and molecular analyses. Plant Breeding, 125:150-155.

4. Ashutosh, Sharma, P. C., Prakash, S. and Bhat, S. R. 2006. Identification of AFLP markers linked to the male fertility restorer gene of CMS (Moricandia arvensis) Brassica juncea and conversion to SCAR marker. Theor. Appl. Genet. 114: 385-392.

5. Ashutosh, Kumar, P., Kumar, D. V., Sharma, P. C., Prakash, S. and Bhat, S. R. 2008. A novel orf108 co-transcribed with the atpA gene is associated with cytoplasmic male sterility in Brassica juncea carrying Moricandia arvensis cytoplasm. Plant and Cell Physiology. 49: 284-289.

Research Support Ongoing Research Projects Sl. No.

Title of the Project Funding Agency Amount (lakhs)

Date of Sanction and Duration

1 Unraveling molecular processes involved in adventive polyembryony towards genetic engineering for the fixation of heterosis

ICAR (NAIP) 669 July, 2007- March, 2012

2 Exploitation of TGMS system in hybrid cotton seed production

ICAR (Technology Mission on Cotton)

15 August 2008 – March 2012

Completed Research Projects (State only major projects of last 3 years): No

Place: New Delhi Signature of Co-PI Date:

38

Name Dr. N. C. Gupta Date of birth 11-11-1977 Male: M SC/ST: NA Designation Scientist (Biotechnology) Institute National Research Centre on Plant Biotechnology Address NRC on Plant Biotechnology, LBS Centre, IARI, New Delhi 12 Telephone 25841787 FAX 25843984 E-mail: [email protected]

Education (Post graduation onwards) & Professional Career Sr. No. University Degree awarded Year Award/Prize/Certificate

1. I.A.R.I, New Delhi M.Sc. (Biochem) 2005 1st

2. I.A.R.I, New Delhi Ph.D. (Biotech) 2011 1st

Position and Employment Name of employer/Institute Post held Period

N.R.C. on Plant Biotechnology, New Delhi Scientist 2009 – till date

Awards and Honours Sl. No

Recognition/ Award

Year Awarding Agency/

Institution

Nature of Award Purpose

1. JRF 2003-2005 ICAR Fellowship M.Sc.

2. JRF/SRF/NET 2005-2010 CSIR Fellowship Ph.D.

Professional Experience and Training Relevant to the Project

Publications (Numbers only) Research papers : 4 Popular articles : Nil Book Chapters/Proceedings : Nil Scientific reviews : 1 Presentation in conferences/ workshop/ symposia : 3 Invited lectures : Nil

Ten Best Publications 1. Gupta N. C.,Sinha S. K., Jolly M., Dubey N. and Sachdev A. (2010). Antisense rna-

mediated inhibition of gmfad2-1 encoding omega-6-desaturase. Indian J. Plant Physiol., 14: 336-343.

2. Kishore K., Sinha S. K., Kumar R., Gupta N. C.,DubeyN. and Sachdev A. (2007). Isolation and characterization of microsomal ω-6 desaturase gene (fad2-1) from soybean. Indian Journal of Experimental Biology, 45: 390-397.

39

3. Kumar R., Sinha S. K., Kishore K., Gupta N. C., Dubey N. and Sachdev A. (2007). Isolation and characterization of fad2-1cDNA sequence from Glycine max L. Indian J. Plant Physiol., 12: 06-12.

Five Best Publications Relevant to the Area of Work: 1. Gupta N. C.,Sinha S. K., Jolly M., Dubey N. and Sachdev A. (2010). Antisense rna-

mediated inhibition of gmfad2-1 encoding omega-6-desaturase. Indian J. Plant Physiol., 14: 336-343.

2. Kishore K., Sinha S. K., Kumar R., Gupta N. C.,DubeyN. and Sachdev A. (2007). Isolation and characterization of microsomal ω-6 desaturase gene (fad2-1) from soybean. IndianJournal of Experimental Biology, 45: 390-397.

3. Kumar R., Sinha S. K., Kishore K., Gupta N. C., Dubey N. and Sachdev A. (2007). Isolation and characterization of fad2-1cDNA sequence from Glycine max L. Indian J. Plant Physiol., 12: 06-12.

Research Support (Externally funded)

Ongoing Research Projects: No externally funded research project (working in inhouse projects only)

Research projects completed: Nil

Place: New Delhi Signature of Co-PI

Date:

40

Name : Dr. Prabhu Kumble Vinod

Designation : Head, Division of Genetics Institute : Division of Genetics, IARI, New Delhi-12 Date of Birth : 20-05-1958 Sex (M/F): Male SC/ST : × Education (Post-Graduation onwards & Professional Career) Sl No.

Institution Place

Degree Awarded

Year Field of Study

1. Banaras Hindu University Varanasi 221 005

M. Sc.(Ag.) 1982 Genetics and Plant Breeding

2. Indian Agricultural Research Institute, New Delhi 110012

Ph.D. 1986 Genetics

Position and Honors

Position and Employment (Starting with the most recent employment) Sl No. Institution

Place Position From (Date) To (date)


Head July 2006 Till date

2. National Phytotron Facility (IARI) New Delhi

Officer-in-Charge January 2003 Till date

3. National Phytotron Facility (IARI) New Delhi

Senior Scientist (PB) & Manager (Biology)

July 1998 Till date

4 National Phytotron Facility (IARI) New Delhi

Manager (Biology) January 1998 Till date

5. IARI, New Delhi Scientist Sr. Scale (PB) April 1992 July 1998 6. IARI RS, Tutikandi, Shimla Scientist Sr. Scale (PB) Oct. 1991 Mar 1992 7. IARI RS Tutikandi, Shimla Scientist S-1 (PB) Oct. 1986 Sept. 1991

Honors/Awards 1. Best Scientist of the Year 2010, Felicitated and honoured by Young Farmers’ Association, Punjab

and Govt. of Punjab in recognition of contribution in effecting a landmark decision by Government of India in enabling recognition of his research contribution in wheat, rice and mustard.

2. ICAR Recognition Award 2008, National Award by ICAR/DARE in recognition of contribution in basmati rice development

3. Fellow, NAAS, A national fellowship from National Academy of Agricultural Sciences in recognition of research on crop improvement

4. Platinum Jubilee Award, National Award by the largest scientific congress in India 5. Distinguished Scientist (Plant Sciences) 2006, Society for Development in Agriculture, Sardar

Vallabh Bhai Patel University of Agriculture & Technology, Meerut Platinum Jubilee Award 2006, Indian Science Congress Association

6. Dr. B. P. Pal Award 2003. Awarded by the IARI, New Delhi in recognition of research in Genetics and Plant Breeding

7. V. S. Mathur Memorial Award 2001-2003. Indian Society of Genetics and Plant Breeding 8. FAO Fellowship 1998. Awarded by the Council for undertaking a 6 month research programme at

the Phytotron under Agriculture and Agri-Food, Canada, Saskatoon Centre, Saskatchewan, Canada.

9. Jawaharlal Nehru Award 1987. Awarded by the Indian Council of Agricultural Research, New Delhi for outstanding post-graduate research in agriculture

41

Professional Experience and Training relevant to the Project Molecular markers for disease resistance in wheat Two recessive genes for resistance working on a complementary basis were identified for the first time and also located on chromosomes 7B and 7D in T. aestivum • An alien leaf rust resistance gene, from Agropyron elongatum that is Lr24 from Agropyron elongatum

located on chromosome 3DS has been tagged with RAPD markers S73700 which is converted to a sequence characterized amplified region (SCAR)

• An alien gene Lr28 from Aegilops speltoides on chromosome 4AL has been tagged with RAPD markers S464700 and S421640 which are converted to SCARs

• An alien gene Lr9 from Aegilops umbellulatum has been tagged and a SCAR marker, a microsatellite marker have been identified linked to the gene.

• The alien gene Lr19 Agropyron elongatum has been tagged with four microsatellite markers on chromosome 7DL

• Molecular maps for genes Lr28, Lr9 and Lr24 have been developed for the first time • Molecular markers were utilized to establish the genetic misidentity and redundancy in the germplasm

of wheat in India • Molecular markers, the RAPD, SCAR and SSR microsatellite were used in tandem to pyramid genes

Lr24 and Lr28 in two wheat backgrounds, HD 2329 ( already fixed to homozygosity) and PBW 343 (in F3 generation) by 2003.

Publications

1. Varieties Released 14 (7 Wheat, 2 Barley, 5 mustard) 2. Research papers in refereed journals 67 3. Symposia/Conference 25 4. Book chapters 12 5. Books 7 6. Popular articles 23 7. Pamphlets/ bulletins 8 8. Thesis (M.Sc / Ph.D) 12

TOTAL 165 Selected peer-reviewed publications (Ten best publications in chronological order) Ragiba M, Prabhu KV and Singh RB. 2004. Location of two complementary recessive genes for

resistance to Helmithosporium leaf blight on chromosomes of hexaploid wheat. Plant Breeding : 123. 407-411.

Cherukuri D. P., S. K. Gupta, A. Charpe, S. Koul, K. V. Prabhu, R. B. Singh and Q. M. R. Haq 2005: Molecular mapping of Aegilops speltoides derived leaf rust resistance gene Lr28 in wheat. Euphytica 143: 19-26.

Dutta M, V. Arunachalam, A. Bandyopadhyay, Prabhu K. V. 1986. Early generation intermating for yield improvement in groundnut (A. hypogaea). Theor. Appl Genet. 71: 662-666.

Sudhir Kumar Gupta, Ashwini Charpe, Sunita Koul, Kumble Vinod Prabhu and Qazi Mohd. Rizwanul Haq. 2005. Development and validation of molecular markers linked to an Aegilops umbellulata derived leaf rust resistance gene Lr9, for marker assisted selection in bread wheat. Genome : 48: 823-830.

Prabhu K V, V. Arunachalam & A. Bandyopadhyay. 1990.Nonparametric approach to multitrait selection for yield in groundnut. Theor. Appl Genet. 80: 223-227

42

Prabhu K V, D J Somers, G. Rakow and R.K. Gugel. 1998. Molecular markers linked to white rust resistance in mustard Brassica juncea. Theor. Appl Genet. 97:. 865-870.129.

Gupta S. K., A. Charpe, S. Koul, Q.M.R. Haque and K. V. Prabhu. 2006. Development and validation of SCAR markers co-segregating with an Agropyron elongatum derived leaf rust resistance gene Lr24 in wheat. Euphytica. (in press).

Daryl J. Somers, Gerhard Rakow, Vinod K. Prabhu and Ken R. D. Friesen. 2001. Identification of a major gene and RAPD markers for yellow seed coat colour in Brassica napus. Genome. 44: 1077-1082.

Gupta S.K, A. Charpe and K. V. Prabhu. 2006. Tagging the Agropyron elongatum derived gene Lr19 with molecular markers in wheat. Theor. Appl. Genet. (Online : DOI 10.1007/s00122-006-0362-7).

S.K. Gupta, A. Charpe, S. Koul, Q.M.R. Haque & K.V. Prabhu. 2006. Development and validation of SCAR markers co-segregating with an Agropyron elongatum derived leaf rust resistance gene Lr24 in wheat. Euphytica. (Online DOI: 10.1007/s10681-006-9113-8).

Basavaraj, S.H., Singh, V.K., Singh, Atul, Singh, Ashutosh Singh, Anita Singh, Sheel Yadav, Ranjith K Ellur, Devinder Singh, Gopala Krishnan, S., Nagarajan, M., Mohapatra, T., Prabhu, K.V. and Singh, A.K. 2010. Marker-assisted improvement of bacterial blight resistance in parental lines of Pusa RH10, a superfine grain aromatic rice hybrid. Molecular Breeding, DOI 10.1007/s11032-010-9407-3. (NAAS ID M029, 7.7)

Anupam Singh, Pallavi, J.K., Promila Gupta, K.V.Prabhu. 2010. Identification of microsatellite marker linked to leaf rust adult plant resistance (APR) gene Lr48 in wheat. Plant Breeding DOI10.1111/J.1439-0523.2010.01820.x. (NAAS ID P040, 7.5)

List maximum of five recent publications relevant to the proposed area of work. Prabhu K V, Sudhir Kumar Gupta, Ashwini Charpe and Sunita Koul. 2004. SCAR marker tagged to the

alien leaf rust resistance gene Lr19 uniquely marking the Agropyron elongatum-derived gene Lr24 in wheat : a revision. Plant Breeding 123: 417-420.

Cherukuri D. P., S. K. Gupta, A. Charpe, S. Koul, K. V. Prabhu, R. B. Singh and Q. M. R. Haq 2005: Molecular mapping of Aegilops speltoides derived leaf rust resistance gene Lr28 in wheat. Euphytica 143: 19-26.

Sudhir Kumar Gupta, Ashwini Charpe, Sunita Koul, Kumble Vinod Prabhu and Qazi Mohd. Rizwanul Haq. 2005. Development and validation of molecular markers linked to an Aegilops umbellulata derived leaf rust resistance gene Lr9, for marker assisted selection in bread wheat. Genome : 48: 823-830.

Gupta S.K, A. Charpe and K. V. Prabhu. 2006. Tagging the Agropyron elongatum derived gene Lr19 with molecular markers in wheat. Theor. Appl. Genet. (Online : DOI 10.1007/s00122-006-0362-7).

S.K. Gupta, A. Charpe, S. Koul, Q.M.R. Haque & K.V. Prabhu. 2006. Development and validation of SCAR markers co-segregating with an Agropyron elongatum derived leaf rust resistance gene Lr24 in wheat. Euphytica. (Online DOI: 10.1007/s10681-006-9113-8).

Basavaraj, S.H., Singh, V.K., Singh, Atul, Singh, Ashutosh Singh, Anita Singh, Sheel Yadav, Ranjith K Ellur, Devinder Singh, Gopala Krishnan, S., Nagarajan, M., Mohapatra, T., Prabhu, K.V. and Singh, A.K. 2010. Marker-assisted improvement of bacterial blight resistance in parental lines of Pusa RH10, a superfine grain aromatic rice hybrid. Molecular Breeding, DOI 10.1007/s11032-010-9407-3. (NAAS ID M029, 7.7)

Anupam Singh, Pallavi, J.K., Promila Gupta, K.V.Prabhu. 2010. Identification of microsatellite marker linked to leaf rust adult plant resistance (APR) gene Lr48 in wheat. Plant Breeding DOI10.1111/J.1439-0523.2010.01820.x. (NAAS ID P040, 7.5)

43

Research Support Ongoing Research Projects Sl No.


Amount (lakhs)


1 Genetic Improvement of Wheat IARI Inhouse 01.04.2009 for 5 years


IARI 616.65 01.04.2009 for 5 years






3. Development of high yielding aromatic/basmati rice varieties and hybrids

IARI Inhouse 01.04.2009 for 5 years

4. Challenge Programme of the Institute: Hybrid development in crops (Wheat, pigeonpea,mustard)

IARI 288.88 01.04.2010 for 5 years



6. Pyramiding drought tolerance in wheat Network project on “Transgenics in Crops (Functional Genomics)”

ICAR 197.00

05.08.2005 for 7 years

7. Genomics and Molecular breeding of linseed/ flax ICAR- Saskatchewan

60.98 XI Plan

9. Network Project on Biotic Stress Resistance in Wheat

DBT 23.00

10. Molecular Breeding for Ug99 Resistance in Wheat ICAR-ACIAR 78.00 11. Network project on Molecular marker assisted

development of biotic stress resistant wheat varieties

DBT 490.07

12. Network project – Molecular markers for broadening the genetic base of stem rust resistance genes effective against strain Ug99, biotic stress rusts.

540.00

13. Development of provitamin A rich indica rice lines through marker assisted backcross breeding (MABB) using high carotenoid golden rice as a donor lines

DBT 99.56

14. Molecular breeding and selection strategies to combine and validate QTLs for improving WUE and heat tolerance of wheat in India

GCP 745.00 March 2011 for 5 years

44

Completed Research Projects (State only major projects of last 3 years) Sl No.


Amount Date of completion



31.03.2009


31.03.2009

3. Molecular Breeding for crop improvement (Brassica)


31.03.2009


DBT 17.02 24.09.2009

5. Development of procedures and protocols for ascertaining the EDV status in rice, wheat, maize and pearlmillet

Deptt. of Agriculture & Coop.

68.00 March, 2009

6. Analysis of Host-Pathogen Interaction in leaf rust infection of wheat: A transcriptomic approach

DBT 13.71 August, 2010

7. Network project on marker assisted selection for leaf rust resistance in wheat

ICAR 20.00 31.03.2008

Place : New Delhi Signature of Co-Principal Investigator Date : 13.12.2011

part i: general information -...

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