Anatomy of Some Dicot Anatomy of Some Dicot Plants in Salaya Plants in Salaya Putthamonthol Putthamonthol NakornpathomNakornpathom

Mahidol Wittayanusorn School, Thailand

IntroductioIntroductionn

Introduction

Each kind of vegetables has its own specific nutrition and benefit. The vegetables under our study are Chilli (Capsicum frutescens L. var. frutescens) , Celery (Apium graveolens L.) , Parsley (Erygium foetidum Linn.) , Dill (Anethum gaveolens L.) and Bitter Cucumber (Momordica charantia L.) .

All of which are abundant and easy to obtain inthe Salaya area and, moreover, the study in the levelof tissue of these plants are not widely known

Introduction Since all the study in anatomy of plants

arealways on the basis of tissue of these plants are not widely known.

Since all the study in anatomy of plants are always on the basis of tissue level study, the data of this project can be used as the basis of other kinds of studies regarding vegetables, such as the study of the relation between the plants’ structure and the ability to collect antioxidant substance.

Materials, ChemicalsMaterials, Chemicalsand Equipmentsand Equipments

Materials

1. Slide

2. Cover glass

3. Aluminium Foil

4. Paper

Chemicals1. Paraffin 8. TBA (Tertiary Butyl

Alcohol)

2. Paraffin oil 9. Absolute ethyl alcohol

3. Xylene 10. FAA 50%

4. 95% Ethanol (Formalin aceto-alcohol)

5. Fast green 11. Haupt’s adhesive

6. Safranin O 12. Distilled water

7. Glycerine 13. Ether

Equipments1. Microtome 9. Microscope

2. Razor blade 10. Oven

3. Vacuum pump 11. Slide warmer

4. Alcohol lamp 12. Camera

5. Thermometer 13. Beaker

6. Paintbrush 14. Staining Jar

7. Dropper 15. Plate

8. Pin 16. Forceps

MethodsMethods

MethodParaffin

methodSelectingFixi

ngDehydrationInfiltr

ationembeddingSectio

ningAffixationStai

ningMounting

Select and cut plant into square pieces, 0.5

cm in length.

Preserve the specimens in Formalin Acetic Acid

(FAA) at least 8 Hr.

Suck the air out of the specimens with vacuum pump until there isn’t

any bubble.

Wash them with ethanol 50%, at least

3 Hr. 3 times.

Steep in TBA 50%

70%

85%

95%

100%At least

12 Hr.

50%70%85%95%100%

Steep in pure TBA at least 12 Hr. 3 times.

Steep in TBA mixed with paraffin oil 1:1 at least

12 Hr. at 60 Celsius degrees.

60o

Steep in paraplast at least 12 Hr. at 60 Celsius degrees. In 3 times.

60o

Eliminate bubble while Embed in paraplast and wait until getting cool.

Cut in shape of cubic and stick on wooden

block. Then cut with rotary microtome.

Mount them with Formalin 3% and haupt’s adhesive

and lay on slide warmer at 40 -50

oC. Then keep at

room temperature for 5 days.

Dip in xylene

5 Minutes

Xylene+ethanol

ether + ethanol Ethan

ol 95% Ethanol 70%

Ethanol 50%

Ethanol 30%

Steep in Safranin-O 1 Hr. Then wash with

water.

Ethanol 95%

Ethanol 70%

Ethanol 50%

Dip in Ethanol 30% 5

Minutes

Absolute Ethanol +Xylene

Absolute Ethanol

Wash with Ethanol 95%

Clove oil

Drop fast green Steep in

Xylene 6 Hr.

Cover them with cover glass by Canada Balsam or permount. Then bake them

in oven.

Clean up the surplus

mountant and keep them in the slide box.

Examine with microscope and analysis the result.

MethodsSearch for information

Select and preserve the plant specimens in FAA

Dehydrate the tissue by preserving it in Tertiary Butyl Alcohol (TBA)

MethodsPreserve the tissue in melting paraplast at 60

oC

Embedding

Cut it with rotary microtome

Stretch the specimens by using 3% formalin and Haupt’s decisive

MethodsStaining with Safrinin O and Fast green

Wash the stain out

Mounting with mounting medium (Permount),Drop it and cover it with cover glass

Results & Results & DiscussionDiscussion

Results

Parsley Leaf

40x

Results

Dill Leaf

100x

Results

Celery Leaf

40x

Results

Chilli Fruit

40x

Results

Bitter Cucumber Fruit

40x

Results

Bitter Cucumber Fruit

40x

Results

Bitter Cucumber Fruit

40x

ConclusionConclusion

Plants which people admire to eat their leafs, celery had larger area of dermal tissue than dill and celery, respectively. While the ratio of ground tissue area and vascular tissue area, celery had much value than celery and dill, respectively. In another group that people admire to eat their fruits, chili had larger area of dermal tissue than bitter cucumber. But bitter cucumber had more value of ratio of ground tissue area and vascular tissue area than chili.

ReferencesConsortium of Thai Medical Schools. 1997. Local Plant. The War Veterans Organization Press, BangkokC. Brickell. 2003. The Royal Horticultural Society A-Z of GARDEN PLANT volume1: A-J. Dorling Kindersley L imited, London. L. Bremness. 1994. HERBS. Dorling Kindersley

Limited, London. L. Bremness. 2000. HERBS. Dorling Kindersley

Limited, London. Faculty, Kasetsart University, Bangkok

References

Prof. Ass. Dr. Prasart Keurmanee. 2006. Plant Tissue Technique. Botany Department, Science Faculty, Kasetsart University, Bangkok

R.F. Evert. 2006. Esau’s Plant Anatomy 3rd edition. John Wiley & Sons, Inc., New

Jersey. Rost T.L., M.G. Barbour, C.R. Stocking, and T.R.

Murphy. Plant Biology. 2nd edition. Thomson Higher Education, USA.

References

Sylvia Lee-Huang, Paul L. Huang, Philip L. Huang, Aldar S. Bourinbaiar, Hao-Chia Chen and Hsiang-Fu Kung. 1995. Inhibition of The Integrase of Human Immunodeficiency Virus (HIV) Type 1 by Anti-HIV Plant Proteins MAP30 and GAP31. M.S. thesis, Department of Biochemistry, New York University School of Medicine, New York, NY 10016; tAmerican BioSciences, New York, NY 10021; INational Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892; and IlLaboratory of Biochemical Physiology, Biological Response Modifiers Program, Division of Cancer Treatment, National Cancer Institute Frederick Cancer Research and Development Center, Frederick, MD. Faculty, Kasetsart University, Bangkok

References

Tiamjai Komkris. 2003. Plant Anatomy. Botany Department, Science Faculty, Kasetsart

University, BangkokUrai Jiramonkholkarn. 2004. Indigenous Vegetables 1. Ammarin Printing and Publishing Ltd., BangkokUrai Jiramonkholkarn. 2004. Indigenous Vegetables 2. Ammarin Printing and Publishing Ltd., Bangkok