1. In Vitro Morphogenetic Responses of Fenugreek and Basil Explants By: Jelisa Thomas Food and Animal Science Mentor: Dr. Leopold Nyochembeng

2. BACKGROUND INFORMATION

3. Interesting Facts FENUGREEK is an herb known for its high medicinal value. it originated from India grown in Europe and Asia Basil This herb is native to India and Persia approximately 150 different species.

4. Fenugreek uses Medically ◦ Soar Throat Culinary ◦ Component in Curry etc. Therapeutically ◦ Oils ◦ Soaps, etc

5. Basil Uses Culinary ◦ flavor used in various cuisines Medically ◦ internally and externally Therapeutically ◦ oil or soaps

6. OBJECTIVES  To develop a protocol for callus production and cell culture on Fenugreek explants  Assess effects of genotype and media on basil plantlet multiplication

7. MATERIALS AND METHODS

8. MEDIA, EXPLANTS PREPARATION, AND CULTURE

9. Seed Germination Germination medium contained  half strength M&S (Murashige and Skoog 1962) salt,  7.5 g/L sucrose  4 g/L agar  pH. 5.5-6.0  Autoclaving at 15psi and 121°C for 20 min. Disinfestations  30% Clorox bleach (1.8% NaOCl) containing 1 drop of Tween 20 under vacuum [600-700mmHg] for 20 minutes.  Rinsed 3 changes of sterile distilled deionize water.  Blotted dry with sterile filter paper and place germination media All cultures were sealed with parafilm and placed in an incubator at 23± 1°C and 16 hr/8 hr light/dark photoperiod.

10. Fenugreek Explants Media o M&S salts o 30 g/L sucrose o A factorial combination of  NAA (0.0, 1.0 mg/L)  Kinetin (0.0, 0.3, 1.0, 3.0 mg/L) o Agar(8 g/L) o pH 5.6-6.0 o autoclaving at 15psi and 121°C for 20 min.

11. Basil seedlings of 5 genotypes (23, 82, 62, 74, and 83) Media M & S salts 30 g/L sucrose a combination of Kinetin (0.1, 1.0, 3.0 mg/L) NAA (0.1 mg/L) Agar(8 g/L). pH 5.6-6.0 Autoclaving at 15psi and 121°C for 20 min.  The seedlings of each basil genotype were evenly distributed in all media.

12. All cultures were sealed with parafilm and placed in an incubator at 23± 1°C and 16 hr/8 hr light/dark photoperiod.

13. Data and Analyses  All cultures were observed weekly for contamination, explants growth and differentiation.  The parameter that were measured included the number of roots formed, shoots, callus production, plantlet production  The data was analyzed by taking the averages of counts.

14. RESULTS AND DISCUSSION

21. CONCLUSION Such responses will help in determining the most suitable protocol for micropropagating these important medicinal plants Choice of explants and the type of genotype used are important criteria for successful application of micropropagation and cell culture techniques in Fenugreek and basil respectively.

22. REFERENCES References Jan Schooley- Ginseng and Medicinal Herbs Specialist/OMAF, “Basil” 2003 http://www.omafra.gov.on.ca/english/crops/ort/herbs/basil.htm accessed 14 July, 2009 http://www.omafra.gov.on.ca/english/crops/ort/herbs/basil.htm James E. Simon, Purdue University, West Lafayette IN 47907-1165, USA, “Basil” 1995 http://www.hort.purdue.edu/newcrop/CropFactSheet/basil.html accessed 15 July, 2009 http://www.hort.purdue.edu/newcrop/CropFactSheet/basil.html Jeanine M. Davis, Extension Horticultural Specialist Department of Horticultural Science North Carolina Cooperative Extension Service, North Carolina State University “Basil” 1997 http://www.ces.ncsu.edu/depts/hort/hil/hil-125.html access 14 July, 2009 http://www.ces.ncsu.edu/depts/hort/hil/hil-125.html S. Elliot “The Facts About Fenugreek Growing and Using Fenugreek” 2009 http://herb-gardens.suite101.com/article.cfm/the_facts_about_fenugreek#ixzz0LrzADCmg http://herb-gardens.suite101.com/article.cfm/the_facts_about_fenugreek#ixzz0LrzADCmg accessed 14 July, 2009 Acram Taji, Prakrash P. Kumar, Prakash Lakshmanan “Invitro plant Breeding” 2002 http://book.google.com/books?id=CQxsDZXHVuYC&dq=stages+of+microprogation&source=gbs _navlinks_s accessed 15 July, 2009 http://book.google.com/books?id=CQxsDZXHVuYC&dq=stages+of+microprogation&source=gbs _navlinks_s

23. ACKNOWLEDGEMENTS Dr. Leopold Nyochembeng Dr. Elisa Moss REU Supporters and Participators Department of Plant and Soil Sciences Department of Food an Animal Sciences