Tissue culture and genetic transformation of hot pepper (Capsicum annum L. var Nok- gouang) mediated by Agrobacterium tumefaciens

Date

4-2008

Degree

Bachelor of Science in Biology

Major Course

Major in Cell Biology

College

College of Arts and Sciences (CAS)

Abstract

To determine the response of hot pepper (Capsicum annuum L. var Nok-gouang) re to tissue culture, different explants were cultured on media containing different auxin-cytokinin combinations for callus growth, root formation, and shoot-bud development. Naphthaleneacetic acid (NAA) was used as auxin and benzylaminopurine (BAP) as cytokinin. Different treatments resulted in different degrees of callus development. The highest percentage for callus induction was obtained in the medium containing 1 mg/L NAA - 2mg/L BAP for main leaf (98.33%), cotyledon with petiole (95%), cotyledon without petiole (95%), and blind leaf explants (91.67%). In 0.5mg/L NAA — lmg/L BAP. 70% callus induction was observed in hypocotyl explants. Statistical analysis revealed that there was a significant difference on the effects of different auxin-cytokinin combinations on callus induction. Percentages of root formation in the cultures treated with different auxin-cytokinin combinations were also determined. In 0.5mg/L NAA — Img/L BAP, 26.67% of the explants in the blind leaf culture developed roots, 13.33% in cotyledon without petiole. and 9% in the main leaf culture. In I mg/L NAA — 2mg/L BAP, 6.67% of the explants in cotyledon with petiole culture developed roots. Shoot-!bud development was not observed in the different auxin-cytokinin combinations. Embryos were transformed using the disarmed A. tumefaciens strain AGL1 with co-integrated vector pCAMBIA3301. The T-DNA contained the phosphinothricin (ppt)- resistance gene as a selectable marker and 13-glucuronidase (GUS) as a reporter gene. A total of 68.75% of the infected explants rooted and survived in the selective medium containing phosphinothricin. When histochemical GUS assay was conducted to validate the presence of ppt-resistance gene, localization of GUS activity in all leaf tissues was absent. Consequently. "false-positives" or "escapes" were obtained. A. tumefaciens strain AGL1 may not be compatible to carry out transformation in C annuum var Nok-gouang.

Language

English

Location

UPLB Main Library Special Collections Section (USCS)

Call Number

Thesis

Document Type

Thesis

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