The artificial inoculation process and shoot regeneration response in tomato (Solanum lycopersicum L.) transformation mediated by Agrobacterium tumefaciens C. for resistance to Tomato Leaf Curl Virus-Taiwan (ToLCV- Tw).

Date

3-2010

Degree

Bachelor of Science in Biology

College

College of Arts and Sciences (CAS)

Adviser/Committee Chair

Adelina A. Barrion

Abstract

The artificial inoculation process in Agrobactersum-mediated transformation evaluated utilizing two tomato lines: CLN1558A and CLN162I L. Non-transformed plants were treated with Yeast Extract Bacto-Peptone (YEP) medium for the control and Agrobacterium-inoculum of optical density (OD600) 0.2, 1.0, and 2.0; whereas transformed plants were inoculated with OD600= 1.0. Symptom severity was scored 17 days post-inoculation for eight weeks. Results showed that symptoms were first produced at OD600 = 0.2 for CLNI558A and were observed in all concentrations in CLN1621L. In the transformed lines CLNI558A and CLN1621L, 12.5% and 13.5%, respectively, first displayed the symptoms. Statistical analysis revealed that the constructs used in the transformed plants was insignificantly different in developing the disease symptoms. Assuming the symptoms displayed were caused by the virus, OD600-0.2 may be used in disease assessment for the non-transformed lines. In terms of the optimum 6-benzyladenine (BA) concentration that will generate the highest mean number of shoots, 1.0, 0.5, and 0.25 mg/L BA were used for tomato explants with constructs pASIR-27 and pSAS-2. Results showed that the highest mean number of shoots was generated in 0.5 mg/L BA (0.87) using pASIR-27 and in 0.25 mg/L BA (0.89) using pSAS-2. Significant differences were found in the BA concentrations in pASIR-27 but none in pSAS-2. Insignificant differences were found in 0.25 and 0.5 mg/L in terms of the construct used; therefore either of these two concentrations may be used in the shooting medium. Optimizing the inoculation process and shoot regeneration concentration would be useful in developing cost effective protocol.

Language

English

Location

UPLB Main Library Special Collections Section (USCS)

Call Number

Thesis

Document Type

Thesis

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