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Identification of Transgenic Rice Plants Harboring HVA1 and Establishment of a System for Propagating It

Author: FangJun
Tutor: YangShiHu
School: Nanjing Agricultural College
Course: Biochemistry and Molecular Biology
Keywords: Transgenic Rice Multiplication Regeneration system Suspension culture Axillary bud Adventitious buds
CLC: S511
Type: Master's thesis
Year: 2003
Downloads: 62
Quote: 0
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Abstract


This experiment to gene gun mediated get the the rice plant regeneration (HVA1 target gene, Bar gene as a selectable marker) material from two aspects. First, the herbicide tolerance of transgenic material detection, molecular detection, field surveys and salt tolerance identification; second is the view of the genetically modified material can not get the seed for the preservation and propagation of genetically modified material, the establishment of the reproductive system of the transgenic plants. 1. Result of transgenic identified: 1.1 turn transgenic plants Basta resistance detection: spraying 4mg / L Basta, a week later, the transgenic plants showed significant resistance, leaves still green plants grew normally, while the control completely yellow leaves plants stunted, finally dead. 1.2 Southern Blot showed that: In the majority of the transgenic lines, a specific band of the expected 1.2kb and HVA1 integrated into the genome in the form of multiple copies. 1.3 field of transgenic plants was observed that: the transgenic plant morphological great variability, the performance of the plants grow shorter, tillering increase, low pollen fertility, selfing and hybridization are not normal seed. Salt tolerance identification of of 1.4 genetically modified material: the whole plant level, the use of transgenic plants contemporary tiller seedlings and tissue culture of asexual reproduction in hydroponic solution tolerance identification, the results show that the salt tolerance of transgenic plants , two weeks after high salt concentration of 0.75% NaCl, transgenic plants could grow, while the basic control plants can not grow. Regeneration medium and rooting medium to transfer genetic material from in vitro level (callus), the the young panicle induction of callus subculture medium, have demonstrated a salt tolerance higher than the control material. The above results show that the HVA1 gene has been integrated into the genome of plants, and to enhance the salt tolerance of rice, it is feasible to use improved crop salt tolerance of. 2. The results: 2.1 transgenic breeding system established transgenic young spike regeneration system. Different cytokinin (6-BA, KT, ZT) the young panicle regeneration, the results show that the best regeneration phase 2mg / L 6-BA. 2.2 The establishment of transgenic material suspension cell lines. In its process to explore the rare elements lanthanum (La 3 ) suspension cells. Found that low concentrations of La 3 (1mg / L) suspension cells promote performance suspension cell growth increased SOD activity increased, decreased MDA (malondialdehyde), renewable improve their ability; while high concentrations of La 3 instead harmful effects of suspension cells. 2.3 stem segments (containing axillary buds) micropropagation system. Of different concentrations of 6-BA on the internode bud rapid propagation. In the induction phase, 20mg/L6-BA to efficiently induce axillary buds and adventitious buds plexus; 10 mg / L 6-BA subculture micropropagation stage of the axillary buds micropropagation effect best, rather than fixed bud cluster 4mg / L 6-BA multiply fastest. Finally rooting stage of 1mg / L IAA1/2MS medium, it is possible to obtain a large number of transgenic seedlings.

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