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Preliminary Study on RNA Interference of Strawberry DHAR Gene

Author: LiuDan
Tutor: GaoQingYu
School: Northeast Agricultural University
Course: Pomology
Keywords: Strawberries DHAR Regeneration System Genetic transformation RNAi
CLC: S668.4
Type: Master's thesis
Year: 2011
Downloads: 48
Quote: 1
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Ascorbic acid plants, not only because it is not normally synthesized AsA few animals (including humans) to provide a rich source of vitamin C, it has a very important physiological functions in the antioxidant effect of plant photosynthesis and growth and metabolism. Therefore, AsA content has become an important indicator to measure the quality of agricultural products. Ascorbic acid reductase dehydroascorbate (DHA) reduced to AsA, recycling of AsA protect cellular components play an important role against oxidative damage. Strawberry high nutritional value, rich in vitamin C. The experiments by RNAi plants build strawberries DHAR gene expression vector to study the role of DHAR in AsA accumulation, the important genes for the vitamin C in the metabolic process to verify DHAR provide a theoretical basis for future high-AsA breeding. The results of this study are as follows: 1. Determination of the vitamin C content of fruit of different maturity and Sen Gala dehydroascorbate reductase activity, the study results show that: the same varieties of strawberries from the mature green stage to the vitamin C content of the full red stage showed a gradual the largest increase between upward trend, full red stage with red ripe stage; Sen Gala dehydrogenation activity of ascorbic acid reductase activity at any time change is also gradually increasing activity was higher in the late growth stage AsA accumulation may mainly in the late growth stage. The the DPS software analysis showed that: and DHAR, activity of the vitamin C content was positively correlated with a correlation coefficient of 0.878, 0.05 level significantly. Initially identified certain positive correlation between the accumulation of strawberry DHAR activity of vitamin C content. Strawberry regeneration system optimization study different hormone combinations adventitious bud induction rate of adventitious bud differentiation and incubated in the dark. The results show that: The Sen Gala leaves induced optimal medium for the MS 3.0mg/L6-BA 0.2mg/L2 ,4-D, the induction rate reached 73.3% in the medium, the average sprouting up 2.2. Cultured for 7 days before dark, and then light train, the higher rate of adventitious buds. Agrobacterium-mediated genetic transformation system optimization strawberry varieties Sen Gala vaccine leaves the material of strawberry genetic transformation system has been optimized. Kanamycin and agents concentration of genetic transformation: 300 mg / L (Cef) sterilization explants highest differentiation rate reached 47.3% adventitious bud differentiation stage, Sen Gala the Km screening pressure is 20mg / L. To further investigate the role of DHAR we do the following research: using RT-PCR method, cloned into strawberry fruit dehydroascorbate enzyme gene DHAR, cDNA fragments; to build dehydroascorbate gene RNAi expression vector Part27-iDHAR; through Agrobacterium-mediated transformation, RNAi plants the expression contained import strawberries Senga La RNAi plants, PCR detection tested positive 7.

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