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The Construction of Monocotyledon Expression Vector with Coptis Japonica 4-hydroxyphenylpyruvate Dioxygenase Gene and the Genetic Transformation of Rice

Author: ZhangLiPing
Tutor: LiangYuLing
School: Hebei University
Course: Botany
Keywords: Rice Cjhppd gene Vector construction Herbicide Transgenic
CLC: S511
Type: Master's thesis
Year: 2009
Downloads: 32
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Abstract

P-hydroxyphenyl pyruvate dioxygenase (HPPD) are the tyrosine catabolic processes the second step the desired enzyme, which catalyzes hydroxyphenyl pyruvate converted into urine black acid. , Urine black acid is a precursor of vitamin E and plastoquinone, so the HPPD indirectly control the synthesis of these two types of substances in plants. The family of three types of herbicides, three ketones, pyrazole and isoxazole class HPPD as target sites, competitive inhibition of the the HPPD enzyme activity, resulting in plants whitened and death. Meanwhile, herbicide-resistant transgenic crops caused widespread interest and concern. The study found that the the Coptis HPPD pyrazole herbicides DTP with some resistance. Rice (Oryza saliva L.) laid the foundation for the study of the model plant as monocots genetic transformation for other gramineous crop gene transformation and gene regulation. The rapid development of rice genome research in the last decade, some varieties of rice the completed sketch sequencing of the whole genome and a large number of stable inheritance of transgenic rice plants obtained through the use of transgenic technology. It was tried to some useful gene into the rice, and used to study the expression in monocot model plant. This study was constructed the monocotyledonous plant expression of of herbicide resistance Cjhppd gene carrier, rice by Agrobacterium-mediated transformation, initially detected by PCR verified Cjhppd gene into Rice. Main results are as follows: Construction of monocot plant expression vector, the vector containing Cjhppd gene fragment, the promoter of the maize ubiquitin UBI, selective marker gene Hpt. With the Nipponbare rice seeds induced callus as receptor material transformation, Agrobacterium tumefaciens-mediated method will Cjhppd gene into rice, co-cultured 189 Nipponbare rice callus three filter subculture, 76 resistant calli resistant callus was 40.2%, the 33 resistant callus regenerated plants resistant plants conversion rate of 17.5%. Extract the transgenic rice genome DNA by PCR test validated Cjhppd gene into Rice. Agrobacterium transformation of rice 33 transgenic plants, including 8 PCR positive number of trees, the positive conversion efficiency of 24.2%. The rice leaf herbicide the DTP resistance analysis preliminary results show that in transgenic rice leaves albino DTP stress are significantly lower than in wild-type rice. Transgenic rice leaf chlorophyll a, chlorophyll b, carotenoid content was significantly higher than the non-transformed wild-type rice. Preliminary judgment, exhibit herbicide resistance by Zhuan Cjhppd transgenic rice.

The Construction of Monocotyledon Expression Vector with Coptis Japonica 4-hydroxyphenylpyruvate Dioxygenase Gene and the Genetic Transformation of Rice

Abstract

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