Rice chlorophyll synthase mutant gene and its application in gene engineering

A technology for mutating genes and chlorophyll is applied in the rice chlorophyll synthase mutant gene Osygl1 and its genetic engineering applications, in the fields of rice chlorophyll synthesis and rice leaf color development, and can solve problems such as preliminary positioning of limited genes.

Inactive Publication Date: 2008-01-02
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
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  • Claims
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AI Technical Summary

Problems solved by technology

At present, although some yellow-green leaf mutants have been obtained in rice, most studies are limited to the preliminary mapping of genes

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1 Molecular Cloning of Rice Chlorophyll Synthase Mutant Gene

[0017] 1. Positioning of the mutant gene ygl1 site

[0018] F 2 Genetic analysis of the populations revealed that the yellow-green leaf phenotype of ygl1 is controlled by a recessive single gene. F derived from ygl1 mutant and Pei'ai 64 2 In the population (12,300 plants), 2741 yellow-leaf recessive individuals were selected for fine mapping of the mutant gene. In the summer of 2005, it was planted in the net room of the Crop Science Research Institute of the Chinese Academy of Agricultural Sciences. The phenotype was identified at the 3-leaf and 1-heart stage, and DNA was extracted at the same time. from F 2 11 individual plants with yellow-green leaves and 34 individual plants with green leaves were randomly selected from the population, PCR amplification was performed on this small population with candidate markers, and then the linkage relationship between candidate markers and mutation sites...

Embodiment 2

[0033] Example 2 Transgenic application test of chlorophyll synthase gene YGL1 and functional verification of chlorophyll synthase mutant gene ygl1

[0034] (1) Construction of cDNA expression vector containing YGL1 coding region

[0035] The hygromycin resistance expression vector pCUbi1390 was cut with Pst I and Spe I (publicly known and public) (provided by Lu Tiegang, a researcher at the Institute of Biology, Chinese Academy of Agricultural Sciences; doctoral thesis of Chinese Academy of Agricultural Sciences, Peng Hao, 2005), and the vector fragment was recovered , spare; by PCR-mediated addition of Pst I / Spe I adapters at both ends of the YGL1 gene cDNA sequence (the adapter primer sequence is SEQ ID NO.25, 26) into the pMD18-T vector, after sequencing confirmation, enzyme digestion, electrophoresis And recover the target fragment, and connect it into the above expression vector to obtain the YGL1 gene recombination vector.

[0036] (2) Agrobacterium-mediated transforma...

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PUM

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Abstract

The invention discloses a nucleotide sequence of rice chlorophyll synthase mutation gene Osygl1 and gene engineering application in the gene engineering domain, which is characterized by the following: the cDNA sequence of Osygl1 is SEQ ID NO.1 and its coded amino acid sequence is SEQ ID NO.2; the gene OsYGL1 is a chlorophyll synthase gene in the rice, which participates the synthesis of chlorophyll a and chlorophyll b to display component expression; the mutation of the gene leads the leaf of rice to become yellow green, which can be genetic mark to identify the kind in the agricultural manufacturing and rice genetic breeding.

Description

technical field [0001] The invention relates to a novel rice chlorophyll synthase mutant gene Osygl1 and its application in genetic engineering, belonging to the field of genetic engineering, and specifically relates to genes for rice chlorophyll synthesis and rice leaf color development. technical background [0002] Plant leaf color is a comprehensive expression of various pigments in chloroplasts. In normal leaves, chlorophyll is dominant and usually appears green. Leaf color variation is a mutation trait with high mutation frequency and easy identification in higher plants. So far, leaf color mutants have been found in almost all higher plants such as rice, wheat, barley, corn, soybean, cotton, tobacco, corn, sunflower, tomato, cucumber, potato, and mulberry. The mutated gene directly or indirectly affects the synthesis and degradation of photosynthetic pigments, especially chlorophyll, resulting in changes in the content and proportion of various pigments, thereby caus...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/52C12N9/00C12N15/82
Inventor 万建民吴自明张欣江玲陈亮明苏宁程治军刘世家刘喜何冰王春明
Owner NANJING AGRICULTURAL UNIVERSITY
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