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Paddy rice photosensitivity semi-rolled leaf (PSL1) gene and application thereof

A light-sensitive, genetic technology, applied in applications, genetic engineering, plant genetic improvement, etc., can solve problems such as genetic research that has not been reported

Active Publication Date: 2015-05-06
CHINA NAT RICE RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The CM2088 mutant is phenotypically similar to the rl15(t) mutant, but its genetic studies have not been reported

Method used

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  • Paddy rice photosensitivity semi-rolled leaf (PSL1) gene and application thereof
  • Paddy rice photosensitivity semi-rolled leaf (PSL1) gene and application thereof
  • Paddy rice photosensitivity semi-rolled leaf (PSL1) gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] 1. Rice material

[0041] Rice (Oryza sativa L.) mutant PSL1 (photosensitivity semi-rolled leaf), the original wild-type material is the japonica rice variety "Nipponbare". Rice (Oryza sativa L.) mutant PSL1 (Semi-rolled leaf 1) was treated with 1% (W / V) concentration of chemical mutagen (ethyl methane sulphonate, EMS) from the japonica variety "Nipponbare". , and then obtained a phenotype-stable mutant PSL1 through extensive screening. Except for the light-sensitive curly leaves of the mutant, other phenotypes are similar to those of the wild type, such as figure 1 shown.

[0042] from figure 1 , we can see that: when in the dark state, the mutant's leaves are in a flat state, when in the light state, the mutant's leaves are in a semi-curled state, and when the light intensity is high, the mutant's leaves It tends to be more polar-curled and cylindrical (that is, the leaves are more curled when the light intensity is 12000lx than when the light intensity is 4000l...

Embodiment 2

[0062] plant transformation

[0063] PCR amplifies a genomic DNA fragment with a full length of 4818bp, including 2018bp of the ATG upstream promoter region of the PSL1 gene, 1713bp of the coding region, and 1087bp of the downstream sequence after the terminator. The primer sequences are 5'-GCGGTACCAAGCCTTATGTTTCTCTGCTTG-3' and 5'- GCTCTAGAGTTGGTTAGGTGATGCTTGTTT-3', this fragment was connected to the pCAMBIA1300 vector to obtain the transformation vector pCAMBIA1300-PSL1.

[0064]This plasmid was transformed into Agrobacterium tumefaciens strain EHA105 by electroporation to transform rice. We used the callus induced by the mature embryos of the mutants, cultured in the induction medium for 2 weeks, and selected the vigorously growing callus as the recipient for transformation. The rice callus was infected with the EHA105 strain containing the binary plasmid vector (pCAMBIA1300-PSL1), co-cultivated in the dark at 25°C for 3 days, and cultured in the light on the selection medi...

Embodiment 3

[0066] Embodiment 3, the method for improving rice leaf shape

[0067] Using the cDNA of the japonica rice variety "Nipponbare" as a template, the primers PSLiF: 5'-GGGGTACCGGACTAGTGCAACTGGCTCGTCTTTCTACC-3' and PSLiR: 5'-CGGGATCCCCGAGCTCCAGGTGACGTTCTCGATGTG-3' were amplified (PCR amplification reaction system and amplification procedure refer to the above) to obtain 412bp PSL1 For the gene fragment, restriction sites Kpn I and Spe I were introduced into the front primer, and restriction sites BamH I and Sac I were introduced into the rear primer. The fragment was ligated into pTCK303 vector in the opposite direction to obtain the interference vector pPSLi. The specific steps are as follows: first, double-digest the fragment and the pTCK303 vector with Spe I and Sac I, perform ligation reaction with T4 DNA ligase overnight at 16°C, transform into DH5a Escherichia coli by heat shock, identify the positive plasmid, and then The plasmid and the fragment were digested with Kpn I a...

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Abstract

The invention belongs to the field of plant gene engineering and in particular relates to a paddy rice photosensitivity semi-rolled leaf (PSL1) gene cloned by a map-based cloning technology, a function for verifying the gene by using a transgenic complementation experiment, and an application of the gene. The gene is used for regulating and controlling the development of bulliform cells and vascular bundles of paraxial surfaces of leaves, influencing the morphological development of the leaf, and adjusting the water transportation rate of the leaves, so that an ideal plant of paddy rice can be molded, a germplasm material for drought-resistant breeding can be creased, and the yield of crops can be improved. In the invention, a protein encoded by the paddy rice photosensitivity semi-rolled leaf (PSL1) gene has an amino acid sequence shown by SEQ ID No:2, and the gene for encoding the protein has a nucleotide sequence shown by SEQ ID No:1.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering. Specifically, the present invention relates to a rice PSL1 gene cloned by map-based cloning technology, and the function of the gene is confirmed by transgene complementation experiments; meanwhile, it also relates to the use of the gene to regulate the development of vesicular cells and vascular bundles on the adaxial surface of leaves, Affecting the morphological development of leaves and adjusting the water transport rate of leaves can shape the ideal plant type of rice, create germplasm materials for drought-resistant breeding, and increase the yield of crops. Background technique [0002] Rice is a monocotyledonous model plant, and the external morphological structure of its leaves consists of blades, leaf sheaths, leaf pillows, ligules and auricles. The leaves include three parts: epidermis, mesophyll and veins. The epidermis is composed of upper and lower epidermal cells, vesicul...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82C12N1/21A01H5/00
CPCC07K14/415C12N15/8261C12N15/8273
Inventor 张光恒钱前王莉徐静侯昕沈年伟粘金沯来凯凯饶玉春曾大力胡江任德勇朱丽郭龙彪董国军高振宇
Owner CHINA NAT RICE RES INST
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