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Rice half-leaf-curl gene SRL10 and application thereof

A gene, rice technology, applied in application, genetic engineering, plant genetic improvement, etc., can solve problems such as complex genetic mechanism

Active Publication Date: 2019-10-18
CHINA NAT RICE RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among these cloned leaf shape regulatory genes, the molecular regulation mechanism mainly involves the regulation of hormone levels, transcription factors, and microRNA levels, and the genetic mechanism is complex.

Method used

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  • Rice half-leaf-curl gene SRL10 and application thereof
  • Rice half-leaf-curl gene SRL10 and application thereof
  • Rice half-leaf-curl gene SRL10 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1: Map-based cloning of the semi-rolled leaf regulation gene SRL10 gene

[0041] 1. Rice material

[0042] Rice (Oryza sativa L.) semi-rolled leaf mutant srl10 was obtained from the japonica rice variety "Wuyunjing 7" after being treated with a chemical mutagen (ethyl methane sulphonate, EMS) at a concentration of 1%, and then mass screening Phenotypically stable semi-rolled leaf mutant srl10. Except for the half-rolled leaves, the mutant's other phenotypes are similar to those of the wild type, such as figure 1 shown.

[0043] 2. Analyze and target groups

[0044] The homozygous semi-rolled leaf mutant srl10 was crossed with the indica variety TN1, F 1 Generation selfing, get F 2 group, F 2 There were 1715 plants in the generation population, including 1306 plants with normal leaves and 409 plants with semi-rolled leaves. The 409 srl10 mutants were selected as the mapping population. At the tillering stage, about 1 gram of young leaves were taken from ...

Embodiment 2

[0058] Example 2: SRL10 Gene Function Complement Verification

[0059] plant transformation

[0060] A genomic DNA fragment with a full length of 8796bp was obtained by PCR segmentation amplification, including 2109bp of the ATG upstream promoter region of the SRL10 gene, a 5394bp coding region, and a 1293bp downstream sequence after the TAA terminator. Using primers 5'-ACGAATTCGAGCTCGGTAATCGGGAGAATGTTTATCATGG-3' and 5'-CTAGAGGATCCCCGGGTACCGAAAATCTGATACCAAATCACC-3' to amplify the first sequence, primers 5'-GGTGATTTGGTATCAGATTTTCTTGTTTGTAAAGGTCATACTGT-3' and 5'-CTAGAGGATCCCCGGGTACCAATCTCTTGCCGGTGAAAT The amplified fragment was connected to the pCAMBIA1300 vector, and then the vector that had been connected to the first segment verified by sequencing was digested with KpnI and then ligated with the second amplified fragment to obtain a genetic transformation vector containing the full-length sequence of the SRL10 genome pCAMBIA1300-SRL10.

[0061] Then, the genetic transformat...

Embodiment 3

[0062] Example 3: Method for improving rice leaf shape by SRL10 gene editing

[0063]The A pYLCRISPR / Cas9-MH / B vector system of the Liu Yaoguang team of South China Agricultural University was used to construct the gene knockout vector. The specific steps are as follows: first, use primers SRL10-U3-F: ggcaGTGTCAATCGGTAAGGGGG and SRL10-U3-R: aaacCCCCCTTACCGATTGACAC to prepare the target linker, and then use the digested pYL gRNA-U3 vector to react with the corresponding linker to prepare the gRNA expression cassette. Then the gDNA expression cassette was amplified by 2 rounds of nested PCR, the gRNA expression cassette prepared in the above step was used as the template for the first round of amplification, and the amplification primers were U-F: 5'-CTCCGTTTTACCTGTGGAATCG-3' and gRNA-R: 5'- CGGAGGAAAATTCCATCCAC-3', the first-round amplification product was diluted 100-fold as the second-round amplification template, and the amplification primers were Uctcg-B1': 5'-TTCAGAggtctcT...

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Abstract

The invention belongs to the field of plant gene engineering, particularly relates to a rice half-leaf-curl gene SRL10 cloned through a map-based cloning technology and determining of the gene function through a transgenic complementation experiment, and meanwhile further relates to gene editing of the gene. The leaf morphology of a plant is improved, the ideal plant type of the rice can be formed, and the yield of crops is improved. The invention discloses a protein for encoding the rice leaf form control gene SRL10. The protein has the amino acid sequence shown in SEQ ID No:2. The inventionfurther meanwhile discloses a gene for encoding the protein. The gene has the nucleotide sequence shown in the 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 semi-rolled leaf gene SRL10 cloned by map-based cloning technology, and the function of the gene is confirmed by transgene complementation experiments; meanwhile, it also relates to the gene editing of the gene to improve the shape of plant leaves, which can Shape the ideal plant type of rice and increase the yield of crops. Background technique [0002] A major challenge in the development of modern agriculture is to meet the growing global agricultural demand. This challenge underscores the urgent need for strategies to sustainably increase food production (Ray et al., 2012). my country is a big rice producer and also a big rice consumer. Rice has experienced two green revolutions and super rice breeding, and its yield has increased significantly. In recent years, my country's rice planting area has been declining year after year...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/46
CPCC07K14/415C12N15/8261
Inventor 张光恒钱前王佳佳徐静周梦玉陈敏敏曾大力胡江朱丽高振宇任德勇郭龙彪董国军陈光沈兰张强
Owner CHINA NAT RICE RES INST
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