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Application of gene in promoting rice grain yield

A rice grain and gene technology, applied in the field of crop molecular genetics and breeding, can solve the problem of not informing the relationship between grain size and yield

Pending Publication Date: 2019-08-16
ZHEJIANG NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no information about the relationship between knockout of Os05g0462000 gene and rice grain size and rice grain yield

Method used

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  • Application of gene in promoting rice grain yield
  • Application of gene in promoting rice grain yield
  • Application of gene in promoting rice grain yield

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1, CRISPR / Cas9 vector construction of Os05g0462000 gene

[0019] According to the nucleotide sequence (SEQ ID NO:1) of the Os05g0462000 gene, two CRISPR / Cas9-edited target sequence sgRNAs were designed through the CRISPR Design program (http: / / crispr.mit.edu / ), and the sequence is sgRNA -1: 5'-GAACAAGGAGGAGTGCATGG-3', sgRNA-2: 5'-GATGTTGGCATGCTTCTCCAGGG-3'; and the sequence of the sgRNA synthesized in a biotechnology company (such as Shanghai Invitrogen).

[0020] Two corresponding vectors were respectively constructed using the CRISPR / Cas9 kit (Biogle, Cat#BGK03), and the method was according to the product instructions.

Embodiment 2

[0021] Rice genetic transformation of embodiment 2, CRISPR / Cas9 vector

[0022] Rice tissue culture and carrier transgene, according to the method of the document "A protocol for Agrobacterium-mediated transformation in rice" (quoted from: Nat Protoc.2006,1(6):2796-2802.), the method constructed in Example 1 The vector is genetically transformed into a wild type rice variety Nipponbare (English name: Nipponbare) to obtain a transgenic rice with Os05g0462000 gene mutation.

Embodiment 3

[0023] Example 3, Sequencing analysis of Os05g0462000 gene mutant strain

[0024] Genomic DNA of transgenic rice was extracted by SDS method: take 0.1 g of rice leaves, grind with liquid nitrogen, add 600 μl of extract (0.1mol / L Tris-Cl pH8.0, 500mmol / L NaCl, 1.25g / L SDS), Incubate at 65°C for 30min. Add 200μl 5mol / L KAC, mix well, and place in ice bath for 30min. Add 500 μl of chloroform, mix well, centrifuge at 10,000 r / min for 5 min, take the supernatant, add 2 / 3 of the supernatant volume isopropanol, mix well, and centrifuge at 12,000 r / min for 5 min. Discard the supernatant, wash the pellet with 70% ethanol, invert to dry, and add 100 μl TE to dissolve the DNA.

[0025] The primer sequences F: 5'-AGATAGATAAGTAAGCAGTGAG-3' and R: 5'-AGCTAGCTCTCGTCGTCGTC-3' of the Os05g0462000 gene were amplified by PCR, and the DNA of the primers was synthesized in a biotechnology company (such as Shanghai Invitrogen). 20μl system for PCR amplification, containing 25~50ng template DNA, ...

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Abstract

The invention relates to a rice grain yield related gene and an application thereof, which belong to the field of crop molecular genetic breeding, in particular, the present invention discloses an application of a gene for promoting the rice grain yield, and a nucleotide sequence of the gene is shown in SEQ ID NO: 2 or SEQ ID NO: 3. The gene can be used to promote the increase of rice grain yield,and can also be used to promote grain size increase of the grain.

Description

technical field [0001] The invention relates to rice grain yield-related genes and applications thereof, and belongs to the field of crop molecular genetics and breeding. Background technique [0002] Rice is an important food crop, and high-yield breeding is an effective guarantee to ensure food security and sustainable agricultural development in my country. Rice yield is closely related to grain weight. Kernel weight refers to the weight of each seed, largely determined by the size of the seed and the degree of filling. Therefore, discovering genes that can increase rice grain weight in rice has great application value for high-yield breeding of rice. [0003] Rice yield is determined by the three factors of tiller number, grain number per panicle and grain weight. The so-called tiller number refers to the number of branches produced by each rice plant in the later stage of development, while the number of spikes refers to the number of grains on each branch, and the g...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/82A01H5/10A01H6/46
CPCC07K14/415C12N15/8261
Inventor 袁俊杰周蒋毅吴悠亦陈析丰马伯军
Owner ZHEJIANG NORMAL UNIVERSITY
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