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Rice gene GSNL4 and application of protein coded by rice gene GSNL4

A protein and rice technology, applied in application, genetic engineering, plant genetic improvement, etc., can solve problems such as negative effects

Pending Publication Date: 2021-06-01
中农常乐(深圳)生物育种技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In order to improve rice yield, breeders usually use polygenic aggregation to seek breakthroughs, but aggregation of multiple yield genes often brings negative effects

Method used

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  • Rice gene GSNL4 and application of protein coded by rice gene GSNL4
  • Rice gene GSNL4 and application of protein coded by rice gene GSNL4
  • Rice gene GSNL4 and application of protein coded by rice gene GSNL4

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] 1. Rice material:

[0037] The original wild type of rice (Oryza sativa L.) grain size and narrow leaf mutant gsnl4 (grain size and narrow leaf4) is the japonica variety Wuyunjing 21 (W21). like figure 1 Shown are the phenotypes of the mutant gsnl4 and the wild-type material. It can be seen from the figure that compared with the wild-type, the rice mutant has narrower leaves, narrower grain width, and shorter grain length.

[0038] The process of obtaining the rice (Oryza sativa L.) grain-type leaf-type mutant gsnl4 is as follows:

[0039] The seeds of japonica rice variety Wuyunjing 21 were mutagenized by EMS and planted in the field. The seeds were collected and planted continuously. From M 2 A narrow-grain and narrow-leaf mutant isolated from the first generation, tentatively named gsnl4 (grain size and narrowleaf4). The mutant is self-crossed for multiple generations to obtain a mutant plant that can be stably inherited.

[0040] The mutant material gsnl4 and th...

Embodiment 2

[0083] Knockout vector construction: eg Figure 4 , respectively design two target sites on the exons of the gene, firstly build into the intermediate carrier gRNA, and then build into the pCAMBIA1300-Cas9 knockout vector.

[0084] Gene editing target sites are:

[0085] gRNA1:AGAACTGGGTCTGGCAGCAC

[0086] gRNA2: ACATGCTCAGACCGCAGGGC

[0087] The gene editing detection primers are:

[0088] C1F: TCTCCTCTTTGCGCACCATT

[0089] C1R: TGGTATTGGACATGTGGGGC

[0090] C2F: CACCTGTAGCTGGAACTTGCT

[0091] C2R: CCCCCACCTGAAAAGTAGGAC

[0092] The construction of GFP vector (such as Figure 5 shown). The primer sequence of the GFP vector is:

[0093] G-F:5'-TACAATTACAGTCGACATGGTGAAGAAGAAAAGAACTG-3'

[0094] G-R: 5'-ATCCTCTAGAGTCGACGCAGTAAAACATGACACG-3';

[0095] The plasmid was transformed into Agrobacterium tumefaciens strain EHA105 by electric shock method, and then the rice callus was transformed by Agrobacterium tumefaciens. We used the callus induced by the immature embryos...

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Abstract

The invention discloses a rice gene GSNL4 (Seq ID No: 1, 2) and an application of a protein coded by the rice gene GSNL4 (Seq ID No: 3), and the gene and the protein are used for regulating and controlling the grain shape and the leaf shape of rice and also used for improving the grain weight and high-yield breeding of the rice. According to the invention, a map-based cloning technology is adopted to clone the GSNL4 gene in rice for the first time, and the function of the gene is identified through a transgenic knockout experiment; through function interpretation of the GSNL4 gene, a foundation is laid for further clarification of a genetic mechanism and an action mechanism of the gene in rice yield formation and creation of high-yield new rice germplasm, and the prospect exploration of the GSNL4 gene in rice high-yield breeding is expanded.

Description

technical field [0001] The invention relates to the field of plant genetic engineering, in particular to the application of a rice gene GSNL4 (grain size and narrow leaf4) and its coded protein. Background technique [0002] Rice is one of the important food crops in my country, and it is also an ideal model plant for monocots. In rice super-high-yield breeding, grain weight is one of the three factors that determine yield. It is generally measured by thousand-grain weight, which is positively correlated with grain size, and grain size can be divided into grain length, grain width and grain thickness. type indicators. Grain shape, as an important shape of the ideal plant shape of rice, plays a vital role in the increase of rice yield. Therefore, the study of rice grain shape has always been a hot spot and focus of domestic and foreign breeders. With the advancement and development of modern molecular biology and marker technologies such as RFLP, RAPD and SSR, more and more...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/82C07K14/415A01H5/10A01H5/12A01H6/46
CPCC07K14/415C12N15/8261Y02A40/146
Inventor 吴世强郭龙彪卢颖莫国雄宋梦秋
Owner 中农常乐(深圳)生物育种技术有限公司