Application of tomato WRKY37 protein in regulation and control of leaf aging resistance of tomatoes and improvement of tomato yield

A technology of WRKY37 and leaf senescence, applied in the field of gene editing, can solve the problems of regulating leaf senescence, achieve great application value, improve the ability of anti-leaf senescence, and increase the effect of fruit yield

Active Publication Date: 2021-09-24
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As the field's understanding of the genetic control of aging continues to deepen, transcription factors of the WRKY, MYB, and NAM / ATAF1 / CUC2 (NAC) families have become regulators of this process (Kim et al., 2018). Studies on the regulation of leaf senescence by the WRKY family of factors have been frequently reported

Method used

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  • Application of tomato WRKY37 protein in regulation and control of leaf aging resistance of tomatoes and improvement of tomato yield
  • Application of tomato WRKY37 protein in regulation and control of leaf aging resistance of tomatoes and improvement of tomato yield
  • Application of tomato WRKY37 protein in regulation and control of leaf aging resistance of tomatoes and improvement of tomato yield

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Experimental program
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Embodiment 1

[0029] Example 1 Knockout of tomato SlWRKY37 gene based on CRISPR-Cas9 system

[0030] 1. Background material preparation

[0031] The variety used in the present invention is tomato Micro Tom. The tomato seeds are surface-sterilized with 3% sodium hypochlorite for 15 minutes, washed with sterile water for 7-8 times after disinfection, and the seeds are sown in MS medium after the seeds are washed. The cotyledons of tomato seedlings were taken as tissue culture materials, the tips of the cotyledons were cut off, and the remaining part was cut into squares of 5mm×5mm.

[0032] 2. Selection of 2 gRNA targets

[0033] Log in to the NCBI website (https: / / www.ncbi.nlm.nih.gov / ) to query the sequence of the tomato S1WRKY37 gene. According to the SlWRKY37 gene sequence, it contains two exons ( figure 1 ), based on the 2gRNA sequence characteristics of CRISPR-Cas9, the selected target is located at the sequence WRKY37-gRNA1 and WRKY37-gRNA2 containing NGG sequence characteristics o...

Embodiment 2

[0049] Example 2 Anti-aging Detection and Application of Transgenic Tomato Lines

[0050] 1. SlWRKY37 gene-edited strain delays aging and improves yield detection

[0051] Taking the tomato Micro Tom wild type as the control group, select the homozygous knockout line slwrky-KO-1slwrky-KO-3 verified by sequencing, and select the wild type with the same growth and the same number, and the gene knockout line slwrky-KO- 1. Seedlings of slwrky-KO-3, 30 strains per strain, were planted in the solar greenhouse and light incubator respectively, and then observed the natural aging status of different strains of SlWRKY37 in the natural growth state, until the whole life cycle of fruit ripening .

[0052] Lighting incubator settings: the light cycle is 16 / 8h (light / dark), the temperature is 25 / 18°C (light / dark), and the light intensity is 125μmol m -2 the s -1 , the relative humidity is about 60%.

[0053] At the later stage of plant growth, compared with the wild-type control group,...

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Abstract

The invention relates to the technical field of gene engineering, in particular to application of tomato WRKY37 protein in regulation and control of leaf aging resistance of tomatoes and improvement of tomato yield. The tomato WRKY37 protein is found to be capable of regulating and controlling the leaf aging resistance of tomatoes and regulating and controlling the fruit yield of the tomatoes. In practical application, the WRKY37 gene in the tomato can be knocked out through a gene editing means, and expression of the WRKY37 protein is inhibited, so that a tomato strain with higher leaf aging resistance is obtained; or the slwrky37-KO plant is hybridized with other tomato varieties to cultivate an anti-aging and high-yield tomato variety. The tomato WRKY37 gene is mutated by utilizing a CRISPR-Cas9 genome site-directed editing system, a high-yield tomato strain is created, the leaf aging resistance of the tomato strain is remarkably improved, the yield is also remarkably improved, and the invention has important application value.

Description

technical field [0001] The invention relates to the technical field of gene editing, in particular to the application of tomato WRKY37 protein in regulating tomato leaf aging resistance and improving tomato yield. Background technique [0002] Aging is a ubiquitous and important physiological phenomenon, which usually refers to the process of the gradual decline of biological organs or the physiological functions of the whole individual and eventually death. Leaf is an important place for plants to synthesize organic compounds using light energy, and it is also one of the most sensitive organs of plant aging (Woo et al., 2019). As the field's understanding of the genetic control of aging continues to deepen, transcription factors of the WRKY, MYB, and NAM / ATAF1 / CUC2 (NAC) families have become regulators of this process (Kim et al., 2018). Studies on the regulation of leaf senescence by the WRKY family of factors have been frequently reported. WRKY transcription factors wer...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C12N15/29C12N15/113A01H5/08A01H5/12A01H6/82
CPCC12N15/8218C12N15/8266C12N15/8261C12N15/113C07K14/415C12N2310/20Y02A40/146
Inventor 张娜王志荣高明赵冰郭仰东王馨曼曹萌董晓南
Owner CHINA AGRI UNIV
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