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Application of tomato wrky37 protein in regulating tomato leaf senescence resistance and improving 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: 2022-07-05
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 regulating tomato leaf senescence resistance and improving tomato yield
  • Application of tomato wrky37 protein in regulating tomato leaf senescence resistance and improving tomato yield
  • Application of tomato wrky37 protein in regulating tomato leaf senescence resistance and improving tomato yield

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Effect test

Embodiment 1

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

[0030] 1. Preparation of background materials

[0031] The variety used in the present invention is tomato Micro Tom. Tomato seeds are surface-sterilized with 3% sodium hypochlorite for 15 minutes, and washed with sterile water for 7-8 times after disinfection. After the seeds are washed, the seeds are sown in MS medium. The cotyledon part of the tomato seedling was taken as tissue culture material, the cotyledon tip was cut off, the remaining part was cut into a 5mm × 5mm square, and the treated cotyledon was placed on the pre-culture medium with the back side up for dark cultivation.

[0032] 2. 2 gRNA target selection

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

Embodiment 2

[0049] Example 2 Detection of anti-aging properties of transgenic line tomato and its application

[0050] 1. SlWRKY37 gene editing lines to delay senescence and improve yield detection

[0051] Taking the wild type of tomato Micro Tom as the control group, the knockout line slwrky-KO-1 slwrky-KO-3, which was verified to be homozygous by sequencing, was selected, and the wild type with the same growth and the same number, the gene knockout line slwrky-KO- 1. The seedlings of slwrky-KO-3, 30 plants per line, were planted in a solar greenhouse and a light incubator respectively, and then the natural senescence status of different lines of SlWRKY37 in the natural growth state was observed 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 125 μmol m -2 s -1 , the relative humidity is about 60%.

[0053] At the later stage of plant growth...

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Abstract

The invention relates to the technical field of genetic engineering, in particular to the application of tomato WRKY37 protein in regulating tomato leaf senescence resistance and improving tomato yield. The present invention finds that tomato WRKY37 protein can regulate the anti-leaf senescence ability of tomato and the fruit yield of tomato. In practical applications, gene editing can be used to knock out the WRKY37 gene in tomato and inhibit the expression of the WRKY37 protein to obtain tomato lines with stronger resistance to leaf senescence; or cross slwrky37‑KO plants with other tomato varieties to breed resistance Senescent and high-yielding tomato varieties. The invention utilizes the CRISPR-Cas9 genome site-directed editing system to mutate the tomato WRKY37 gene to create a high-yield tomato line, the resistance to leaf senescence of the tomato line is significantly improved, the yield is also significantly improved, and 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 senescence 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 function of the entire individual and eventually death. Leaf is an important site for plants to synthesize organic compounds using light energy, and is also one of the most sensitive organs for plant senescence (Woo et al., 2019). With the increasing understanding of the genetic control of aging in the field, 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 are frequently reported. WRKY transcription factors were ...

Claims

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Application Information

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Patent Type & Authority Patents(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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