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Gene for increasing tomato fruit weight and number of ventricles and regulation and control method thereof

A gene regulation and tomato technology, applied in the field of bioengineering, can solve problems such as enhancing tomato stress resistance

Pending Publication Date: 2021-10-01
HEFEI UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Studies in Arabidopsis have shown that LCD may be involved in the process of drought resistance in Arabidopsis. Mutations in the LCD1 gene in tomato lead to early ripening of tomato fruits. However, the phenotype of tomato plants with high expression of LCD1 has not been reported yet, so it is speculated that the LCD1 gene is highly expressed , resulting in endogenous H 2 Increased S content may enhance stress resistance of tomato

Method used

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  • Gene for increasing tomato fruit weight and number of ventricles and regulation and control method thereof
  • Gene for increasing tomato fruit weight and number of ventricles and regulation and control method thereof
  • Gene for increasing tomato fruit weight and number of ventricles and regulation and control method thereof

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Effect test

Embodiment 1

[0036] Embodiment 1: A kind of gene that increases tomato fruit weight and ventricle number and its regulation method

[0037] A gene regulation method for increasing the weight of tomato fruit and the number of ventricles, by obtaining the complete coding sequence of the LCD1 gene from the amplified tomato, connecting the coding sequence to the high vector pBI121, transforming the cotyledon of tomato by using the Agrobacterium infection method, and obtaining the transgene Tomato plants: Compared with non-transformed tomatoes, the average weight of transgenic tomato fruits increased by 82.17%, and the number of ventricles increased by 1-2.

[0038] A gene regulation method for increasing the weight of tomato fruit and the number of ventricles is characterized in that the specific implementation method includes the following steps.

[0039] (1) Amplification of the coding sequence of tomato LCD1

[0040] The sequence LOC101258894 of the tomato cysteine ​​desulfhydrylase gene L...

Embodiment 2

[0070] Embodiment 2: A kind of gene that increases tomato fruit weight and ventricle number and its regulation method

[0071] A gene for increasing tomato fruit weight and ventricle number, the nucleotide sequence of the gene is shown in Seq No.1.

[0072] A gene regulation method for increasing tomato fruit weight and ventricle number, comprising the following steps:

[0073] Step 1: using tomato cDNA as a template, performing PCR amplification with upstream primer-F and downstream primer-R, and obtaining the target gene fragment after the PCR amplification product is purified;

[0074] Upstream primer-F: 5'- tacggtacctctagaggatccTAATCCTAAATGGAACCGGC-3';

[0075] Downstream primer-R: 5'- aacatcgtaaggatactcgagTTCTGAGTGAAGCATCTTAC-3';

[0076] Step 2: Carry out double enzyme digestion on the vector pBI121 with the restriction endonuclease BamHI and the restriction endonuclease XhoI, and obtain the linear pBI121 vector after the digestion product is purified;

[0077] Step 3...

Embodiment 3

[0088] Embodiment 3: A kind of gene that increases tomato fruit weight and ventricle number and its regulation method

[0089] A gene for increasing tomato fruit weight and ventricle number, the nucleotide sequence of the gene is shown in Seq No.1.

[0090] A gene regulation method for increasing tomato fruit weight and ventricle number, comprising the following steps:

[0091] Step 1: using tomato cDNA as a template, performing PCR amplification with upstream primer-F and downstream primer-R, and obtaining the target gene fragment after the PCR amplification product is purified;

[0092] Upstream primer-F: 5'- tacggtacctctagaggatccTAATCCTAAATGGAACCGGC-3';

[0093] Downstream primer-R: 5'- aacatcgtaaggatactcgagTTCTGAGTGAAGCATCTTAC-3';

[0094] Step 2: Carry out double enzyme digestion on the vector pBI121 with the restriction endonuclease BamHI and the restriction endonuclease XhoI, and obtain the linear pBI121 vector after the digestion product is purified;

[0095] Step 3...

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Abstract

The invention relates to a gene for increasing tomato fruit weight and number of ventricles and regulation and control method thereof. The regulation method comprises the following steps: step 1, using tomato cDNA as a template, carrying out PCR amplification by using an upstream primer-F and a downstream primer-R, and purifying a PCR amplification product to obtain a target gene segment; step 2, carrying out double enzyme digestion on the vector pBI121 by using restriction enzyme BamHI and restriction enzyme XhoI, and purifying an enzyme digestion product to obtain a linear vector pBI121; step 3, subjecting the target gene segment and a linear pBI121 vector to recombination, and obtaining an LCD1-pBI121 plasmid; step 4, transforming an EHA105 agrobacterium competent cell by using the LCD1-pBI121 plasmid; step 5, infecting tomato cotyledons with the EHA105 agrobacterium tumefaciens containing the LCD1-pBI121 plasmid, and obtaining transgenic positive seedlings. The average weight of the fruit is 7.815 g. LCD1 high expression plant fruit 4 ventricles and higher ventricles take up about 35%.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and relates to a gene for increasing tomato fruit weight and ventricle number and a regulation method thereof. Background technique [0002] Tomato (scientific name: Solanum lycopersicum ), that is, tomato, is a plant of the tubular flower order, Solanaceae, and tomato genus. Tomato fruit is rich in nutrients, rich in lycopene, flavonoids, vitamin C and other nutrients. Tomatoes are widely cultivated in my country. How to increase the weight of tomato fruits and increase the yield of tomatoes is still a problem at present. Searching for new genes controlling fruit size plays an important role in tomato molecular breeding. [0003] Tomato fruit size or weight is an important agronomic trait of tomato, which has been affected by artificial selection during tomato evolution. Currently, multiple genes have been found to control fruit size. For example, mutations in ENO, an AP2 / ERF transcr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/60C12N15/84A01H5/08A01H6/82
CPCC12N9/88C12N15/8205C12N15/8261C12Y404/01001
Inventor 张华胡康棣姚改芳孙红叶孙忱彭湘君赵玉琪宋慧慧李立霞
Owner HEFEI UNIV OF TECH