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Tomato bacterial wilt resistance gene Slalpha-KGDH E2 and application thereof

One-KGDHE2, tomato bacterial wilt technology, applied to tomato bacterial wilt resistance gene Slα-KGDHE2 and its application field, can solve problems such as difficult large-scale promotion and application, limited breeding progress, time-consuming and labor-intensive costs, and achieve important research value , increased gene expression, and broad application prospects

Active Publication Date: 2021-04-09
ZHEJIANG UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, high-efficiency, low-toxicity and non-polluting chemical or biological fungicides that can be applied to production have not been developed so far. Although some methods such as paddy and dry crop rotation can effectively control the development of bacterial wilt, most of them are time-consuming, labor-intensive and costly. Difficult to promote and apply on a large scale
At present, the most effective way to prevent and control bacterial wilt is to use modern molecular technology to speed up the breeding and promotion of disease-resistant varieties, but the progress of tomato bacterial wilt-resistant molecular breeding in my country is very limited.

Method used

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  • Tomato bacterial wilt resistance gene Slalpha-KGDH E2 and application thereof
  • Tomato bacterial wilt resistance gene Slalpha-KGDH E2 and application thereof
  • Tomato bacterial wilt resistance gene Slalpha-KGDH E2 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Genetic analysis of resistance to bacterial wilt of tomato 'ZRS-7'

[0038] The F1 generation was obtained by crossing the resistant variety 'ZRS-7' with stable and extreme disease-resistant traits and the susceptible variety 'HTY-9' as parents. The results showed that the F1 generation was susceptible to bacterial wilt, but the disease index was lower compared with the susceptible parents; 2 A total of 156 plants were counted, and the resistance and susceptibility traits were separated. The disease rate was 72.22% and the disease index was 58.8%. The onset time of most of the susceptible plants of the F2 generation was about 2 days later than that of the susceptible parents; some of the F2 generation plants were resistant to the disease, the onset time was delayed, and symptoms began to appear at 10dpi. Due to the complexity of the genetic law of bacterial wilt resistance, it was determined that the resistance to bacterial wilt of 'ZRS_7' tomato was recessiv...

Embodiment 2

[0039] Example 2 Localization of the main resistance gene of 'ZRS-7' tomato bacterial wilt resistance

[0040] 25 highly disease-resistant and highly susceptible plants were selected from the F2 generation segregation population, and the genomic DNA was extracted and mixed in equal amounts to construct a disease-resistant pool (R-pool) and a disease-susceptible pool (S-pool) respectively. -7' and 'HTY-9' were the two parental pools plus the disease-resistant pool and the susceptible pool, a total of four samples were subjected to whole-genome sequencing. Through BSA-seq analysis, compare the difference of SNP-index and Indel-index between the offspring and the reference parent and the distribution of ΔSNP / Indel-index between the two offspring, and perform regional positioning of target traits based on SNP and Indel markers, taking into account the distribution of differential loci The degree of difference with Δindex locates the main resistance gene in the candidate interval o...

Embodiment 3

[0041] Example 3 Screening, Prediction and Verification of Candidate Genes

[0042] Based on the preliminary positioning results, combined with transcriptome sequencing for further screening of resistant genes. The stems of the two parents and the highly resistant and susceptible plants in the F2 generation were sampled and RNA was extracted for library construction and transcriptome sequencing. The expression of the 12 genes obtained by screening in transcriptome sequencing (such as figure 2 Shown in A and B), further use fluorescent quantitative PCR to verify the candidate genes, combined with SNP mutation types, gene function annotation and KEGG pathway analysis, and target one of the candidate genes Slα-KGDH E2 (Solyc12g005080), which is involved in Synthetic and metabolic pathways of various secondary metabolites, which play a key role in the tricarboxylic acid cycle. Slα-KGDH E2 has a C→T mutation in the promoter region, which is predicted to form a MIKC-type MADS tra...

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Abstract

The invention discloses a tomato bacterial wilt resistance gene Slalpha-KGDH E2 and application thereof. The Slalpha-KGDH E2 codes an alpha-ketoglutarate dehydrogenase E2 subunit, the nucleotide sequence of the gene is shown as SEQ ID NO: 1, and the protein sequence coded by the gene is shown as SEQ ID NO: 2. Reduction of the expression quantity of the gene or partial base mutation can enable tomato plants to lose resistance to tomato bacterial wilt. After ralstonia solanacearum is inoculated into disease-resistant plants, the expression quantity of the Slalpha-KGDH E2 gene is remarkably increased compared with that of a control group; and functional verification shows that the Slalpha-KGDH E2 gene has a key positive regulation effect on the resistance of tomatoes to ralstonia solanacearum, the resistance of tomatoes to bacterial wilt can be improved by overexpression of the Slalpha-KGDH E2 gene, and a new way is provided for breeding of new tomato disease-resistant varieties, so that the Slalpha-KGDH E2 gene has good application value.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, and in particular relates to tomato bacterial wilt resistance gene S1α-KGDH E2 and application thereof. Background technique [0002] Bacterial wilt is a bacterial vascular tissue disease caused by Ralstonia solanacearum E.F.Smith. It is mainly transmitted through soil, infecting plants through the root system and then entering the xylem, causing wilting of leaves and even death of the whole plant. Ralstonia solanacearum is one of the most destructive plant pathogenic bacteria in the world. It mainly occurs in tropical, subtropical and some warm temperate regions. It has a wide geographical distribution, many physiological races and strong pathogenicity. There are more than 200 kinds of plants in more than 50 families, including main crops such as vegetables, potatoes, tobacco, etc., resulting in severe reduction or even failure of vegetable crops, which can be called "plant cancer". In ...

Claims

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Application Information

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IPC IPC(8): C12N15/53C12N9/02C12N15/82A01H5/00A01H6/82
CPCC12N9/0008C12N15/8281C12Y102/04002
Inventor 卢钢刘红燕徐成翠刘悦宋建伟余小林
Owner ZHEJIANG UNIV
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