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Potato StLecRK-VI.1 and StTET8 genes and use thereof in improving late blight resistance

A technology of potato late blight and stlecrk-vi.1, applied in the direction of plant genetic improvement, application, genetic engineering, etc., can solve the problems of late blight resistance and other problems that have not been seen in potato, and achieve the effect of improving late blight resistance.

Active Publication Date: 2021-07-23
HUAZHONG AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Problems in the prior art: Breeding varieties with broad-spectrum resistance to late blight is the main breeding goal of potato disease-resistant breeding
The lectin receptor kinase gene is a potential ideal target, but there are no reports about the regulation of StLecRK-VI.1 and StTET8 genes on potato late blight resistance

Method used

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  • Potato StLecRK-VI.1 and StTET8 genes and use thereof in improving late blight resistance
  • Potato StLecRK-VI.1 and StTET8 genes and use thereof in improving late blight resistance
  • Potato StLecRK-VI.1 and StTET8 genes and use thereof in improving late blight resistance

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Experimental program
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Effect test

Embodiment 1

[0039] This embodiment provides the potato StLecRK-VI.1 gene, including:

[0040] 1. The potato StLecRK-VI.1 gene, whose CDS sequence is shown in SEQ ID No. 1 in the sequence table, contains 1998 bp nucleotides.

[0041] 2. The potato StTET8 gene, whose CDS sequence is shown in SEQ ID No. 2 in the sequence table, contains 801 bp of nucleotides.

Embodiment 2

[0043] This embodiment provides a method for cloning the potato StLecRK-VI.1 gene and constructing a plant interference vector, including:

[0044] 1. Expression analysis of StLecRK-VI.1 gene under late blight stress

[0045] Using fluorescent quantitative PCR (qRT-PCR) technology, it was found that a lectin receptor like kinase gene (GenBank number, XP_006341207.2) with unknown function in potato responds to the induction of late blight pathogen and is resistant to late blight. Induced 24-hour down-regulated expression in sexual materials, which means that the gene may be involved in late blight resistance ( figure 1 ). StEF1α was used as an internal reference gene, the forward primer was 5'-ATTGGAAACGGATATGCTCCA-3', and the reverse primer was 5'-TCCTTACCTGAACGCCTGTCA-3'. The forward specific primer of StLecRK-IV.1 is 5'-CGATTTCACCTACAACGGGT-3', and the reverse specific primer is 5'-GACCCTCCTGCAATTGTGAA-3'. Use the recommended reaction system of BlasTaqTM2×qPCR MasterMix (...

Embodiment 3

[0054] This example provides the application of interfering with the expression of the potato StLecRK-VI.1 gene to improve resistance to late blight, including:

[0055] 1. Potato genetic transformation

[0056] Potato tuber thin slices were used as explants, and the potato variety was 'E Potato 3' (hereinafter referred to as 'E3'). The transformation method is Agrobacterium transformation method, specifically refer to Si et al. (2003). Potato chips and Agrobacterium were co-cultured on the P1 medium for 2 days, and then the resistant shoots were cultivated on the screening medium P2 (containing kan). Rooting culture was carried out in the rooting medium P3 containing screening antibiotics (kan) and cephalosporin (cef), and finally whole plants were obtained. See Table 1 for transgenic medium.

[0057] Table 1 Potato transgenic medium

[0058]

[0059] 2. Detection of transgenic positive strains

[0060] Cut 1-2 small leaves of the seedlings to be tested in the ultra-c...

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Abstract

The invention provides potato StLecRK-VI.1 and StTET8 genes and use thereof in improving a late blight resistance. The potato late blight resistance can be improved by interfering with the lectin receptor kinase StLecRK-VI.1 gene in potatoes. No obvious phenotypic change exists in interference of the gene in potatoes. The StLecRK-VI.1 interacts with a protein StTET8 capable of positively regulating and controlling a late blight resistance, and the protein expression amount of the interacting protein StTET8 can be reduced. Therefore, the potato late blight resistance can be improved through interference of the StLecRK-VI.1 gene and (or) overexpression of the StTET8 gene.

Description

technical field [0001] The invention belongs to the field of potato late blight resistance gene mining, in particular to potato StLecRK-VI.1 and StTET8 genes and their application in improving late blight resistance. Background technique [0002] Potato is the fourth largest food crop in the world after rice, wheat and corn. It is an extremely important grain and vegetable dual-purpose crop in my country. The planting area and output of potatoes in China rank first in the world, accounting for about a quarter. Potatoes play a very important role in ensuring the food security of the world and our country. Late blight is a serious global disease of potato, known as potato "cancer". Late blight resistance is an important goal of potato breeding, and the breeding of durable resistant varieties has always been a difficult problem for breeders at home and abroad. [0003] At present, potato late blight resistance breeding in the world relies heavily on the main resistance genes...

Claims

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Application Information

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IPC IPC(8): C12N15/54C12N9/12C12N15/29C12N15/82A01H5/00A01H6/82
CPCC12N9/1205C12N15/8282C07K14/415
Inventor 田振东郭磊戚烨通周晶
Owner HUAZHONG AGRI UNIV
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