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StSci proteins that alter self-incompatibility in diploid potato material

A potato and diploid technology, applied in the field of genetics and breeding, can solve the problems of inability to recognize S-RNase, distant kinship, stolon lengthening, etc., and achieve the effect of solving the obstacle of self-incompatibility

Active Publication Date: 2021-12-03
AGRI GENOMICS INST CHINESE ACADEMY OF AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, a specific SLF protein can only relieve the cytotoxicity of one or several S-RNases, and certainly cannot recognize the S-RNase produced by its own pistil, so diploid potato self-fertilization is incompatible
[0005] The existing methods for creating self-compatible diploid potato materials mainly use a self-compatible diploid wild potato S.chacoence as the male parent, and cross pollination with other diploid potato materials The main disadvantage of this method is that the genetic relationship between the wild potato and the diploid cultivated potato is far away, and the hybrid offspring will introduce many wild undesired traits, such as smaller potato pieces, potato meat, etc. Increased solanine content, elongated stolons, etc., require multiple generations of backcrossing to eliminate these undesired wild traits, which consumes a lot of time and human resources

Method used

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  • StSci proteins that alter self-incompatibility in diploid potato material
  • StSci proteins that alter self-incompatibility in diploid potato material
  • StSci proteins that alter self-incompatibility in diploid potato material

Examples

Experimental program
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Effect test

Embodiment 1

[0092] Example 1 StSCI gene mapping

[0093] The self-incompatible diploid material PI225689 was pollinated with the pollen of the diploid potato material RH89-039-16, and the seeds were harvested as the F1 generation. The self-compatibility traits of the F1 generation plants were investigated, and half of the plants were self-compatible and self-incompatible, indicating that the self-compatibility gene in RH89-039-16 was in a heterozygous state, while RH89- 039-16 showed self-compatibility traits, which indicated that the gene controlling the self-compatibility of RH89-039-16 was dominant, and this dominant gene was named StSCI gene. The leaves of 40 self-compatible plants and 40 self-incompatible plants were selected from the F1 generation and mixed to extract genomic DNA, and the genomic DNA of the two groups of mixed pools was sequenced.

[0094] The sequencing results were analyzed, and the StSCI gene was preliminarily located in the 2.5Mb interval at the end of chromoso...

Embodiment 2

[0099] Example 2 StSCI gene verification

[0100] In order to verify that the PGSC0003DMG400016861 gene is the self-compatibility-induced gene in RH89-039-16, pCAMBIA1301 was used as the backbone vector to construct a promoter region containing 1245 bp upstream of the start codon of the PGSC0003DMG400016861 gene in RH89-039-16 and a 3352 bp The plant binary vector containing the full-length region of the gene, with the help of the Agrobacterium transformation system, transferred the PGSC0003DMG400016861 gene in RH89-039-16 together with its promoter into the original self-incompatible diploid potato material S15 In -65, complementary transgenic plants were obtained. At the same time, an overexpression vector with CaMV35S as the promoter linked to the coding region of the PGSC0003DMG400016861 gene was constructed, and transferred into the self-incompatibility material PI 225689 by the same method.

[0101] At the same time, using CRISP / Cas9 technology, a gene knockout vector o...

Embodiment 3

[0102] Example 3 A method for producing self-compatible diploid potatoes

[0103] Using Agrobacterium LBA440, the pCAMBIA1301 vector carrying the nucleotide sequence encoding the StSCI protein (SEQ ID NO.2) and the promoter (SEQ ID NO.3) was transferred into a self-incompatible of diploid potatoes. It was found that the introduction of the above-mentioned pCAMBIA1301 vector made the StSCI gene in the diploid potato express the StSCI protein, and then transformed the self-incompatible diploid potato into self-compatible.

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Abstract

The present invention provides a StSCI protein that changes the self-incompatibility of diploid potato materials, the amino acid sequence of the StSCI protein comprises or consists of the following sequence: 1) the amino acid sequence shown in SEQ ID NO.1; or , 2) a functional homologous sequence having at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO.1; or 3) adding, deleting, replacing one or more in the amino acid sequence shown in SEQ ID NO.1 A protein having multiple (eg, 1-10) amino acids and having self-incompatibility inhibiting activity. The invention has the advantage that the StSCI protein can inhibit the cytotoxic effect of various types of S-RNase, has heredity, fundamentally overcomes the defect of diploid potato self-incompatibility, and is conducive to the realization of high-generation pure diploid potato. Breeding of inbred lines.

Description

technical field [0001] The invention relates to the technical field of genetic breeding, in particular to a StSCI protein for changing the self-incompatibility of diploid potato materials. Background technique [0002] Diploid potato seed breeding refers to the cultivation of high-generation inbred lines with basically homozygous genomes through continuous multi-generation selfing, and then crossing the inbred lines with each other to produce F1 generation hybrid seeds with heterosis, which is achieved by using the first generation of hybrid seeds Potato planting scheme. Compared with traditional tetraploid tuber breeding, diploid seed breeding has the following outstanding advantages: First, the parents of traditional tetraploid breeding contain four sets of chromosomes that are highly heterozygous, the genetic background is complex, and the frequency of excellent traits is low , the breeding efficiency is low, and it is difficult to breed good varieties. As the diploid p...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/113C12N15/11C12N15/82A01H5/00A01H5/10A01H1/02A01H6/82
CPCC07K14/415C12N15/8287C12N15/8216A01H1/02A01H6/827A01H5/04A01H1/022C12Q1/6895C12Q2600/13
Inventor 黄三文马玲尚轶张春芝李灿辉
Owner AGRI GENOMICS INST CHINESE ACADEMY OF AGRI SCI
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