Method for overcoming diploid potato self-incompatibility

A potato and diploid technology, applied in the field of biotechnology and genetic breeding

Active Publication Date: 2019-05-14
AGRI GENOMICS INST CHINESE ACADEMY OF AGRI SCI +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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  • Method for overcoming diploid potato self-incompatibility
  • Method for overcoming diploid potato self-incompatibility
  • Method for overcoming diploid potato self-incompatibility

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Embodiment 1

[0073] Embodiment one: a kind of method for overcoming diploid potato self-incompatibility provided by the invention, comprises the steps:

[0074] (1) In potato self-incompatibility determining gene S-RNase gene S p3 and S p4 A target fragment is selected in the first exon region; in the step (1), the target fragment is located on the target gene S-RNase, and one chain of the target fragment has a nucleic acid sequence structure as shown in SEQ ID No.1. One strand of the target fragment has a nucleic acid sequence structure as shown in SEQ ID No.1. For example, wherein the target fragment is located on the target gene S-RNase, one strand of the target fragment has a 5'-(N) X -NGG-3' structure, (N) X Represents a base sequence {N with number X 1 ,N 2 ...N X}, N 1 ,N 2 ...N X Each in represents any one of the bases A, G, C, and T, and N in NGG is any one of the bases A, G, C, and T.

[0075] (2) According to the nucleic acid sequence of the target fragment obtained in...

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Abstract

The invention discloses a method for overcoming diploid potato self-incompatibility. The method comprises the steps of 1, selecting a target fragment; 2, constructing a gene target shooting recombinant vector; 3, achieving function deletion mutation of a S-RNase gene inside a cell; 4, regenerating a plurality of potato plants; 5, specifically amplifying a DNA block containing the target fragment of the S-RNase gene in the regeneration plant; 6, selecting the regeneration plant edited by the S-RNase gene; 7, further screening out a diploid gene editing strain from the gene edited plant; 8, performing expanding propagation and planting on the gene edited strain, and performing self-incompatibility phenotype identification on the blooming stage; 9, amplifying product sequencing on the harvested self-compatible plant later generations, and detecting heredity and separation of the edited target gene for later generations. The method has the advantages that the simple, accurate and efficiency method is found for overcoming diploid potato self-incompatibility.

Description

technical field [0001] The invention belongs to the technical fields of biotechnology and genetic breeding, and specifically relates to a method for overcoming self-incompatibility of diploid potatoes. Background technique [0002] Diploid potato is a self-incompatible type of gametophyte, and its pollen tube can germinate on the stigma and extend into the style, but its growth is subsequently inhibited. This trait is controlled by the S-RNase gene, whose expression inhibits pollen tube elongation, making it difficult to obtain inbred lines in potatoes. [0003] Previous studies found that the tobacco S-RNase gene was only slightly expressed in the flower bud to bud stage, but a higher S-RNase protein enrichment was detected in the flowering stage. Our research on pistil protein in potato has also reached a similar conclusion, which creates the possibility for potato to overcome self-incompatibility through self-fertilization at the bud stage. However, selfing at the bud s...

Claims

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Application Information

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IPC IPC(8): C12N15/82C12N9/22A01H5/00A01H6/82C12Q1/6895
CPCC12Q1/6895C12N9/22C12N15/8287C12N15/8289C12N15/827C12N15/8213C12Q2600/156C12Q2600/13A01H6/827
Inventor 黄三文张春芝彭真叶明旺
Owner AGRI GENOMICS INST CHINESE ACADEMY OF AGRI SCI
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