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A method to overcome self-incompatibility in diploid potato

A potato and diploid technology applied in the field of biotechnology and genetic breeding to achieve the effects of short breeding cycle, strong operability and low fruit setting rate

Active Publication Date: 2022-06-03
AGRI GENOMICS INST CHINESE ACADEMY OF AGRI SCI +1
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  • Claims
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  • A method to overcome self-incompatibility in diploid potato
  • A method to overcome self-incompatibility in diploid potato
  • A method to overcome self-incompatibility in diploid potato

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Embodiment 1

[0073] Embodiment 1: a method for overcoming the self-incompatibility of diploid potato provided by the present invention, comprising the following steps:

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

[0075] (2) according to the nucleic acid sequence of the target fragment obtai...

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Abstract

A method for overcoming diploid potato self-incompatibility disclosed by the invention comprises (1) selecting a target segment; (2) constructing a recombinant vector for gene targeting; (3) realizing the functional loss of the intracellular S-RNase gene Mutation; (4) regenerating several potato plants; (5) specifically amplifying the DNA segment of the S-RNase gene in the regenerated plant containing the target fragment; (6) selecting the regenerated plant whose S-RNase gene was edited; (7) The selected gene-edited plants were further screened for diploid gene-edited lines; (8) the selected gene-edited lines were propagated and colonized, and the self-compatibility phenotype was identified at the flowering stage; (9) the harvested Sequencing of amplification products of offspring of self-compatibility plants to detect inheritance and segregation of edited target genes in offspring. The invention has the advantage of finding a simple, precise and efficient method for overcoming the self-incompatibility of diploid potatoes.

Description

technical field [0001] The invention belongs to the technical field of biotechnology and genetic breeding, in particular to a method for overcoming the self-incompatibility of diploid potatoes. Background technique [0002] Diploid potato is a gametophyte self-incompatible type, and its pollen tube can germinate on the stigma and elongate 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 potato. [0003] Previous studies have found that tobacco S-RNase gene is only slightly expressed in the stage from flower bud to bud stage, but a higher enrichment of S-RNase protein was detected in the flowering stage. Our study on potato pistil protein also came to a similar conclusion, which created a possibility for potato to overcome self-incompatibility through selfing at the bud stage. However, selfing at bud stage has higher requirem...

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Application Information

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Patent Type & Authority Patents(China)
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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