Method for breeding self-compatible potatoes
Disclosed is a method for breeding self-compatible potatoes, including the following steps: (1) selecting a self-compatible potato variety material and referring to it as PG6359, and cloning the S-RNase gene of PG6359 through the transcriptome sequencing method; and (2) obtaining two full-length sequences of the S-RNase gene from the cloned S-RNase gene in step (1) and referring to them as S s11 and S s12 respectively, and after carrying out an artificial self-pollination for the variety material PG6359, selecting the variety material having the genotype of S s11 S s11 from the offspring as the female parent, and selecting a self-incompatible material as the male parent, and then obtaining a self-compatible F 1 generation by hybridization. The invention overcomes the self-incompatibility of diploid potatoes, and does not require the introduction of any wild potato gene fragments, thereby avoiding linkage drag, and providing a basis for the rapid creation of a diploid potato inbred line.
1. A method for breeding self-compatible potatoes, comprising the following steps:
(1) selecting a self-compatible potato variety material and referring to it as a variety material PG6359, and cloning the S-RNase gene of the variety material PG6359 through the transcriptome sequencing method; wherein the transcriptome sequencing method comprises: firstly extracting RNA by utilizing the variety material PG6359, and performing the transcriptome sequencing by the sequencing platform to obtain 2 Gb of sequencing data; de novo assembling the transcriptome data by Trinity software, and calculating the expression of each transcript by RNASeq by Expectation Maximization (RSEM) software; then performing the sequence alignment search tool by utilizing the known S-RNase protein sequence in the potato reference genome, and selecting the sequence with an alignment reliability E value less than 1E-5 and an expression level Fragments Per Kilobase of exon model per Million mapped fragments (FPKM) value greater than 200 as a candidate sequence of the S-RNase gene; designing amplification primers to amplify the full length of the S-RNase gene of variety material PG6359, and determining its expression by qPCR; and
(2) obtaining two full-length sequences of the S-RNase gene from the cloned S-RNase gene in step (1) and referring to them as S s11 and S s12 respectively, wherein the gene sequence of S s11 is shown in SEQ ID NO:1, and the gene sequence of S s12 is shown in SEQ ID NO:2; and after carrying out an artificial self-pollination for the variety material PG6359, selecting the variety material having the genotype of S s11 S s11 from the offspring as the female parent and referring to it as material A, and selecting a self-incompatible material as the male parent and referring to it as material B, then obtaining a self-compatible F 1 generation by hybridization; performing genotype detection for the F 1 generation to confirm that the F 1 generation contains the S s11 gene, and detecting that individuals of the F 1 generation are self-compatible after self-pollination of the F 1 generation.
2. The method for breeding self-compatible potatoes according to claim 1 , wherein the individual of the F 1 generation is used as a female parent in step (2), and the self-incompatible material B is used as a male parent to perform backcross; then performing genotype detection for the resulting BC 1 generation material to obtain the individual containing S s11 gene as the female parent, and continuing to backcross with the self-incompatible material B; after multiple generations of backcrossing, then performing another generation of self-crossing.
3. The method for breeding self-compatible potatoes according to claim 1 , wherein the sequence of the upstream primer of the amplification primers is shown in SEQ ID NO:3, and the sequence of the downstream primer of the amplification primers is shown in SEQ ID NO:4.
4. A method for generating self-compatible potatoes, comprising performing hybridization by using a potato plant comprising the polynucleotide shown in SEQ ID NO:1 as the first parent, so that the offspring comprises the polynucleotide, wherein the S-RNase genotype of the female parent is S s11 S s11 .
5. A method for generating self-compatible potatoes, comprising performing hybridization by using the potato plant, or a potato plant produced by a plant part, a tuber, a tuber part, a seed or a plant cell thereof comprising SEQ ID NO:1 as the first parent, so that the offspring comprises the polynucleotide, wherein the S-RNase genotype of the female parent is S s11 S s11 .
6. The method for generating self-compatible potatoes according to claim 5 , wherein the second parent is a self-incompatible material.
7. The method according to claim 6 , further performing backcross by using the offspring comprising the polynucleotide the first parent, and the self-incompatible material as the second parent, so as to obtain a self-compatible material having the genetic background of the second parent.