CN-118879731-B - Application of qCTS11.2 gene in regulating cold tolerance of rice

Abstract

The invention relates to application of qCTS11.2 gene in regulating rice cold tolerance, and a method for improving rice cold tolerance, which comprises the steps of replacing original qCTS11.2 gene in a genome of a starting rice variety by using low-function qCTS11.2 gene or knocking out qCTS11.2 gene from the genome of the starting rice variety. The invention discovers a novel cold tolerance related gene qCTS11.2, provides theoretical basis and gene resources for cold tolerance improvement and breeding of rice, and can be used for cross breeding and transgenic breeding.

Inventors

• ZHOU LEI
• CHEN JUNXIAO
• XU HUASHAN
• LI CHANGYAN
• WANG HUIYING
• SHI SHAOJIE
• WU YAN
• YOU AIQING
• WU BIAN
• DU JINGHUA
• FU MINGHUI
• Li sanhe
• LIU KAI
• ZHA WENJUN
• ZHANG SIYUE

Assignees

• 湖北省农业科学院粮食作物研究所

Dates

Publication Date

20260508

Application Date

20240808

Priority Date

20240605

Claims (2)

1. 1. A method for improving cold tolerance of rice is characterized by comprising the step of knocking out qCTS11.2 genes from a genome of a starting rice line, wherein the starting rice line is Japanese sunny, and the qCTS11.2 gene sequence is shown as SEQ ID NO. 1.
2. 2. The method of claim 1, wherein the qts 11.2 gene is knocked out of the genome of the starting rice line by gene editing.

Description

Application of qCTS11.2 gene in regulating cold tolerance of rice Technical Field The invention relates to the field of rice molecular breeding, in particular to application of qCTS11.2 gene in regulating cold tolerance of rice. Background Rice (Oryza sativa L.) is one of three large food crops worldwide, and plays a vital role in guaranteeing the world food safety. Rice originates in tropical and subtropical areas and is more sensitive to low temperature stress than other crops. The rice may suffer cold injury in various growth periods, such as slow emergence, slow tillering, late maturation and reduced grain weight due to low temperature in the vegetative growth period, and reduced glume flowers and fruiting rate due to low temperature in the reproductive growth period, especially in the booting period, and finally reduced yield. Wherein, the low temperature in the seedling stage is the natural disaster with the highest occurrence frequency and the greatest influence on the yield of early rice and regenerative rice. Therefore, developing the research of cold resistance of rice, improving the cold resistance of rice, and has great significance for promoting the yield increase and stable yield of rice in China and guaranteeing the national grain safety. In the research, a recombinant inbred line genetic population is constructed by Japanese sunny and WD16343, the BSA-seq is utilized to carry out QTL positioning on the cold tolerance character, qCTS11.2 is predicted to be a candidate gene for cold tolerance of rice according to the function annotation analysis of the candidate gene, a transgenic material is constructed and verified, and meanwhile, the systematic evolution and expression characteristics of the gene are analyzed, so that theoretical basis and gene resources are provided for cultivating excellent rice varieties resisting low-temperature cold damage. Disclosure of Invention In the research, we find that two qCTS11.2 gene haplotypes exist in rice, one of the haplotypes can endow the rice with cold tolerance, and further, we find that the content tolerance of transgenic rice can be improved after the qCTS11.2 gene is knocked out. Based on the research, the invention provides application of qCTS11.2 gene in regulating cold tolerance of rice. The invention also provides a method for improving the cold tolerance of rice, which comprises the step of replacing the original qCTS11.2 gene in the genome of the original rice variety by using the low-function qCTS11.2 gene or knocking out the qCTS11.2 gene from the genome of the original rice variety. In a specific embodiment, the method comprises the steps of: And (3) crossing the rice line containing the low-function qCTS11.2 gene with the rice line containing the normal-function qCTS11.2 gene, and using the low-function qCTS11.2 gene as a screening mark to ensure that the target rice line only contains the low-function qCTS11.2 gene. In a specific embodiment, the rice line containing the low-function qts 11.2 gene is NIP, or contains the qts 11.2 gene identical to NIP. In a specific embodiment, the low-function qCTS11.2 gene sequence is shown in SEQ ID NO. 1. In a specific embodiment, the qcts11.2 gene is knocked out from the genome of the starting rice line by gene editing. The invention discovers a novel cold tolerance related gene qCTS11.2, provides theoretical basis and gene resources for cold tolerance improvement and breeding of rice, and can be used for cross breeding and transgenic breeding. Drawings FIG. 1 shows the selection of cold tolerance related gene qCTS11.2 in rice seedling stage. Wherein A is a cold-resistant phenotype diagram of NIP and WD16343, B is a schematic diagram of RI L group construction, C is a comparison of survival rates of RI L group and Japanese and WD16343 after 4 ℃ cold treatment, D is distribution of delta (SNP-i ndex) values on each chromosome, E is a fine genetic map of a cold-resistant gene, F is qRT-PCR for detecting expression quantity of qCTS11.2 after different cold treatment time of Japanese and WD16343 seedlings, P value is obtained by double-tail t test, error line, s.e.m. Fig. 2 is a CRISPR/Cas9 knockout qcts11.2 increasing rice seedling cold tolerance. Wherein A is a CRISPR/Cas9 knockout qCTS11.2 schematic diagram, the upper part is a gene structure qCTS11.2 of qCTS11.2, the lower part shows the sequence variation of the knockout family, and B is the cold-resistant phenotype of the qCTS11.2 knockout family and NIP. C is qCTS11.2 knockout family and NIP survival rate under 4 ℃ cold treatment conditions. Significance was found from the double tail t test, error bars, s.e.m, repeat number n=3. Detailed Description The principles and features of the present invention are described below with reference to the drawings, the examples are illustrated for the purpose of illustrating the invention and are not to be construed as limiting the scope of the invention. 1. Screening of candidate Gene qCTS11.2 We constructe