CN-118726404-B - Application of qCTS11.1 gene in regulating cold tolerance of rice

CN118726404BCN 118726404 BCN118726404 BCN 118726404BCN-118726404-B

Abstract

The invention relates to application of qCTS11.1 gene in regulating rice cold tolerance, and a method for improving rice cold tolerance, which comprises the steps of replacing original qCTS11.1 gene in a genome of a starting rice variety by using low-function qCTS11.1 gene or knocking out qCTS11.1 gene from the genome of the starting rice variety. The invention discovers a novel cold tolerance related gene qCTS11.1, 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

YOU AIQING
WANG MENGJING
XU HUASHAN
LI CHANGYAN
WANG HUIYING
SHI SHAOJIE
WU YAN
WU BIAN
ZHOU LEI
DU JINGHUA
Li sanhe
LIU KAI
ZHENG XINGFEI
ZHA WENJUN
CHEN JUNXIAO

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. A method for improving cold tolerance of rice is characterized by comprising the step of knocking out qCTS11.1 gene from genome of a starting rice line, wherein the starting rice line is Japanese sunny, and the qCTS11.1 gene sequence is shown as SEQ ID NO. 1.
2. The method of claim 1, wherein the qts 11.1 gene is knocked out of the genome of the starting rice line by gene editing.

Description

Application of qCTS11.1 gene in regulating cold tolerance of rice Technical Field The invention relates to the field of rice molecular breeding, in particular to application of qCTS11.1 gene in regulating cold tolerance of rice. Background Rice (Oryza sativa) is the most dominant ration crop in our country, and more than half of the population takes rice as main food. Since rice originates from tropical and subtropical areas, the rice is more sensitive to low-temperature stress than other crops, the low-temperature stress can cause the growth of rice seedlings to be blocked, physiological metabolism to be disturbed, and when serious, the rice leaves can be yellow, withered and even dead, so that the yield and quality are greatly reduced. Therefore, developing the research of cold resistance of rice and excavating excellent cold resistance genes has very important theoretical and practical 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, qCTS11.1 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 qCTS11.1 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 qCTS11.1 gene is knocked out. Based on the research, the invention provides application of qCTS11.1 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 qCTS11.1 gene in the genome of the original rice variety by using the low-function qCTS11.1 gene or knocking out the qCTS11.1 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 qCTS11.1 gene with the rice line containing the normal-function qCTS11.1 gene, and using the low-function qCTS11.1 gene as a screening mark to ensure that the target rice line only contains the low-function qCTS11.1 gene. In a specific embodiment, the rice line containing the low-function qts 11.1 gene is NIP, or contains the qts 11.1 gene identical to NIP. In a specific embodiment, the low-function qCTS11.1 gene sequence is shown in SEQ ID NO. 1. In a specific embodiment, the qcts11.1 gene is knocked out from the genome of the starting rice line by gene editing. The invention discovers a novel cold tolerance related gene qCTS11.1, 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 qCTS11.1 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 qCTS11.1 expression quantity after different cold treatment time of Japanese and WD16343 seedlings is detected by qRT-PCR. The P value is obtained from the two-tailed t-test, error bars, s.e.m. Fig. 2 is a CRISPR/Cas9 knockout qcts11.1 increasing rice seedling cold tolerance. Wherein A is a CRISPR/Cas9 knockout qCTS11.1 schematic diagram, the upper part is a gene structure qCTS11.1 of qCTS11.1, the lower part shows the sequence variation of the knockout family, B is the cold-resistant phenotype of the qCTS11.1 knockout family and NIP, and C is the survival rate of the qCTS11.1 knockout family and NIP under the 4 ℃ cold treatment condition. 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 qCTS11.1 The method selects parent materials including Nippon and WD16343 with obvious difference of rice seedling stage cold resistance phenotype, wherein the Nip shows stronger seedling stage cold resistance relative to WD16343 (figure 1A), utiliz