CN110407922B - Rice cold-resistant gene qSCT11 and application thereof - Google Patents

Abstract

The invention provides a rice cold-resistant gene qSCT11 and application thereof, wherein the nucleotide sequence of the gene is shown as SEQ ID NO.1, and the nucleotide sequence of cDNA of the gene is shown as SEQ ID NO. 2. According to the invention, the survival rate of rice with the overexpression qSCT11 gene is found to be obviously higher than that of wild rice under low-temperature stress; through constructing rice with RNAi inhibiting qSCT11 gene expression and rice with qSCT11 gene knockout, the survival rate of transgenic rice strains is obviously lower than that of wild rice strains after low-temperature stress is discovered. The invention also provides application of the rice cold-resistant gene qSCT11 in rice cold-resistant breeding.

Description

Rice cold-resistant gene qSCT11 and application thereof

Technical Field

The invention belongs to the technical field of plant genetic engineering, and particularly relates to a rice cold resistance gene qSCT11 and application thereof.

Background

Rice is one of the most important food crops in the world and provides 21% of the worldwide per capita caloric intake. Rice is sensitive to cold stress due to its origin in tropical and subtropical regions. In high latitude and high altitude areas, cold stress severely restricts the growth and production of rice, and is one of the most important abiotic stress factors causing the loss of rice yield and limiting the rice planting distribution. Seedling death and pollen abortion at the booting stage are two major factors that lead to yield loss under cold stress.

Molecular genetic research shows that the cold resistance of rice is a quantitative character which is jointly regulated and controlled by various genetic factors and environments. The genetic basic research of the cold resistance of rice is still in the initial stage. Many Quantitative Trait Loci (QTLs) have been located and cloned, such as qLTG3-1, Ctb1, COLD1, qCT-9, CTB4a, qPSR10, and HAN 1. Among them, natural variation of COLD1, CTB4a, qPSR10 and HAN1 genes has been proved to enhance the adaptability of rice to low-temperature environment.

However, the natural cold tolerance allele is still rarely found, the cold tolerance allele of rice is mainly derived from natural variation, and the location and cloning of the gene are helpful for people to understand the genetic basis of the cold tolerance of rice, but the successful application of the gene in breeding practice is still quite limited. Therefore, a new cold-resistant gene is explored, the cold resistance of the rice is improved, the production safety of the rice is maintained, the planting area of the rice is expanded to a geographical area with lower temperature, and the method has important application value for breeding of cold-resistant varieties of the rice and research on cold-resistant molecular basis.

Disclosure of Invention

The invention aims to provide a rice cold-resistant gene and application thereof.

The invention firstly provides a protein coded by a rice cold resistance gene qSCT11, and the amino acid sequence of the protein is as follows:

a) an amino acid sequence shown as SEQ ID NO. 3; or

b) The amino acid sequence shown in SEQ ID NO.3 is formed into an amino acid sequence with the same function by replacing, deleting and/or adding one or more amino acid residues.

The rice cold-resistant gene qSCT11 provided by the invention is as follows:

a) the nucleotide sequence is shown as SEQ ID NO.1 in the sequence table; or

b) The nucleotide sequence shown in SEQ ID NO.1 is used for replacing one or more nucleotides to obtain the nucleotide sequence for coding the same functional protein; or

c) The CDS sequence of the rice cold-resistant gene qSCT11 is shown in SEQ ID No. 2.

The invention also provides a biological material containing the rice cold-resistant gene qSCT11, wherein the biological material is an expression cassette, a vector, a host cell or a recombinant bacterium.

The invention provides application of the rice cold resistance gene qSCT11 or protein coded by the gene or biological material containing the gene in improving the cold resistance of plants.

The invention provides application of the rice cold-resistant gene qSCT11 or protein coded by the gene or biological material containing the gene in cultivating cold-resistant transgenic plants.

The invention provides application of the rice cold-resistant gene qSCT11 or protein coded by the gene or a biological material containing the gene in improvement of plant cold-resistant germplasm resources.

The invention provides application of the rice cold-resistant gene qSCT11 or protein coded by the gene or biological material containing the gene in improving the low-temperature survival rate of plants.

In the above applications, the plant includes, but is not limited to, rice, wheat, soybean, sorghum, millet, barley, and corn.

Furthermore, the invention also provides a method for improving the cold resistance of rice, which is to express or over-express the cold resistance gene qSCT11 of the rice by a transgenic, hybridization, backcross, selfing or asexual propagation method.

Specifically, in the method, the transgene comprises introducing a recombinant expression vector containing a rice cold-resistant gene qSCT11 into rice by using a Ti plasmid, a plant virus vector, direct DNA transformation, microinjection, a gene gun, conductance and an agrobacterium-mediated method to obtain a transgenic rice strain.

According to the invention, the survival rate of rice with the overexpression qSCT11 gene is found to be obviously higher than that of wild rice under low-temperature stress; through constructing rice with RNAi inhibiting qSCT11 gene expression and rice with qSCT11 gene knockout, the survival rate of transgenic rice strains is obviously lower than that of wild rice strains after low-temperature stress is discovered. The invention also provides application of the rice cold-resistant gene qSCT11 in rice cold-resistant breeding.

Drawings

FIG. 1 is a graph showing the results of relative expression levels of qSCT11 gene in example 2.

FIG. 2 is a comparison of plant phenotypes before and after low temperature treatment in example 2.

FIG. 3 is a graph showing the results of the survival rate of seedlings in example 2.

FIG. 4 is a graph showing the results of the relative expression levels of qSCT11 gene in example 3.

FIG. 5 is a comparison of plant phenotypes before and after low temperature treatment in example 3.

FIG. 6 is a graph showing the results of the survival rate of seedlings in example 3.

FIG. 7 is the result of the target site sequencing of the qSCT11 knockout plant in example 4.

FIG. 8 is a comparison of plant phenotypes before and after low temperature treatment in example 4.

FIG. 9 is a graph showing the results of the survival rate of seedlings in example 4.

FIG. 10 is a chromosome structure diagram of the near isogenic NIL-NIP genotype in example 5.

FIG. 11 is a graph showing the results of relative expression levels of qSCT11 gene in example 5.

FIG. 12 is a comparison of plant phenotypes before and after low temperature treatment in example 5.

FIG. 13 is a graph showing the results of the survival rate of seedlings in example 5.

Detailed Description

The following examples are intended to further illustrate the present invention but should not be construed as limiting the invention. Modifications or substitutions to methods, procedures, or conditions of the invention may be made without departing from the spirit and scope of the invention.

Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art; the biochemical reagents and materials used in the examples are all commercially available or known in the art. The following examples were repeated 3 times and the results averaged.

EXAMPLE 1 cloning of Gene qSCT11

1. DNA of a rice variety Nipponbare is extracted, Polymerase Chain Reaction (PCR) is carried out by using primers 5'-CCCAGAAGAGTAACCT AT-3' and 5'-TGAGCAACTGAGCAAT-3', and the obtained PCR product is sequenced to obtain a gene sequence of a rice gene qSCT11, wherein the gene sequence consists of 5446 basic groups and is shown as SEQ ID NO. 1. The PCR procedure was as follows, pre-denaturation at 94 ℃ for 5 min; 35 cycles (denaturation at 94 ℃ for 30 seconds; annealing at 55 ℃ for 30 seconds; extension at 72 ℃ for 6 minutes) and extension at 72 ℃ for 7 minutes.

2. RNA of a rice variety Nipponbare leaf is extracted and is reversely transcribed into cDNA, PCR is carried out by primers 5'-ACA CGCC TACACTCAACA-3' and 5'-AGCAACACCTCACCTCC-3', the size of an amplification product is 922bp (containing 91bp non-coding sequences), the obtained PCR product is subjected to sequencing analysis to obtain a coding sequence (CDS) of a gene qSCT11, the CDS consists of 831 basic groups, and the nucleotide sequence is shown in SEQ ID NO: 2. The PCR procedure was as follows: pre-denaturation at 94 ℃ for 5 min; 30 cycles (denaturation at 94 ℃ for 30 seconds; annealing at 55 ℃ for 30 seconds; extension at 72 ℃ for 1 minute) and extension at 72 ℃ for 7 minutes.

3. The coding sequence (CDS) was translated using Primer 3 software (http:// frodo.wi. mit. edu /) to obtain an amino acid sequence encoding 276 amino acids, which sequence is the amino acid sequence shown in SEQ ID NO. 3.

The primers are synthesized by Shanghai, and the sequence determination is performed by Shanghai. DNA and RNA extraction, PCR and reagent formulation refer to J. SammBruke, et al, molecular cloning, A laboratory Manual, third edition, Jindong goose, et al, (Shi), scientific Press, 2002.

Example 2 overexpression of qSCT11 Gene to improve Cold tolerance of Rice at seedling stage

1. Construction of overexpression vectors

According to the cDNA sequence of the rice gene qSCT11, the full-length coding sequence shown in SEQ ID NO.2, homologous recombination joints at two sides of a KpnI enzyme cutting site of a pC1301S vector are respectively introduced into an upstream primer and a downstream primer, and the design primers are as follows:

OX-F:5′-atgatgatgataaaggtaccatggaaaatgtaacagaaaaag-3′

OX-R:5′-ctagaggatccccgggtacctgctctcccccccagaaatgac-3′

PCR was performed using the cDNA clone obtained in example 1 as a template and the primers OX-F and OX-R, and the product size was 868 bp. The PCR procedure was as follows: pre-denaturation at 94 ℃ for 5 min; 30 cycles (denaturation at 94 ℃ for 30 seconds; annealing at 55 ℃ for 30 seconds; extension at 72 ℃ for 1 minute) and extension at 72 ℃ for 7 minutes. The qSCT11 coding region fragment separated by PCR amplification is connected to KpnI cut pC1301S vector by homologous recombination. And after sequencing comparison, transforming the DNA into agrobacterium EHA105 to obtain the qSCT11 gene overexpression vector.

2. Obtaining of overexpression transgenic plants

The qSCT11 gene overexpression vector is transferred into indica rice 9311 through agrobacterium-mediated genetic transformation. The T0 generation transgenic plant is obtained through selective culture, differentiation, rooting and seedling hardening. All transgenic materials are propagated to obtain T1 generation materials with stable inheritance. For a T1 generation transgenic line, if the T2 generation plants obtained by sampling the T1 generation transgenic line are all transgenic plants, the T1 generation transgenic line is a homozygous transgenic line. 3 homozygous transgenic lines OX-1, OX-2, OX-3 were selected for subsequent analysis.

3. Identification of overexpression transgenic plants

(1) Identification of expression level

Taking 2-leaf stage seedlings, extracting total RNA, carrying out reverse transcription to obtain cDNA, and carrying out fluorescence real-time quantitative PCR. The primers for detecting the expression level of the qSCT11 gene are as follows:

RT-F:5′-ACCGCTTCCGCCTCCCTCCCAT-3′；

RT-R:5′-CTGCTCGCAGGTGAAGAGGAGCC-3′。

the reagent used for the quantitative analysis was FastStart Universal SYBR Green Master (ROX). The instrument used was a real-time fluorescent quantitative PCR instrument ViiA7 from Applied Biosystems, USA. The Ubqtin gene serves as an internal reference.

The relative expression of qSCT11 gene in the overexpressed material is shown in figure 1. In FIG. 1, WT represents rice variety 9311, and OX-1, OX-2, and OX-3 represent qSCT11 gene overexpression transgenic lines. In the qSCT11 gene overexpression transgenic line, the relative expression of the qSCT11 gene is obviously higher than that of rice 9311.

(2) Cold-resistant phenotype identification

Seeds are taken and soaked for 2 days at the temperature of 30 ℃, germination is carried out for 1 day, then 36 germinated seeds are sown in a black small square box (the length, the width and the height are both 6cm) filled with nutrient soil, the seeds are cultured to the 2-leaf stage in an illumination incubator (the illumination time is 16h/d and the temperature is 30 ℃), and the seeds are photographed before treatment. And (3) placing the 2-leaf stage seedlings in a low-temperature incubator at 4 ℃ for treatment for 36h, then placing the seedlings back in an illumination incubator for recovery of culture for 1 week, photographing and counting the survival rate. Each material was tested in 3 replicates.

The photographs before and after the treatment are shown in FIG. 2. In FIG. 2, WT indicates rice variety 9311, and OX-1, OX-2, and OX-3 indicate qSCT11 gene overexpression transgenic plants, with a scale of 5 cm. After low temperature stress, the growth condition of the qSCT11 gene overexpression transgenic plant is obviously superior to that of rice 9311.

The survival results are shown in figure 3. In FIG. 3, WT indicates rice variety 9311, and OX-1, OX-2, and OX-3 indicate qSCT11 gene overexpression transgenic plants. After low temperature stress, the survival rate of the qSCT11 gene overexpression transgenic plant is obviously higher than that of rice 9311. The average survival rates were: WT: 36.32%, OX-1: 67.36%, OX-2: 73.36%, OX-3: 70.29 percent.

Example 3 RNAi inhibition of qSCT11 Gene expression reduces Cold tolerance of Rice at seedling stage

1. Construction of RNAi expression vectors

According to the cDNA sequence of the rice gene qSCT11, the full-length coding sequence shown in SEQ ID NO.2 is compared with the whole genome sequence, a specific sequence of 407bp is selected, and the shown nucleotide sequence is SEQ ID NO. 4. And homologous recombination joints of KpnI enzyme cutting sites and SpeI enzyme cutting sites on a pDS1301 vector are respectively introduced to an upstream primer and a downstream primer, and the design primers are as follows:

RNAi-1F:5′-agggcgcgcctgcaggtaccccgcctccaccgcttccgcctccct-3′

RNAi-1R:5′-tccacatcgcgataggtacctcatgctctcccccccagaaatgac-3′

RNAi-2F:5′-aggactctagacccactagtccgcctccaccgcttccgcctccct-3′

RNAi-2R:5′-gcagacccgggaggactagttcatgctctcccccccagaaatgac-3′

using the cDNA clone obtained in example 1 as a template, PCR was performed using primers RNAi-1F and RNAi-1R, and the product size was 447 bp. The PCR procedure was as follows: pre-denaturation at 94 ℃ for 5 min; 30 cycles (denaturation at 94 ℃ for 30 seconds; annealing at 55 ℃ for 30 seconds; extension at 72 ℃ for 30 seconds) and extension at 72 ℃ for 7 minutes. And connecting the fragment amplified and separated by the PCR to a pDS1301 vector digested by KpnI through homologous recombination to obtain an intermediate vector. After sequencing and correction, PCR was performed using the cDNA clone obtained in example 1 as a template and primers RNAi-2F and RNAi-2R, and the product size was 447 bp. The PCR procedure was as follows: pre-denaturation at 94 ℃ for 5 min; 30 cycles (denaturation at 94 ℃ for 30 seconds; annealing at 55 ℃ for 30 seconds; extension at 72 ℃ for 1 minute) and extension at 72 ℃ for 7 minutes. The PCR amplified fragment was ligated to the SpeI digested intermediate vector by homologous recombination. And after sequencing comparison, transforming the DNA into agrobacterium EHA105 to obtain a qSCT11 gene RNAi expression vector.

2. Obtaining of RNAi transgenic plants

The qSCT11 gene RNAi expression vector is transferred into japonica rice middle flower 11 through agrobacterium-mediated genetic transformation. The T0 generation transgenic plant is obtained through selective culture, differentiation, rooting and seedling hardening. All transgenic materials are propagated to obtain T1 generation materials with stable inheritance. For a T1 generation transgenic line, if the T2 generation plants obtained by sampling the T1 generation transgenic line are all transgenic plants, the T1 generation transgenic line is a homozygous transgenic line. 3 homozygous transgenic lines RNAi-1, RNAi-2, and RNAi-3 were selected for subsequent analysis.

3. Identification of RNAi transgenic plants

(1) Identification of expression level

Taking 2-leaf stage seedlings, extracting total RNA, carrying out reverse transcription to obtain cDNA, and carrying out fluorescence real-time quantitative PCR. The primers for detecting the expression level of the qSCT11 gene are as follows:

RT-F:5′-ACCGCTTCCGCCTCCCTCCCAT-3′；

RT-R:5′-CTGCTCGCAGGTGAAGAGGAGCC-3′。

the reagent used for the quantitative analysis was FastStart Universal SYBR Green Master (ROX). The instrument used was a real-time fluorescent quantitative PCR instrument ViiA7 from Applied Biosystems, USA.

The Ubqtin gene serves as an internal reference.

The relative expression of qSCT11 gene in the RNAi material is shown in figure 4. In FIG. 4, WT represents flower 11 in the rice variety, and RNAi-1, RNAi-2 and RNAi-3 represent qSCT11 gene RNAi transgenic lines. In the RNAi transgenic strain of the qSCT11 gene, the relative expression level of the qSCT11 gene is obviously lower than that of the rice middle flower 11.

(2) Cold-resistant phenotype identification

Seeds are taken and soaked for 2 days at the temperature of 30 ℃, germination is carried out for 1 day, then 36 germinated seeds are sown in a black small square box (the length, the width and the height are both 6cm) filled with nutrient soil, the seeds are cultured to the 2-leaf stage in an illumination incubator (the illumination time is 16h/d and the temperature is 30 ℃), and the seeds are photographed before treatment. And (3) placing the 2-leaf stage seedlings in a low-temperature incubator at 4 ℃ for processing for 96h, then placing the seedlings back in an illumination incubator for restoring the culture for 1 week, photographing and counting the survival rate. Each material was tested in 3 replicates.

The photographs before and after the treatment are shown in FIG. 5. In FIG. 5, WT shows flower 11 in the rice variety, and RNAi-1, RNAi-2, and RNAi-3 show qSCT11 gene RNAi transgenic plants with a scale of 5 cm. After low temperature stress, the growth condition of the RNAi transgenic plant of the qSCT11 gene is obviously worse than that of the rice middle flower 11.

The survival results are shown in figure 6. In FIG. 6, WT shows flower 11 in the rice variety, and RNAi-1, RNAi-2, and RNAi-3 show qSCT11 gene RNAi transgenic plants. After low temperature stress, the survival rate of the RNAi transgenic line of the qSCT11 gene is obviously lower than that of the rice middle flower 11. The average survival rates were: 77.12%, RNAi-1: 36.98%, RNAi-2: 40.98%, RNAi-3: 24.35 percent.

Example 4Crispr knock-out qSCT11 Gene reduces Cold tolerance of Rice at seedling stage

1. Construction of a Crispr knockout vector

According to the gene sequence of the rice gene qSCT11, see the full-length sequence of SEQ ID NO.1, a target site for knocking out the qSCT11 gene is designed through a website http:// skl.scau.edu.cn/targettdisign, and the designed target site is as follows: 5'-AATCGAGTCAGAACTGAGAG-3' are provided. Introducing homologous recombination linkers at two sides of a KpnI enzyme cutting site of a pCXUN-CAS 9 vector into an upstream primer and a downstream primer respectively, and designing the primers as follows:

U3-F:5′-cccctttcgccaggggtaccgtaattcatccaggtctccaag-3′

U3-R:5′-tacgaattcgagctcggtaccgctgtgccgtacgacggtacg-3′

Crispr-F:5′-aatcgagtcagaactgagaggttttagagctagaaatagcaagtta-3′

Crispr-R:5′-ctctcagttctgactcgattgccacggatcatctgcacaactc-3′

OsU3 is used as a template, and PCR is firstly carried out by using a primer U3-F and Crispr-R to obtain a PCR product 1, wherein the size of the product is 476 bp. The PCR procedure was as follows: pre-denaturation at 94 ℃ for 5 min; 30 cycles (denaturation at 94 ℃ for 30 seconds; annealing at 55 ℃ for 30 seconds; extension at 72 ℃ for 30 seconds) and extension at 72 ℃ for 7 minutes. And then OsU3 is used as a template, and a PCR product 2 is obtained by using a primer U3-R and Crispr-F, and the size of the product is 368 bp. The PCR procedure was as follows: pre-denaturation at 94 ℃ for 5 min; 30 cycles (denaturation at 94 ℃ for 30 seconds; annealing at 55 ℃ for 30 seconds; extension at 72 ℃ for 30 seconds) and extension at 72 ℃ for 7 minutes. And finally, performing PCR by using a mixture of the PCR product 1 and the PCR product 2 as a template and using primers U3-F and U3-R to obtain a final PCR product with the size of 824 bp. The PCR procedure was as follows: pre-denaturation at 94 ℃ for 5 min; 30 cycles (denaturation at 94 ℃ for 30 seconds; annealing at 55 ℃ for 30 seconds; extension at 72 ℃ for 1 minute) and extension at 72 ℃ for 7 minutes. The final product isolated by PCR amplification was ligated into KpnI-digested pCXUN _ CAS9 vector by homologous recombination. And after the sequencing comparison is correct, the vector is transformed into agrobacterium EHA105, and the Crispr knockout vector of the qSCT11 gene is obtained.

2. Acquisition of Crispr knockout plants

The gene Crispr knockout vector of qSCT11 is transferred into the japonica rice middle flower 11 through agrobacterium-mediated genetic transformation. The T0 generation transgenic plant is obtained through selective culture, differentiation, rooting and seedling hardening. All transgenic materials were propagated to obtain homozygous mutant and stably inherited T1 generation material without cas9 protein residue. 3 homozygous transgenic lines Crispr-1, Crispr-2 and Crispr-3 were selected for subsequent analysis.

3. Identification of Crispr knockout plants

1) Genotyping

And (3) taking seedlings in the 2-leaf stage, and extracting DNA. PCR was performed using primers sq-F and sq-R, and the amplification product size was 474 bp. Sequencing the obtained PCR product, and analyzing the condition that the target site of the qSCT11 gene is edited. The PCR procedure was as follows, pre-denaturation at 94 ℃ for 5 min; 32 cycles (denaturation at 94 ℃ for 30 seconds; annealing at 55 ℃ for 30 seconds; extension at 72 ℃ for 30 seconds) and extension at 72 ℃ for 7 minutes. The primer sequences used were as follows:

sq-F：5′-ATTACCTTCTTTTTGAGCTGCCATG-3′

sq-R：5′-GATGAGGCCATTAGCATTGAAAATT-3′

the results of sequencing the target sites of the Crispr knockout material are shown in FIG. 7. In FIG. 7, WT indicates flower 11, Crispr-1, Crispr-2 and Crispr-3 in the rice variety, and qSCT11 gene Crispr knockout line. In the Crispr knockout strain of the qSCT11 gene, the qSCT11 gene is homozygously mutated at a target site.

2) Cold-resistant phenotype identification

Seeds are taken and soaked for 2 days at the temperature of 30 ℃, germination is carried out for 1 day, then 36 germinated seeds are sown in a black small square box (the length, the width and the height are both 6cm) filled with nutrient soil, the seeds are cultured to the 2-leaf stage in an illumination incubator (the illumination time is 16h/d and the temperature is 30 ℃), and the seeds are photographed before treatment. And (3) placing the 2-leaf stage seedlings in a low-temperature incubator at 4 ℃ for processing for 96h, then placing the seedlings back in an illumination incubator for restoring the culture for 1 week, photographing and counting the survival rate. Each material was tested in 3 replicates.

The photographs before and after treatment are shown in FIG. 8. In FIG. 8, WT indicates flower 11, Crispr-1, Crispr-2 and Crispr-3 in the rice cultivar, and qSCT11 gene Crispr knockout plants are indicated on a scale of 5 cm. After low temperature stress, the growth condition of the Crispr knockout plant of the qSCT11 gene is obviously worse than that of the rice middle flower 11.

The survival results are shown in figure 9. In FIG. 9, WT shows flower 11 in the rice cultivars, Crispr-1, Crispr-2, and Crispr-3 show the Crispr knockout plant of the qSCT11 gene. After low-temperature stress, the survival rate of the Crispr knockout plant of the qSCT11 gene is obviously lower than that of the rice middle flower 11. The average survival rates were: WT: 56.77%, Crispr-1: 35.45%, Crispr-2: 37.23%, Crispr-3: 37.04 percent.

Example 5 application of Rice Gene qSCT11 in improving Cold resistance of Rice variety 9311

1. Near isogenic line construction

Taking 9311 as a receptor parent and Nipponbare as a donor parent, hybridizing 1 time with the receptor parent 9311 for 4 generations continuously, combining with molecular marker for auxiliary selection, finally obtaining a near isogenic line with qSCT11 replaced by Nipponbare and genetic background 9311, and naming the near isogenic line as NIL-NIP. The genotype was checked by re-sequencing and it was found that the whole genome contained 4.5Mb of the Nipponbare introgression fragment only at qSCT 11. The NIL-NIP genotype of the near isogenic line is shown in FIG. 10.

2. Identification of expression level

NIL-NIP and NIL-9311 are treated at low temperature (4 ℃) for 0h, 3h and 24h of seedlings in 2-leaf stage, total RNA is extracted, cDNA is obtained by reverse transcription, and fluorescence real-time quantitative PCR is carried out. The primers for detecting the expression level of the qSCT11 gene are as follows:

RT-F:5′-ACCGCTTCCGCCTCCCTCCCAT-3′；

RT-R:5′-CTGCTCGCAGGTGAAGAGGAGCC-3′。

the reagent used for the quantitative analysis was FastStart Universal SYBR Green Master (ROX). The instrument used was a real-time fluorescent quantitative PCR instrument ViiA7 from Applied Biosystems, USA. The Ubqtin gene serves as an internal reference.

The relative expression level of qSCT11 gene in the near isogenic line is shown in FIG. 11. In FIG. 11, after low temperature treatment for 0h and 24h, there was no significant difference in the relative expression of qSCT11 gene in NIL-NIP and NIL-9311, and after low temperature treatment for 3h, the relative expression of qSCT11 gene in NIL-NIP was significantly higher than that of NIL-9311.

3. Cold-resistant phenotype identification

Seeds are taken and soaked for 2 days at the temperature of 30 ℃, germination is carried out for 1 day, then 36 germinated seeds are sown in a black small square box (the length, the width and the height are both 6cm) filled with nutrient soil, the seeds are cultured to the 2-leaf stage in an illumination incubator (the illumination time is 16h/d and the temperature is 30 ℃), and the seeds are photographed before treatment. And (3) placing the 2-leaf stage seedlings in a low-temperature incubator at 4 ℃ for treatment for 4h, then placing the seedlings back in the illumination incubator for 1 week of recovery culture, photographing and counting the survival rate. Each material was tested in 3 replicates.

The photographs before and after treatment are shown in FIG. 12. In FIG. 12, the growth status of NIL-NIP was significantly better than NIL-9311 after low temperature stress.

The survival results are shown in figure 13. In FIG. 13, the survival rate of NIL-NIP after low temperature stress was significantly higher than NIL-9311. The average survival rates were: NIL-NIP: 42.78 percent and NIL-9311:13.62 percent.

Although the invention has been described in detail hereinabove by way of general description, specific embodiments and experiments, it will be apparent to those skilled in the art that many modifications and improvements can be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.

Sequence listing

<110> university of agriculture in Huazhong

<120> rice cold-resistant gene qSCT1 and application thereof

<130> KHP191113835.2

<160> 24

<170> SIPOSequenceListing 1.0

<210> 1

<211> 5446

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

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cccagaagag taacctataa aaatacactt gacggctcga ggatccaatt ttccaacgga 60

tggtctatga tctctgacaa aacaagtaca cccaaacact tttggaggaa caatgaattc 120

attcttccca aacaccattt catatggagt cttcatccca agtattcgtg atggcatgcg 180

attgatgaga tatgttgctg tcatgactgc ctcactccat agaaactttg gtacattcat 240

cgtatacatg agtgaacgag ttacctccag aatatgttga tttttcctct cagctactcc 300

gttctgtgga ggtgtgtcag cacaagaagt ttgatgtact attccttctg ctgagagaaa 360

actcctaaat tcatcattca catattctcc cccattatcg ctcctaataa ctttaacaca 420

agtgtttaat tgattcctaa cccaagcata gaaattcttg aaacatttca atacttcact 480

cttgtgtctc atgagatata tccaagtcat tcgtgagtgg caatcaataa aggtgacaaa 540

gtactttgct ccacttaccg aagcaatagg agaagtccat acatcagagt gaaccaacac 600

aaatggcaaa atgcttcgaa gtcctctgct tatgtatgtg gttcgtgtat gttttccata 660

ctcgcatgca tcacacatta atttgatttt gtccacctta ctcatctcat caggaaatac 720

cttagacata gtatcaaagg acatatgtcc caattgacaa tgttttagaa tcaatttagc 780

ctcctcttca cttgtgctag ctaacaaagc actacatagt gctttgtccg tccctcttct 840

atctagatac cacaacccat tatgcctaac gccggttcca agctttcttc cagttcttct 900

ctcctcaatt agacaattct ctcggtcaat agtaattcga caatccatat gatcaaccaa 960

cgagcttacg gatactaggt taacaggaaa agagggagca tataaaactg acgacaatgt 1020

aattgatgga gtacatttga ctgttcccac tcccctcaca tgtcgtaaag ttccgtcggc 1080

agtttgaatt gtttctttaa atgatgactg aaatggagta tatgatgtaa actcactcga 1140

cattcctgta acatgtctag atgctccgga gtctaatatc caagttgagt taggtacctg 1200

cataggatgt gcgagattaa caaagttgcc aagcactgtt gttttctcct tcactttctg 1260

aattcttccc cttatttctt caacttcttc agccaatttc tcaagtattt gctcaattcc 1320

ttgaatttcc cccatggcag atcaaccaaa tcttctagga tactagccaa atcaattaat 1380

aggagtaaca acactactcc tcagtttctc accaaaaaca gaacagcagc agtagcacta 1440

cttcctccca gcagcactac ttttcagttt cccccaaatc agcacaacag cagcagcaag 1500

ccaagctctc gtactcagtt tcttccccaa atcagaacaa cagcaacagc aaaccaagct 1560

tccgcttcac ctccaaatcc tcaccttctc aagtagcagc agcaccagca acagcaccac 1620

caaagtcacc accactggaa cggacaacaa tagccaatag caccaaatcc ttctctgttg 1680

atcctacagc caacagctgc gcgtctctca gccgctgtgc tccctacagc ttgtgctgca 1740

ccaaatcctt gcgcagctca accagcgagc gtgcgccctg ctcctccttc acgctgctcg 1800

cgtcgagaac ctctgcagca gccgtcgccg cctccgccgt caacactgcc gctccgcctc 1860

cgctgtccga cgtcgccgct ccgcctcagc cgtcgacgtc gccgcagaac ccctctggtc 1920

acagcgccgc cgccgactag ggttcgcgtc ggcgaggtcg ccggtgaccc agctctgata 1980

ccatgtagtt ttgtgatttt tcctctagaa gcttcacttc atttcctttc tcatctacta 2040

tacatataca ttggtggaat gggccctatt gggctgaaat acacacgcct acactcaaca 2100

gttaaagatg gaaaatgtaa cagaaaaagg ttttggagtt accaaggtgc caattgatca 2160

ttatttcttg tgatacattt cacttgtccc atggcatcta gttccagcag gggtgacatt 2220

tgcaatatta tactgatgtt ctaggtgttc acaatagagc tcaaatggca cagcatattc 2280

caactcttag gtaactttgt gtatgtggtg tgctagtgca gaatgatatc cgtaaaaatc 2340

cactatttag acaacagttc catgagatgt gtgcaaaagt tggagtagac cctctggctt 2400

cgaataaagg agcttgggca gaacttctag ggattggtga cttctactat gaattgggtt 2460

ggtaacttaa tttaagctat cttatttatt ttgatcctgg aagtttcttt cttttattat 2520

tggaccattt atgttctgct tagcatctgt gtctattgca ccaggagttc agattgttga 2580

catatgcata gcaaccagag cgaccaacgg tggcttgatt gacttgctag atctccgtaa 2640

actcctgtgt cagaagagga aagctgatct tggatcctta acatcagatg attgtttgcg 2700

tgcaataagt aagctgaagg caagtatttt aaggaattct acagctacat atttttttgg 2760

aatcttataa ttgcattcat gccagtttta caagatcgca ctcgaatcga ttaattcttg 2820

gacagatctg ttaattcttc tgatccttcg cttgtgaact ctcaggtcct tggtagtggt 2880

tttgaagtaa tttctgttgg gaagaaaaag cttgtacgtt ctgttcctac agaactgaat 2940

aaagatcaca atgggatact tgagctagct caggtttgta atcacagctt ttaatagtgt 3000

ataatactac agtacactta acaagtatgt ggatgctgtc ataaccttat catgcttgca 3060

ccgaacaagt atgtggatgg tgtataacct ttggttggaa gtcgtaaagc cctatatatt 3120

ccacataaac acttgtttta tcatgcttat ttcagtcaac atagagacat agcacaaaac 3180

tatgttaagc caaagctttc aaattgctgc cttgctcaat ctgtcacctt gccacaatat 3240

gaccatacct gcccagttat aacatcattc acatgcaggc tgaaggtttt gtcacagtag 3300

aacaagtcaa gagaaagttc tcgtggtcaa ctggccgtgc catcgatgtc cttgaaactt 3360

tgctcaaggt aaatcctgct cctgcatttc cactctttag gccacctttg tgcttcattt 3420

ccgtgcccaa agaaaagaaa actgacttcc tttgatgcaa gttatgtatt ggtgattaga 3480

attctctctg ttgaatccac gcttaagcca ttgcaggatt accttctttt tgagctgcca 3540

tgtataaata aagtactttt cttatgtgca tctctattga cacttgttat gaacgtcttt 3600

ggcatctata tgcaactctg atctctatta cacaaacagg aagggcttgc tatgatcgac 3660

gatgggcaca gggatggcaa gcgcagatac tggttcccgt gcgccactct cagttctgac 3720

tcgattggtg ccgatgccaa gtcatgacct gttccactcc attgttgtca cctgtatcct 3780

tttttccatt ggttcatttc tgtacagctc tggaagaatc acgagatagc aatacacttt 3840

cagtgctggg ctgccggcaa aaatgttaca ttgtgattcg tgcatcaggc aggattctga 3900

acgtatttgt actagtggtc tatactatac tagttcttct ttcacgaaat tctactcgta 3960

attgtaaatt ttcaatgcta atggcctcat ctttcctcat attttcgctt atgtttatgc 4020

catcagctag aatttgaatt tttcaacttt aaatttagag ttgattttga gggttttttt 4080

cattgaagtt tatttccagc cacgtatata aaaattctat ttataaatta tttttcgttt 4140

gtaaatatgg gtattccacg ccgtaagttg ctaatgtgag caagagaaat gctcgcaata 4200

aaaatgatta ataaaaacca tattcattcg gcccacgggc ctcagccctc aaggcttcta 4260

ccaatgacaa ttgacagccg gacaggagta attggcaaga agaaataaaa gggaaaaaag 4320

aggcggaaat tcatttttgg gaaatggcca ccacagaggc gaagcgccgg cggtgattcc 4380

tcctccccta ctccggcggc cgccgccgac tcgtcgttac cgtcgctaga tctcatccct 4440

gacatagccc gccgcttgac gtccctggag gatttcttct ccctgcgcgc ctcctaccgc 4500

gcgctcctcc cggcgttgcg ccgcctcctc gcctcgcagt ccccgctcct cctcgtctcc 4560

ctctacccct ccttctccga ggcgctcttc cacccgcgcc tccgccgcct ccaccgcttc 4620

cgcctccctc ccatgggggc accacctgcc cccctcacgc tacaccctcc tctacgccca 4680

cggcttcctc accaccgccg ccaacaacta cccgccaagg ctcctcttca cctgcgagca 4740

gctccgcctc cccaaggtct tcgcaccctt ctcccgcgtc atcctcacgg cggacctcct 4800

cgtcgtcatc ttcttggccg gccgggccac cgtccagcac tgccaccccg gggacgcgct 4860

ctggcgcgtg gcctccgccc ctgcacccca agtgttcgac gatttgatct ctgtcaacgg 4920

caccctctac gcgctggttg gcctccgtct tgccacgctc gagctgtcgg agagttcact 4980

ggagctgtca tttctggggg ggagagcatg atgacgcgaa caggccggag ggggatcatc 5040

ggttcatgct ccgggagtgt ggaggtgagg tgttgctcat cagcgtggag catgaggaga 5100

gggttgtgta ccgcgtgttc cgttgggtgt ctgacaaaag gaagtgggag atgatcacca 5160

acttgggcgg gcggtcgcta tttcttggct tggatggatt tgcagcttgc gttgatgatc 5220

atccaggggt tcgagggggc tgggccacgc ttgggtgagt ggcatgagta ttccttggct 5280

gatggaatct gtgatgtctg caatgctgac tatccaggct caccacccct gaacaaacag 5340

ttcactcatc agaccatcag tttggatctt ccccagcttg tgctgatgca agcctgagaa 5400

gggttcatcc tagatcaatg ttgaagtcct attgctcagt tgctca 5446

<210> 2

<211> 831

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 2

atggaaaatg taacagaaaa aggttttgga gttaccaagg tgttcacaat agagctcaaa 60

tggcacagca tattccaact cttaggtaac tttgtgtatg tggtccttgg tagtggtttt 120

gaagtaattt ctgttgggaa gaaaaagctt gtacgttctg ttcctacaga actgaataaa 180

gatcacaatg ggatacttga gctagctcag gctgaaggtt ttgtcacagt agaacaagtc 240

aagagaaagt tctcgtggtc aactggccgt gccatcgatg tccttgaaac tttgctcaag 300

gaagggcttg ctatgatcga cgatgggcac agggatggca agcgcagata ctggttcccg 360

tgcgccactc tcagttctga ctcgattggt gccgatgcca agcgctcttc cacccgcgcc 420

tccgccgcct ccaccgcttc cgcctccctc ccatgggggc accacctgcc cccctcacgc 480

tacaccctcc tctacgccca cggcttcctc accaccgccg ccaacaacta cccgccaagg 540

ctcctcttca cctgcgagca gctccgcctc cccaaggtct tcgcaccctt ctcccgcgtc 600

atcctcacgg cggacctcct cgtcgtcatc ttcttggccg gccgggccac cgtccagcac 660

tgccaccccg gggacgcgct ctggcgcgtg gcctccgccc ctgcacccca agtgttcgac 720

gatttgatct ctgtcaacgg caccctctac gcgctggttg gcctccgtct tgccacgctc 780

gagctgtcgg agagttcact ggagctgtca tttctggggg ggagagcatg a 831

<210> 3

<211> 276

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 3

Met Glu Asn Val Thr Glu Lys Gly Phe Gly Val Thr Lys Val Phe Thr

1 5 10 15

Ile Glu Leu Lys Trp His Ser Ile Phe Gln Leu Leu Gly Asn Phe Val

20 25 30

Tyr Val Val Leu Gly Ser Gly Phe Glu Val Ile Ser Val Gly Lys Lys

35 40 45

Lys Leu Val Arg Ser Val Pro Thr Glu Leu Asn Lys Asp His Asn Gly

50 55 60

Ile Leu Glu Leu Ala Gln Ala Glu Gly Phe Val Thr Val Glu Gln Val

65 70 75 80

Lys Arg Lys Phe Ser Trp Ser Thr Gly Arg Ala Ile Asp Val Leu Glu

85 90 95

Thr Leu Leu Lys Glu Gly Leu Ala Met Ile Asp Asp Gly His Arg Asp

100 105 110

Gly Lys Arg Arg Tyr Trp Phe Pro Cys Ala Thr Leu Ser Ser Asp Ser

115 120 125

Ile Gly Ala Asp Ala Lys Arg Ser Ser Thr Arg Ala Ser Ala Ala Ser

130 135 140

Thr Ala Ser Ala Ser Leu Pro Trp Gly His His Leu Pro Pro Ser Arg

145 150 155 160

Tyr Thr Leu Leu Tyr Ala His Gly Phe Leu Thr Thr Ala Ala Asn Asn

165 170 175

Tyr Pro Pro Arg Leu Leu Phe Thr Cys Glu Gln Leu Arg Leu Pro Lys

180 185 190

Val Phe Ala Pro Phe Ser Arg Val Ile Leu Thr Ala Asp Leu Leu Val

195 200 205

Val Ile Phe Leu Ala Gly Arg Ala Thr Val Gln His Cys His Pro Gly

210 215 220

Asp Ala Leu Trp Arg Val Ala Ser Ala Pro Ala Pro Gln Val Phe Asp

225 230 235 240

Asp Leu Ile Ser Val Asn Gly Thr Leu Tyr Ala Leu Val Gly Leu Arg

245 250 255

Leu Ala Thr Leu Glu Leu Ser Glu Ser Ser Leu Glu Leu Ser Phe Leu

260 265 270

Gly Gly Arg Ala

275

<210> 4

<211> 407

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 4

ccgcctccac cgcttccgcc tccctcccat gggggcacca cctgcccccc tcacgctaca 60

ccctcctcta cgcccacggc ttcctcacca ccgccgccaa caactacccg ccaaggctcc 120

tcttcacctg cgagcagctc cgcctcccca aggtcttcgc acccttctcc cgcgtcatcc 180

tcacggcgga cctcctcgtc gtcatcttct tggccggccg ggccaccgtc cagcactgcc 240

accccgggga cgcgctctgg cgcgtggcct ccgcccctgc accccaagtg ttcgacgatt 300

tgatctctgt caacggcacc ctctacgcgc tggttggcct ccgtcttgcc acgctcgagc 360

tgtcggagag ttcactggag ctgtcatttc tgggggggag agcatga 407

<210> 5

<211> 18

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 5

cccagaagag taacctat 18

<210> 6

<211> 16

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 6

tgagcaactg agcaat 16

<210> 7

<211> 18

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 7

acacgcctac actcaaca 18

<210> 8

<211> 17

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 8

agcaacacct cacctcc 17

<210> 9

<211> 42

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 9

atgatgatga taaaggtacc atggaaaatg taacagaaaa ag 42

<210> 10

<211> 42

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 10

ctagaggatc cccgggtacc tgctctcccc cccagaaatg ac 42

<210> 11

<211> 22

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 11

accgcttccg cctccctccc at 22

<210> 12

<211> 23

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 12

ctgctcgcag gtgaagagga gcc 23

<210> 13

<211> 45

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 13

agggcgcgcc tgcaggtacc ccgcctccac cgcttccgcc tccct 45

<210> 14

<211> 45

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 14

tccacatcgc gataggtacc tcatgctctc ccccccagaa atgac 45

<210> 15

<211> 45

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 15

aggactctag acccactagt ccgcctccac cgcttccgcc tccct 45

<210> 16

<211> 45

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 16

gcagacccgg gaggactagt tcatgctctc ccccccagaa atgac 45

<210> 17

<211> 22

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 17

accgcttccg cctccctccc at 22

<210> 18

<211> 23

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 18

ctgctcgcag gtgaagagga gcc 23

<210> 19

<211> 42

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 19

cccctttcgc caggggtacc gtaattcatc caggtctcca ag 42

<210> 20

<211> 42

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 20

tacgaattcg agctcggtac cgctgtgccg tacgacggta cg 42

<210> 21

<211> 46

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 21

aatcgagtca gaactgagag gttttagagc tagaaatagc aagtta 46

<210> 22

<211> 43

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 22

ctctcagttc tgactcgatt gccacggatc atctgcacaa ctc 43

<210> 23

<211> 25

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 23

attaccttct ttttgagctg ccatg 25

<210> 24

<211> 25

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 24

gatgaggcca ttagcattga aaatt 25

Claims (23)

1. The application of the protein coded by the rice cold-resistant gene qSCT11 in improving the cold-resistant capability of plants; the amino acid sequence of the protein coded by the rice cold-resistant gene qSCT11 is shown in SEQ ID No. 3.

2. The application of the rice cold resistance gene qSCT11 in improving the cold resistance of plants; the nucleotide sequence of the rice cold-resistant gene qSCT11 is shown in SEQ ID NO. 1.

3. The application of the expression cassette containing the rice cold-resistant gene qSCT11 in improving the cold-resistant capability of plants; the nucleotide sequence of the rice cold-resistant gene qSCT11 is shown in SEQ ID NO. 1.

4. The application of a vector containing a rice cold resistance gene qSCT11 in improving the cold resistance of plants; the nucleotide sequence of the rice cold-resistant gene qSCT11 is shown in SEQ ID NO. 1.

5. The application of the recombinant bacterium containing the rice cold-resistant gene qSCT11 in improving the cold-resistant capability of plants; the nucleotide sequence of the rice cold-resistant gene qSCT11 is shown in SEQ ID NO. 1.

6. The application of the protein coded by the rice cold-resistant gene qSCT11 in culturing cold-resistant transgenic plants; the amino acid sequence of the protein coded by the rice cold-resistant gene qSCT11 is shown in SEQ ID No. 3.

7. The application of the rice cold-resistant gene qSCT11 in culturing cold-resistant transgenic plants; the nucleotide sequence of the rice cold-resistant gene qSCT11 is shown in SEQ ID NO. 1.

8. The application of the expression cassette containing the rice cold-resistant gene qSCT11 in culturing cold-resistant transgenic plants; the nucleotide sequence of the rice cold-resistant gene qSCT11 is shown in SEQ ID NO. 1.

9. The application of the vector containing the rice cold-resistant gene qSCT11 in culturing cold-resistant transgenic plants; the nucleotide sequence of the rice cold-resistant gene qSCT11 is shown in SEQ ID NO. 1.

10. The application of the recombinant strain containing the rice cold-resistant gene qSCT11 in culturing cold-resistant transgenic plants; the nucleotide sequence of the rice cold-resistant gene qSCT11 is shown in SEQ ID NO. 1.

11. The application of the protein coded by the rice cold-resistant gene qSCT11 in the improvement of plant cold-resistant germplasm resources; the amino acid sequence of the protein coded by the rice cold-resistant gene qSCT11 is shown in SEQ ID No. 3.

12. The application of the rice cold-resistant gene qSCT11 in the improvement of plant cold-resistant germplasm resources; the nucleotide sequence of the rice cold-resistant gene qSCT11 is shown in SEQ ID NO. 1.

13. The application of the expression cassette containing the rice cold-resistant gene qSCT11 in the improvement of plant cold-resistant germplasm resources; the nucleotide sequence of the rice cold-resistant gene qSCT11 is shown in SEQ ID NO. 1.

14. The application of the vector containing the rice cold-resistant gene qSCT11 in the improvement of plant cold-resistant germplasm resources; the nucleotide sequence of the rice cold-resistant gene qSCT11 is shown in SEQ ID NO. 1.

15. The application of the recombinant strain containing the rice cold-resistant gene qSCT11 in the improvement of plant cold-resistant germplasm resources; the nucleotide sequence of the rice cold-resistant gene qSCT11 is shown in SEQ ID NO. 1.

16. The application of the protein coded by the rice cold-resistant gene qSCT11 in improving the low-temperature survival rate of plants; the amino acid sequence of the protein coded by the rice cold-resistant gene qSCT11 is shown in SEQ ID No. 3.

17. The application of the rice cold-resistant gene qSCT11 in improving the low-temperature survival rate of plants; the nucleotide sequence of the rice cold-resistant gene qSCT11 is shown in SEQ ID NO. 1.

18. The application of the expression cassette containing the rice cold-resistant gene qSCT11 in improving the low-temperature survival rate of plants; the nucleotide sequence of the rice cold-resistant gene qSCT11 is shown in SEQ ID NO. 1.

19. The application of the vector containing the rice cold-resistant gene qSCT11 in improving the low-temperature survival rate of plants; the nucleotide sequence of the rice cold-resistant gene qSCT11 is shown in SEQ ID NO. 1.

20. The application of the recombinant bacterium containing the rice cold-resistant gene qSCT11 in improving the low-temperature survival rate of plants; the nucleotide sequence of the rice cold-resistant gene qSCT11 is shown in SEQ ID NO. 1.

21. The use according to any one of claims 1 to 20, wherein the plant is rice, wheat, soybean, sorghum, millet, barley, maize.

22. A method for improving the cold resistance of rice is characterized in that the rice expresses or over expresses a rice cold resistance gene qSCT11 by a transgenic, hybridization, backcross, selfing or asexual propagation method, and the nucleotide sequence of the rice cold resistance gene qSCT11 is shown as SEQ ID No. 1.

23. The method of claim 22, wherein said transgene comprises introducing a recombinant expression vector comprising rice cold tolerance gene qSCT11 into rice using Ti plasmid, plant viral vector, direct DNA transformation, microinjection, gene gun, conductance, agrobacterium-mediated methods to obtain transgenic rice lines.

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