CN-119752945-B - Stain-resistant gene Di19 and application thereof in rape stain-resistant breeding

Abstract

The invention belongs to the technical field of genetic engineering, and particularly discloses a stain-resistant gene Di19 and application thereof in rape stain-resistant breeding, wherein the Di19 gene of Russian mustard is transferred into rape for the first time by using a transgenic technology, and the obtained transgenic rape has obviously improved stain resistance compared with wild rape. Therefore, the invention provides a new gene resource for the breeding of the stain-resistant rape.

Inventors

• ZHENG MING
• LIU NIAN
• HUA WEI

Assignees

• 中国农业科学院油料作物研究所

Dates

Publication Date

20260512

Application Date

20250121

Priority Date

20250103

Claims (5)

1. 1. The application of the gene Di19 in improving the resistance of rape is characterized in that the coding sequence of the gene is shown as SEQ ID NO. 1.
2. 2. The application of the encoding protein of the gene Di19 in improving the resistance of rape is characterized in that the amino acid sequence of the protein is shown as SEQ ID NO. 2.
3. 3. The application of the expression vector containing the gene Di19 and the recombinant bacterium in improving the resistance of rape is characterized in that the coding sequence of the gene is shown as SEQ ID NO. 1.
4. 4. A method for improving the stain resistance of rape is characterized in that a Di19 gene is expressed in rape, and the coding sequence of the gene is shown as SEQ ID NO. 1.
5. 5. A breeding method of a stain-resistant rape is characterized by comprising the steps of constructing a plant expression vector containing a Di19 gene, transferring the Di19 gene into rape by an agrobacterium transformation method, and continuously screening to obtain homozygous rape of which the Di19 gene is transferred, wherein the coding sequence of the Di19 gene is shown as SEQ ID NO. 1.

Description

Stain-resistant gene Di19 and application thereof in rape stain-resistant breeding Technical Field The invention belongs to the technical field of genetic engineering, and particularly relates to a stain-resistant gene Di19 and application thereof in rape stain-resistant breeding. Background Rape is used as the oil crop with the largest planting area in China, and the rape seed oil accounts for more than 50% of the oil yield of the domestic oil crop. However, at present, global climate warming causes extreme weather increase such as overcast and rainy days, strong rainfall and the like, so that rape is often subjected to waterlogging stress. Meanwhile, the Yangtze river basin and the Huang-Huai-Hai plain are two major regions of rape in China, and under the rice-oil and rice-oil ship mode promoted and implemented in the Yangtze river basin, the winter rape varieties sensitive to the waterlogging are mainly planted, so that the water content of the rice field is seriously exceeded and the rape is extremely easily stressed by the waterlogging in rainy days concentrated in autumn and winter. The current research shows that the waterlogging stress can cause the rape to suffer serious hypoxia stress and ion poisoning, seriously influence the development of underground root systems of the rape and the growth of overground plants, and simultaneously can also reduce the disease resistance of the rape, the lodging tendency and the oil content of seeds, and seriously restrict the development of the rape industry. Therefore, the cultivation of the stain-resistant rape variety is suitable for the condition of excessive water in rape planting areas, which is helpful for promoting the high-quality development of rape industry in China and has important strategic significance for guaranteeing the safety of national 'oil bottles'. However, genetic improvement of the stain-resistant rape varieties in China is slow at present, available excellent stain-resistant germplasm resources are few, and stable QTL intervals and available functional gene loci are lacking. Therefore, the excavation of excellent resistance genes of the rape and the analysis of the regulation mechanism thereof can provide excellent gene loci and germplasm resources for the resistance breeding of the rape, and the acceleration promotes the more accurate and effective genetic improvement of the resistance rape. Disclosure of Invention The invention provides a gene Di19 for improving the resistance of rape, wherein the coding sequence of the Di19 gene is shown as SEQ ID NO.1, and the amino acid sequence of the coding protein is shown as SEQ ID NO. 2. In another aspect, the invention provides a method for improving the resistance of rape by over-expressing the Di19 gene in rape, comprising constructing a plant expression vector (such as plant expression vector pCAMBIA 1305) containing the Di19 gene, and transferring the Di19 gene into rape by agrobacterium transformation. The invention also provides a breeding method of the stain-resistant rape, which comprises the steps of constructing a plant expression vector containing a Di19 gene, transferring the Di19 gene into the rape by an agrobacterium transformation method, and continuously screening to obtain homozygous transgenic Di19 gene rape. Compared with the prior art, the method has the beneficial effects that the Di19 gene of the Russian mustard is transferred into the rape for the first time by utilizing the transgenic technology, and compared with the non-transformed wild rape, the obtained transgenic rape has obviously improved resistance in germination period. Drawings FIG. 1 shows the results of protein homology alignment of Di 19. FIG. 2 is a schematic diagram of construction of recombinant plasmid pCAMBIA1305-Di 19. FIG. 3 shows the growth state of rape seedlings after waterlogging stress. WT means wild-type rape and OE Di19 means Di19 transgenic rape. FIG. 4 shows the germination rate of rape after a waterlogging stress. WT means wild-type rape and OE Di19 means Di19 transgenic rape. Detailed Description For further explanation of the present invention, the stain-resistance gene Di19 provided by the present invention and its application in rape stain-resistance breeding will be described in detail below with reference to the accompanying drawings and examples, but it is only some, but not all, examples of the present invention, and those skilled in the art can obtain other examples according to the present invention without inventiveness, and these examples are all within the scope of the present invention. Unless otherwise indicated, all technical means used in the examples described below are conventional means well known to those skilled in the art, and all materials reagents used are commercially available. In a specific embodiment of the invention, brassica napus (Brassica napus l.) is used as Brassica napus 862 (a laboratory-harvested spring rape line), and the escherichia coli DH5 a compete