CN-118360295-B - Perennial ryegrass heat-resistant gene LpHSP-10

Abstract

The invention discloses a perennial ryegrass heat-resistant gene LpHSP-10, which is remarkably up-regulated in expression under high temperature stress and reaches a peak value at 12 hours, the nucleotide sequence of the gene is shown as SEQ ID NO.1, and the gene LpHSP-10 is knocked out by a gene editing technology to cultivate a heat-resistant ryegrass variety. The invention discovers a gene LpHSP-10 which is more sensitive to heat stress based on the whole genome association analysis and the transcriptome sequencing result. The ryegrass heat-resistant gene LpHSP-10 provided by the invention can improve the heat resistance of ryegrass by regulating and controlling the expression of heat stress related genes and then performing gene knockout, is beneficial to cultivating ryegrass varieties suitable for growing in hotter areas in the south, and simultaneously reduces the breeding workload, shortens the breeding scale, shortens the breeding period, improves the breeding efficiency and accelerates the breeding of the heat-resistant ryegrass varieties.

Inventors

• NIE GANG
• BI MING
• ZHANG XINQUAN
• ZHOU JIE
• LIU YANG
• YANG ZHONGFU
• Yan Sanbo
• HUANG LINKAI
• FENG GUANGYAN

Assignees

• 四川农业大学

Dates

Publication Date

20260512

Application Date

20240520

Claims (1)

1. 1. The application of the perennial ryegrass thermosensitive gene LpHSP-10 is characterized in that the thermosensitive property of the arabidopsis is improved by over-expressing the gene LpHSP-10 in the arabidopsis, and the nucleotide sequence of the gene LpHSP-10 is shown as SEQ ID NO. 1.

Description

Perennial ryegrass heat-resistant gene LpHSP-10 Technical Field The invention belongs to the field of genetic engineering with enhanced heat resistance of plants, and relates to a heat-resistant gene LpHSP-10 of perennial ryegrass, a protein coded by the heat-resistant gene, an amplification primer, an overexpression vector and application of the gene LpHSP-10. Background Perennial ryegrass (Lolium perenne L.) belongs to the genus Lolium of Gramineae, has the characteristics of developed fibrous roots, more tillers, high lawn forming speed, strong disease and pest resistance and the like, is an important pioneer grass seed for lawn planting, and is high in regeneration capacity and yield after being mowed, and is an important cultivated pasture and green manure crop. Perennial ryegrass are widely planted in temperate regions of the world as excellent grassland mixed sowing pasture, lawn greening landscape grass and ecological restoration improvement grass seeds. However, as typical cold season grasses and turf grasses, perennial ryegrass are sensitive to high temperatures in warm, cool and humid climates, and are difficult to go over the summer in southern areas. High temperature stress is an important factor limiting perennial ryegrass growth and development, and with global warming, the growth and development of perennial ryegrass can be greatly affected by the occurrence of extremely high temperatures. How to actively cope with the threat of global warming to the survival of perennial ryegrass, how to improve the heat resistance of perennial ryegrass is explored, and the problem of the limitation of popularization and application of the perennial ryegrass as pasture grass and turf grass is particularly important. Disclosure of Invention In view of the above, the invention aims to provide a perennial ryegrass heat resistance gene LpHSP-10. Another objective of the present invention is to provide a protein encoded by gene LpHSP-10. The invention further aims at providing a primer pair for cloning the ryegrass heat-resistant gene LpHS P-10. The fourth object of the invention is to provide an over-expression vector containing the ryegrass heat-resistant gene LpHSP-10. The invention aims at providing a method for improving the heat resistance of perennial ryegrass. The sixth object of the present invention is to provide a method for constructing an Arabidopsis transgenic plant using the gene LpHSP-10. The seventh object of the present invention is to provide a method for verifying the heat resistance of gene Lp HSP70-10 by said transgenic plant. The eighth object of the present invention is to provide a use of the gene LpHSP, which is useful for the production of a polypeptide. The invention aims at providing a breeding method of perennial ryegrass. The inventor continuously reforms and innovates through long-term exploration and trial and repeated experiments and efforts, and the technical scheme provided by the invention is that the perennial ryegrass heat-resistant gene LpHSP-10 is provided, the expression of the gene is obviously up-regulated under high temperature stress and reaches a peak value at 12 hours, and the nucleotide sequence of the gene is shown as SEQ ID NO. 1. The invention also provides a protein coded by the gene LpHSP-10, and the amino acid sequence of the protein is shown as SEQ ID NO. 2. The invention also provides a primer pair for cloning the ryegrass heat-resistant gene LpHSP-10, wherein the base sequence of the primer pair is as follows: An upstream primer F5'-ATGTCGGGCAAGGGCGAAGGTCCGG-3'; and the downstream primer R is 5'-TTAGTCGACCTCCTCAATCTTGGGC-3'. The invention also provides an over-expression vector containing the ryegrass heat-resistant gene LpHSP-10, wherein the full-length ryegrass heat-resistant gene LpHSP-10 is inserted into the vector, and the over-expression vector LpHSP-10 is obtained by containing a CaMV 35S promoter. The invention also provides a method for improving the heat resistance of perennial ryegrass, which is characterized in that the gene LpHSP-10 is knocked out, and the expression quantity of the gene is analyzed by transcriptome sequencing and real-time fluorescent quantitative PCR technology. The invention also provides a construction method of the arabidopsis transgenic plant by using the gene LpHSP-10, the transgenic plant is obtained through agrobacterium-mediated transformation and screening, and heat resistance verification is carried out under the high-temperature condition. The invention also provides a method for verifying the heat resistance of the gene LpHSP-10 by using the transgenic plant, and the heat resistance of the transgenic plant is evaluated by measuring indexes including chlorophyll fluorescence, conductivity and chlorophyll content. The invention also provides application of the gene LpHSP-10, namely knocking out the gene LpHS P-10 to improve heat resistance of plants, and the gene is used for cultivating new heat-resistant plant va