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KR-102963512-B1 - OsBGAL9 gene and use thereof for promoting rice growth and increasing stress tolerance

KR102963512B1KR 102963512 B1KR102963512 B1KR 102963512B1KR-102963512-B1

Abstract

The present invention relates to the OsBGAL9 gene for promoting rice growth and increasing stress tolerance, and the use thereof. Transgenic plants overexpressing the rice-derived OsBGAL9 gene of the present invention have strong tolerance to high and low temperature stress and have the effect of being resistant to rice blast and bacterial leaf blight. In addition, they have the effect of promoting plant stem growth. Therefore, the OsBGAL9 gene is expected to be very useful for the development of transgenic plants with increased stress tolerance and promoted plant growth. Since plants with strong stress tolerance and promoted plant stem growth can be obtained by using this gene, it is expected to contribute significantly to increasing the yield of economic crops.

Inventors

  • 전종성
  • 보티수안규
  • 황빗중
  • 이찬희

Assignees

  • 경희대학교 산학협력단

Dates

Publication Date
20260513
Application Date
20230814
Priority Date
20230518

Claims (11)

  1. A recombinant vector for increasing stress tolerance or promoting growth in plants, comprising a gene encoding rice-derived OsBGAL9 ( Oryza sativa beta-galactosidase 9) represented by the nucleotide sequence of SEQ ID NO. 1, The above stress is biological and non-biological stress, and A recombinant vector for increasing stress tolerance or promoting growth of plants, wherein the biological stress is blast or bacterial leaf blight.
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  6. In paragraph 1, A recombinant vector for increasing stress tolerance or promoting growth in plants, characterized in that the above-mentioned abiotic stress is high temperature or low temperature stress.
  7. In a transgenic plant in which a gene encoding rice-derived OsBGAL9 ( Oryza sativa beta-galactosidase 9), represented by the nucleotide sequence of SEQ ID NO. 1, is introduced or overexpressed, and the plant's stress tolerance is increased or growth is promoted, The above stress is biological and non-biological stress, and The above biological stress is a transgenic plant, which is rice blast or bacterial leaf blight.
  8. In Paragraph 7, A transgenic plant characterized by being one or more selected from the group consisting of rice, barley, wheat, rye, corn, sugarcane, oats, and onions.
  9. Seeds of the transgenic plant of Paragraph 7.
  10. A method for increasing stress tolerance or promoting growth of a plant, comprising the step of introducing or overexpressing a gene encoding rice-derived OsBGAL9 ( Oryza sativa beta-galactosidase 9) represented by the nucleotide sequence of SEQ ID NO. 1 in a plant, wherein The above stress is biological and non-biological stress, and A method for increasing plant stress tolerance or promoting growth, wherein the biological stress mentioned above is blast disease or bacterial leaf blight.
  11. A method for producing a transgenic plant with increased stress tolerance or promoted growth, comprising the step of introducing a recombinant vector containing a gene encoding rice-derived OsBGAL9 ( Oryza sativa beta-galactosidase 9), represented by the nucleotide sequence of SEQ ID NO. 1, into a plant to transform it; The above stress is biological and non-biological stress, and A method for producing a transgenic plant with increased stress tolerance or promoted growth, wherein the biological stress is rice blast or bacterial leaf blight.

Description

OsBGAL9 gene and use thereof for promoting rice growth and increasing stress tolerance The present invention relates to the OsBGAL9 gene for promoting rice growth and increasing stress tolerance, and the use thereof. Rice ( Oryza sativa ) is one of the world's most important crops for seed production, but it is susceptible to over 60 types of diseases, and depending on the year, severe damage can lead to a 50–90% reduction in yield. Furthermore, as the average global temperature rises due to increased greenhouse gases from industrialization and deforestation, abnormal climate phenomena are gradually intensifying. Consequently, crop yields are decreasing, placing the country in a situation where there is a high likelihood of food insecurity. Such damage has emerged as the biggest obstacle to securing a stable food supply for humanity. For example, in Korea, an outbreak of rice blast disease in the Tongil variety in 1978 caused serious problems in terms of food security, leading to the import of large quantities of rice from abroad and becoming a social issue for a considerable period. To address these issues, plantlets with enhanced stress tolerance gene function were produced; however, while stress tolerance increased, plant growth was inhibited. Similarly, promoting plant growth resulted in a weakening of stress tolerance. Accordingly, the inventors discovered OsBGAL9 for the development of transgenic plants with increased stress tolerance and promoted plant growth, and completed the present invention as it was determined that the gene has very high agricultural utility value as it promotes rice growth while simultaneously regulating complex resistance to the environment and pathogens. Figure 1 shows the expression of OsSPL7 and OsBGAL9 in WT (Wild Type), OsSPL7-OX, and Osspl7 plants in one embodiment of the present invention. A and B used rice ACTIN as an internal control. C and D used rice UBQ5 as an internal control. (A), (C) Relative expression levels of OsSPL7. (B), (D) Relative expression levels of OsBGAL9. Figure 2 is a figure showing the phylogenetic tree and sequence analysis of OsBGAL9 in one embodiment of the present invention. (A) Diagram of the OsBGAL9 gene (Black box: exon, Line: intron, Red triangle: HSE binding site). (B) Phylogenetic relationship between OsBGAL9 and other family GH35 proteins. (C) Protein sequence of OsBGAL9. FIG. 3 is a figure showing the identification of the OsSPL7 binding site in the OsBGAL9 promoter in one embodiment of the present invention. (A) Diagram of two HSE binding sites in the OsBGAL9 promoter. (B) Schematic diagram of a structure used for transcriptional activity analysis in rice protoplasts. (C) Relative LUC/GUS activity in transformed protoplasts. (D) is a Y1H analysis showing that OsSPL7 binds to the OsBGAL9 promoter. (E) is an EMSA analysis for detecting OsSPL7 binding to the p-type HSE in the OsBGAL9 promoter. FIG. 4 is a figure showing the production of an OsBGAL9 loss of function and overexpression line in one embodiment of the present invention. (A) Diagram of the CRISPR/Cas9 target and T-DNA insertion site in OsBGAL9. (B) Diagram of the OsBGAL9 overexpression structure. (C) Sequence analysis of the target site in the Osbgal9 mutant generated by CRISPR/Cas9. (D) Relative OsBGAL9 expression levels in HY (Wild Type) and T-DNA insertion loss plants Osbgal9-t. (E) shows the relative OsBGAL9 expression levels in overexpressed and CRISPR/Cas9 loss-of-function plants determined by RT-qPCR. Figure 5 is a figure showing the phenotype of OsBGAL9 overexpression and Osbgal9 loss plant in one embodiment of the present invention. (A) Growth phenotypes of OsBGAL9-OX and Osbgal9 plants grown in paddy fields at the mature stage. (B) Plant height of DJ (Wild Type), OsBGAL9-OX, and Osbgal9 at the mature stage. (C) Internode shape of DJ(Wild Type), OsBGAL9-OX and Osbgal9. (D) Length between each node and relative contribution between each node to the total stem length. (E) Representative longitudinal section of the second node. (F) Cell length of the second node observed in the longitudinal section. Figure 6 is a figure showing the phenotype of an Osbgal9-t T-DNA mutant plant compared with a wild-type control HY in one embodiment of the present invention. (A) Growth phenotypes of Osbgal9-t and HY plants grown in a paddy field at the mature stage. (B) Plant height of Osbgal9-t and HY plants. (C) Internode morphology of HY (Wild Type) and Osbgal9-t. (D) Relative contribution between each node to the total stem length of HY (Wild Type) and Osbgal9-t plants. (E) Representative longitudinal section of the second node. (F) Cell length of the second node observed in the longitudinal section. FIG. 7 is a figure showing the histochemical GUS staining of OsBGAL9pro:GUS plants in one embodiment of the present invention. (A) Tissue stained at the seedling stage. (B) Tissue stained at the flowering stage. (C) Relative GUS and OsBGAL9 transcriptome levels in different tissues. (D) Ti