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CN-121975838-A - HIS mutant protein and application thereof

CN121975838ACN 121975838 ACN121975838 ACN 121975838ACN-121975838-A

Abstract

The invention provides an HIS mutant protein and application thereof in plant breeding, in particular to the HIS mutant protein, which can not only endow plants with herbicide resistance, but also reduce plant height and improve yield after being over-expressed in plants, and has very wide application prospects in biological breeding.

Inventors

  • XIE HONGTAO

Assignees

  • 山东舜丰生物科技有限公司

Dates

Publication Date
20260505
Application Date
20260105
Priority Date
20250114

Claims (13)

  1. Use of a HIS1 (HPPD INHIBITOR SENSITIVE 1) mutein for the preparation of a plant with improved traits selected from the group consisting of the following, or for improving plant traits, wherein said HIS1 mutein comprises a mutation at amino acid position 242 corresponding to the amino acid sequence shown in SEQ ID No.1 compared with the amino acid sequence of the parent HIS1 protein - Any one or more of the following: The yield is improved; The number of fruit pods is increased; the number of seeds is increased; increasing the grain weight; The plant height is reduced; increased resistance to HPPD-inhibiting herbicides.
  2. 2. The use according to claim 1, wherein the HIS1 mutein is introduced into a plant to prepare or to modify a plant.
  3. 3. The use according to claim 2, wherein the HIS1 mutein is introduced in a plant such that the HIS1 mutein is overexpressed in a plant.
  4. 4. The use according to claim 1, characterized in that it comprises a step of treating the plant with an HPPD-inhibiting herbicide.
  5. 5. The use according to claim 4, wherein the treatment is spraying of the plant with an HPPD inhibiting herbicide.
  6. 6. The use according to any one of claims 1 to 5, wherein the plant is soybean.
  7. 7. Use of a nucleic acid molecule encoding a HIS1 mutein according to any one of claims 1 to 6 or a biological material comprising said nucleic acid molecule for the preparation of a plant with improved traits or for improving plant traits.
  8. 8. A method for improving a plant, or a method for preparing a plant with improved traits, comprising the step of introducing into said plant the HIS1 mutein of any one of claims 1 to 6, wherein said improved traits are selected from the group consisting of - Any one or more of the following: The yield is improved; The number of fruit pods is increased; the number of seeds is increased; increasing the grain weight; The plant height is reduced; increased resistance to HPPD-inhibiting herbicides.
  9. 9. The method of claim 8, comprising the step of overexpressing the HIS1 mutein in a plant cell, plant seed, plant tissue, plant part of the plant.
  10. 10. The method according to any one of claims 8-9, wherein the method comprises the steps of obtaining a plant that overexpresses the HIS1 mutein and growing the plant.
  11. 11. The method of claim 10, further comprising the step of treating the plant with an HPPD-inhibiting herbicide.
  12. 12. The method of claim 11, wherein the plant is treated with the HPPD inhibiting herbicide 10 days after sowing the plant.
  13. 13. A method of preparing a hybrid plant, the method comprising crossing a first plant with a second plant to obtain the hybrid plant, wherein the first plant is a plant prepared by the method of any one of claims 8-12.

Description

HIS mutant protein and application thereof The present application claims priority from chinese patent application CN202510052734.2, with application date 2025, 1, 14. The present application incorporates the entirety of the above-mentioned chinese patent application. Technical Field The invention belongs to the field of agricultural genetic engineering, and particularly relates to an HIS mutant protein and application thereof in plant breeding. Background Soybeans are a source of major human oils and proteins, one of the world's most important commercial crops, and a major source of human vegetable oils and vegetable proteins. With the improvement of living standard and the improvement of dietary structure, the demand of high-quality soybean oil is growing, and the cultivation of high-yield high-quality soybean becomes one of the important targets of soybean breeding. HIS1 (HPPD INHIBITOR SENSITIVE 1), gene number LOC_Os02g17940 in rice, which codes for 351 amino acids of Fe (II)/2-ketoglutarate dependent oxygenase (Fe (II)/2-oxoglutarate (2 OG) -DEPENDENT OXYGENASES), the 242 th mutation of the rice HIS1 protein can enhance herbicide resistance, especially resistance to HPPD-inhibiting herbicides. In order to further investigate the use of HIS1 and its muteins for herbicide resistance and trait improvement, osHIS muteins were expressed in soybean in order to obtain a high quality soybean variety. Disclosure of Invention The invention provides application of HIS1 mutant protein in plant property improvement, and after the HIS1 mutant protein is over-expressed in a plant, the plant height of the plant is reduced, and the yield is improved. In one aspect, the invention provides the use of a HIS1 (HPPD INHIBITOR SENSITIVE 1) mutein in the preparation of a plant with improved traits, or in the improvement of plant traits. In another preferred embodiment, the HIS1 mutein comprises a mutation at amino acid position 242 corresponding to the amino acid sequence depicted in SEQ ID No.1 compared to the amino acid sequence of the parent HIS1 protein. In one embodiment, the parent HIS1 protein is derived from rice. In another preferred embodiment, the parent HIS1 protein or wild type HIS1 protein is encoded by the HIS1 gene, which is numbered LOC_Os02g17940 in rice, encoding Fe (II)/2-oxoglutarate-dependent oxygenase (Fe (II)/2-oxoglutarate (2 OG) -DEPENDENT OXYGENASES), and the amino acid is as shown in SEQ ID No. 1. In another preferred embodiment, the mutation is an insertion, deletion or substitution of an amino acid. In another preferred embodiment, the mutein is a HIS1 herbicide resistance/tolerance protein, in particular resistance/tolerance to HPPD inhibitor herbicides. In one embodiment, the amino acid position 242 is S. In one embodiment, the amino acid at position 242 is mutated to a non-S amino acid, e.g., A, V, G, Q, F, W, Y, D, N, E, K, M, T, C, P, H, R, I, L, preferably G. In one embodiment, the amino acid at position 242 is mutated to G. In one embodiment, the HIS1 mutein is selected from any one of the following groups I-III: I. HIS1 mutant protein obtained by generating mutation at 242 th amino acid site of amino acid sequence shown in SEQ ID No. 1; II. A HIS1 mutein having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the HIS1 mutein of I and retaining herbicide resistance activity; III has the mutation site described in I compared with the HIS1 mutein described in I, and has the sequence of substitution, deletion or addition of one or more amino acids including substitution, deletion or addition of 1,2,3, 4, 5, 6, 7, 8, 9 or 10 amino acids compared with the HIS1 mutein described in I, and retains the activity of herbicide resistance. In another preferred embodiment, the HIS1 mutein further comprises additional mutation sites corresponding to one or more of positions 327, 283, 111, 145, 75, 218, 322, 5, 80 and 89 of the amino acid sequence shown in SEQ ID No.1, which additional mutation sites are capable of maintaining or enhancing the tolerance or resistance of the mutein to HPPD-inhibiting herbicides or increasing the applicability of the HIS1 mutein to herbicides. In another preferred embodiment, the parent HIS1 protein is derived from a monocot and/or dicot. In another preferred embodiment, the parent HIS1 protein is derived from one or more plants selected from the group consisting of Gramineae, leguminosae, chenopodiaceae, and Brassicaceae. In another preferred embodiment, the parent HIS1 protein is derived from one or more plants selected from the group consisting of Arabidopsis thaliana, rice, tobacco, maize, sorghum, barley, wheat, millet, soybean, tomato, potato, quinoa, lettuce, canola, cabbage, strawberry. In another preferred embodiment, the parent HIS1 protein is derived from rice, wherein the rice comprises indica rice and japonica rice, and