CN-121975816-A - Method for promoting plant seeds to grow

Abstract

The invention provides a method for promoting plant seeds to grow. Specifically, the invention discovers that the DBR1 gene regulates the size of plant seeds for the first time, and the DBR1 gene of the plant is mutated by means of random mutagenesis, so that the activity of DBR1 protein is reduced, and the seed enlargement can be promoted. In addition, the invention also provides a method for screening mutation capable of promoting plant seed to grow and a plant with improved related characters.

Inventors

• ZHENG BINGLIAN
• LI LEI
• GOU CHUANJIE
• GE HAORAN
• YANG WENYA

Assignees

• 复旦大学

Dates

Publication Date

20260505

Application Date

20260204

Claims (10)

1. 1. A method of promoting plant seed enlargement, the method comprising: 1) Mutating the DBR1 gene or DBR1 protein of the plant; 2) Introducing a mutant DBR1 gene or DBR1 protein into the plant, or 3) Contacting the plant with a DBR1 protein inhibitor, Wherein the mutation reduces the activity of the DBR1 protein or reduces the expression of the DBR1 protein.
2. 2. The method of claim 1, wherein the plant is a plant capable of producing seeds.
3. 3. The method of claim 1, wherein the plant comprises an arabidopsis, soybean, maize, rice, wheat, horticultural plant.
4. 4. The method of claim 1, wherein the mutation increases the cumulative amount of lasso RNA.
5. 5. The method of claim 1, wherein the mutation increases the cumulative amount of lasso RNA by 5% -50%, preferably by 5% -25%, more preferably by 5% -15%.
6. 6. A transgenic plant, characterized in that the transgenic plant is (i) a plant into which a mutation of the DBR1 gene or DBR1 protein is introduced, or (ii) a plant into which a mutation of the DBR1 gene or DBR1 protein is introduced; Wherein the mutation reduces the activity of the DBR1 protein or reduces the expression of the DBR1 protein.
7. 7. A plant with improved traits, which is a plant treated by the method of claim 1.
8. 8. The use of a substance selected from the group consisting of a gene editing agent for mutating a DBR1 gene or a DBR1 protein, a mutant DBR1 gene, a mutant DBR1 protein, or a DBR1 protein inhibitor for promoting plant seed enlargement.
9. 9. The use according to claim 8, wherein the substance is a composition comprising: 1) A gene editing agent for mutating DBR1 gene or DBR1 protein, a mutant DBR1 gene, a mutant DBR1 protein, or a DBR1 protein inhibitor, and 2) An agronomically acceptable carrier.
10. 10. A method of screening for a mutation that promotes plant seed enlargement, the method comprising: A. Randomly mutagenizing a plant to obtain a mutated plant; B. Detecting the content of DBR1 protein and/or lasso RNA in said mutated plant, and C. Compared with non-mutated plants, mutated plants with reduced DBR1 protein content and/or increased lasso RNA content are selected, and the sequence of the DBR1 gene and/or DBR1 protein is detected, thereby obtaining mutations which can promote plant seed enlargement.

Description

Method for promoting plant seeds to grow Technical Field The invention belongs to the field of agriculture, and in particular relates to a method for promoting plant seeds to grow. Background Plant seed size is one of the most important yield traits. According to the prior studies, mature seeds of angiosperms consist of three parts, embryo, endosperm and seed coat, which are developed from the zygote, the central cell that completes fertilization and the beads in the maternal tissue, respectively. Thus, plant seed size is co-regulated by maternal and zygotic tissue. For plant seed size regulation networks, several pathways have been identified to determine seed size by affecting endosperm size and maternal tissue growth, including the IKU pathway, ubiquitin-proteasome pathway, G protein signaling pathway, MAPK signaling pathway, plant hormones, and transcriptional regulators. At present, research and report on regulating genes of plant seed size are limited, so that development of a cultivation method for regulating seed size is urgently needed. Disclosure of Invention The invention aims to provide a method for promoting plant seeds to grow. In a first aspect of the present invention there is provided a method of promoting plant seed enlargement, the method comprising: 1) Mutating the DBR1 gene or DBR1 protein of the plant; 2) Introducing a mutant DBR1 gene or DBR1 protein into the plant, or 3) Contacting the plant with a DBR1 protein inhibitor, Wherein the mutation reduces the activity of the DBR1 protein or reduces the expression of the DBR1 protein. In another preferred example, the plant comprises an arabidopsis, soybean, maize, rice, wheat, horticultural plant. In another preferred embodiment, the plant is a plant capable of producing seed. In another preferred embodiment, the DBR1 gene encodes a lasso RNA debranching enzyme. In another preferred embodiment, the mutation reduces the activity of the DBR1 protein or reduces the expression of the DBR1 protein. In another preferred embodiment, the mutation increases the cumulative amount of lasso RNA. In another preferred embodiment, the mutation increases the cumulative amount of lasso RNA by 5% -50%, preferably by 5% -25%, more preferably by 5% -15%. In another preferred embodiment, the means for introducing mutations comprises gene editing or random mutagenesis. In another preferred embodiment, the mutation comprises a point mutation, an insertion, a deletion, a frame shift mutation or an amino acid substitution. In another preferred embodiment, the mutation comprises a single nucleotide substitution of the wild-type DBR1 gene sequence. In another preferred embodiment, the mutation comprises a single amino acid substitution of the amino acid sequence of the wild-type DBR1 protein. In another preferred embodiment, the mutation comprises a mutation of amino acid 69 of the amino acid sequence of the soybean DBR1 protein from Ser (serine) to Phe (phenylalanine). In another preferred embodiment, the mutation comprises a mutation of amino acid 169 of the amino acid sequence of the soybean DBR1 protein from Asp (aspartic acid) to Asn (asparagine). In another preferred embodiment, the mutation comprises a mutation of amino acid 135 of the amino acid sequence of the DBR1 protein of Arabidopsis from Gly (glycine) to Arg (arginine). In another preferred embodiment, the mutant DBR1 protein is a DBR1 protein having reduced activity compared to the wild-type DBR1 protein. In another preferred embodiment, the mutant DBR1 gene encodes a DBR1 protein having reduced activity or reduced expression compared to the wild-type DBR1 gene. In another preferred embodiment, the gene sequence of the wild soybean DBR1 is shown in SEQ ID NO. 1. In another preferred embodiment, the CDS sequence of the wild-type soybean DBR1 is shown in SEQ ID NO. 2. In another preferred embodiment, the amino acid sequence of wild-type soybean DBR1 is shown in SEQ ID NO. 3. In another preferred embodiment, the gene sequence of the wild type Arabidopsis DBR1 is shown in SEQ ID NO. 8. In another preferred embodiment, the CDS sequence of the wild type Arabidopsis DBR1 is shown in SEQ ID NO. 9. In another preferred embodiment, the amino acid sequence of the wild type Arabidopsis DBR1 is shown in SEQ ID NO. 10. In another preferred embodiment, the CDS sequence of the mutant soybean DBR1 is shown in SEQ ID NO.4 or 6. In another preferred embodiment, the amino acid sequence of the mutant soybean DBR1 is shown in SEQ ID NO.5 or 7. In another preferred embodiment, the CDS sequence of the mutant Arabidopsis DBR1 is shown in SEQ ID NO. 11. In another preferred embodiment, the amino acid sequence of the mutant Arabidopsis DBR1 is shown in SEQ ID NO. 12. In another preferred embodiment, the DBR1 protein inhibitor is an inhibitor that reduces DBR1 protein activity or reduces DBR1 protein expression. In another preferred embodiment, the DBR1 protein inhibitor is selected from the group consisting of a nucleotide, a