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CN-116024225-B - Novel gene for increasing gramineous plant seed yield and application thereof

CN116024225BCN 116024225 BCN116024225 BCN 116024225BCN-116024225-B

Abstract

The invention provides a novel gene for increasing the grain yield of gramineous plants and application thereof. Disclosed are a novel GRAS11 and its downstream molecule EXPB, which are positively regulated genes for the yield or endosperm quality of gramineous plant seeds, which are related to the regulation of plant yield traits or endosperm quality, and whose loss of function results in reduced yield and quality, but which have no effect on other plant traits. The invention provides a new way for improving the plant characteristics.

Inventors

  • WU YONGRUI
  • WANG JIECHEN
  • JI CHEN

Assignees

  • 中国科学院分子植物科学卓越创新中心

Dates

Publication Date
20260512
Application Date
20211027

Claims (9)

  1. 1. A method for improving yield and endosperm quality of a gramineous plant, comprising up-regulating GRAS11 in a plant, wherein the improving yield of a gramineous plant comprises increasing grain weight, increasing grain width and increasing grain thickness; The upregulation includes driving expression of exogenous GRAS11 in the plant with an endosperm-specific promoter that is a 27-kD gamma-prolamin promoter; The amino acid sequence of the GRAS11 polypeptide is shown as SEQ ID NO. 2, and the gramineous plant is corn.
  2. 2. The method of claim 1, wherein the method comprises (1) transforming a plant organ or tissue with a nucleic acid encoding a GRAS11 exogenous nucleic acid to obtain a plant tissue or organ transformed with a nucleic acid encoding the polypeptide, and (2) regenerating a plant from the plant tissue or organ transformed with a nucleic acid encoding a polypeptide of the invention obtained in step (1).
  3. 3. The method of claim 2, comprising (s 1) providing an agrobacterium carrying an expression vector comprising a nucleic acid encoding GRAS11, (s 2) contacting a plant tissue or organ with the agrobacterium of step (s 1) such that the nucleic acid encoding the polypeptide is transferred into and integrated into the chromosome of the plant cell, (s 3) selecting a plant tissue or organ into which the nucleic acid encoding GRAS11 is introduced, and (s 4) regenerating the plant from the plant tissue or organ of step (s 3).
  4. 4. The method of claim 1, wherein up-regulating GRAS11 expression or activity in the plant further comprises subjecting the obtained plant to selfing, crossing, or introspection to obtain a progeny plant.
  5. 5. The method of claim 1, wherein said GRAS11 modulates downstream molecule EXPB, wherein the amino acid sequence of said EXPB polypeptide is as shown in SEQ ID NO. 4, and wherein said EXPB increases endosperm thickness and average cell width of endosperm cells of Gramineae and expands endosperm cell volume.
  6. 6. A method for increasing endosperm thickness and average cell width of endosperm cells and expanding endosperm cell volume of gramineous plant comprises up-regulating expression or activity of GRAS11 downstream molecule EXPB in plant, wherein the amino acid sequence of polypeptide of GRAS11 is shown as SEQ ID NO. 2, the amino acid sequence of polypeptide of EXPB is shown as SEQ ID NO. 4, and EXPB expands endosperm cell volume, and the gramineous plant is maize.
  7. 7. The GRAS11 is used for (a) improving the yield of gramineous plants and (b) improving the endosperm quality of gramineous plants, wherein (a) comprises the steps of improving the weight of seeds, improving the width of the seeds and improving the thickness of the seeds, and (b) comprises the steps of increasing the thickness and the width of endosperm and promoting the expansion and the enlargement of endosperm cells, the amino acid sequence of the polypeptide of the GRAS11 is shown as SEQ ID NO. 2, and the gramineous plants are corn.
  8. 8. The GRAS11 and the application of a downstream molecule EXPB15 thereof are used as molecular markers for identifying properties of gramineous plants, wherein the properties comprise yield properties and endosperm properties, the yield properties comprise seed weight, seed width and seed thickness, the endosperm properties comprise endosperm thickness and width and endosperm cell expansion and enlargement, the amino acid sequence of polypeptide of the GRAS11 is shown as SEQ ID NO. 2, the amino acid sequence of polypeptide of the EXPB is shown as SEQ ID NO. 4, and the gramineous plants are corn.
  9. 9. A method for directionally selecting or identifying gramineae plants with high yield and high endosperm quality comprises identifying GRAS11 and its downstream molecules EXPB in a test plant, wherein if the expression or activity of GRAS11 and its downstream molecules EXPB in the test plant is higher than the average value of the expression or activity of GRAS11 and its downstream molecules EXPB in the plant, the gramineae plants with high yield and high endosperm quality are obtained, the yield is high, the yield comprises increased grain weight, increased grain width and increased grain thickness, the endosperm quality is high, the endosperm quality comprises increased endosperm thickness and width and promoted endosperm cell expansion and increase, the amino acid sequence of a polypeptide of GRAS11 is shown as SEQ ID NO.2, the amino acid sequence of a polypeptide of EXPB15 is shown as SEQ ID NO.4, and the gramineae plants are corn.

Description

Novel gene for increasing gramineous plant seed yield and application thereof Technical Field The invention belongs to the fields of biotechnology and botanics, and particularly relates to a novel gene for increasing the grain yield of gramineous plants and application thereof. Background In gramineae plants, corn has stronger drought resistance, cold resistance, barren resistance and excellent environmental adaptability compared with rice and wheat, so that the planting area of corn is high in crops. However, the yield per unit area of the corn is obviously lower than that of rice and the like, so that the research on corn germplasm resources is promoted, and the development of a new variety of high-yield corn has great potential. The theoretical yield of the corn is determined by the number of ears and the grain weight, and the grain weight is determined by four indexes of the length, width and thickness of the grains and the fullness of the grains. Corn ears are closely linked among three seed-examining indexes of tight arrangement, mutual extrusion, grain length, grain width and grain thickness and are greatly influenced by the number of ears, so that the corn yield related QTL positioning is slow relative to the progress of rice. In 2019, the agricultural crop in China assembled a three-generation genome by using a tropical small maize variety (SK) as a material and applying Pacbio sequencing technology, and located a quantitative genetic locus qHKW1 for simultaneously controlling grain type and grain weight through recombinant inbred line, cloned ZmBAM d gene, which codes CLV1/BAM receptor kinase protein, positively regulates grain type and grain weight (Yang et al, 2019). In 2020, the quantitative trait locus qKW, which encodes a dynw motif pentapeptide repeat protein, was cloned by the chinese farmer, and involved in C-U editing of chloroplast NADH dehydrogenase-like complex subunit ndhB, affecting maternal photosynthesis, and thus affecting maize filling (Huang et al 2020). Corn grain weight-related gene resources obtained through QTL positioning are limited, and the adoption of a transgenic means for improving the corn grain yield is another way. At present, two genes are mainly utilized for improving corn yield by a transgenic method, namely 1, improving efficiency of transferring carbohydrates into grains in corn, such as over-expression ZmGIF/Mn 1 (Li et al, 2013), and 2, improving corn endosperm starch synthesis capability, such as over-expression of AGPase size subunits or E.coli AGPase (Boehlein et al, 2014;Tuncel and Okita,2013). Endosperm (Endosperm) generally refers to the nutritive tissue formed by the fusion of sperm and polar nuclei of a angiosperm during double fertilization, also known as endosperm. Such tissues are not gametophytes nor sporophytes, and their chromosomal ploidy is generally triploid, an important component of the seeds of many plants (e.g., gramineae). At present, the gene resources utilized by transgenes are mainly concentrated on carbohydrate transport and storage, but from the standpoint of endosperm development, endosperm cell volume increase is a passive process. The synergic regulation and control of endosperm cell expansion and stored substance transport synthesis are beneficial to obtaining higher-yield germplasm resources. Therefore, from this point of view, it is urgent in the art to excavate a target gene that can achieve efficient regulation. Disclosure of Invention The invention aims to provide a novel gene for increasing the grain yield of gramineous plants and application thereof. In a first aspect of the invention, there is provided a method of increasing yield or endosperm quality in a gramineous plant comprising up-regulating the expression or activity of GRAS11 or a downstream molecule EXPB thereof in the plant. In one or more embodiments, the GRAS11 includes homologues thereof, preferably homologues thereof in gramineae plants. In one or more embodiments, EXPB includes homologs thereof, preferably homologs thereof in gramineae. In one or more embodiments, the GRAS11 comprises a protein or gene. In one or more embodiments, EXPB includes a protein or gene. In one or more embodiments, the GRAS11 indirectly acts to regulate the downstream molecule EXPB via transcription factors that interact with it. In one or more embodiments, the increasing yield of the gramineous plant includes increasing grain weight (grain weight), increasing grain width (grain width), and increasing grain thickness (grain thickness). In one or more embodiments, the improving the endosperm quality of the grass plant includes increasing the endosperm thickness and/or width, promoting endosperm cell expansion and/or increase (including width increase). In one or more embodiments, the "promoting endosperm cell expansion and/or augmentation" includes an advance in cell expansion and/or augmentation over time. In one or more embodiments, the "promoting endosperm cell expansion and/or