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CN-121992006-A - Gene SCE for regulating heat resistance, and application thereof in heat stress response pathway

CN121992006ACN 121992006 ACN121992006 ACN 121992006ACN-121992006-A

Abstract

The invention provides a gene SCE for regulating heat resistance, a heat stress response path and application thereof. The invention discloses a novel gene SCE, which codes polypeptide with important biological functions, also discloses a TT1-SCE-Hsp heat stress response path participated by the SCE, and application of the gene SCE in improving plant heat resistance or yield traits under heat stress. The invention has important significance for genetic improvement of plant stress resistance.

Inventors

  • LIN HONGXUAN
  • YU HONGXIAO
  • CAO YINGJIE
  • SHAN JUNXIANG
  • YE WANGWEI
  • DONG NAIQIAN

Assignees

  • 中国科学院分子植物科学卓越创新中心

Dates

Publication Date
20260508
Application Date
20241108

Claims (13)

  1. 1.A method for improving heat tolerance or yield under heat stress in a plant comprising: down-regulating SCE in plants to increase heat tolerance or yield under heat stress in plants, or Regulating interaction of TT1, SCE and Hsp in a TT1-SCE-Hsp heat stress response path participated by SCE in the plant, enhancing protein folding mediated by Hsp, and further improving heat resistance of the plant or yield under heat stress; Wherein the SCE comprises SCE1 or SCE2.
  2. 2. The method of claim 1, wherein the increasing yield under heat stress in the plant comprises increasing survival rate, increasing grain weight and/or increasing fruiting rate, or The Hsp-mediated protein folding includes correcting misfolding to correct folding or folding of unfolded protein, preferably folding of the protein to its native state; preferably, the Hsp comprises Hsp24.1 or Hsp40.
  3. 3. The method of claim 1, wherein modulating the TT1-SCE-Hsp heat stress response pathway comprises: (a) Upregulation of TT1 to promote interaction of TT1 with SCE, promote TT 1-mediated degradation of SCE, and thereby increase plant heat tolerance or yield under heat stress, or (B) The interaction of SCE and Hsp is inhibited to reduce SUMO modification of Hsp and increase accumulation of Hsp, thereby enhancing Hsp-mediated protein folding and further improving heat resistance of plants or yield under heat stress.
  4. 4. The method of claim 3, wherein downregulating the SCE comprises downregulating expression, activity or stability thereof, preferably comprising knocking out or silencing a gene encoding the SCE in the plant, inhibiting the activity of the SCE, or promoting ubiquitination of the SCE, preferably comprising gene editing with a CRISPR system to knock out the gene encoding the SCE, knocking out the gene encoding the SCE by homologous recombination, subjecting the SCE to a loss-of-function mutation in a plant containing the SCE, silencing the SCE with an interfering molecule that specifically interferes with expression of the gene encoding the SCE, upregulating TT1 to promote ubiquitination of the SCE mediated thereby, or Upregulation of TT1 includes upregulation of expression, activity or stability thereof, including transferring a coding gene of TT1 or an expression construct or vector containing the coding gene into a plant, performing a functional gain mutation on TT1, promoting TT1 expression with an expression-enhancing promoter or a tissue-specific promoter, or promoting TT1 expression with an enhancer; preferably, SCE1 or SCE2 down regulates one of them when SCE is down regulated.
  5. 5. The method of claim 4, wherein SCE is down-regulated with a reagent that directs the editing of the CRISPR gene or is capable of forming the sgRNA, preferably the reagent that forms the sgRNA comprises primers of the sequences shown in SEQ ID NO. 17 and SEQ ID NO. 18, primers of the sequences shown in SEQ ID NO. 19 and SEQ ID NO. 20, and primers of the sequences shown in SEQ ID NO. 39 and SEQ ID NO. 40.
  6. 6. Use of SCE, TT1-SCE-Hsp heat stress responsive pathways comprising same or regulatory molecules thereof for increasing plant heat tolerance or yield under heat stress, preferably increasing plant yield under heat stress comprising increasing survival, increasing grain weight and/or increasing fruiting rate, The regulatory molecule is SCE down-regulating molecule which can improve heat resistance or yield under heat stress of plant, or The regulatory molecule is TT1 up-regulating molecule which promotes interaction of TT1 and SCE, thereby promoting TT 1-mediated SCE degradation and further improving heat resistance or yield under heat stress of plants, or The regulatory molecule is an SCE and Hsp interaction inhibition molecule, which reduces SUMO of Hsp and increases accumulation of Hsp, thereby enhancing Hsp-mediated protein folding and further improving heat resistance of plants or yield under heat stress; Wherein the SCE comprises SCE1 or SCE2.
  7. 7. The use of claim 6, wherein the SCE downregulating molecule comprises an agent that knocks out or silences SCE, an agent that inhibits SCE activity, an agent that reduces SCE stability, preferably a CRISPR gene editing agent, homologous recombination agent or site-directed mutation agent for SCE, an agent that mutates SCE with loss of function, an interfering molecule that specifically interferes with expression of a gene encoding SCE, an agent that upregulates TT1 to promote ubiquitination degradation of SCE mediated thereby, more preferably an agent that downregulates SCE comprising an sgRNA that is based on CRISPR gene editing or an agent that forms the sgRNA, wherein the agent that forms the sgRNA comprises a primer of the sequences set forth in SEQ ID NO:17 and SEQ ID NO:18, a primer of the sequences set forth in SEQ ID NO:19 and SEQ ID NO:20, a primer of the sequences set forth in SEQ ID NO:39 and SEQ ID NO:40, or The TT1 up-regulating molecule comprises transferring coding gene of TT1 or expression construct or vector containing the coding gene into plants, carrying out functional mutation on TT1, promoting TT1 expression by using an expression enhanced promoter or a tissue specific promoter, or promoting TT1 expression by using an enhancer.
  8. 8. The method of any one of claims 1 to 5 or the use of any one of claims 6 to 7, wherein the plant is or the SCE is from a plant comprising the group consisting of gramineae, leguminous, cruciferae, solanaceae, preferably the plant is or the SCE is from a plant comprising the group consisting of rice (Oryza sativa), maize (Zea mays), millet (SETARIA ITALICA), barley (Hordeum vulgare), wheat (Triticum aestivum), millet (Panicummiliaceum), sorghum (Sorghum bicolor), rye (SECALE CEREALE), oat (AVENA SATIVA L), brachypodium (Brachypodium distachyum), soybean (Glycine max), potato (Solanum tuberosum), tomato (Solanum lycopersicum), capsicum (Capsicum annuum), canola (Brassica napus), arabidopsis thaliana (Arabidopsis thaliana).
  9. 9. The method according to any one of claims 1 to 5 or the use according to any one of claims 6 to 7, wherein the amino acid sequence of the SCE polypeptide is selected from the group consisting of (i) a polypeptide of the amino acid sequence shown as SEQ ID No. 4 or 8, (ii) a polypeptide derived from (i) having the regulatory trait function by substitution, deletion or addition of one or more amino acid residues of the amino acid sequence shown as SEQ ID No. 4 or 8, (iii) a polypeptide having the amino acid sequence homology of 80% or more with the amino acid sequence shown as SEQ ID No. 4 or 8, and (iv) an active fragment of the polypeptide of the amino acid sequence shown as SEQ ID No. 4 or 8, or (v) a polypeptide formed by adding a tag sequence or an enzyme cleavage site sequence to the N-terminus of the polypeptide of the amino acid sequence shown as SEQ ID No. 4 or 8, or by adding a signal peptide sequence to the N-terminus thereof.
  10. 10. Use of SCE or a TT1-SCE-Hsp heat stress responsive pathway comprising the same in plants as a molecular marker for identifying heat tolerance or yield under heat stress in plants or as a molecular marker for targeted screening of plants, preferably yield traits under heat stress including survival, grain weight and/or fruiting rate.
  11. 11. A method of selecting or identifying a plant for heat tolerance or yield under heat stress, comprising identifying expression or sequence characteristics of SCE in a test plant, or identifying a TT1-SCE-Hsp heat stress response pathway in a plant, wherein the test plant is a plant with high heat tolerance or yield under heat stress if SCE is low or not expressed, and wherein the test plant is a plant with low heat sensitivity or yield under heat stress if SCE is high expressed.
  12. 12. A method for screening a substance for improving heat resistance or yield under heat stress of a plant, comprising (1) adding a candidate substance to a system expressing SCE, and (2) detecting the system, observing the expression, activity or stability of SCE therein, and if the expression, activity or stability is improved, indicating that the candidate substance is a substance useful for improving heat resistance or yield under heat stress of a plant.
  13. 13. A method for screening a substance for regulating heat resistance or yield under heat stress in a plant, comprising (1) adding a candidate substance to a system expressing a TT1-SCE-Hsp heat stress response pathway, (2) detecting the system, observing the expression, activity or interaction of TT1-SCE-Hsp heat stress response pathway protein therein, and indicating that the candidate substance is a substance useful for improving heat resistance or yield under heat stress in a plant if it promotes the interaction of TT1 with SCE or inhibits the interaction of SCE with Hsp; preferably, the promoting TT1 interaction with SCE comprises promoting TT 1-mediated degradation of SCE; preferably, inhibiting the interaction of SCE with Hsp comprises reducing SCE-mediated SUMO modification of Hsp, increasing accumulation of Hsp.

Description

Gene SCE for regulating heat resistance, and application thereof in heat stress response pathway Technical Field The invention belongs to the fields of botanic and molecular biology, and particularly relates to a gene SCE for regulating heat resistance, a heat stress response path and application thereof. Background Global warming poses a significant threat to food safety and human development, and the global average air temperature has risen by 1.25 ℃. This increase is expected to exceed 1.5 ℃ in the next decade. Rice (Oryza sativa) is a staple food crop of global importance, and the yield of rice decreases by about 3.2% whenever the global air temperature rises by 1 ℃. High temperature stress refers to the phenomenon that plants are subjected to a high temperature environment exceeding the proper temperature range for normal growth in the growth process, so that the growth and development of the plants are inhibited or damaged. In recent years, as the emission of greenhouse gases increases, the global air temperature continues to rise, and the high temperature environment has an important influence on the life cycle of plants and agricultural production. Under high temperature stress, the physiological and biochemical processes of plants are affected to different extents. Plants sense and transmit high temperature signals under high temperature stress through a variety of cellular signal transduction pathways. Plants under high temperature stress trigger a range of gene expression changes that are regulated by a variety of signal transduction pathways. Research shows that the heat shock protein is a stress protein induced by plants under high temperature stress and has the function of protecting cell structures and functions. In addition, plants can regulate gene expression under high temperature stress by regulating transcription factors, miRNAs and the like so as to cope with high temperature environments. With the increasing global population, improving the heat resistance and adaptability of crops has become a major issue. Therefore, high-temperature resistant rice germplasm resources are needed to be deeply excavated, the regulation and control mechanism of heat resistance of rice is systematically researched, novel and innovative crop germplasm is cultivated, and high and stable crop yield is realized. The molecular mechanism of plant response to heat stress and tolerance is deeply analyzed, and important theoretical basis and gene resources can be provided for cultivating new varieties of crops with heat resistance. Although numerous scientific studies are currently focused on the investigation of plants against heat stress regulatory networks, including upstream sensors, core regulatory factors, and downstream response factors, the art has heretofore lacked effective regulatory targets. Disclosure of Invention The invention aims to provide a gene SCE for regulating heat resistance, a heat stress response path and application thereof. In a first aspect of the invention there is provided a method of increasing yield under heat tolerance or stress in a plant comprising down-regulating SCE (SUMO E2 binding enzyme) in a plant, thereby increasing yield under heat tolerance or stress in a plant, or modulating TT1-SCE-Hsp (small heat shock protein) interactions involved in SCE in a plant heat stress response pathway, increasing accumulation of Hsp (preventing degradation thereof), enhancing Hsp-mediated protein folding, thereby increasing yield under heat tolerance or stress in a plant, wherein the SCE comprises SCE1 or SCE2. In one or more embodiments, the increasing yield under heat stress in a plant includes increasing survival, increasing grain weight, and/or increasing fruiting rate. In one or more embodiments, the Hsp-mediated protein folding includes correcting misfolding to correct folding or folding of unfolded protein, preferably such that the protein folds into its native state. In one or more embodiments, the Hsp comprises Hsp24.1 or Hsp40. In one or more embodiments, modulating the TT1-SCE-Hsp heat stress response pathway includes (a) upregulating TT1 to promote TT1 interaction with SCE (TT 1-SCE module), promote TT 1-mediated degradation of SCE (via ubiquitin degradation pathway), and thereby increase plant heat tolerance or yield under heat stress, or (b) inhibiting SCE interaction with Hsp to reduce SUMO modification of Hsp, increase accumulation of Hsp (prevent degradation thereof), thereby enhancing Hsp-mediated protein folding, and thereby increase plant heat tolerance or yield under heat stress. In one or more embodiments, downregulating SCE comprises downregulating its expression, activity or stability, preferably comprising knocking out or silencing a gene encoding SCE in a plant, inhibiting the activity of SCE, or promoting ubiquitination degradation of SCE, preferably comprising gene editing with a CRISPR system to knock out a gene encoding SCE, knocking out a gene encoding SCE by