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CN-121472315-B - Application of calcium sensing protein ShCBL gene

CN121472315BCN 121472315 BCN121472315 BCN 121472315BCN-121472315-B

Abstract

The invention provides an application of a calcium sensing protein ShCBL gene, belonging to the field of biotechnology, wherein the application is used for increasing the content of proline and superoxide dismutase in sugarcane or rice and reducing the content of malondialdehyde by over-expressing or over-expressing the ShCBL gene in the sugarcane or rice so as to improve the cold resistance. Researches show that when ShCBL a is super-expressed or over-expressed in plants, cold stress related physiological indexes such as proline, superoxide dismutase (SOD), malondialdehyde (MDA) and the like can be obviously influenced, so that the cold resistance of rice and sugarcane is enhanced. ShCBL1 is used as a positive regulator for cold resistance regulation and control, and can be used for molecular breeding of cold-resistant varieties.

Inventors

  • WANG JUNGANG
  • YU GUANGRUN
  • FENG CUILIAN
  • LIN JISHAN
  • ZHAO TINGTING
  • WU RUIJIE
  • ZHANG XINGTAN
  • LI XIAOFENG

Assignees

  • 中国热带农业科学院三亚研究院
  • 中国热带农业科学院热带生物技术研究所

Dates

Publication Date
20260508
Application Date
20260108

Claims (10)

  1. 1. An application of ShCBL gene in improving cold tolerance of rice, which is characterized in that the application improves the cold tolerance by over-expressing ShCBL gene in the rice; the sequence of ShCBL gene is shown as SEQ ID NO. 1.
  2. 2. The use of ShCBL gene according to claim 1 to improve cold tolerance in rice, wherein the use is to transform rice with a recombinant vector comprising ShCBL1 gene or a strain comprising ShCBL1 gene to over-express ShCBL gene in rice.
  3. 3. The use of ShCBL gene according to claim 2 for improving cold tolerance in rice, wherein said recombinant vector comprising ShCBL gene is obtained by ligating said ShCBL gene to an over-expression vector.
  4. 4. The use of ShCBL gene according to claim 2 for improving cold tolerance in rice, wherein the strain comprising ShCBL gene is agrobacterium comprising ShCBL gene or escherichia coli comprising ShCBL1 gene.
  5. 5. A method for improving cold tolerance of rice, which is characterized in that ShCBL gene is over-expressed in the rice, and the cold tolerance of plants is improved by enhancing the content and/or activity of ShCBL1 protein in the plants; the sequence of ShCBL gene is shown as SEQ ID NO. 1.
  6. 6. The method for improving cold tolerance of rice according to claim 5, wherein the method comprises transforming rice with a recombinant vector comprising ShCBL gene or a strain comprising ShCBL gene to overexpress ShCBL gene in rice.
  7. 7. The method for improving cold tolerance of rice according to claim 6, wherein the recombinant vector comprising ShCBL gene is obtained by ligating ShCBL1 gene to an over-expression vector.
  8. 8. The method for improving cold tolerance of rice according to claim 6, wherein the strain containing ShCBL1 gene is agrobacterium containing ShCBL1 gene or escherichia coli containing ShCBL1 gene.
  9. 9. A rice breeding method is characterized in that the rice with enhanced cold resistance is obtained by over-expressing ShCBL gene in rice; the sequence of ShCBL gene is shown as SEQ ID NO. 1.
  10. 10. The rice breeding method according to claim 9, wherein the rice is transformed with a recombinant vector comprising ShCBL gene or a strain comprising ShCBL1 gene to overexpress ShCBL gene in the rice.

Description

Application of calcium sensing protein ShCBL gene Technical Field The invention belongs to the technical field of biology, in particular to the fields of plant stress biology and molecular breeding, and particularly relates to application of a calcium sensing protein ShCBL gene. Background Low temperature stress is an important environmental factor limiting the growth of various crops, and low temperature can cause damage to cell membranes, aggravate accumulation of active oxygen, inhibit photosynthetic efficiency and finally affect crop yield. Existing studies show that Ca 2+ has a central role in plant stress signal transduction, wherein CBL (Calcineurin B-like) protein is a key Ca 2+ sensing factor and can regulate a stress response network in cooperation with CIPKs. Calcineurin B-type Proteins (Calcineurin B-Like Proteins, CBLs) serve as plant-specific calcium ion sensors, occupy a central position in the stress signaling pathway, accurately decode calcium signals through specific interactions with downstream target Proteins and initiate corresponding stress-tolerant response mechanisms. Under drought stress, arabidopsis AtCBL/9 is combined with CIPK, and potassium ion channels are regulated to maintain cell homeostasis, and an ABA signal channel is activated to promote stomatal closure so as to regulate drought tolerance. Wheat TaCBL enhances the osmotic regulation by increasing the proline content. In salt and alkali stress, atCBL (SOS 3) and CIPK24 form an SOS complex, so that sodium ion transport proteins are driven to excrete or compartmentalize Na+ into vacuoles, and rice OsCBL10 and soybean GmCBL1 also enhance salt tolerance through the mechanism. The ShCBL gene is a member of the CBL family identified in XTT22 of sugarcane material, and similar to its homologous gene in rice, there is potential to be involved in regulation of salt stress and osmotic stress. However, the function of the ShCBL gene in regulating plant cold tolerance has not been reported before. Disclosure of Invention The invention provides application of a calcium sensing protein ShCBL gene aiming at the problems. In order to achieve the above purpose, the technical scheme adopted by the invention is as follows: Use of ShCBL gene to improve cold tolerance of plants, wherein said use increases proline and superoxide dismutase content in sugarcane or rice and reduces malondialdehyde content by over-expressing or over-expressing ShCBL gene in sugarcane or rice, thereby improving cold tolerance; The sequence of ShCBL gene is shown as SEQ ID NO. 1; Detecting the content of stress-resistant physiological indexes such as proline, superoxide dismutase, malondialdehyde and the like of a sugarcane ShCBL1 over-expression plant (ShCBL-OE) and a negative control plant (WT-XTT 22), and confirming the cold resistance of the sugarcane ShCBL-OE; The expression of the cold response marker gene of rice ShCBL1 over-expression plants (ShCBL-OE) and negative control plants (WT-ZH 11) is detected to confirm the cold resistance of rice ShCBL-OE. Further, the application is to transform sugarcane or rice by utilizing a recombinant vector containing ShCBL gene or a strain containing ShCBL1 gene, so that ShCBL gene is over-expressed or over-expressed in the sugarcane or rice; Specifically, the application is that the cDNA of sugarcane leaf is used as a template for PCR amplification, a ShCBL gene full-length sequence is obtained by cloning, and the agrobacterium transformation method is used for over-expressing or over-expressing ShCBL gene in rice or sugarcane materials to obtain an over-expressed plant; The method comprises the steps of carrying out 4 ℃ cold treatment on rice ShCBL1 over-expression plants (ShCBL-OE) and negative control plants (WT-ZH 11), carrying out-5 ℃ cold treatment on sugarcane ShCBL1 over-expression plants (ShCBL-OE) and negative control plants (WT-XTT 22), determining ShCBL1 to positively regulate the cold resistance of rice and sugarcane in seedling stage by comparing the phenotype difference of cold resistance between ShCBL over-expression plants (ShCBL 1-OE) and negative control plants before and after cold treatment, extracting total RNA of leaves, carrying out reverse transcription to obtain cDNA, and quantitatively analyzing ShCBL1 gene expression quantity in ShCBL-OE and negative control plants by using real-time fluorescence to confirm the reliability of phenotype data. Further, the recombinant vector containing ShCBL1 gene is obtained by connecting ShCBL1 gene to over-expression vector; the recombinant vector containing ShCBL gene is ShCBL gene over-expression vector. Further, the strain containing ShCBL1 gene is agrobacterium containing ShCBL1 gene or escherichia coli containing ShCBL gene; the strain containing ShCBL gene is ShCBL gene over-expression strain. A method for improving cold tolerance of plants, which comprises the steps of over-expressing or super-expressing ShCBL gene in sugarcane or rice, increasing the