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CN-121975820-A - Alfalfa MsCML gene for regulating and controlling salt tolerance and application thereof

CN121975820ACN 121975820 ACN121975820 ACN 121975820ACN-121975820-A

Abstract

The invention discloses a alfalfa MsCML gene for regulating and controlling salt tolerance and application thereof. The nucleotide sequence of MsCML69,69 gene is shown as SEQ ID NO.1, and the corresponding amino acid sequence is shown as SEQ ID NO. 2. The invention clones cDNA sequence of calmodulin-like gene MsCML-69 from alfalfa, connects the obtained MsCML-69 gene with plant expression vector, constructs recombinant expression vector, transforms plant leaf with the constructed recombinant expression vector, and finally cultures into transgenic plant. The result shows that the over-expression of the gene can obviously improve the salt tolerance of the plant. The MsCML gene discovery provides gene resources for the cultivation of new alfalfa stress-resistant germplasm, and improves the application range of the gene, and the application of the gene has important production practice significance.

Inventors

  • YU SHUHAN
  • PAN SHUYIN
  • ZHANG LU
  • MAO YINGYING
  • TAN WENYUAN

Assignees

  • 浙江农林大学

Dates

Publication Date
20260505
Application Date
20260407

Claims (10)

  1. 1. A alfalfa MsCML gene for regulating and controlling salt tolerance is characterized in that the nucleotide sequence is shown as SEQ ID NO. 1.
  2. 2. The protein coded by the alfalfa MsCML gene for regulating salt tolerance according to claim 1, wherein the amino acid sequence of the protein is shown as SEQ ID NO. 2.
  3. 3. A biological material comprising the MsCML gene of claim 1, wherein said biological material is an expression cassette, a recombinant expression vector, or a recombinant microorganism.
  4. 4. The biomaterial according to claim 3, wherein the recombinant expression vector is obtained by inserting MsCML gene according to claim 1 between Nco I and BstE II cleavage sites of pCAMBIA3301 vector.
  5. 5. Use of MsCML gene according to claim 1, protein according to claim 2, or biological material according to claim 3 for increasing salt tolerance in plants or for growing transgenic plants with increased salt tolerance.
  6. 6. The use according to claim 5, wherein overexpression of the alfalfa MsCML gene of claim 1 in a plant of interest, said plant of interest being alfalfa, increases the salt tolerance of the plant.
  7. 7. The use according to claim 6, wherein the target plant is alfalfa.
  8. 8. A method for improving the salt tolerance of alfalfa, which is characterized in that the alfalfa MsCML gene according to claim 1 is overexpressed in alfalfa to improve the salt tolerance of alfalfa.
  9. 9. The method according to claim 8, wherein the process of overexpressing the alfalfa MsCML69,69 gene in alfalfa comprises constructing an overexpression vector of the alfalfa MsCML69,69 gene according to claim 1, and transferring the overexpression vector into alfalfa by an agrobacterium-mediated method to obtain transgenic alfalfa with improved salt tolerance.
  10. 10. The method of claim 9, wherein the alfalfa is alfalfa.

Description

Alfalfa MsCML gene for regulating and controlling salt tolerance and application thereof Technical Field The invention belongs to the field of plant genetic engineering, and particularly relates to a alfalfa MsCML gene for regulating and controlling salt tolerance and application thereof. Background Abiotic stress of drought, salt and alkali, cold injury, heat injury and the like can obviously inhibit the growth and development process of plants, greatly reduce the yield of crops, and influence the normal growth state and the product quality of forest, fruit trees, flowers and gardening ornamental plants. Therefore, cultivating a plant variety with excellent stress tolerance is one of the core targets of sustainable development of the planting industry, and identifying key stress tolerance genes and improving the stress tolerance capability of plants based on the key stress tolerance genes has important practical significance and necessity for achieving the target. The plant evolved from calmodulin (CaMs), calmodulin-like (CMLs), ca 2+ signal decoding system composed of Calpain (CDPKs) and calmodulin b protein and calmodulin phosphatase interacting protein kinase complex (CBLs-CIPKs) and the like, the receptors regulate and control downstream target (Day I, Reddy V, Ali G, Reddy A. (2002). Analysis of EF-hand-containing proteins in Arabidopsis. Genome Biology 3, research0056.1).CMLs through Ca 2+ dependent conformational change to sense Ca 2+ rapid and transient change and combine with the Ca 2+, after combining, CMLs undergoes conformational change and further combines with and activates downstream target protein, thereby regulating and controlling downstream physiological and biochemical reaction, and a great deal of literature report about realizing signal transmission (Tian W, Wang C, Gao Q, Li L, Luan S. (2020). Calcium spikes, waves and oscillations in plant development and biotic interactions[J]. Nature Plants 6, 750-759). shows that CMLs is regulated and controlled in plant growth and development, Stress response and hormone signaling play a key role in a plurality of physiological processes. For example, overexpression PvCML9 enhances the salt tolerance (Yang M, Zhou B, Song Z, Tan Z, Liu R, Luo Y, Guo Z, Lu S. (2024). A calmodulin-like protein PvCML9 negatively regulates salt tolerance. Plant Physiology and Biochemistry 210, 108642).AtCML10 of seashore paspalum (Paspalum vaginatum O. Swartz) by mediating the production of Arabidopsis ascorbic acid through interaction with phosphomannose mutase (AtPMM), thereby modulating Arabidopsis stress response (Cho K, Nguyen H, Kim S, Shin J, Cho D, Hong S, Shin J, Ok S. (2016). CML10, a variant of calmodulin, modulates ascorbic acid synthesis. New Phytologist 209, 664-78). tea tree (CAMELLIA SINENSIS) CsCML16, CsCML18-2, csCML38 and CsCML42 for low temperatures, Both salt and drought respond (Ma Q, Zhou Q, Chen C, Cui Q, Zhao Y, Wang K, Arkorful E, Chen X, Sun K, Li X. (2019). Isolation and expression analysis of CsCML genes in response to abiotic stresses in the tea plant (Camellia sinensis). Scientific Reports 9, 8211).MtCML42 to mediate the accumulation of CBF transcription factors and raffinose to positively regulate the protein interaction between cold resistance (Sun Q, Huang R, Zhu H, Sun Y, Guo Z. (2021). A novel Medicago truncatula calmodulin-like protein (MtCML42) regulates cold tolerance and flowering time. Plant Journal 108, 1069-82).MsCML10 and MsGSTU and MsFBA6 of medicago sativa (Medicago truncatula) and activate them, thereby maintaining the steady state of ROS at low temperature, promoting the accumulation of sugar and finally improving the cold resistance (Yu S, Wu J, Sun Y, Zhu H, Sun Q, Zhao P, Huang R, Guo Z. (2022). A calmodulin-like protein (CML10) interacts with cytosolic enzymes GSTU8 and FBA6 to regulate cold tolerance. Plant Physiology 190, 1321-33). of medicago sativa that the CML39 gene of tomato is obviously induced at high temperature on the transcription level (Ding et al, 2020). further studies have shown that overexpression of SlCML39 reduces the tolerance of arabidopsis to high temperature stress during germination and seedling growth phases (Ding H, Qian Y, Fang Y, Ji Y, Sheng J, Ge C. (2021). Characteristics of SlCML39, a tomato calmodulin-like gene, and its negative role in high temperature tolerance of Arabidopsis thaliana during germination and seedling growth. International Journal of Molecular Sciences 22, 11479). Alfalfa (Medicago sativa L.) is one of the most important leguminous grasses in the world, and has the characteristics of high biomass, good nutrition quality, wide cultivation range and the like. Soil salinization is an important factor affecting the growth of alfalfa and limiting the yield of the alfalfa, so that the related genes of salt tolerance of the alfalfa are excavated and regulated, and the target genes can be provided for the breeding of salt tolerance molecules of the alfalf