CN-121992031-A - Application of ramie BnWRKY gene in increasing cadmium accumulation in plant roots and enhancing cadmium resistance of plants

Abstract

The invention discloses an application of a ramie BnWRKY gene in increasing cadmium accumulation in plant roots and enhancing cadmium resistance of plants, belonging to the technical field of plant genetic engineering. The invention provides application of a ramie BnWRKY gene in increasing cadmium accumulation in plant roots and enhancing cadmium tolerance of plants, wherein a CDS nucleotide sequence of the ramie BnWRKY gene is shown as SEQ ID NO. 1. The invention makes clear the specific functions of the ramie BnWRKY gene in cadmium accumulation and cadmium resistance regulation for the first time, and has the advantages of enhancing the cadmium resistance of plants after over-expression in the plants, obviously increasing the cadmium accumulation of plant root systems and reducing the cadmium accumulation of overground parts.

Inventors

• CHEN PING
• WU DUQING
• YANG ZHEN
• YAN SHIWEI
• GU YIZHOU
• ZHU AIGUO
• CHEN KUNMEI
• GAO GANG
• CHEN JIA
• WANG XIAOFEI
• CHEN JIKANG
• ZHAO HAOHAN

Assignees

• 中国农业科学院麻类研究所

Dates

Publication Date

20260508

Application Date

20260331

Claims (10)

1. 1. The application of the ramie BnWRKY gene in increasing cadmium accumulation in plant roots and enhancing cadmium resistance of plants is characterized in that the CDS nucleotide sequence of the ramie BnWRKY gene is shown as SEQ ID NO. 1.
2. 2. The use according to claim 1, wherein the amino acid sequence of the protein encoded by the ramie BnWRKY gene is shown in SEQ ID NO. 2.
3. 3. The use according to claim 1, wherein increased cadmium accumulation and increased cadmium tolerance in plant roots is achieved by up-regulating the expression level of BnWRKY gene in plants or increasing the content of protein encoded by BnWRKY gene in plants.
4. 4. Use according to any one of claims 1 to 3, wherein the plants comprise arabidopsis thaliana and ramie.
5. 5. A primer pair for amplifying the ramie BnWRKY gene according to claim 1, wherein the base sequences of the primer pair are respectively shown as SEQ ID NO.3 and SEQ ID NO. 4.
6. 6. A method for increasing cadmium accumulation in plant roots and enhancing cadmium resistance of plants, which is characterized in that the ramie BnWRKY gene according to claim 1 is introduced into a receptor plant to obtain a transgenic plant which over-expresses BnWRKY gene, and the cadmium accumulation amount in the plant roots is increased and the cadmium resistance is enhanced.
7. 7. The method of claim 6, wherein an over-expression vector comprising the BnWRKY gene is constructed, the over-expression vector is transformed into agrobacterium, and the BnWRKY gene is introduced into a recipient plant by agrobacterium-mediated inflorescence dip-dyeing.
8. 8. The method according to claim 6, wherein the over-expression vector is obtained by ligating BnWRKY gene to pEGOEP S-G418 between XbaI and BamHI cleavage sites by homologous recombination based on pEGOEP S-G418, and the primer pair base sequences for amplifying BnWRKY gene fragment for homologous recombination are shown in SEQ ID NO.5 and SEQ ID NO. 6.
9. 9. The method according to any one of claims 6 to 8, further comprising screening the obtained transgenic plants to identify the primers with the base sequences shown in SEQ ID NO.7 and SEQ ID NO. 8.
10. 10. The use of the gene BnWRKY of ramie according to claim 1 for cultivating plant species for the remediation of cadmium-contaminated soil.

Description

Application of ramie BnWRKY gene in increasing cadmium accumulation in plant roots and enhancing cadmium resistance of plants Technical Field The invention relates to the technical field of plant genetic engineering, in particular to application of a ramie BnWRKY gene in increasing cadmium accumulation in plant roots and enhancing cadmium resistance of plants. Background Cadmium (Cd) is an unnecessary heavy metal element in the plant growth process, and is also an extremely toxic environmental pollutant. In addition, cd can be enriched in human body step by step through a food chain, thus the Cd is a serious threat to human health. Ramie (Boehmeria nivea L.) is a special perennial bast fiber crop in China, has developed root system, rapid growth, strong heavy metal tolerance and accumulation capacity, does not enter a food chain, and is considered as an ideal crop for repairing cadmium-polluted soil. The field test for restoring Cd contaminated soil shows that the Cd content of the soil is reduced from 1.8 mg/kg to 0.8-1.0 mg/kg after 4 years of ramie planting, the reduction range is 44% -55%, and the Hunan province has also brought the ramie into a related scheme for adjusting the production structure of heavy metal severely contaminated farmland, so that the method is one of the first-choice restoration crops. However, the special ramie variety suitable for repair is lacking at present, and new ramie varieties with high fiber yield, excellent quality, strong cadmium resistance and high accumulation are urgently needed to be cultivated. The molecular breeding method can accelerate the breeding process of ramie which is a perennial crop, but the lack of available Cd tolerance related genes seriously affects the development of molecular breeding work of a variety special for Cd pollution repair. WRKY Transcription Factors (TFs) are among the largest and most important families of transcription factors in plants, the N-terminus of which contains a conserved WRKYGQK domain, playing a vital role in plant growth and development and in coping with biotic and abiotic stresses. A great deal of researches show that the WRKY protein regulates and controls the tolerance of cadmium from multiple dimensions such as absorption, transportation, compartmentalization, chelant synthesis regulation and antioxidation systems of heavy metal Cd. PyWRKY71 to regulate the removal of Reactive Oxygen Species (ROS), improve the resistance of plants to cadmium, pyWRKY to regulate the expression of Glutathione (GSH) and Phytochelatin (PC), increase the chelation of Cd 2+ to reduce the toxicity of Cd 2+, and ZmWRKY64 and AtWRKY33 can regulate the expression of cadmium transport proteins HMA2 and NRAMP1, reduce the inflow of Cd and improve the tolerance of plants. There is no report in ramie that the WRKY family genes play a specific role in cadmium stress. The excavation of the gene for increasing cadmium accumulation and enhancing cadmium tolerance has important theoretical and practical significance for cultivating the cadmium repair plant variety. Disclosure of Invention The invention aims to overcome the technical defects, and provides application of a ramie BnWRKY gene in increasing cadmium accumulation in plant roots and enhancing cadmium tolerance of plants, which solves the technical problems of how to increase cadmium accumulation and enhance cadmium tolerance of plants in the prior art. In order to achieve the technical purpose, the technical scheme of the invention provides application of a ramie BnWRKY gene in increasing cadmium accumulation in plant roots and enhancing cadmium resistance of plants, wherein a CDS nucleotide sequence of the ramie BnWRKY gene is shown as SEQ ID NO. 1. In any embodiment, the amino acid sequence of the protein encoded by the ramie BnWRKY gene is shown as SEQ ID NO. 2. In any embodiment, increased cadmium accumulation and increased cadmium tolerance in plant roots is achieved by up-regulating the expression level of BnWRKY gene in the plant, or increasing the content of protein encoded by BnWRKY gene in the plant. In any embodiment, the plant comprises arabidopsis thaliana and ramie. In addition, the invention also provides a primer pair for amplifying the ramie BnWRKY gene, and the base sequences of the primer pair are respectively shown as SEQ ID NO.3 and SEQ ID NO. 4. In addition, the invention also provides a method for increasing cadmium accumulation in plant roots and enhancing cadmium tolerance of plants, the ramie BnWRKY gene is introduced into a receptor plant to obtain a transgenic plant over-expressing BnWRKY gene, and the cadmium accumulation amount in the roots of the transgenic plant is increased and the cadmium tolerance is enhanced. In any embodiment, an over-expression vector containing the BnWRKY gene is constructed, the over-expression vector is transformed into agrobacterium, and the BnWRKY gene is introduced into a recipient plant by agrobacterium-mediated inflorescence dip-d