CN-118995794-B - Method for improving recovery capability of kiwi fruits after high-temperature stress and application of method

Abstract

The invention provides a method for improving the recovery capacity of kiwi fruits after high-temperature stress, namely, the HSFA2-2 gene in kiwi fruit plants is over-expressed by a genetic engineering method, and the nucleotide sequence of the HSFA2-2 gene is shown as SEQ ID NO. 1. The invention also provides application of the method in cultivation of a novel kiwi fruit variety resistant to high temperature stress. Meanwhile, the invention also provides a cultivation method of the novel kiwi fruit variety resistant to high temperature stress and the novel kiwi fruit variety cultivated based on the method. The novel kiwi fruit variety disclosed by the invention has the advantages that when subjected to strong high-temperature stress at 45 ℃ for 4 hours, the edge of the leaf is curled, but the leaf can be restored to a stretched state within one day after the high-temperature stress is eliminated, the phenomenon of drying up does not occur, the stem section is also restored to a straight growth state, and the plant restoration capability is obviously improved.

Inventors

• LIU XIAOFEN
• WU SHI
• SU WENYUE
• YIN XUEREN

Assignees

• 安徽农业大学

Dates

Publication Date

20260512

Application Date

20240911

Claims (7)

1. 1. A method for improving the recovery capacity of kiwi fruits after high-temperature stress is characterized in that the HSFA2-2 gene in kiwi fruit plants is over-expressed by a genetic engineering method, and the nucleotide sequence of the HSFA2-2 gene is shown as SEQ ID NO. 1.
2. 2. The method for improving the recovery capacity of kiwi fruits after high temperature stress according to claim 1, wherein the HSFA2-2 gene is used for encoding a heat shock transcription factor, and the amino acid sequence is shown as SEQ ID NO. 2.
3. 3. The method for improving the recovery capacity of kiwi fruits after high temperature stress according to claim 1, wherein the expression product of the HSFA2-2 gene positively regulates the recovery capacity of kiwi fruits after high temperature stress.
4. 4. An application of the method for improving the recovery capacity of kiwi fruits after high temperature stress according to any one of claims 1-3 in cultivation of new varieties of kiwi fruits resistant to high temperature stress.
5. 5. A cultivation method of a novel kiwi fruit variety resistant to high temperature stress is characterized by firstly cloning a heat shock transcription factor AcHSFA-2 by utilizing a PCR technology, wherein the nucleotide sequence of HSFA2-2 is shown as SEQ ID NO.1, then carrying a PCR product of AcHSFA-2 into an expression vector for plant genetic transformation after agarose gel electrophoresis, separation and purification, then transforming a recombinant expression vector containing AcHSFA2-2 into an agrobacterium EHA105 strain, finally transforming AcHSFA-2 into a kiwi fruit plant by utilizing an agrobacterium mediated kiwi fruit leaf disc method, and screening a kiwi fruit material successfully transformed to obtain the novel kiwi fruit variety resistant to high temperature stress required by a target.
6. 6. The method for cultivating a novel high-temperature stress-resistant kiwi fruit variety according to claim 5, wherein the expression level of a heat shock transcription factor HSFA2-2 in the novel high-temperature stress-resistant kiwi fruit variety is improved by 75-129 times.
7. 7. The method for cultivating the novel high-temperature stress resistant kiwi fruit variety according to claim 5, wherein the novel high-temperature stress resistant kiwi fruit variety normally grows in an environment without high temperature stress, edges of leaves curl under the high temperature stress, but the leaves can be restored to a stretched state within one day after the high temperature stress, and stem segments are also restored to a straight growth state.

Description

Method for improving recovery capability of kiwi fruits after high-temperature stress and application of method Technical Field The invention relates to the fields of plant molecular biotechnology and genetic engineering, in particular to a method for improving the recovery capacity of kiwi fruits after high-temperature stress and application thereof. Background The change in global ambient temperature increases the frequency of extremely high temperature events, resulting in high temperature stresses that greatly limit plant growth. The kiwi fruit is a perennial vine fruit tree originated in China, the fruit is rich in vitamin C and various microelements, the nutritional value is high, and the kiwi fruit is known as the king of the fruit. The planting area and the annual yield of the Chinese goosebeery in China are 18.26 ten thousand hm 2 and 219.7 ten thousand tons respectively, which are 67.9% and 50.5% of the total world, and are important fruit trees for promoting the income of agricultural rural areas in the first world and 2019. However, kiwi fruits are heat-labile and extremely high in summer often cause yield and quality reduction of fruits and even death of trees. Therefore, the cultivation of the novel kiwi fruit variety which is resistant to high temperature stress and can still recover the growth capacity after being subjected to high temperature stress has important significance. According to the existing research progress, when high temperature stress is encountered, plants rapidly produce various physiological changes in response to the stress, and the harm of the high temperature stress to growth is reduced. Heat shock transcription factors (Heat shock transcription factors, HSF) are the family of transcription factors most closely related to high temperature stress response in plants. Under high temperature stress, the gene expression level of a large number of heat shock transcription factors is obviously increased, so that the high temperature stress is responded. The number of HSF members in kiwi fruits, and only 4 members of the HSFA2 subgroup, designated AcHSFA-1/-2/-3/-4, were identified, only AcHSFA-1 was functional. And the physiological indexes of the kiwi fruits responding to different intensity heat stress change differently, which indicates that the regulation and control of the heat stress response mechanism are finer. Therefore, based on the functions of different heat shock transcription factors, the novel kiwi fruit variety with enhanced recovery capacity after being subjected to high temperature stress is cultivated by combining the kiwi fruit genetic transformation technology, and the method has great feasibility and great industrial significance. Disclosure of Invention The technical problem to be solved by the invention is to provide a method for improving the recovery capacity of kiwi fruits after high-temperature stress by modifying the heat shock transcription factor HSFA2-2 of kiwi fruits and application thereof. The invention adopts the following technical scheme to solve the technical problems: A method for improving the recovery capacity of kiwi fruits after high temperature stress is characterized in that the HSFA2-2 gene in kiwi fruit plants is over-expressed by a genetic engineering method, and the nucleotide sequence of the HSFA2-2 gene is shown as SEQ ID NO. 1. As one of the preferable modes of the invention, the HSFA2-2 gene is used for encoding a heat shock transcription factor, and the amino acid sequence is shown as SEQ ID NO. 2. As one of the preferable modes of the invention, the expression product of the HSFA2-2 gene positively regulates the recovery capacity of the kiwi fruits after high temperature stress. The application of the method for improving the recovery capability of the kiwi fruits after high-temperature stress in the cultivation of novel kiwi fruits varieties resistant to high-temperature stress. A method for culturing a novel kiwi fruit variety resistant to high temperature stress is characterized by firstly cloning a heat shock transcription factor AcHSFA-2 by utilizing a PCR technology, wherein the nucleotide sequence of HSFA2-2 is shown as SEQ ID NO.1, then carrying a PCR product of AcHSFA-2 into an expression vector (any commercial expression vector) for plant genetic transformation after agarose gel electrophoresis, separation and purification, then transforming a recombinant expression vector containing AcHSFA2-2 into an agrobacterium EHA105 strain by a method such as electric shock or liquid nitrogen freeze thawing, finally transforming AcHSFA-2 into kiwi fruit plants by using an agrobacterium-mediated kiwi fruit leaf disc method, and screening kiwi fruit materials successfully transformed, thus obtaining the novel kiwi fruit variety resistant to high temperature stress required by a target. As one of the preferable modes of the invention, the expression quantity of the heat shock transcription factor HSFA2-2 in the novel kiwi fru