Kiwi fruit germplasm material resistant to extreme high-temperature growth conditions and cultivation method

A technology for high-temperature growth and kiwifruit, applied in biochemical equipment and methods, chemical instruments and methods, and botanical equipment and methods, etc., can solve the problems of decreased pollen germination rate, increased reactive oxygen species, decreased antioxidant enzyme activity, etc. The effect of easy promotion and application, reasonable design and convenient operation

Pending Publication Date: 2022-03-29
ZHEJIANG UNIV
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  • Application Information

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Problems solved by technology

High temperature stress leads to the increase of reactive oxygen species in plant cells, the decrease of antioxidant enzyme activity and the decrease of pollen germination rate, which seriously affects the yield and quality of crops

Method used

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  • Kiwi fruit germplasm material resistant to extreme high-temperature growth conditions and cultivation method

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Experimental program
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Effect test

Embodiment 1

[0013] (1) Research methods

[0014] 1. High temperature stress treatment of common kiwifruit plants: select vigorous common kiwifruit tissue culture seedlings grown in 1 / 2MS rooting medium, and inoculate 3 plants in each bottle as three biological replicates. The plants were placed in a constant temperature oven at 50°C to simulate natural high temperature stress conditions for 2 hours. Then the plants were re-placed in a tissue culture room at 20°C, cultured normally, and the phenotype changes were observed.

[0015] (2) Research results

[0016] After 2 hours of extreme high temperature treatment at 50°C, the leaves of common kiwifruit plants wilted and withered, and some leaves fell off; within 9 days after the high temperature treatment was eliminated, the plants were still in a state of wilting, withering, sluggish growth, and basically lost vitality (attached figure 1 c and 1d).

Embodiment 2

[0018] (1) Research methods

[0019] 1. Cloning of heat shock transcription factor AcHsfA2-1

[0020] The heat shock transcription factor AcHsfA2-1 was cloned by using ordinary PCR technology, using the cDNA of the leaf of Actinidia sinensis 'Donghong' as a template, and applying the primers whose sequences are SEQ NO.3 and SEQ NO.4. PCR system includes 15μL MaxBuffer, 1.2μL SEQ NO.3 primer, 1.2μL SEQ NO.4 primer, 0.6μL dNTP Mix (10mM each), 0.6μL Max Super-Fidelity DNA Polymerase, 1 μL cDNA template, 10.4 μL nuclease-free water for a total volume of 30 μL. The PCR program was: 5min at 95°C, 38 thermal cycles (95°C for 15s, 58°C for 15s, 72°C for 90s), 5min at 72°C, and terminated.

[0021] 2. Expression vector construction

[0022] After agarose gel electrophoresis, separation, and purification of the above PCR products, they are loaded into an expression vector that can be used for plant genetic transformation, which can be any commercial expression vector. The connec...

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Abstract

The invention provides a kiwifruit germplasm material with strong resistance to an extreme high-temperature growth condition and a cultivation method. The kiwifruit germplasm material is obtained by over-expressing a heat shock transcription factor HsfA2-1. The kiwifruit germplasm material has extremely high resistance in the aspect of high temperature stress resistance, can continuously resist extreme high temperature stress of 50 DEG C for at least 2 hours, and can continuously and normally grow after being stressed. According to the invention, a heat shock transcription factor AcHsfA2-1 in Donghong kiwi fruit is taken as an example, and the cultivation method of the germplasm material is elaborated. The application of other genes, of which the homology with the nucleotide sequence or amino acid sequence of the AcHsfA2-1 is higher than 90%, in the aspect is within the protection range of the invention. The cultivation method is reasonable in design, convenient to operate and easy to popularize and apply.

Description

technical field [0001] The invention belongs to the fields of plant molecular biotechnology and genetic engineering, and relates to a kiwifruit germplasm material resistant to extreme high temperature growth conditions and a cultivation method. Background technique [0002] Plants are colonized in the soil and cannot avoid environmental stress by moving, so the ability to resist environmental stress is very important for plant growth and development. With the intensification of global warming, the damage of high temperature stress to plant growth and development is becoming more and more serious, and enhancing high temperature resistance is becoming more and more important for the survival and reproduction of plants. High temperature stress leads to the increase of reactive oxygen species in plant cells, the decrease of antioxidant enzyme activity and the decrease of pollen germination rate, which seriously affects the yield and quality of crops. Statistics from China and I...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/84A01H5/00A01H6/00C07K14/415C12N15/29
CPCC12N15/8205C12N15/8271C07K14/415
Inventor 殷学仁沈任佳王文球刘晓芬
Owner ZHEJIANG UNIV
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