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Applications of At-ACA8 gene of arabidopsis thaliana for enhancing plant stress resistance and regulating plant growing development

A technology of transgenic plants, Arabidopsis, applied to angiosperms/flowering plants, botanical equipment and methods, applications, etc., to achieve the effect of improving the ability of plants to resist adversity

Inactive Publication Date: 2012-08-22
KUNMING INST OF BOTANY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The research on At-ACA8 gene mainly focuses on the amino acid composition and autoinhibitory function of the N-terminal autoinhibitory protein domain, and its cell biological role in regulating plant growth and development has not been reported.

Method used

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  • Applications of At-ACA8 gene of arabidopsis thaliana for enhancing plant stress resistance and regulating plant growing development
  • Applications of At-ACA8 gene of arabidopsis thaliana for enhancing plant stress resistance and regulating plant growing development
  • Applications of At-ACA8 gene of arabidopsis thaliana for enhancing plant stress resistance and regulating plant growing development

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Screening of At-ACA8 gene homozygous mutant aca8 and detection of gene expression:

[0041] (1) Screening of At-ACA8 gene homozygous mutant aca8:

[0042] The Arabidopsis thaliana T-DNA insertion mutant (seed number is CS859889) of the At-ACA8 gene (sequence listing is shown in Table 1, Table 2, and Table 3), and the homozygous mutant strain is identified by PCR, such as Figure 2A As shown, the primers used are as follows: 5'-end primer: 5'-CTTCAACGCCTTTGTTCTCTG-3'; 3'-end primer: 5'-CCAGTAATCCAAAGTTGCTCG-3'; LBpROK2: 5'-ATTTTGCCGATTTCGGAA C-3', further sequencing verification found that the The mutant originates from the insertion of T-DNA into exon 34, resulting in gene mutation. The result is as Figure 2B As shown, the identified CS889 were all homozygous.

[0043] (2) Semi-quantitative RT-PCR identification of At-ACA8 gene transcription level in leaves of Col-0 and mutant aca8:

[0044] Total RNA was extracted from leaves of Arabidopsis thaliana seedlings grow...

Embodiment 2

[0174] Stress resistance of mutant aca8 to low temperature stress:

[0175] (1) Low temperature stress phenotype test of wild-type plants and mutant aca8:

[0176] The Col-0 and aca8 seeds were sterilized and sowed in MS medium containing 1.5% sucrose, vernalized at 4°C for 3 days, and then germinated and grew normally (temperature 22°C, humidity 50%-60%, light Cycle 10h light / 14h dark), the aseptic seedlings after 14 days of cultivation were subjected to low temperature freezing and thawing experiments. The treatment conditions are: cold acclimation at 4°C for 3 days, rapid cooling to -6°C for 3.5h-4h, heating to 4°C for recovery for one day, and then normal culture at 22°C. Col-0 can be observed after about 3-5 days Phenotypic differences from aca8. Such as Figure 3A As shown, the survival rate of the sterile seedlings of aca8 was higher after low temperature treatment, and the results showed that the sterile seedlings of mutant aca8 were more resistant to low temperatur...

Embodiment 3

[0181] Stress resistance of mutant aca8 to high temperature stress:

[0182] (1) High temperature stress phenotype test of wild-type plants and mutant aca8:

[0183] The Col-0 and aca8 seeds were sterilized and sown in MS medium containing 1.5% sucrose concentration, vernalized at 4°C for 3 days, and then germinated and grew normally (temperature 22°C, humidity 50%-60%, photoperiod 10h of light / 14h of darkness), the aseptic seedlings after 14 days of cultivation were subjected to high-temperature heat tampering and heat shock treatment, the conditions were: heat taming at 38°C for 2h, rest at room temperature 22°C for 1h, heat up to 45°C for 3-4h, and then Placed at 22°C to resume culture. After about 3-5 days, the phenotypic difference between wild-type Col-0 and mutant aca8 can be observed, such as Figure 4A As shown, the survival rate of aca8 sterile seedlings after high temperature treatment is higher, indicating that the mutant aca8 sterile seedlings are more resistant...

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Abstract

The invention provides an application of an arabidopsis thaliana calcium ion transported-ATP (Adenosine Tri-Phosphate) enzyme gene At-ACA8. The application comprises an application of the gene on aspects of regulating plant growing development and resisting adverse circumstance stress, and an application of the gene in an aspect of an adverse circumstance stress resisting transgenic plant variety. A T-DNA (Transforming-Deoxyribonucleic Acid) inserting mutant aca8 of the At-ACA8 gene obtained from an arabidopsis thaliana biological resource center is utilized as a target and an experiment result confirms that the mutant has obvious resistances on adverse circumstances of low temperature, high temperature, penetration compelling and the like; a seed germination phase is sensitive to whether sucrose exists or not; the germination of seeds is seriously inhibited under the condition of not adding the sucrose and a growing phase is more sensitive to the raising of the concentration of the sucrose; the seed germination phase is sensitive to a hormone ABA (Abscisic Acid); and less ABA is applied to obviously inhibiting the germination of the seeds. The invention discloses responding functions of regulating the plant growing development and resisting stress of the calcium ion conduction system and provides a good-quality gene for the variety improvement of the plants; the invention represents a new method and a mechanism which are used for improving the resisting adverse circumstance resisting capability and have good resisting variety and important economic meanings and application values.

Description

Technical field: [0001] The invention belongs to the field of plant genetic engineering, and in particular relates to the application of the Arabidopsis calcium ion transport-ATPase protein gene At-ACA8, and relates to the use of the mutant aca8 of the gene At-ACA8 in improving plant resistance to low temperature, high temperature and osmotic stress And hormone response and the application of regulating plant growth and development. At the same time, it also involves the use of the gene At-ACA8 in cultivating transgenic plants that are resistant to various stresses, sensitive to no-sucrose and high-concentration sucrose, and sensitive to the hormone ABA in the seed germination stage Variety applications. Background technique: [0002] The growth and development of plants are closely related to the environment in which they exist. In nature, due to various reasons such as different geographical locations, climatic conditions, and human activities, various adversities are cau...

Claims

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Application Information

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IPC IPC(8): C12N15/82A01H5/00
Inventor 李唯奇张洁张旭东王瑞萍
Owner KUNMING INST OF BOTANY - CHINESE ACAD OF SCI
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