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Application of Arabidopsis thaliana At-ACA8 gene in plant stress tolerance and regulation of plant growth and development

A technology for plant growth and transgenic plants, which is applied in the application field of Arabidopsis thaliana At-ACA8 gene in plant stress resistance and regulation of plant growth and development, so as to achieve the effect of improving the ability of plants to resist stress

Inactive Publication Date: 2013-05-29
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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  • Application of Arabidopsis thaliana At-ACA8 gene in plant stress tolerance and regulation of plant growth and development
  • Application of Arabidopsis thaliana At-ACA8 gene in plant stress tolerance and regulation of plant growth and development
  • Application of Arabidopsis thaliana At-ACA8 gene in plant stress tolerance and regulation of plant growth and development

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Screening of At-ACA8 gene homozygous mutant aca8 and detection of gene expression:

[0042] (1) Screening of At-ACA8 gene homozygous mutant aca8:

[0043] The Arabidopsis thaliana T-DNA insertion mutant (seed number is CS859889) of the At-ACA8 gene (sequence list is shown in Table 1, Table 2, and Table 3), and the homozygous mutant strain was 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.

[0044] (2) Semi-quantitative RT-PCR identification of At-ACA8 gene transcription levels in leaves of Col-0 and mutant aca8:

[0045] Total RNA was extracted from leaves of Arabidopsis thaliana seedlings grown...

Embodiment 2

[0175] Stress resistance of mutant aca8 to low temperature stress:

[0176] (1) Low temperature stress phenotype test of wild-type plants and mutant aca8:

[0177] 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 normally (temperature 22°C, humidity 50%-60%, light Cycle 10h light / 14h dark), the aseptic seedlings after 14 days of culture 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. like 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 temperature stress.

[01...

Embodiment 3

[0182] Stress resistance of mutant aca8 to high temperature stress:

[0183] (1) High temperature stress phenotype test of wild-type plants and mutant aca8:

[0184] The Col-0 and aca8 seeds were sterilized and sown 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%, 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. 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 to high temperature, and the recovery ability after heat shock is stronger than that of the wild type.

[0185] At the same time, high...

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Abstract

The invention provides application of Arabidopsis thaliana calcium ion transfer-ATP enzyme gene At-ACA8, which comprises application of the gene in regulation of plant growth and development and stress tolerance, and application of the gene in cultivation of stress-tolerant transgenic plant species. By using At-ACA8 gene T-DNA insertion mutant aca8 obtained from the Arabidopsis thaliana biological resource center as the object, the experimental result proves that the mutant has obvious stress tolerance to low temperature, high temperature, infiltration and the like; the mutant is very sensitive to the existence of sucrose in the seed germination stage, the seed germination can be seriously inhibited under the conditions of not adding sucrose, and the mutant is very sensitive to the increase of the sucrose concentration in the growth stage; and the mutant is sensitive to hormone ABA in the seed germination stage, so the addition of a small amount of ABA can obviously inhibit the seed germination. The calcium ion signal transduction system provided by the invention has the response functions of regulating the plant growth and development and inhibiting stress, provides a high-quality gene for improving plant species, describes a new method and mechanism for enhancing plant stress tolerance and cultivating species with favorable tolerance, and has important economic meaning and application value.

Description

[0001] This application is a divisional application of the patent application "Application of Arabidopsis thaliana At-ACA8 gene in enhancing plant stress resistance and regulating plant growth and development" (application number: 2012100123229, application date: January 16, 2012). Technical field: [0002] 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 applica...

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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