Application of atga2ox1 gene in regulating stress resistance of plants and method for cultivating stress-resistant plants

A plant stress resistance and stress resistance technology, applied in the field of plant bioengineering, can solve problems such as reduced survival rate, and achieve the effects of high success rate, simple operation, and improved stress resistance ability

Active Publication Date: 2020-04-03
JILIN ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Spraying exogenous gibberellin on Arabidopsis ga1-3 mutant material found that its survival rate under salt stress was reduced to 54.5%

Method used

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  • Application of atga2ox1 gene in regulating stress resistance of plants and method for cultivating stress-resistant plants
  • Application of atga2ox1 gene in regulating stress resistance of plants and method for cultivating stress-resistant plants
  • Application of atga2ox1 gene in regulating stress resistance of plants and method for cultivating stress-resistant plants

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] Embodiment 1 Plant expression vector construction and genetic transformation of maize

[0072] Clone the full-length cDNA of the AtGA2ox1 gene (GENE BANK accession number: AT1g78440), connect it into the monocotyledonous plant expression vector pCAM-UGN, obtain the expression vector containing the AtGA2ox1 gene, and name it pCAM-UGN-GA2ox, such as figure 1 shown. Agrobacterium-mediated method was used to infect HiII maize immature embryos to obtain transgenic maize plants.

Embodiment 2

[0073] The identification of embodiment 2 transgenic plants and the expression analysis of exogenous gene

[0074] Identification of transgenic plants:

[0075] Total DNA was extracted from leaves of transgenic plants by CTAB method. Design PCR amplification primers according to the AtGA2ox1 gene sequence, as shown in the following table:

[0076] name Numbering Primer sequence (5'-3') AtGA2ox-445F SEQ ID NO.1 GGTTCGGGTCCACTATTT AtGA2ox-445R SEQ ID NO.2 CTATGCCTCACGCTCTTG

[0077] The PCR reaction system is:

[0078] Reagent Volume (μL) Taq Buffer(10×) 2.0 dNTP (2.5mM) 2.0 AtGA2ox-445F (10μM) 1.5 AtGA2ox-445R (10μM) 1.5 Taq DNA polymerase 0.2 wxya 2 o

to 20.0

[0079] The PCR reaction conditions are:

[0080]

[0081] The amplified product was subjected to 1% agarose gel electrophoresis, and the PCR electrophoresis result was observed under an ultraviolet gel imager.

[0082] E...

Embodiment 3

[0086] Phenotypic Analysis of Example 3 Corn Plant Drought Stress

[0087] Choose corn plants grown in a greenhouse at a temperature of 27°C±2°C, potted (10×10cm), peat soil: vermiculite (5:1, v / v), and grown for 21 days under natural light conditions (each genotype Each 10 strains) were subjected to drought stress treatment, and watering was stopped for 9 days. The maize leaves on the 9th day after drought stress were taken, frozen in liquid nitrogen, and stored in a -80°C refrigerator for subsequent determination of relevant physiological indicators. The amount of substrate and moisture used for each independent experimental material was kept the same.

[0088] In this example, to further evaluate whether the overexpression of the AtGA2ox1 gene can improve the drought stress resistance of maize plants, three independent transgenic maize lines (1#, 2#, 4#) were randomly selected for relevant experimental analysis. Corn plants at the seedling stage of 21 days were selected, ...

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Abstract

The invention provides an application of the AtGA2ox1 gene in regulating plant stress resistance and a method for cultivating stress-resistant plants, and relates to the technical field of plant bioengineering. The invention proves through experiments that the Arabidopsis gibberellin 2 oxidase gene AtGA2ox1 Can regulate plant stress resistance. Therefore, the application of the AtGA2ox1 gene, the recombinant vector comprising the AtGA2ox1 gene, the expression cassette comprising the AtGA2ox1 gene, the cells comprising the AtGA2ox1 gene or the recombinant bacteria comprising the AtGA2ox1 gene can effectively adjust the stress resistance of the plant and improve the plant's own ability to resist stress. The method for cultivating stress-resistant plants provided by the invention has simple operation and high success rate, and provides a new method and idea for future plant breeding, especially for improving the stress-resistant ability of plants.

Description

technical field [0001] The invention relates to the technical field of plant bioengineering, in particular to an application of AtGA2ox1 gene in regulating stress resistance of plants and a method for cultivating stress-resistant plants. Background technique [0002] Maize is an important food and economic crop in the world, and its growth is often disturbed by the external environment. Drought is currently one of the main factors affecting the growth of maize. Maize yields are estimated to be reduced by 15%-20% each year due to drought, and this trend increases as droughts become more frequent and severe. Therefore, how to improve the drought resistance of maize has become the focus of attention. [0003] Gibberellins are a class of plant hormones that exist widely. Its metabolic mode and action mechanism have been studied thoroughly, and most of the genes related to it in the model plant Arabidopsis have been cloned. Recent studies have found that when plants respond to...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/82A01H5/00A01H6/46
CPCC12N15/8271C12N15/8273
Inventor 郝东云刘相国陈子奇景海春柳青李楠尹悦佳刘洋韩四平
Owner JILIN ACAD OF AGRI SCI
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