Application of ZTL protein and coding gene thereof to regulating and controlling ABA tolerance of plant

A technique for encoding genes and transgenic plants, which is applied in the application field of ZTL protein and its encoding genes in regulating plant tolerance to ABA, can solve the problems of no biological clock function, etc., and achieve the effect of improving genetic characteristics

Inactive Publication Date: 2015-05-20
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The mRNA of FKF1 is circadian clock regulated, but it does not have any cir...

Method used

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  • Application of ZTL protein and coding gene thereof to regulating and controlling ABA tolerance of plant
  • Application of ZTL protein and coding gene thereof to regulating and controlling ABA tolerance of plant
  • Application of ZTL protein and coding gene thereof to regulating and controlling ABA tolerance of plant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Embodiment 1, the acquisition and identification of ZTL transgenic plants

[0056] The ZTL gene involved in this example is derived from Arabidopsis thaliana, and its sequence in the Arabidopsis genome is shown in sequence 1 in the sequence table, sequence 1 consists of 3164 nucleotides, of which the 416th Position -1245 is an intron sequence; the cDNA sequence of the ZTL gene is shown in sequence 2 in the sequence table, and sequence 2 is composed of 2334 nucleotides, wherein the 171-2000th position is a coding sequence (ORF); sequence 1 Both Sequence 2 and Sequence 2 encode the protein shown in Sequence 3 in the Sequence Listing, and Sequence 3 consists of 609 amino acid residues.

[0057] 1. Construction of recombinant expression vector pRTL2-ZTL

[0058] The total RNA of Arabidopsis wild-type (C24 ecotype) was extracted, and cDNA was obtained after reverse transcription. Using the obtained cDNA as a template, carry out PCR amplification with primer 1 and primer 2,...

Embodiment 2

[0108] Example 2, ZTL transgenic plant tolerance analysis test to ABA

[0109] ABA can inhibit the growth of plant seedlings. With the increase of exogenous ABA concentration, the inhibition of Arabidopsis wild type (C24 ecotype) was enhanced, and the growth of its main root and leaves would be affected to varying degrees. The experiment was repeated three times.

[0110] 1. Seedling growth experiment

[0111] With Arabidopsis wild-type (C24 ecotype), the ZTL gene point mutant ztl-1 and the T 3 The generation homozygous ZTL transgenic line OE, and the control plant obtained in Example 1 transformed into the pRTL2 empty vector were used as experimental materials. The seeds of each experimental material were sown on MS medium containing different concentrations of ABA (0 μM, 0.8 μM and 1.2 μM) (80-100 seeds were sown for each experimental material). After 3 days of low-temperature stratification at 4°C, they were moved into a light incubator, and the growth of each Arabidops...

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Abstract

The invention discloses application of ZTL protein and a coding gene thereof to regulating and controlling the ABA tolerance of a plant. The application provided by the invention is the following application of the protein shown in sequence 3: (a1) regulating and controlling the ABA tolerance of the plant; or (a2) breeding a plant variety with improved ABA tolerance. According to the invention, the plant hormone ABA can be used as a selective herbicide, of which the selectivity depends on the ABA sensitivity of the plant, the ABA sensitivity or tolerance of the plant is adjusted by utilizing ZTL, high ZTL-gene expression and herbicide-resistant transgenic crops are obtained by means of gene engineering, and selective weeding is realized. The invention meets the requirements of sustainable agricultural development and has important practical values and market prospects in studying ABA-tolerant molecular mechanisms of plants, improving hereditary characters, developing environment-friendly and pollution-free weeding methods and ways and the like.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to the application of a ZTL protein and its coding gene in regulating plant tolerance to ABA. Background technique [0002] The plant hormone ABA plays an important regulatory role in various stages of plant development, including embryonic maturation, seed development and seedling growth. ABA is also a key hormone for plants to adapt to stress, such as drought, high salinity and low temperature stress. The mechanism of ABA signal transduction has been extensively studied, and many components related to ABA signal transduction have been found, including ABA receptors. The possible G protein-coupled receptor GCR2 and the GPCR-like G proteins GTG1 and GTG2 have been reported as candidate ABA plasma membrane receptors. However, whether GCR2 is involved in the regulation of ABA-regulated seed germination and post-germination growth is still controversial. GTGS is a positive regulator of AB...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01G13/00
CPCA01G13/00C07K14/415C12N15/8274
Inventor 张大鹏于泳涛塞尔希·波尔托莱斯·科梅拉斯王小芳
Owner TSINGHUA UNIV
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