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Wheat gene tafus3 and its application

A technology of wheat and genes, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve the problems of separation, cloning, ear germination resistance, and less stress resistance.

Inactive Publication Date: 2019-06-18
HENAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

People have isolated FUS3 gene from Arabidopsis thaliana, barley, indica rice, rape and other plants and conducted preliminary research on its function. There are few studies on the reverse reaction

Method used

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  • Wheat gene tafus3 and its application
  • Wheat gene tafus3 and its application
  • Wheat gene tafus3 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1: Cloning process of wheat gene TaFUS3

[0026] (1) The wheat gene TaFUS3 was isolated from the panicle-resistant variety "Huaimai 0360" and the easy-spinger variety "Zhongyou 9507". The total RNA was extracted from the grains of the above two varieties at 21, 28 and 35 days after flowering, and then the mRNA was further isolated. The mRNA was transcribed into cDNA by reverse transcription, and the cDNA was used as a template for gene cloning.

[0027] (2) A pair of degenerate primers (upstream primer P1: 5′-WGGGAGAATTGTTCTYCCRAA-3′ and downstream primer P2: 5′-ARTACCTSACRTTSGCTACYG-3′) were designed according to the conserved functional domains of the plant FUS3 gene registered in GenBank. A cDNA fragment with a molecular weight of about 470bp was obtained by PCR technology. The amplified product was recovered and cloned on the pMD18-T Eseay vector. After sequencing, sequence analysis proved that this cDNA fragment was a part of the TaFUS3 gene. On the bas...

Embodiment 2

[0036] Example 2: The role of wheat TaFUS3 gene in seed development and dormancy

[0037] The role of the wheat gene TaFUS3 in seed development: Select the wheat variety "Zhongyou 9507" which is easy to germinate and have mature, plump and uniform grains of uniform size. After sterilized, put them in a petri dish to germinate. After germinating for 10 days, move them to the soil to grow. During the whole growth cycle The embryos of mature seeds, the primary roots and buds 1 day after germination, the roots and leaves, young spikes 10 days after germination, the seeds 25 days after flowering, the embryos and endosperm 25 days after flowering were used as materials for the spatiotemporal expression characteristics of gene development : The relative expression level of wheat TaFUS3 gene was analyzed by real-time fluorescent quantitative PCR analysis method, and the results were as follows: figure 1 Shown: TaFUS3 gene is mainly expressed specifically in wheat embryos at 25 days af...

Embodiment 3

[0041] Embodiment 3: The effect of wheat TaFUS3 gene in stress resistance response

[0042] Select the mature, plump and uniform seeds of the wheat variety "Zhongyou 9507", disinfect them with 70% alcohol for 1 minute, wash them with sterile water for 3 times, and use 0.1% HgCl 2 Disinfect for 6 minutes, rinse with sterile water for 4-5 times, divide into four parts on average, and place them in Petri dishes (A), (B), (C) and (D) with two layers of sterile filter paper respectively for stress deal with. Among them, 8mL NaCl (150mmoL / L) solution was added to Petri dish (A), 8mL abscisic acid (ABA, 30μmoL / L) solution was added to Petri dish (B), and 8mL mannitol (300mmoL / L) was added to Petri dish (C). ) solution, cultivated in an artificial climate chamber (the natural light time is 12 hours / day, and the climate chamber temperature is 20° C.). Add 8 mL of sterilized water to the petri dish (D) and place it in a light incubator at 32°C (the light time and climate chamber tempe...

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Abstract

The invention relates to the technical field of plant genetic engineering, in particular to a wheat stress resistance gene TaFUS3 and its application. The wheat stress-resistance-related gene TaFUS3 of the present invention is isolated from the ear-resistant germination variety "Huaimai 0360" and the easy-ear germination variety "Zhongyou 9507". The full-length cDNA of the gene is 1461 bp, including 47 bp of 5 ′-non-coding region; 484 bp 3′-non-coding region; 906 bp coding region, terminator, and 28 bp polyadenylation tail, the protein encoded by the gene is a full-length protein consisting of 301 amino acids Composed of B3 domain-containing proteins. The wheat stress-resistance-related gene TaFUS3 of the present invention can be applied to seed development and dormancy, stress-resistance response, panicle germination resistance, stress-resistant breeding, gene chip production, and the design and synthesis of new stress-resistance-related genes has high practical application value.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, and in particular relates to a wheat gene TaFUS3 and its application. Background technique [0002] Plant transgenic research began in the late 1970s and early 1980s. In 1983, Zambryski obtained the first transgenic plant in the world. In 1985, Horsch et al. created the leaf disc method in Agrobacterium transformation. Since then, agricultural Bacteria-mediated method, gene gun method, electroporation method, PEG-mediated method, pollen tube channeling method, microinjection method, low-energy ion beam-mediated method, etc. The number of successfully transgenic species has continued to expand, involving more than 120 species of plants in 35 families. Among them, the transgenic plants obtained by the Agrobacterium-mediated method accounted for more than 85% of the total number of transgenic plants, and the transgenic plants obtained by the biolistic method accounted for about 10%...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C07K14/415A01H5/00A01H6/46C40B40/08
CPCC07K14/415C12N15/8267C12N15/8271C40B40/08
Inventor 任江萍王新国孟晓丹
Owner HENAN AGRICULTURAL UNIVERSITY
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