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Grain development related protein TaGSR1 and application thereof in wheat plant type breeding

A technology related to protein and protein, applied in the field of plant genetic engineering and molecular breeding, to achieve broad market application prospects, improve lodging resistance, and improve the effect of reasonable close planting

Active Publication Date: 2021-10-22
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

After searching, there is no report on the coding sequence of the wheat TaGSR1 gene, its encoded grain development-related protein TaGSR1, and the application of the protein TaGSR1 in regulating wheat plant type breeding

Method used

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  • Grain development related protein TaGSR1 and application thereof in wheat plant type breeding
  • Grain development related protein TaGSR1 and application thereof in wheat plant type breeding
  • Grain development related protein TaGSR1 and application thereof in wheat plant type breeding

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Construction of the expression vector

[0042] Tagsr1 coding sequence clone

[0043] In order to obtain the coding sequence of Tagsr1 proteins, the sequence of Tagsr1 gene was amplified in the wheat variety stone. The primers used were TAGSR1-F and Tagsr1-R, and the PCR reaction system was: KOD-FX NEO BUFFER: 25 μL, DNTP ( 2mm): 10 μL, Tagsr1-F (10 μm): 1.5 μL, Tagsr1-F (10 μm): 1.5 μL, stone new 828GDNA (50 ng / μL): 1 μL, KOD-FX NEO: 1μL, DDH 2 O Supple 50 μL. The PCR reaction procedure was: 98 ° C predetermitability 2min, 98 ° C denaturation 12SEC; 58 ° C for 20 SEC, 68 ° C extends 25sec, and reacted 35 cycles; 68 ° C resilience 5 min.

[0044] PCR results figure 1 As shown, the lane 1 is Transgen 2K Plus Marker, the lane 2 is the Tagsr 1 purpose strip.

[0045] PCR products are sent to Qingdao Oyster Oxi Biotechnology Co., Ltd. Sequencing, the Tagsr1 gene in the stone new 828 wheat is the known sequence similarity of the Ensembl Plants database (http: / / plants....

Embodiment 2

[0054] Example 2 Genodes of Generation and Identification

[0055] 1. Tagsr1 genetically modified future generation

[0056] Experimental Example 1 constructs a binary expression vector PLGY02-Tagsr1 transformed into Agrobacterium Eha105 sensitive cells, specifically: add recombinant double-firing carrier PLGY02-TAGSR1 to Agrobacterium Eha105 sensation to ice bath 5 min, liquid nitrogen quick freeze 5min 37 ° C heat shock reaction 5 min, after ice bath, no anti-LB, put into 28 ° C shaker recovery 2H, then apply a coater to LB (containing rifum, streptomycin and kanamycin) On the plate, at 28 ° C inverted culture was cultured to the cloning length. Picking monoclonal inoculated in LB culture solutions containing corresponding resistance, rock, 160 rpm, and incubated for 24 hours in 28 ° C.

[0057] JW1 wheat seeds of about 15 days after taking the powder, peeling the young embryo. Agrobacterium suspension was taken from 1 ml of bacteria in 1.5 ml of centrifuge tubes, and 1.4 uL ace...

Embodiment 3

[0063] Example 3 After the wheat expressed Tagsr1 gene, the representative identification

[0064] Wild-type receptor varieties JW1, hypotensed transgenic wheat OE-TAGSR1 and TAGSR1 and its homologous copy of the homogence strain Tagsr1-Ko were planted in the artificial climulation room of Qingdao Campus, Shandong University, cultivating conditions: light 16h, Darkness 8 hours; daytime temperature 22 ° C, night temperature 16 ° C; humidity 40% -50%; CO 2 The concentration is 500 ppm-700 ppm.

[0065] Analysis of the length, width and thousand grains of the grain length, width and thousand grains of the JW, OE-Tagsr1 and Tagsr1-KO.

[0066] Compared with wild-type JW1, the length and width of the seed grain and width of the Overexpression strain were significantly reduced, while the mutant strain Tagsr1-Ko grains increased significantly, and the width of the grain had no significant change (in Table 2, Image 6Indicated. Measuring the grains of the grains showed that the OE-Tagsr1 o...

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Abstract

The invention discloses an application of a grain development related protein TaGSR1 or an encoding gene thereof in wheat plant type breeding. The amino acid sequence of the grain development related protein TaGSR1 is as shown in SEQ ID NO.1, and the nucleotide sequence of the coding gene of the grain development related protein TaGSR1 is as shown in SEQ ID NO.2. The application in wheat plant type breeding refers to the application of regulating and controlling wheat plant height or the application of regulating and controlling included angles of wheat leaves. The relationship between the wheat TaGSR1 gene and the plant height and leaf included angle of wheat is defined, and it is verified that the plant height reduction and leaf included angle reduction of wheat can be achieved through high expression of TaGSR1. According to the invention, rapid breeding of ideal plant types of wheat is realized by utilizing a genetic engineering technology, a feasible method is provided for improving lodging resistance and rational close planting of wheat, and the application has important breeding application value and wide market application prospect.

Description

Technical field [0001] The present invention relates to a grain development-related protein and its applications, and more particularly to a grain development-related protein Tagsr1 or a use of genes thereof in wheat strain; belonging to plant gene engineering and molecular breeding technology. Background technique [0002] Wheat is the world's second largest grain crop, providing 20% ​​carbohydrates and 23% protein. Human food safety issues are facing a reduction in cultivated land, global warming, population continuous growth and environmental pollution intensification, and how to improve wheat production is important for food security under limited resource conditions. Wheat's "green revolution" reduces the high wheat strains, increasing the anti-challenging capacity of wheat, greatly increasing the output of wheat, which also demonstrates the importance of the crop ideal plant type to yield. [0003] The rapeseous lactone (BR) acts as a sixth largestytone of plants, plays a v...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/84A01H5/10A01H6/46
CPCC07K14/415C12N15/8261
Inventor 白明义吕金洋樊敏李根英韩超
Owner SHANDONG UNIV
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