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Method for regulating and controlling seed size by using ERECTA gene

A technology of gene regulation and transgenic seeds, applied in the field of gene technology and botany, can solve the problem of undiscovered seeds of ERECTA gene, achieve the effect of significant regulation effect, ensure accuracy, and avoid the interference of false positives

Inactive Publication Date: 2017-09-19
HEBEI NORMAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing research on the ERECTA gene has not found that it is involved in the regulation of seed size

Method used

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  • Method for regulating and controlling seed size by using ERECTA gene
  • Method for regulating and controlling seed size by using ERECTA gene
  • Method for regulating and controlling seed size by using ERECTA gene

Examples

Experimental program
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Effect test

Embodiment

[0025] The present invention takes Arabidopsis thaliana as an example to carry out overexpression of ERECTA gene in Arabidopsis thaliana.

[0026] 1. Construction of transformation vector

[0027] First, design the following primers according to the Arabidopsis ERECTA gene (gene number AT2G26330, see SEQ ID No.1 in the sequence table):

[0028] Forward primer SEQ ID No.2: 5'-CACCATTCAAAAATCCCTCTCCAACATCATAA-3';

[0029] Reverse primer SEQ ID No. 3: 5'-CTCACTGTTCTGAGAAATAACTTGTCCA-3'.

[0030] The DNA in the leaves of Arabidopsis Col-0 was extracted as a template, and the above primers were used for PCR amplification; the high-fidelity enzyme Phusion DNApolymerase (Thermo Lot 00242505) was used for amplification, and the reaction system (50 μl) was as follows:

[0031]

[0032] The cycle program was: pre-denaturation at 98°C for 1 min; denaturation at 98°C for 10 s, annealing at 58°C for 30 s, extension at 72°C for 5 min and 30 s, 35 cycles; extension at 72°C for 10 min; s...

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Abstract

The invention discloses a method for regulating seed size by using the ERECTA gene. By cloning the 2037bp promoter of the ATG upstream of the ERECTA gene and the 5526bp ERECTA gene, the Gateway method is used to construct a recombinant vector, and gERECTA can be efficiently and quickly cloned into a binary On the vector pGWB4; use Anti-GFP to detect the expression of gERECTA-GFP fusion protein by Western Blot to ensure the rapid and accurate detection of the expression product. Positive seedlings are screened to homozygous T3 generation to ensure the accuracy of the results and avoid the interference of false positives . The control effect of the invention is remarkable, the result is accurate and reliable, the control method is simple and convenient to operate, can be formulated, and is easy to popularize and apply.

Description

technical field [0001] The invention relates to the fields of gene technology and botany, in particular to a method for using ERECTA gene to regulate seed size. technical background [0002] Plant seeds are mainly composed of three parts: embryo, endosperm and seed coat, so the size of the seed is regulated by the embryo, endosperm and integument. The regulation of seed size is an important issue in developmental biology. In evolution, seed size is an important selective trait. At the same time, in agricultural production, seed size is an important agronomic trait, which can be used for molecular breeding of crops. And provide a theoretical basis for improved varieties. In recent years, some genes involved in the regulation of seed size have been reported at home and abroad, but the determination mechanism of seed size is not very clear. Therefore, the research on the regulation of plant seed size has important theoretical and practical significance. [0003] The ERECTA ge...

Claims

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Application Information

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IPC IPC(8): C12N15/82A01H5/00
CPCC12N15/8261
Inventor 汤文强蔡兴卜吴雪丹
Owner HEBEI NORMAL UNIV
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