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Soybean vernalization gene GmVRN1 and colonizing method and application thereof

A cloning method and gene technology, applied in the field of soybean vernalization gene GmVRN1 and its cloning

Inactive Publication Date: 2015-09-23
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although vernalization is not required for soybean flowering, comparative genomics studies have shown that genes of the vernalization pathway are preserved in the soybean genome, the functions of which have been poorly studied

Method used

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  • Soybean vernalization gene GmVRN1 and colonizing method and application thereof
  • Soybean vernalization gene GmVRN1 and colonizing method and application thereof
  • Soybean vernalization gene GmVRN1 and colonizing method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Cloning of Example 1 Soybean GmVRN1 Gene

[0028] Selected Huachun No. 5 soybean seeds treated with 10% H 2 o 2 After surface disinfection for 1 min, rinse with distilled water, transfer to river sand and place in the dark for germination. Three days later, the uniform seedlings were selected and transferred to 1 / 2 Hoagland nutrient solution for further cultivation under the conditions of 12h light / 12h darkness and 50% humidity. The sampling period was the four-leaf stage, and the sampling time was 10 hours after the light began. The sampling site was the aboveground part of the plant. RNA was extracted after quick freezing in liquid nitrogen, and the reaction was carried out by primer F: 5-CGCGGATCCATGAGAGACTTTTCATTTCAT-3; primer R: 5-CGGGGTACCCTAATGTGGTGCAC-3 Transcription PCR, recovery by TaKaRa gel recovery and purification kit, the recovered product was connected to the sequencing vector, transformed into Escherichia coli, and then carried out PCR identification ...

Embodiment 2

[0031] Example 2 Analysis of GmVRN1 Responses to Different Lighting

[0032] Under short-day conditions, the expression level of GmVRN1 remained stable at first, and at 21 days ( figure 2 ), its expression peaked and then began to decline. Under long-day conditions, the expression level of GmVRN1 gradually increased, and its expression level also reached the peak at 21 days, and then began to decline. The difference is that the expression level of GmVRN1 under long-day conditions is higher than that under short-day conditions at 21 days. It shows that GmVRN1 responds to different light conditions.

Embodiment 3

[0033] Embodiment 3 Genetic transformation and screening of Arabidopsis

[0034] Treatment before transformation of Arabidopsis thaliana: when the main moss of Arabidopsis thaliana grows to 5-6cm, cut off the whole inflorescence at the base of the inflorescence to remove the dominance of its apex. After 1 week, 4-6 new lateral mosses will grow on the axillary buds. When the lateral moss inflorescences form flower buds and partially bloom or form 1-2 siliques, they can be used for transformation. Before transformation, the grown horns need to be cut off Put the stored Agrobacterium liquid or plate-preserved Agrobacterium on the plate containing antibiotics for activation; pick the activated monoclonal positive Agrobacterium containing the target gene into 5mL of YEP liquid medium containing antibiotics, 28 ℃, 250rpm overnight activation; pour the above-mentioned liquid into a beaker, quickly invert the plant to be transformed into the liquid, submerge the inflorescence for 2 mi...

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Abstract

The invention discloses a soybean vernalization gene GmVRN1 and a colonizing method and application thereof and belongs to the technical field of resources and environment. Related genes of vernalization approaches in a soybean genome is explored through comparative genomics, real-time PCR technology is applied to screening of vernalization approach gene difference expression of AtDREB1A genetically-modified soybean, the soybean vernalization gene GmVRN1 is colonized, and application authentication is performed on a flowering phase regulating and controlling function of the soybean vernalization gene GmVRN1 by converting arabidopsis.

Description

technical field [0001] The invention belongs to the technical field of resources and environment, and in particular relates to a soybean vernalization gene GmVRN1 and its cloning method and application. Background technique [0002] Soybean originated in China and has been one of the main grain and oil crops in my country since ancient times. Soybean grains are rich in protein and fat, and are an important source of vegetable protein and edible oil, so they have great economic value. In recent years, with the development of molecular biology and molecular genetics, the study of plant developmental biology has made great progress. It has become the goal of researchers to gradually reveal the essence of flower development at the molecular level. Therefore, the mining and functional exploration of new genes related to flowering will not only help to analyze the flower development process or response pathways, but also have far-reaching significance for crop improvement. [00...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C12N15/29C12N15/84A01H5/00
Inventor 张秀香年海吕菁马启彬孟醒宋瑞凤
Owner SOUTH CHINA AGRI UNIV
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