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A precocious gene eam.z and its molecular marker snp595 and its application

A molecular marker and early-maturing technology, applied in the field of molecular biology, can solve problems such as prominent stubble gaps, multiple cropping, and short harvest intervals in multi-cropping systems, and achieve the effect of improving selection efficiency

Active Publication Date: 2021-04-27
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Our country has a lot of people and little land. In order to improve land use efficiency and increase crop yield, multi-cropping system has been widely implemented all over the country. However, the multi-cropping system has short harvest intervals and the contradiction between stubble is prominent. If the variety matures late, this contradiction will be exacerbated, resulting in double cropping. vicious circle of crops

Method used

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  • A precocious gene eam.z and its molecular marker snp595 and its application
  • A precocious gene eam.z and its molecular marker snp595 and its application
  • A precocious gene eam.z and its molecular marker snp595 and its application

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Embodiment 1

[0032] Example 1 Molecular marker SNP595

[0033] 1) The present invention uses the chemical mutagen EMS to treat the barley variety Hua 30, and obtains an early-maturing mutant Zhezaomai No. 1 with stable traits through multiple generations of screening and identification, and clones the early-maturing gene eam.z by gene mapping and other methods, and studies It was found that the base of the eam.z gene at 595bp in the coding region of the wild-type barley gene HORVU3Hr1G114970 changed from A to T, forming a stop codon TAG, which led to the premature termination of the encoded protein and a premature phenotype. The point is different from the previously reported barley early maturity gene mutation site ( figure 1 ). Utilizing the mutation site, the present invention designs a KASPar marker, which can quickly and accurately detect whether the barley material contains the eam.z mutation gene.

[0034] The phenotype of the early-maturing mutant Zhezaomai 1 was as follows: the...

Embodiment 2

[0044] Example 2 Molecular marker identification eam.z genotype

[0045] Methods for identifying eam.z genotypes using molecular markers:

[0046] 1) DNA extraction: Take 100 mg of barley leaves, and use the CTAB method to extract DNA. The specific steps are as follows:

[0047]① Grind the sheared leaves in liquid nitrogen and put them into 1.5ml centrifuge tubes, add 600 μL of CTAB to each tube and place in a warm water bath at 65°C for 50-60 minutes (take out every 10 minutes and shake gently);

[0048] ②Add 600 microliters of chloroform / isoamyl alcohol (24:1) and mix well for 15 minutes. Slowly, weigh in pairs, and after balancing, centrifuge at 9600 rpm for 10 minutes at 4°C;

[0049] ③Pipe the supernatant to another 1.5ml new centrifuge tube, add 2 times the volume of absolute ethanol and mix slightly (precipitate in -20°C refrigerator for 30-60min);

[0050] ④ Pick out the precipitate with a pipette tip, put it in a 1.5ml centrifuge tube, wash it twice with 70% ethanol...

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Abstract

The invention discloses an early-maturing gene eam.z and its molecular marker SNP595 and its application. It is obtained by changing the base at 595 bp of the wild-type barley gene HORVU3Hr1G114970 coding region from A to T, and its nucleotide sequence is as shown in SEQ ID NO.1 shown; the present invention also provides a molecular marker SNP595 based on the eam.z gene mutation site, which is selected from the primer pair shown in SEQ ID NO.5-7. Using the marker to carry out the assisted selection of the eam.z gene for early-maturing breeding can guarantee 100% accuracy and speed up the progress of early-maturing barley breeding.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and relates to a precocious gene eam.z and its molecular marker SNP595 and applications. Background technique [0002] Our country has a lot of people and little land. In order to improve land use efficiency and increase crop yield, multi-cropping system has been widely implemented all over the country. However, the multi-cropping system has short harvest intervals and the contradiction between stubble is prominent. If the variety matures late, this contradiction will be exacerbated, resulting in double cropping. A vicious circle of crops. Therefore, breeding early-maturing barley varieties is of great significance for making full use of light and heat resources and improving the multiple cropping index (Sun Dongfa et al., 2006). In addition, winter barley is mainly planted in my country. In the late stage of barley growth and development, it is often affected by dry and hot wind, caus...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C07K14/415C12Q1/6895C12Q1/6858C12N15/11
CPCC07K14/415C12Q1/6858C12Q1/6895C12Q2600/13C12Q2600/156C12Q2531/113C12Q2563/107
Inventor 华为朱靖环王其飞郭刚刚徐延浩
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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