Method for identifying maize intercross species by using molecular marker

A technology of molecular markers and corn, which is applied in the field of identification of corn intercross species using molecular markers, can solve the problems of inability to identify corn intercross species, and achieve the effect of low cost and easy operation

Active Publication Date: 2013-10-09
JIANGSU ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The object of the present invention is to provide a method for using InDel molecular markers to identify corn intercross species in view of the problem that the current molecular marker detection method for the purity of corn varieties cannot identify corn intercross species

Method used

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  • Method for identifying maize intercross species by using molecular marker
  • Method for identifying maize intercross species by using molecular marker
  • Method for identifying maize intercross species by using molecular marker

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Embodiment Construction

[0027] The embodiment of the present invention uses maize inbred lines B73 and Mo17 as parents and crossed crossed species as materials. The present invention is based on the identification of said crossed species.

[0028] Materials and Methods

[0029] 1.1 Material

[0030] The tested materials were maize inbred lines B73 and Mo17, orthogonal species B73×Mo17 (BM) and reverse cross species Mo17×B73 (MB).

[0031] extract

[0032] B73, Mo17, BM, MB leaf DNA and BM, MB endosperm (5 samples each) DNA extraction was performed in accordance with the relevant steps of Karroten Plant Genomic DNA Extraction Kit (Nanjing Sage Technology Co., Ltd.), and the concentration of the extracted DNA samples was determined and 0.8 % Agarose gel to check the quality.

[0033] Amplification and agarose gel electrophoresis

[0034] The PCR reaction volume is 25 μl, which contains 2 mmol L -1 MgCl 2 ,100 μmol L -1 dNTP, 0.2 μmol L -1 Primer, 1 U Taq enzyme, 50 μg template DNA, topped with 20 μ...

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Abstract

The invention relates to method for identifying maize intercross species by using a molecular marker, which is characterized by comprising the following steps of: extracting the endosperm, leaf blade and parental leaf blade DNA of a maize intercross species to be inspected; developing an InDel marker and screening out a codominant marker among parents according to the sequence difference among parent genomes; analyzing the quality of amplification products of leaf blade DNA allelic genes of the intercross species to determine the molecular marker of the maize intercross species; searching the PCR exponential amplification phase of the marker, clearly expressing the cycle number of the amplification products on agarose gel, and amplifying the endosperm and leaf blade DNA of the intercross species in the same batch; quantitatively analyzing an electrophoretic band to obtain the measured value of relative amount of amplification products of leaf blade and endosperm allelic genes of the intercross species; obtaining the theoretical value of relative amount of amplification products of endosperm allelic genes according to the measured value of relative amount of amplification products of leaf blade allelic genes of the intercross species; and counting the measured value and the theoretical value of relative amount of amplification products of endosperm allelic genes of the intercross species to be inspected by using a Chi-square test, calculating the probability P, and judging the intercross species.

Description

technical field [0001] The invention relates to a method for identifying intercross species, in particular to a method for identifying intercross species of maize by using molecular markers. Background technique [0002] Corn is an important food crop in my country and even in the world, and plays an important role in agricultural production. Whether corn can be produced normally is related to the safety of my country's grain production. Seed purity is one of the important indicators of seed quality and the main basis for seed grading. There are many main reasons for the reduction of corn seed purity in production: insufficient isolation conditions, incomplete emasculation, and mechanical mixing during harvesting [Xue Yanying, Chen Xingkui, Fan Yan, Chen Xiaodan. SSR molecular marker technology in hybrid maize Application in Seed Purity Identification. Miscellaneous Cereal Crops, 2007, 27(1): 6-7]. The resulting seed hybridization methods are also various, including hybri...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 蒋璐张体付赵涵
Owner JIANGSU ACAD OF AGRI SCI
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