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Functional marker of maize recessive nuclear male sterility mutant gene ms1 and applications thereof

A technique for mutating genes and nuclear males, applied in the field of functional markers of the recessive nuclear male sterility gene ms1 in maize, can solve problems such as lagging identification period, incomplete linkage between marker traits and sterility, and difficulty in marker trait identification

Inactive Publication Date: 2018-12-07
UNIV OF SCI & TECH BEIJING +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, since the discovery of maize recessive genetic sterile materials, people have used traditional breeding techniques to explore and study various marker traits of the genetic sterile gene, such as using the close linkage relationship between marker traits and sterility, The grain color marker system method, the yellow-green seedling linkage marker method, and the multi-filament linkage marker system have been developed. However, due to the incomplete linkage between marker traits and sterility, difficulties in the identification of marker traits, and delays in the identification period, these methods and attempts have been in the field. It has not been widely used in corn production

Method used

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  • Functional marker of maize recessive nuclear male sterility mutant gene ms1 and applications thereof
  • Functional marker of maize recessive nuclear male sterility mutant gene ms1 and applications thereof
  • Functional marker of maize recessive nuclear male sterility mutant gene ms1 and applications thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0033] Example 1: Obtaining maize male sterile mutant material

[0034] The test material of the present invention is the corn B73 inbred line Ms1 The sterile strain obtained by gene mutation is obtained by the method of transposon insertion. The obtained sterile lines showed complete non-pollen male sterility, such as figure 1 As shown, the specific manifestations are: the anthers become smaller, whitish, and the pollen cannot be stained by iodine-potassium iodide. The CMS material is named ms1-alb .

Embodiment 2

[0035] Example 2: ms1 Mutation site analysis and development of functional markers

[0036] male sterile material ms1-alb yes Ms1 caused by point mutations. Such as figure 2 As shown, the mutant gene ms1 The coding region of the wild type did not have any difference, but in ms1 A 3980bp transposon sequence DTA-ZM00023 was inserted at the -51 site upstream of the gene start codon.

[0037] According to the position and sequence of the transposon insertion, a series of primers targeting the wild type can be developed using primer design software. Ms1 and mutant ms1 Specific functional co-segregation molecular markers of genes. Primer sequences contained within the inserted transposon sequence DTA-ZM00023 or across transposon boundaries can be used as ms1 Molecular markers of mutant genes. The primer sequence of the PCR product flanking the transposon, or the primer sequence of the PCR product spanning the entire transposon, can not be effectively amplified because ...

Embodiment 3

[0038] Example 3: Development of functional markers ms1-IN1, ms1-IN2 and Ms1-1, Ms1-2

[0039] In the present invention, for ms1 For the mutation site, Primer5.0 software was used for primer design, and four pairs of functional molecular markers were developed: ms1-IN1, ms1-IN2 and Ms1-1, Ms1-2. Among them, ms1-IN1 and ms1-IN2 can specifically detect nuclear male sterile materials ms1-alb mutant gene ms1 . Ms1-1 and Ms1-2 are wild type Ms1 Gene detection markers. The schematic diagram of the position of each labeled amplification primer is shown in image 3 As shown in A, each labeled amplification primer is ms1 The corresponding specific position in the gene sequence is as follows image 3 As shown in B, the primer sequence corresponding to the marker name and the size of the amplification product are as follows image 3 C shown.

[0040] The mutant functional marker ms1-IN1 includes the first primer ms1-IN-1F and the second primer ms1-IN-1.2R, which can be used in...

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Abstract

The invention relates to a functional marker of a maize nuclear male sterility gene ms1, a functional marker developing method and applications of the functional marker. According to the mutation siteof the mutant gene ms1, the functional marker which comprises two functional markers ms1-IN1 and ms1-IN2 is designed to ensure that a maize mutant ms1-alb and the mutant gene ms1 in a maize sterile material transferred from mutant can be specifically detected; and by utilizing the ms1-IN1, the ms1-IN2 or other similar functional markers and functional markers Ms1-1, Ms1-2 or other similar functional markers of a corresponding wild type allele Ms1, the situation of a nuclear male sterility gene Ms1 allele in a sample can be quickly identified through PCR (Polymerase Chain Reaction) detection and agarose gel electrophoresis detection. By utilizing the functional marker designed by the invention, the allele genotypes of the maize nuclear male sterility gene ms1 or a fertile gene Ms1 can be accurately detected and marked; the functional marker can be used for identification of male sterile strains and screening of target single stains; and the functional marker has important application value in maize male sterile strain breeding, sterile hybrid seed production and molecular marker-assisted selection and has important significance for improving maize cross breeding efficiency and seedproduction efficiency, guaranteeing seed production quality and the like.

Description

technical field [0001] Generally, the present invention is in the fields of crop molecular breeding, molecular biology and genetic engineering. Concretely relates to a kind of maize recessive male sterility gene ms1 Functional flags, functional flag development methods and applications. Background technique [0002] Plant male sterility (male sterility, MS) refers to the abnormal development of male organs in higher plants, unable to produce functional male gametes (pollen), but the normal development of female organs, which can receive normal male gametes for fertilization and fruiting, and can The phenomenon in which sterility is passed on to offspring. [0003] Maize is an important food crop, and its male sterile materials are of great value in the study of maize male flower development mechanism, genetic breeding application, heterosis research and application. Maize male sterility can be divided into three types according to the differences in sterility inheritance ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 万向元吴锁伟安学丽谢科李金萍刘欣洁柳双双张春霞董振营田有辉
Owner UNIV OF SCI & TECH BEIJING
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