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Functional markers of maize recessive kernel male sterile mutant gene ms33 and application of functional markers

A mutated gene, ms33-in1 technology, which can be used in the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc. difficulties, etc.

Active Publication Date: 2019-01-08
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 markers of maize recessive kernel male sterile mutant gene ms33 and application of functional markers
  • Functional markers of maize recessive kernel male sterile mutant gene ms33 and application of functional markers
  • Functional markers of maize recessive kernel male sterile mutant gene ms33 and application of functional markers

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1: Obtaining maize male sterile mutant material

[0045] The test material of the present invention is the normal fertile inbred line of maize Ms33 The sterile strains obtained by gene mutation are obtained by transposon insertion, including two mutant strains, ms33-6029 and ms33-6038. The obtained sterile lines showed complete non-pollen male sterility, respectively as figure 1 , figure 2 As shown, the specific manifestations are: the anthers become smaller, whitish, and the pollen cannot be stained by iodine-potassium iodide.

Embodiment 2

[0046] Example 2: ms33 Mutation site analysis and development of functional markers

[0047] male sterile material ms33-6029 genetic mutation in Ms33 At the +223 site of the gene, there is a 1387bp transposon insertion, and its mutant gene is named ms33-6029 .

[0048] male sterile material ms33-6038 The gene mutation contains two sites, one is a T-G point mutation at +571 and a subsequent deletion of 5 bases at +572-+576; the other is a 732bp transposon after the 3'-UTR insert. The mutant gene named ms33-6038 .

[0049] mutant gene ms33-6029 and ms33-6038 The schematic diagram of the mutation site is shown in image 3 shown.

[0050] 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. Ms33 and mutant ms33 Specific functional co-segregation molecular markers of genes. Primer sequences designed for the inserted transposon sequence or PCR products spa...

Embodiment 3

[0051] Example 3: Development of functional markers ms33-IN1, ms33-IN2, ms33-IN3, ms33-IN4 and ms33-IN5

[0052] In the present invention, for ms33-6029 and ms33-6038 Using Primer5.0 software for primer design, a total of 5 pairs of functional molecular markers were developed: ms33-IN1, ms33-IN2, ms33-IN3, ms33-IN4 and ms33-IN5. Among them, ms33-IN1 and ms33-IN2 can specifically detect nuclear male sterile materials ms33-6029 mutant gene ms33- 6029 , two labeled amplification primers in ms33 Schematic diagram of the position in the gene, in ms33 The corresponding specific position in the gene sequence and the primer sequence corresponding to the marker name and the size of the amplification product are as follows: Figure 4 A, 4B and 4C are shown. ms33-IN3, ms33-IN4 and ms33-IN5 can specifically detect nuclear male sterile materials ms33-6038 mutant gene ms33-6038, The schematic diagram of the position of the amplification primers, the amplification primers are ...

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Abstract

The invention relates to functional markers of a maize kernel male sterile gene ms33 as well as a development method and application of the functional markers. The invention designs five functional markers includes, but is not limited to, the functional markers ms33-IN1 and ms33-IN2 for a mutant ms33-6029 as well as ms33-IN3, ms33-IN4, ms33-IN5 for the mutant ms33-6038, and the functional markerscan be used for specifically detecting the maize mutant ms33-6029 or ms33-6038 as well as the mutant gene ms33 in a maize sterile material transformed therefrom. By utilizing the five functional markers or other similar functional markers involved in the invention, alleles of the kernel male sterile gene ms33 or a fertile gene Ms33 in a sample can be accurately and rapidly identified by PCR and agarose gel electrophoretic detection; the functional markers can be used for identification of male sterile lines and screening of target individuals, have important application values in maize male sterile line cultivation, sterile hybrid seed production and molecular marker-assisted selection, and are of great significance to improvements on maize cross breeding and seed production efficiency, guarantee of the quality of seed production and other aspects.

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 ms33 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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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 万向元董振营安学丽谢科朱涛涛吴锁伟田有辉王彦博刘欣洁李金萍
Owner UNIV OF SCI & TECH BEIJING