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Molecular mark for detection of Zea mays L. opaque mutant 5512G, and applications of same

A mutant and transparent technology, which is applied in the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of declining agricultural traits, no observation of 5512G mutant resistance to insects and diseases, etc., to achieve accurate High performance and easy operation

Inactive Publication Date: 2014-12-03
SHANGHAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, the 5512G mutant has not been observed to have adverse agricultural traits common to o2 mutants, such as decreased resistance to insects and diseases

Method used

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  • Molecular mark for detection of Zea mays L. opaque mutant 5512G, and applications of same
  • Molecular mark for detection of Zea mays L. opaque mutant 5512G, and applications of same
  • Molecular mark for detection of Zea mays L. opaque mutant 5512G, and applications of same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: Using 5512G Mutant (5512G / 5512G) to Cultivate Dominant High Lysine Maize

[0034] Since the 5512G mutant has a 45.70% increase in lysine (lys) content compared to its corresponding wild type, see Figure 4 , so high-lysine maize can be bred by crossing this gene with other elite varieties. By crossing the 5512G mutation homozygote (5512G / 5512G) with other corns with excellent production traits, a semi-dominant high-lysine heterozygote can be obtained, and high-lysine commercial corn can be obtained directly. Recessive homozygotes for dominant high lysine corn kernels can also be isolated in the F2 generation. Recessive homozygous through multi-generation selfing, recurrent selection, selection of seeds with good grain development and high lysine content generation by generation, and finally obtain stable recessive homozygous, high lysine inbred lines or populations, that is, high Lysine corn varieties.

Embodiment 2

[0035] Example 2: Obtaining co-dominant molecular markers and determining the linkage relationship with the 5512G gene

[0036] See the phenotype map of maize kernel homozygous mutant (5512G / 5512G), heterozygous (5512G / +) and wild type (+ / +) figure 1 . F1 (5512G / +) was obtained by crossing homozygous mutant (5512G / 5512G) with homozygous wild type (+ / +), and then selfing to obtain F2 population, see figure 2 . Genotype segregation appeared in the F2 population, including + / +, 5512G / + and 5512G / 5512G three genotypes, and the grains with + / + genotype were all non-Opaque wild type grains, with 5512G / + genotype The grains are all semi-Opaque heterozygous grains, and the grains with 5512G / 5512G genotype are Opaque mutant types.

[0037] The present invention extracts the genomic DNA of homozygous mutants, wild-types and individual plants of the F2 population through the constructed F2 population, uses the method of map-based cloning, and obtains the polymorphism and compares it ...

Embodiment 3

[0045] Example 3: Polymorphism identification of molecular markers among different parents

[0046] Genomic DNA of 5512G / 5512G mutant, wild type and inbred lines widely used in various breedings was extracted, and the polymorphism among different varieties was analyzed by PCR reaction. The results show that 4.9 has polymorphisms in 5512G and CIMBL62, CIMBL146, By807, Ry732, P178, B73, SC55, JY01, Liao5263, TY5, 7327, GEMS14 (see attached Figure 8 ), 5.7 has polymorphisms in 5512G with CIMBL1, CIMBL62, CIMBL146, Zi330, S22, chuan48-2, CA47, Z2018F, 05WN230, JY01, XZ698, Liao5263, TY5, Zheng28, DH29, B77, GEMS53 (see attached Figure 9 ). This result proves that molecular markers 4.9 and 5.7 can accurately detect the existence of 5512G gene in common populations with multiple breeding backgrounds.

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Abstract

The invention relates to a molecular mark of a high lysine gene in Zea mays L., and applications of the same. The molecular mark is InDel molecular mark-4.9 and 5.7 which both can be amplified through a PCR reaction and detected through an agarose gel electrophoresis, see base sequences represented by SEQ ID No: 1. By using the molecular mark in complete linkage with 5512G gene, genotype identification can be carried out on DNA of any organization of Zea mays L. in any period, and it is detected whether the 5512G in each individual plant of cross breeding filial generation exists or not in a heterozygosis or homozygous state Simultaneously, by using the molecular mark, the mutant and 22 domestic mainly-breeding parents can be distinguished, and the existence of 5512G can be detected accurately in any breeding background populations. The detection method is high in accuracy and simple in operation, and provides an important technology means for breeding assisted by DNA molecular mark of 5512G.

Description

technical field [0001] The invention relates to a molecular marker for detecting the maize opaque phenotype mutant 5512G and its application. technical background [0002] Maize (Zea mays L.) is one of the three major food and feed crops. According to FAO statistics, in the ten years from 2001 to 2010, the total output of corn in the world ranked first among cereal crops, and the gap between the total output of the other two major cereal crops widened year by year; 34%, while wheat and rice accounted for 27% and 28%, respectively. As the "king of feed", corn plays a pivotal role in the development of animal husbandry and breeding, which puts forward higher requirements for the improvement of the nutritional content and protein quality of corn. [0003] However, ordinary corn grains lack essential amino acids, such as lysine and tryptophan, and the protein quality is poor. Long-term single consumption will cause severe malnutrition. Therefore, scientists and breeding exper...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 宋任涛吴悄陈逸涵王冠朱杰唐远平梅冰
Owner SHANGHAI UNIV
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