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Lysinecorn SSR (simple sequence repeat) molecular marker auxiliary selecting and breeding method

A molecular marker-assisted, high-lysine technology, applied in the field of high-lysine corn SSR molecular marker-assisted selection breeding, can solve the problems of long breeding cycle, high cost of lysine determination, low efficiency, etc., and achieve the goal of breeding cycle Short, speed up the breeding process, high efficiency effect

Inactive Publication Date: 2011-08-17
HENAN ACAD OF AGRI SCI
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Problems solved by technology

but o 2 The gene is a recessive gene. Conventional breeding methods transfer common inbred lines into QPM near-isogenic lines. One generation of backcross must be selfed. More than 5 generations, and finally 3 to 4 generations of self-crossing, the breeding cycle is very long, the cost of lysine determination is high, and the efficiency is low

Method used

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  • Lysinecorn SSR (simple sequence repeat) molecular marker auxiliary selecting and breeding method
  • Lysinecorn SSR (simple sequence repeat) molecular marker auxiliary selecting and breeding method
  • Lysinecorn SSR (simple sequence repeat) molecular marker auxiliary selecting and breeding method

Examples

Experimental program
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Effect test

Embodiment 1

[0031] Example 1 SSR marker screening

[0032] o 2The polymorphisms of the three microsatellite markers Phi057, Phi112, and umc1066 among the 12 inbred lines are shown in figure 1 . It can be seen from the figure that at o 2 Among the three primers in the gene, primer 5 (umc1066) can distinguish Zheng 58, Chang 7-2, Ye 478 and two o 2 Gene donor inbred lines CA339 and CA042. While the other 2 primers could not be distinguished at the same time. Primer 3 (Phi057), no polymorphism between No. 1 (Zheng 58) and No. 11 (CA339); between No. 6 (Chang 7-2), No. 9 (Ye 478) and No. 8 (CA042) All have good polymorphisms, but the No. 8 band type is not clear. Primer 4 (Phi112), there is no polymorphism between No. 1 (Zheng 58) and No. 11 (CA339); there is a good polymorphism between No. 6 (Chang 7-2) and No. 8 (CA042) , There is no polymorphism between No. 9 (tuck 478) and No. 8 (CA042). Primer 5 (umc1066) has good polymorphism between No. 1 (Zheng 58) and No. 11 (CA339), and poly...

Embodiment 2

[0034] Example 2 Using SSR Molecular Marker Backcross Breeding to Create Near-Isogenic Lines

[0035] F 1 ;Second season: the recurrent parent was backcrossed with common corn, and BC was obtained 1 f 1 ;Third season: sow BC in ear rows 1 f 1 , each group selects 100 individual plants with good agronomic traits and genetic background close to the corresponding reincarnated parents for sampling, extracts DNA, detects genotypes, and obtains the successfully transferred o 2 Gene single plant, continue to backcross to get BC 2 f 1 ;Fourth season: sow BC in panicle rows 2 f 1 Similarly, each group selects 100 individual plants with better agronomic traits and genetic background close to the corresponding reincarnated parents for sampling, extracts DNA, and detects genotypes, and obtains the ones that have been successfully transferred into o 2 single plant of the gene. Then self-cross 2 generations. A total of 1 generation of crossing, 2 generations of backcrossing and 2 ...

Embodiment 3

[0040] Example 3 Field test of transformed materials and identification of genotype SSR molecular markers

[0041] It has been carried out in the experimental farm of Henan Academy of Agricultural Sciences since 2000, and one generation is multiplied in Hainan every year. A random block design was adopted, with single-row plots, the row length of each plot was 3.5m, the row spacing was 0.6m, the plant spacing was 0.2m, and each row had 18 plants. From the seedling stage, the comprehensive agronomic traits were observed in the field, and the genotype SSR molecular markers of the transformed materials were identified.

[0042] Directly observe the external shape of corn endosperm and count various structural types. The grain endosperm structure of high lysine near-isogenic lines transformed from common maize inbred lines changed greatly, but the endosperm types and textures of different maize germplasms were different. In Zheng 584 and 478 of the horse-tooth type, there are on...

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Abstract

The invention provides a lysinecorn SSR (simple sequence repeat) molecular marker auxiliary selecting and breeding method. By combining a backcross transforming method with a molecular marking method, the method successfully constructs the lysinecorn isogenic gene systems QZ58, Q478 and QC72. After the SSR molecular marker auxiliary selecting and breeding method is used the mono-clones which contain the O2 gene can be detected by SSR molecular marker in the O2 gene after one generation is backcrossed, and the second generation can be backcrossed without selfing, so that the breeding is short in period, low in cost, and high in efficiency. Aiming at different breeding target requirements, the method is used for performing the gene type detection to the backcross offspring seedling leaves; and the method is not influenced by the environment conditions and combines with south propagating and generation adding, so that the target genes are transferred into a good material within a shorter time.

Description

technical field [0001] The invention relates to the field of crop breeding, in particular to a high-lysine corn SSR molecular marker-assisted selection breeding method. Background technique [0002] Since Mertz et al. discovered in 1964 that the opaque-2 gene could significantly increase the content of lysine and tryptophan in corn kernels, breeders from all over the world have invested in high-quality protein corn breeding. There are a large number of excellent and high-quality protein corn varieties used in production practice. This is because the germplasm resources of high-quality protein corn are relatively poor, and it is difficult to directly combine single-cross varieties. Most breeders use germplasm with high lysine to transform common corn inbred lines into A near-isogenic line with high lysine. However, practice has proved that these near-isogenic lines transformed from common maize have undergone a series of changes, such as a decrease in bulk density, an incre...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 铁双贵岳润清齐建双王延召朱卫红卢彩霞
Owner HENAN ACAD OF AGRI SCI
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