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Maize phosphate starvation responses intron length polymorphism marker for corn

A length polymorphism and intron technology, applied in the field of molecular biology and genetic breeding, can solve the problem of high cost, and achieve the effect of low cost, strong applicability and simple operation

Inactive Publication Date: 2014-04-02
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most SSR markers are located in intergenic regions
In recent years, due to the development of genome research, a large number of InDels and SNPs have been developed into molecular markers, but the detection of such markers is mainly carried out by direct sequencing, Taqman probes, SNaPshot, DNA chip technology and other methods, and the cost is high

Method used

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  • Maize phosphate starvation responses intron length polymorphism marker for corn
  • Maize phosphate starvation responses intron length polymorphism marker for corn

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 ZmCRE1 Gene Intron Length Polymorphism Marker Development

[0029] 1. Genomic DNA extraction

[0030] Genomic DNA was extracted from 30 seedling leaves of maize inbred lines (Table 1, provided by the Maize Research Institute of Sichuan Agricultural University) using the CTAB method.

[0031] serial number Inbred pedigree serial number Inbred pedigree 01 178 American hybrid 78599 selection line 16 Qi205 (VeiAi141×ZhongXi017)×Population70 02 9782 american hybrid 17 Dan599 American hybrid 78599 selection line 03 B73 BSSS group selection system 18 Chang7-2 Huang Zaosi×Wei 95×S901 04 MO17 CL187-2×C103 19 Dan340 Bone Brigade 9 × Pale Corn (Radiation) 05 CML166 P66C1F215-4-1-2-BB-2-BBB 20 S37 SUWAN1 group 06 (CML-306)-B SINT.AM.TSR-19-1-2-3-1-BB-f 21 Jiao51 Guizhou Local Breeding Lines 07 (CML-473)-B P31C4S5B-23-#-#-4-BBBB 22 Dan598 American hybrid 78599 selectio...

Embodiment 2

[0045] Example 2 ZmAMYB5 gene intron length polymorphic marker development

[0046] 1. Extract genomic DNA, same as Example 1.

[0047] 2. PSR-ILP specific primer design

[0048] The gene sequence and cDNA sequence of the ZmAMYB5 gene (GRMZM2G058310) were downloaded in the MaizeGDB database. After Blast sequence alignment and sequence analysis, specific primers were designed for the second intron of ZmAMYB5 gene. The upstream and downstream primers were designed respectively in the 2nd exon region and the 3rd exon region spanning the 2nd intron, and after specific primer screening and optimization, the upstream primer ZmAMYB5-ILP-F and downstream primer ZmAMYB5-ILP-R were obtained ,details as follows:

[0049]ZmAMYB5-ILP-F: 5’ TCTTCTACACCAACCGGAGTG 3’

[0050] ZmAMYB5-ILP-R: 5’ AGTCCCACCTCAATGTCCAC 3’

[0051] 3. PCR amplification and detection

[0052] 3.1 PCR reaction system Same as Example 1.

[0053] 3.2 PCR reaction procedure

[0054] Pre-denaturation at 94°C for ...

Embodiment 3

[0056] Example 3 ZmMGD2 Gene Intron Length Polymorphism Marker Development

[0057] 1. Extract genomic DNA, same as Example 1.

[0058] 2. PSR-ILP specific primer design

[0059] The gene sequence and cDNA sequence of the ZmMGD2 gene (GRMZM2G178892) were downloaded in the MaizeGDB database. After Blast sequence alignment and sequence analysis, specific primers were designed for the 8th intron of ZmMGD2 gene. The upstream and downstream primers were respectively designed in the 8th exon region and the 9th exon region spanning the 8th intron, and after specific primer screening and optimization, the upstream primer ZmMGD2-ILP-F and downstream primer ZmMGD2-ILP-R were obtained ,details as follows:

[0060] ZmMGD2-ILP-F: 5’ CTGGACCAGGTACCATTGCT 3’

[0061] ZmMGD2-ILP-R: 5’ CGGGCAACAAGACTAGCAG 3’

[0062] 3. PCR amplification and detection

[0063] 3.1 PCR reaction system, same as Example 1.

[0064] 3.2 PCR reaction procedure

[0065] Pre-denaturation at 94°C, 5 min; denat...

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Abstract

The invention discloses a maize phosphate starvation responses intron length polymorphism marker. Specific primers, namely PSR-ILP markers, are designed in exon areas on two sides of a cross-intron of a phosphate starvation responses gene. PCR detection, polyacrylamide gel electrophoresis and silver-stain development are carried out in 30 skeleton inbred lines widely applied to production and breeding of corn in China, fragments with different amplified lengths are obtained in different inbred lines, and the silver-stain development is carried out on the PSR-ILP markers in different inbred lines. The molecular marker which is simple to operate, low in cost, and strong in applicability, is provided for QTL positioning of the corn phosphate starvation responses gene and auxiliary breeding of the molecular marker, and a foundation is laid for efficient breeding of phosphate in maize.

Description

technical field [0001] The invention relates to the fields of molecular biology and genetic breeding, in particular to the intron length polymorphism marker of the corn low phosphorus stress response gene. Background technique [0002] As the largest multi-purpose crop such as feed, grain, and industrial raw materials, corn plays a huge role in promoting the development of my country's industry and people's lives. Among them, the inbred line 178 is a typical low-phosphorus-resistant material, 9782, Zheng58, Chang7-2, etc. are typical low-phosphorus-sensitive materials, and the others are mostly intermediate materials. [0003] The depth of research on molecular genetics of phosphorus utilization is extremely disproportionate to its important position in crops, and it lags far behind model plants such as rice, tomato and soybean. The genetic mechanism of plant Phosphate Starvation Responses (PSR) is very complex, and a phosphorus deficiency molecular response network regulat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11
Inventor 高世斌吴玲刘丹何春萌荣廷昭刘海岚苏顺宗卢艳丽曹墨菊张素芝周树峰聂治柳雅珍闫冰
Owner SICHUAN AGRI UNIV
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