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SRAP (sequence-related amplified polymorphism) molecular marker method for detecting rice mitochondrion karyotype

A molecular marker, mitochondrial technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of lack of specificity and no application value, and achieve the effect of simple operation, improved efficiency and wide application range.

Inactive Publication Date: 2012-07-18
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Molecular Plant Breeding, 2004, 2(2): 223-228), lacks specificity, and has no application value in actual production

Method used

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  • SRAP (sequence-related amplified polymorphism) molecular marker method for detecting rice mitochondrion karyotype
  • SRAP (sequence-related amplified polymorphism) molecular marker method for detecting rice mitochondrion karyotype
  • SRAP (sequence-related amplified polymorphism) molecular marker method for detecting rice mitochondrion karyotype

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Using the SRAP molecular marker to detect rice mitochondrial karyotype to identify rice cytoplasmic male sterile line mitochondrial karyotype, a set of SRAP (Sequence-related Amplification Polymorphism) molecular marker method for qualitative detection of rice mitochondrial karyotype, the specific steps are as follows:

[0039] A. Mitochondrial SRAP molecular marker PCR primer design:

[0040] The PCR primers for mitochondrial sequence-specific SRAP molecular markers are listed in the table below, and its parameters include the length of the amplified fragment and the annealing temperature.

[0041]

[0042] B. Material management and DNA extraction:

[0043] Select 10 different types of rice cytoplasmic male sterile lines: Jihexi 41A, Gang 46A, II-32A, Xieqingzao A, Maxie A, K17A, Chujing 23A, Baoyuan A, Zhenshan 97A and Yuetai A( Figure 1~2 1-10 are numbered in order respectively). When it grew to two leaves and one heart, 2 grams of seedlings were taken from ea...

Embodiment 2

[0058] Using the SRAP molecular marker to identify the rice mitochondrial karyotype to detect the mitochondrial karyotype of rice conventional varieties, a SRAP molecular marker method for detecting the rice mitochondrial karyotype, the specific operation is as follows:

[0059] A. Mitochondrial SRAP molecular marker PCR primer design:

[0060] The PCR primers for mitochondrial sequence-specific SRAP molecular markers are designed in the following table, and its parameters include the length of the amplified fragment and the annealing temperature.

[0061]

[0062] B. Material management and DNA extraction:

[0063] Select 20 copies of farm materials: SA-Phi-1, SA-Ind-1, SA-Cam-1, SA-Nep-1, SA-Ban-1, SA-Ban-1, SA-Ind-2, SA-Ind-3, SA-Ban-2, SA-Ind-4, SA-Pak-2, Gla-Cam-1, SA-Cam-2, SA-Mya-1, Gla-Gui-1, Rhi- Sri-1, Niv-Sri-1, SA-Ban-3, NR-Mya-1, Niv-Ban-1 ( image 3 1-20 as the code number in sequence). When it grew to two leaves and one heart, 2 grams of seedlings were ta...

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Abstract

The invention discloses an SRAP (sequence-related amplified polymorphism) molecular marker method for detecting rice mitochondrion karyotype. The SRAP molecular marker method includes steps: firstly, designing an SRAP marker primer and characteristic parameters of the SRAP marker primer; secondly, building a PCR (polymerase chain reaction) amplification system, selecting a rice cytoplasmic male sterile line material, leading the rice cytoplasmic male sterile line material to grow until the rice cytoplasmic male sterile line material has two leaves and a core, taking seedlings from the materials, grinding the seedlings in liquid nitrogen into powder and extracting the total DNA (deoxyribose nucleic acid); thirdly, creating a PCR amplification system, and leading the standard PCR amplification reaction system to be 25 mul; and fourthly, realizing statistic analysis for a detection result of mitochondrion karyotype molecules. The SRAP molecular marker method is based on rice mitochondrion genome heritable variation, and a set of molecular markers with rich polymorphism and flexibility in detection is created by widely screening mitochondrion repeat regions. Rice varieties and geneticresources are quickly and accurately detected by an experimental means. The method is simple and convenient in operation, is stable and reliable, can be used for identifying different cytoplasmic male sterile lines and the rice varieties and has the advantages of accuracy, high efficiency and speediness, and efficiency of identifying the sterile lines and new varieties is improved.

Description

technical field [0001] The present invention relates to the technical field of molecular marker detection, more specifically to a method for qualitatively detecting mitochondrial kenotypes in rice sterile lines and germplasm resources, which is applicable to the identification and detection of various conventional rice varieties, hybrid rice combinations and sterile lines . Background technique [0002] Rice mitochondria are inherited maternally, so the source of rice cytoplasm can be traced by mitochondrial markers. Although the mitochondrial mutation rate is considered to be slower than that of the nucleus, the intergenic regions of the mitochondria are prone to recombination or exchange of genomes in cells, so the mutations are also very rich. This difference can serve as a specific molecular signature of the mitochondrial genome at the genetic level. Mitochondrial karyotypes can be marked by using mitochondrial molecular tags, so different rice varieties can be effecti...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 李绍清谢红卫钱明娟朱英国
Owner WUHAN UNIV