Molecular marking method used for rice cytoplasm identification

A molecular marker and cytoplasmic technology, applied in the field of molecular markers for rice cytoplasmic identification, can solve the problems of cumbersome and complicated operations, time-consuming and labor-intensive operations, and achieve the effect of overcoming cumbersome and complicated operations, simple equipment, and reliable and stable results

Active Publication Date: 2014-07-16
江西省超级水稻研究发展中心
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Problems solved by technology

[0006] In related technologies, for rice, RFLP molecular markers are generally used to study its evolution analysis and kinship; however, RFLP molecular markers require a series of operations such as enzyme digestion, gel electrophoresis, membrane transfer, and radiolabeled probe hybridization. , the whole operation is cumbersome, time-consuming and labor-intensive

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  • Molecular marking method used for rice cytoplasm identification
  • Molecular marking method used for rice cytoplasm identification
  • Molecular marking method used for rice cytoplasm identification

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Embodiment 1

[0039] In the embodiment of the present invention, a molecular marker method for identification of rice cytoplasm is provided, including the following steps, please refer to figure 1 :

[0040] Step 101: analyze and compare the mitochondrial genomes of CW, Lead, Nipponbare and 9311, and screen out non-conserved MS sequences;

[0041] CW, Lead, Nipponbare and 9311 are the four rice varieties whose mitochondrial genome sequencing has been completed so far. 9311 is a typical indica rice variety. The CW cytoplasm is derived from Chinese common wild rice W1 (Oryza rufipogon), and the LD cytoplasm is derived from the indica rice variety Lead in Myanmar. In rice, the largest mitochondrial genome is CW (559,045bp), while LD is only 434,745bp. Synteny analysis shows that Nipponbare and 9311 are relatively conservative, while CW and LD differ from the other two sequenced rice genomes in sequence composition and arrangement Therefore, there are a large number of specific sequences (MS) ...

Embodiment 2

[0057] The molecular marker method for rice cytoplasmic identification of the present embodiment comprises the following steps:

[0058] 1. Analyze and compare the mitochondrial genomes of CW, Lead, Nipponbare and 9311, and screen out non-conserved MS sequences;

[0059] In this step, specifically, the mitochondrial genomes of CW, Lead, Nipponbare and 9311 were analyzed and compared using BLAST2 software, and 55 non-conserved MS sequences were screened out.

[0060] BLAST2 software is accurate for the comparison of mitochondrial genomes of different varieties of rice, and 55 non-conserved MS sequences can ensure the number of corresponding primers, thereby increasing the probability of high polymorphism in subsequent amplification.

[0061] 2. Design corresponding molecular marker PCR primers according to the MS sequence;

[0062] The above-mentioned steps are the same as steps 101-102 of the first embodiment, and will not be repeated here.

[0063] 3. Extract the total DNA ...

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Abstract

The invention relates to the field of rice utilization, in particular to a molecular marking method used for rice cytoplasm identification. The molecular marking method comprises the steps that CW, Lead, Nipponbare and 9311 mitochondria genomes are analyzed and compared, and non-conservative MS sequences are screened out; corresponding molecular marking PCR primers are designed according to the MS sequences; total DNA of different kinds of rice is extracted; the total DNA is used as a template, PCR amplification is conducted with the molecular marking PCR primers as primers and amplification products are obtained; electrophoresis detection is conducted on the amplification products, the polymorphism primers causing amplification differences are screened out and target strips obtained by amplifying the polymorphism primers are obtained; the target strips obtained by amplifying the polymorphism primers are counted, and a cluster graph is established so as to conduct genetic relationship analysis. The molecular marking method used for rice cytoplasm identification is simple and convenient to operate, simple in needed equipment and reliable and stable in result.

Description

technical field [0001] The invention relates to the field of rice utilization, in particular to a molecular marker method for identifying rice cytoplasm. Background technique [0002] Due to the recombination of the mitochondrial genome, the sequence of the plant mitochondrial genome is very different. A comparative analysis of the intergenic regions of the mitochondrial genomes of any two plant species (even homologous species) reveals that most of the intergenic regions are not conserved. . Therefore, plant mitochondrial genomes have their own specific sequences (Mitochondrial-specific sequences, referred to as MS). [0003] For example, comparing the mitochondrial genomes of wheat Ks3 and Km3, only 85.2% of the sequences are conserved sequences, and Ks3 contains 11.38% of specific sequences that Km3 does not have. Compared with the NCBI database, 7.3% of the Ks3 sequences are new sequences, most of which Specific sequences are located in intergenic regions where the rat...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q2565/125
Inventor 谢红卫钱明娟蔡耀辉颜满莲毛凌华李永辉聂元元颜龙安
Owner 江西省超级水稻研究发展中心
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