Molecule technical method for identifying indica/japonica type of Yunnan rice old variety of Yunnan rice old variety

A molecular technology, rice technology, applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of time-consuming and labor-intensive, indica-japonica differentiation difficult to distinguish, achieve high accuracy, simple and easy identification method, The effect of avoiding red tape

Inactive Publication Date: 2009-04-15
YUNNAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method requires planting the materials in the field and doing a lot of field observations and records, which is time-consuming and labor-intensive, especially when faced with a large number of rice seed resources.
Moreover, it is difficult to distinguish some ol

Method used

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  • Molecule technical method for identifying indica/japonica type of Yunnan rice old variety of Yunnan rice old variety
  • Molecule technical method for identifying indica/japonica type of Yunnan rice old variety of Yunnan rice old variety
  • Molecule technical method for identifying indica/japonica type of Yunnan rice old variety of Yunnan rice old variety

Examples

Experimental program
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Effect test

Example Embodiment

[0020] Embodiment one

[0021] Thirty rice seed resources from high altitude in Yunnan were identified by "morphological index method" and molecular identification method to identify the indica and japonica characteristics of the materials, and the identification results of the two methods were compared to investigate the reliability of the molecular identification method.

[0022] For molecular identification, 5 seeds of the material to be identified were germinated at 20°C, 5 days after germination, the leaves were taken to extract the total DNA of rice, the DNA concentration of the sample was measured and the DNA was quantitatively diluted to 25ng / ul, and then PCR reaction was carried out with the specific primer JI; 2% agarose (containing EtBr0.1ug / ml) gel electrophoresis separation; PCR amplification products and electrophoresis results were detected on an ultraviolet transilluminator, and the indica and japonica characteristics of the material were identified according to...

Example Embodiment

[0032] Embodiment two

[0033] The indica and japonica characteristics of the materials were identified by "morphological index method" and molecular identification method respectively, and the identification results of the two methods were compared to investigate the reliability of the molecular identification method.

[0034] For molecular identification, 5 seeds of the material to be identified were germinated at 35°C, 5 days after germination, the leaves were taken to extract the total DNA of rice, the DNA concentration of the sample was measured and the DNA was quantitatively diluted to 25ng / ul, and then PCR reaction was performed with specific primer JI; 2% agarose (containing EtBr0.1ug / ml) gel electrophoresis separation; PCR amplification products and electrophoresis results were detected on an ultraviolet transilluminator, and the indica and japonica characteristics of the material were identified according to the electrophoresis results. There are two products with mo...

Example Embodiment

[0043] Embodiment three

[0044] 120 accessions of rice seed resources from low altitude in Yunnan were identified by "morphological index method" and molecular identification method to identify the indica and japonica characteristics of the materials, and the identification results of the two methods were compared to investigate the reliability of the molecular identification method.

[0045] For molecular identification, 5 seeds of the material to be identified were germinated at 30°C, 5 days after germination, the leaves were taken to extract the total DNA of rice, the DNA concentration of the sample was measured and the DNA was quantitatively diluted to 25ng / ul, and then PCR reaction was performed with specific primer JI; 2% agarose (containing EtBr0.1ug / ml) gel electrophoresis separation; PCR amplification products and electrophoresis results were detected on an ultraviolet transilluminator, and the indica and japonica characteristics of the material were identified accord...

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Abstract

The invention relates to a method of a molecular authenticating technique for the indica-japonica of old species of Yunnan Rice, which belongs to the breeding field in the technical field of agricultural biotechnology. The technical proposal is as follows: five grains of the material seeds to be authenticated are taken for germinating under the temperature of 20 to 35 DEG C; after the seeds geminate for five days, the leaves are taken for extracting the total DNA of the rice; the concentration of the DNA is detected and the DNA is quantificationally diluted to 25ng/ul; a specific primer JI designed according to the sequence of the specific long-arm section of the 10th chromosome of the rice is used for carrying out PCR reaction; gel electrophoresis is carried out on the 2 percent agarose (containing EtBr0.1ug/ml); the PCR product is detected on a UV transilluminator; and the indica-japonica characteristics of the material are authenticated according to the electrophoresis result. The indica type rice has two PCR products with the molecular weights being respectively 280bp and 380bp; and the japonica rice has one PCR product with the molecular weight of 280bp, thus authenticating the indica-japonica characteristics of the old species of Yunnan Rice. The method has the effects of being quickly and effectively authenticating the indica-japonica characteristics of the old of Yunnan Rice.

Description

technical field [0001] The invention relates to a molecular technology method for identifying indica and japonica rice varieties of old Yunnan rice varieties, which belongs to the field of agricultural biotechnology, and more specifically belongs to the field of breeding. Background technique [0002] my country is one of the world-recognized origin centers of cultivated rice in Asia, one of the countries with the richest rice seed resources and one of the earliest rice-growing countries in the world. Due to its low latitude and large altitude difference (the lowest point is 76 meters above sea level and the highest point is 6,800 meters), Yunnan has obvious three-dimensional climate characteristics. Its geographical and climate diversity has bred rich and diverse rice seed resources. As the center of the richest diversity of ecological and genetic resources, it constitutes one of the most important gene banks of cultivated rice in Asia. [0003] At present, more than 6,000...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 谭亚玲谭学林洪汝科郭艾陈丽娟黄大军张忠林金寿林
Owner YUNNAN AGRICULTURAL UNIVERSITY
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