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A dna fingerprint of potato and its acquisition method and its special primers

A technology of DNA fingerprinting and potato is applied in the field of DNA fingerprinting of potato and its acquisition and its special primers, which can solve the problems of chaotic varieties, characters easily affected by environmental conditions, multiple species of the same species, etc., so as to improve the accuracy and efficiency, The effect of shortening the breeding cycle and enriching fingerprints

Inactive Publication Date: 2017-06-13
广西壮族自治区农业科学院经济作物研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the lack of systematic and perfect identification methods for potato varieties in the early days, there were multiple phenomena of the same species and multiple species with the same name, and the confusion of varieties caused great losses to potato production.
Potato variety identification has historically been based on traditional morphological traits that are susceptible to environmental conditions
In addition, the parents of potato varieties bred in my country are basically tetraploid cultivars introduced from Europe in the 1950s and 1960s. difficulty

Method used

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  • A dna fingerprint of potato and its acquisition method and its special primers
  • A dna fingerprint of potato and its acquisition method and its special primers
  • A dna fingerprint of potato and its acquisition method and its special primers

Examples

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Effect test

Embodiment 1

[0031] Example 1 Utilizes special primers to obtain the DNA fingerprint of potato

[0032] Operation process: extraction of potato genomic DNA → PCR amplification → agarose gel electrophoresis → EB staining → obtaining a clear DNA fingerprint → statistical analysis results.

[0033] The specific method and process are as follows:

[0034] 1. DNA extraction

[0035] DNA was extracted from potato samples using the modified specific CTAB method (cetyltrimethylammonium bromide method).

[0036] Include the following sub-steps:

[0037] (1) Take fresh potato leaves, take 0.3-0.5g of each variety, cut them into pieces with scissors, put them into a mortar and add 1g of PVP (polyvinylpyrrolidone) to the mortar, and pour in enough Liquid nitrogen was used to fully grind the leaf samples into powder.

[0038] (2) Transfer all the powder samples to a 2ml centrifuge tube that has been preheated at 65°C with 1.0ml of CTAB extract in advance, then add 20μl of β-mercaptoethanol, and war...

Embodiment 2

[0064] Application of Example 2 Potato DNA Fingerprint

[0065] The potato DNA fingerprint spectrum that embodiment 1 obtains is carried out cluster analysis, obtains cluster dendrogram, as Figure 8 As shown, it can be seen that when the threshold of genetic distance is about 0.55, 9 potato varieties can be divided into 2 groups. The first large group includes 7 potato varieties including Jizhangshu No. 8, Zhongshu No. 13, Zhongshu No. 14, Feiwu Ruita, Zhongshu No. 6, Zhongshu No. 8 and Zhongshu No. 17; the second largest group Only two varieties, Zhongshu No. 11 and Zhongshu No. 7, are included. When the threshold of genetic distance is about 0.36, the first large group can be divided into two subgroups. When the threshold of genetic distance is about 0.30, it can be further divided.

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Abstract

The invention discloses DNA fingerprints of a potato and an obtaining method and a special primer thereof. The special primer for obtaining the DNA fingerprints of the potato is composed of sequences SEQ ID NO:1-7 in a sequence table. The obtaining method of the DNA fingerprints of the potato comprises the following steps that the genome DNA of the potato is extracted; the genome DNA is used as a template, and PCR amplification is conducted through the special primer; agarose gel electrophoresis separation is conducted on a PCR product, and the DNA fingerprints of the potato are displayed through EB staining. The obtained DNA fingerprints of the potato are high in polymorphism, abundant and lay the technical foundation for improving the accuracy and efficiency of potato selective breeding, shortening a breeding period and conducting potato molecular marker-assisted breeding.

Description

technical field [0001] The invention relates to a DNA fingerprint of potato and its acquisition method and special primers. Background technique [0002] Potato is a perennial herbaceous plant belonging to the family Solanaceae, and its tubers are edible. It is the fourth most important food crop in the world. my country is the largest potato production country in the world, and potato breeding in my country is mainly based on inter-variety or inter-specific hybridization. According to incomplete statistics, from the 1940s to 2000, more than 170 potato varieties were bred in China. In recent years, with the acceleration of potato breeding in my country, the number of varieties bred has increased rapidly. From 2000 to 2007, there were nearly 110 varieties approved at the provincial level or above. However, due to the lack of systematic and perfect identification methods for potato varieties in the early stage, the phenomenon of multiple names of the same species and multipl...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6858C12Q1/6895C12Q2531/113C12Q2565/125
Inventor 熊发前刘俊仙蒋菁唐秀梅贺梁琼黄志鹏李忠韩柱强钟瑞春唐荣华何海旺唐洲萍
Owner 广西壮族自治区农业科学院经济作物研究所