Molecular specificity mark primer for authenticating mushroom strains 241 and 241-4 and detecting method

A technology of labeling primers and specificity, applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc. Difficult to achieve fast and accurate identification and other problems, to achieve the effect of short detection time, high accuracy and simple method

Inactive Publication Date: 2010-05-26
ZHEJIANG FORESTRY ACAD
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Problems solved by technology

Because these methods are susceptible to the impact of environmental factors and physiological conditions, the strains with less difference in morphological characteristics, especially some mushroom related species such as 241 and 241-4, Cr02 and Cr04, Shenxiang 2, Shenxiang 4 and Shenxiang 6, 939-9 and 939-6 are difficult to achieve rapid and accurate identification
[0004] In addition, due to the genetic characteristics of asexual reproduction of shiitake mushroom strains, the same species with different names, different species with the same name, false species, and inferior species are common in the strain market. In addition, the degradation of strain quality has not only greatly affected It harms the interests of shiitake mushroom producers and breeders, and also greatly affects the sustainable and healthy development of my country's edible fungus industry

Method used

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  • Molecular specificity mark primer for authenticating mushroom strains 241 and 241-4 and detecting method

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Embodiment 1

[0046](1) Mycelia culture and genomic DNA extraction: transfer the low-temperature-preserved shiitake mushroom strains (the production strains of shiitake mushroom 241 and 241-4 are provided by Qingyuan County Edible Fungi Research Center) to PDA slant (PDA slant medium: Take 200 grams of peeled potatoes, cut them into small pieces, add 1000 ml of water to boil for 30 minutes, filter out the potato pieces, add water to the filtrate to make up to 1000 ml, add 20 grams of glucose, 15 grams of agar, dissolve and pack, and sterilize at 121 °C After cooling for 30 minutes, make a slant) and incubate at 25°C. After 14 days, receive 100ml of PD liquid medium (PD liquid medium: take 200 grams of peeled potatoes, cut into small pieces, add 1000ml of water and boil for 30 minutes, filter out the potato pieces, add water to the filtrate to 1000ml, add glucose 20 gram, sterilized at 121°C for 30 minutes, cooled to obtain PD liquid medium), cultured with shaking at 100 rpm / min at 25°C for ...

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Abstract

The invention provides a molecular specificity mark primer for authenticating mushroom strains 241 and 241-4 and a detecting method for differentiating and authenticating the mushroom strains 241 and 241-4 by using same. The sequence of an upstream primer is 5'-GGTTCTGCTCCTTGTGACTG-3', and the sequence of a downstream primer is 5'-TCTGCTCTTCAGATGCAAGCT-3'. Compared with convectional morphology detection, antagonism tests and fruiting tests, the invention has the advantages of short detection time and high accuracy. The detection time in the method is only 1 to 2 days, but the time of the convectional antagonism tests is at least two weeks, and the time of the fruiting tests is at least three months.

Description

(1) Technical field [0001] The invention relates to a molecular-specific marker primer capable of distinguishing mushroom-producing close relatives 241 and 241-4, and a method for distinguishing and identifying mushroom-producing strains 241 and 241-4 using the primer. (2) Background technology [0002] Lentinula edodes (Berk.) Pegler is loved by domestic and foreign consumers because of its delicious taste and important food and medicinal value. It is an edible fungus with the largest commercial production scale in my country and second only to Agaricus bisporus in the world. . China is the first country to cultivate shiitake mushrooms, and the current output of shiitake mushrooms has reached 70% of the world. [0003] Lentinus edodes strains are the most important means of production in the production of shiitake mushrooms. At present, there are more than 100 kinds of shiitake mushrooms used in commercial cultivation in China. Different shiitake mushroom strains have diff...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12N15/11
Inventor 李海波吴学谦付立忠魏海龙吴庆其贺亮程俊文
Owner ZHEJIANG FORESTRY ACAD
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