Extracting method of RNA of rubber tree powdery mildew

A powdery mildew fungus and extraction method technology, applied in the biological field, can solve the problems of inability to obtain powdery mildew spores in a uniform state, lack of powdery mildew tissue culture collection and RNA extraction methods, and inability to carry out functional gene expression research and other problems.

Inactive Publication Date: 2015-06-10
HAINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The improved Trizol method collects and extracts powdery mildew is actually a mixture of spores and hyphae tissue RNA, and it is impossible to obtain spores of powdery mildew with uniform germination and appress

Method used

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  • Extracting method of RNA of rubber tree powdery mildew
  • Extracting method of RNA of rubber tree powdery mildew
  • Extracting method of RNA of rubber tree powdery mildew

Examples

Experimental program
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Example Embodiment

[0029] Example 1

[0030] (1) Select the 7-10-day-old leaves of the grafted seedlings of the rubber 7-33-97 strain as the culture O.heveae Materials, fresh conidia of HO-73 at 24 h after inoculation for 12 days were used as the source of experimental bacteria. Gently shake the leaves 24 hours before inoculation to shake off the old spores, and use the new 24-hour-old conidia as the inoculation material to ensure the uniformity of the spore viability and the development of the infection structure in the experiment; place the Parafilm (BEMIS product from the United States) film on the bottom to have a moisturizing effect. In the fresh-keeping box (40 cm×30 cm) of filter paper, use a soft brush to brush the powdery mildew spores on the rubber blade on the surface of the Parafilm film, the culture temperature is 22 °C, the relative humidity is 50%~80%, and the fresh-keeping box is sealed in a dark place. save;

[0031] (2) After transferring to the surface of the Parafilm membr...

Example Embodiment

[0037] Example 2

[0038] (1) Select the 7-10-day-old leaves of the grafted seedlings of the rubber 7-33-97 strain as the culture O.heveaeMaterials, fresh conidia of HO-73 at 24 h after inoculation for 10 d were used as the source of experimental bacteria. Gently shake the leaves 24 hours before inoculation to shake off the old spores, and use the new 24-hour-old conidia as the inoculation material to ensure the uniformity of the spore viability and the development of the infection structure in the experiment; place the Parafilm (BEMIS product from the United States) film on the bottom to have a moisturizing effect. In a fresh-keeping box (40 cm × 30 cm) of filter paper, use a soft brush to brush the powdery mildew spores on the rubber blade on the surface of the Parafilm film, the culture temperature is 21 °C, the relative humidity is 50%~80%, and the fresh-keeping box is sealed in a dark place. save;

[0039] (2) 24 hours after transferring to the surface of the Parafilm ...

Example Embodiment

[0045] Example 3

[0046] (1) Select the 7-10-day-old leaves of the grafted seedlings of the rubber 7-33-97 strain as the culture O.heveae Materials, fresh conidia of HO-73 at 24 h after inoculation for 10 d were used as the source of experimental bacteria. Gently shake the leaves 24 hours before inoculation to shake off the old spores, and use the new 24-hour-old conidia as the inoculation material to ensure the uniformity of the spore viability and the development of the infection structure in the experiment; place the Parafilm (BEMIS product from the United States) film on the bottom to have a moisturizing effect. In a fresh-keeping box (40 cm × 30 cm) of filter paper, use a soft brush to brush the powdery mildew spores on the rubber blade on the surface of the Parafilm film, the culture temperature is 21 °C, the relative humidity is 50%~80%, and the fresh-keeping box is sealed in a dark place. save;

[0047] (2) After being transferred to the surface of Parafilm and cul...

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Abstract

The invention relates to the field of biotechnology and discloses an extracting method of RNA of rubber tree powdery mildew. The powdery mildew is induced to germinate on hydrophobic media and form appressoria, different growth periods of the powdery mildew are convenient to distinguish, and the purpose of extracting the RNA in a grading mode is achieved. By extracting the RNA of the rubber tree powdery mildew at different periods, it is displayed that the concentration of the RNA and integrity meet the Illumina transcriptome library requirement through quantitative determination, 18SrRNA genes are amplified through RT-PCR, and a foundation is further provided for transcriptome sequencing analysis and gene expression research.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for extracting RNA of rubber tree powdery mildew fungus. Background technique [0002] Rubber tree powdery mildew is caused by rubber powder spores of the genus Oidium heveae B.A. Steinmann) infection, which is widely distributed in rubber planting areas around the world, and is also the focus of early spring prevention and control of "two diseases and one insect" in rubber planting areas in my country. Powdery mildew of rubber tree is an obligate parasite, which cannot be cultured in vitro in artificial medium, and is difficult to genetically manipulate. More research focuses on cytological observation and control. At present, no rubber tree strain resistant to powdery mildew has been cultivated for large-scale production. About The molecular experimental system for the functional verification of the pathogenic gene of powdery mildew of rubber tree has not been established...

Claims

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Application Information

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IPC IPC(8): C12N15/10C12R1/645
Inventor 缪卫国梁鹏刘文波郑服丛林春花
Owner HAINAN UNIVERSITY
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