Glycoprotein elicitor of magnaporthe grisea in rice leaves and its purification method

A rice blast fungus and elicitor technology, which is applied in the field of purification of rice blast fungus glycoprotein elicitor, can solve the problems of complex genetic background, single blast resistant variety, severe susceptibility, etc., and achieves high induction activity, low price and easy The effect of cultivation

Inactive Publication Date: 2008-02-13
SOUTH CHINA AGRI UNIV
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Problems solved by technology

However, due to the complex genetic background of Magnaporthe grisea, it is easy to mutate, and the blast-resistant varieties are relatively simplistic, and the genetic homogeneity of the main varieties makes it difficult for disease-resistant breeding to keep up with the mutation speed of the pathogenic race of Magnaporthe oryzae. Often results in a new blast-resistant rice variety being severely susceptible 3-5 years after planting in a large area

Method used

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  • Glycoprotein elicitor of magnaporthe grisea in rice leaves and its purification method
  • Glycoprotein elicitor of magnaporthe grisea in rice leaves and its purification method
  • Glycoprotein elicitor of magnaporthe grisea in rice leaves and its purification method

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Embodiment Construction

[0034] The activated rice blast fungus mycelium was cultured in liquid (liquid medium: yeast extract 5g; glucose monohydrate 22g; supplemented with distilled water to 1000mL.), 25 ℃ constant temperature shaking culture for 8-10 days, double-layer gauze filtration, double Repeated washing with distilled water 3 times, frozen at -20 ℃. Thaw the frozen mycelium at 15°C-25°C, then freeze at -20°C, and freeze and thaw repeatedly 5-7 times to fully break the mycelial cells. The freeze-thawed mycelium (5mL / g) was suspended with double distilled water, and homogenized repeatedly for 5-7 times with a high-speed tissue masher (DS-1 tissue masher) at 12,000 r / min. The homogenate was stirred at 4°C for 10-12 hours with a magnetic stirrer, centrifuged at 10,000g for 30min, and the supernatant was taken. The supernatant was filtered with filter paper, and the obtained filtrate was ultra-filtered with PM-10 membrane with a high-purity nitrogen pressure of 0.45MPa. The obtained concentrated ...

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Abstract

The present invention first obtains coarse exciton with rice blast germ mycelium as material and through culturing, freezing-thawing, homogenating, centrifuging and PM-10 membrance ultrafiltering, and then obtain the new rice blast germ glucoprotein exciton through further freezing-thawing and serial chromatographic purifications. The rice blast germ glucoprotein exciton has molecular weight 49.08 kDa, isoelectric point 6.01 and silver staining electrophoresis purity. Mass spectroscopic identification shows that the exciton is rice blast germ MG05155.4 open reading frame coded hypothetical protein MG05155.4. The exciton has stable property, relatively high induction on Phe deaminase activity in non-compatible interactive rice line and lowest effective concentration of 1.5 nmol / L for inducing rice peroxidase. The present invention has excellent application foreground in the biological prevention and control of rice blast.

Description

technical field [0001] The invention relates to the field of plant protection and biotechnology, in particular to the purification of a novel glycoprotein elicitor of M. oryzae. Background technique [0002] Rice blast disease is caused by the ascomycete Magnaporthe grisea (Hebert) Barr (its asexual generation is Pyricularia grisea (Cooke) Sacc.), which is widely distributed in countries and regions where rice is cultivated. It is one of the three major diseases of rice, which seriously limits the high yield, stable yield and high quality of rice. Rice blast occurs in varying degrees throughout the year, and the output is generally reduced by 10-20% in epidemic years, and more than 50% in severe cases. Since the 1990s, the annual incidence of rice blast in my country has exceeded 3.8 million hectares, and the average annual loss of rice has reached hundreds of millions of kilograms. [0003] At present, the main control measures for rice blast are selection of disease-resi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/195C07K1/34
Inventor 纪春艳王振中
Owner SOUTH CHINA AGRI UNIV
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