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

A technology of blast fungus and purification method, which is applied to the preparation method of peptides, chemical instruments and methods, bacterial peptides, etc., can solve the problems of complex genetic background, single blast-resistant variety, and severe disease susceptibility, and achieve high induction activity, Inexpensive, easy-to-grow effects

Inactive Publication Date: 2006-01-18
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

[0024]The activated blast fungus mycelium was cultured in liquid (liquid medium: 5g of yeast extract; 22g of glucose monohydrate; supplemented with distilled water to 1000mL.), 25 ℃ constant temperature shaking culture for 8-10 days, double-layer gauze filter, double-layer gauze Repeated washing with distilled water 3 times, frozen at -20 ℃. The frozen mycelium was thawed at 15°C-25°C, then frozen at -20°C, repeated freezing and thawing 5-7 times, and the mycelial cells were fully broken. Suspend the frozen-thawed mycelia (5mL / g) in double distilled water, and homogenize repeatedly 5-7 times with a high-speed tissue masher (DS-1 tissue masher) at 12,000r / min. Stir the homogenate at 4°C for 10-12 hours with a magnetic stirrer, centrifuge at 10,000 g for 30 min, and take the supernatant. The supernatant was filtered with filter paper, and the resulting filtrate was ultrafiltered with PM-10 membrane under high-purity nitrogen pressure of 0.45 MPa. The obtained concentrated solut...

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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 fields of plant protection and biotechnology, in particular to the purification of a new glycoprotein elicitor of blast fungus oryzae. Background technique [0002] Rice blast disease is caused by the Ascomycete Magnaporthe grisea (Hebert) Barr (its asexual generation is Pyricularia grisea (Cooke) Sacc.), and 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 different degrees all the year round, and the production is generally reduced by 10-20% in popular years, and more than 50% in severe cases. Since the 1990s, the annual occurrence area of ​​rice blast in my country has been more than 3.8 million hectares, and the average annual loss of rice has reached hundreds of millions of kilograms. [0003] At present, rice blast is mainly controlled by breeding r...

Claims

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Application Information

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IPC IPC(8): C07K14/195C07K1/34
Inventor 纪春艳王振中
Owner SOUTH CHINA AGRI UNIV
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