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Clone, expression and application of antimicrobial protein HAS1 encoding gene

A technology that encodes genes and antibacterial proteins is applied in the fields of application, genetic engineering, and plant genetic improvement, and can solve problems such as sugarcane smut that has not yet been seen

Active Publication Date: 2013-04-10
INST OF TROPICAL BIOSCI & BIOTECH CHINESE ACADEMY OF TROPICAL AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

So far, there have been no reports on the use of microorganisms and their biocontrol agents to successfully control sugarcane smut

Method used

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  • Clone, expression and application of antimicrobial protein HAS1 encoding gene
  • Clone, expression and application of antimicrobial protein HAS1 encoding gene
  • Clone, expression and application of antimicrobial protein HAS1 encoding gene

Examples

Experimental program
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Effect test

Embodiment 1

[0038] 1. Screening and identification of antagonistic bacteria of plant pathogenic fungi

[0039] Screening of bacterial strains: Weigh 5 grams of rhizosphere soil (the rhizosphere soil of sugarcane plants in the plot where sugarcane smut occurs seriously in Lingao County, Hainan Province, my country), put it into a triangular flask with 100ml sterile water, and put it on a shaker. After shaking vigorously for 30 minutes, let stand for 15 minutes. Take 1ml and dilute to 10 -4 、10 -5 、10 -6 Concentration gradient, take 0.1ml from each concentration gradient suspension, and spread it on LB medium with a sterile stick (the formula is: each liter contains 10g of peptone, 5g of yeast powder, 10g of NaCl, 12g of agar powder, pH7.0~7.2) On the plate, cultured upside down in a constant temperature incubator at 30°C, observed continuously for 3 days, picked strains with different colony shapes, streaked again, purified and stored for later use. Coat the teliospores of sugarcane smu...

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Abstract

The invention discloses a clone, an expression and an application of an antimicrobial protein HAS1 encoding gene. The nucleotide sequence of the antimicrobial protein HAS1 encoding gene is shown in SEQIDNO.1, and the amino acid sequence is shown in SEQIDNO.2. The sequences are obtained by taking a biocontrol strain HAS as a research object, performing the antibacterial mechanism, taking genome DNA (deoxyribonucleic acid) of the strain HAS as a template, using a specific expression primer to perform PCR (polymerase chain reaction) amplification to obtain the full expression sequences of the encoding gene, then, constructing a prokaryotic expression vector to express the sequences, and at last taking a sugar cane smut virus as a target to carry out an expression product bacteriostasis experiment. The experiment results show that the expression product has the same capability of resisting against the sugar cane smut virus. The online comparison shows that the antimicrobial protein is a novel one; and encoding the gene sequences of the antimicrobial protein can provide a new material for the future breeding for disease resistance.

Description

technical field [0001] The present invention relates to a new antibacterial protein produced by microorganisms that has antibacterial activity against plant pathogenic bacteria, and more specifically relates to the identification and application of a new antibacterial protein derived from microorganisms. The invention is based on the fact that the biocontrol strain HAS has good inhibitory effect on sugarcane smut. Background technique [0002] Sugarcane smut is caused by Ustilago scitaminea Sydow. A fungal disease caused by Basidiomycetes Ustilago. The disease was first discovered in Natal, South Africa in 1877, and was subsequently reported in most countries in Asia and Africa in the Eastern Hemisphere. It occurred in Argentina in the Americas in 1940, and then the disease also occurred in the Western Hemisphere. From the 1970s to the 1990s, Hawaii, Florida, Morocco, Australia and other countries and regions also reported the occurrence of the disease one after another....

Claims

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Application Information

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IPC IPC(8): C12N15/31C12N15/70C07K14/32A01N47/44A01P3/00A01H5/00
Inventor 熊国如伍苏然赵更峰冯翠莲张树珍
Owner INST OF TROPICAL BIOSCI & BIOTECH CHINESE ACADEMY OF TROPICAL AGRI SCI
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