CN107955796B - Red silk ribbon attached to an official seal or a medal aspergillus Q-1 and its application in termite control - Google Patents

Red silk ribbon attached to an official seal or a medal aspergillus Q-1 and its application in termite control Download PDF

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CN107955796B
CN107955796B CN201810001542.9A CN201810001542A CN107955796B CN 107955796 B CN107955796 B CN 107955796B CN 201810001542 A CN201810001542 A CN 201810001542A CN 107955796 B CN107955796 B CN 107955796B
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aspergillus
termite
silk ribbon
official seal
ribbon attached
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CN107955796A (en
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罗兰
袁忠林
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Qingdao Agricultural University
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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14Fungi; Culture media therefor
    • C12N1/145Fungal isolates
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12RINDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00Microorganisms ; Processes using microorganisms
    • C12R2001/645Fungi ; Processes using fungi
    • C12R2001/66Aspergillus
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N63/00Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
    • A01N63/30Microbial fungi; Substances produced thereby or obtained therefrom
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14Fungi; Culture media therefor

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Abstract

The invention discloses red silk ribbon attached to an official seal or a medal aspergillus (Aspergillus nomius) Q-1, it is preserved on December 7th, 2017: China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number is CGMCC NO.14994, preservation address are as follows: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3.Institute of Microorganism, Academia Sinica.Red silk ribbon attached to an official seal or a medal aspergillus (Aspergillus nomius) Q-1 provided by the invention to termite have it is pathogenic, can be used for the prevention and treatment of termite.The spore concentration of red silk ribbon attached to an official seal or a medal aspergillus Q-1 is 106When, processing termite 20d corrected mortality is 74.32%.The bacterial strain is expected to play a significant role in the prevention and treatment of termite.

Description

Red silk ribbon attached to an official seal or a medal aspergillus Q-1 and its application in termite control
Technical field
The present invention relates to microorganisms technical fields, and in particular to application of the red silk ribbon attached to an official seal or a medal aspergillus Q-1 in termite control.
Background technique
A kind of ancient one of five major pest of social insect and the world of termite, distribution is wide, and harm is serious.China It is one of serious country of termite damage, wherein 40% land is distributed.Termite can destroy the timber structure of building, damage water Library dykes and dams, farm-forestry crop of causing harm, moth eat buried cable means of transportation etc..It was counted according to 2007, China is every year because of termite harm Loss is up to 1,500,000,000 yuan or more.
It is still one of current major measure using Agro-chemicals control termite.Drug of the China for termite control is poisoned with poison Tick, fenvalerate, Biphenthrin, Permethrin, imidacloprid etc., currently on the market with more drug be mainly Biphenthrin, Imidacloprid.Chemical agent have it is effective, at low cost, easy to operate, but also to environment and human health while killing termite May have potential risks and influence.
Biological control termite is environmentally friendly, can be to avoid chemical prevention bring a series of problems.Existing Su Yun at present The multiple-microorganisms such as golden bacillus, Serratieae, pseudomonas aeruginosa, muscardine, green muscardine fungus and aspergillus flavus have termite Pathogenic relevant report.Bacteria agent or preparation can reduce or substitute the use of chemical agent, to both reach prevention and prevent The purpose of termite is controlled, and avoids or reduces chemical agent and uses the threat brought to environment and human health.
Summary of the invention
Red silk ribbon attached to an official seal or a medal aspergillus (Aspergillus nomius) Q-1 provided by the present invention, was preserved on December 7th, 2017: China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number are CGMCC NO.14994, preservation address Are as follows: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3.
Red silk ribbon attached to an official seal or a medal aspergillus (Aspergillus nomius) Q-1 provided by the invention to termite have it is pathogenic, can be used for white The prevention and treatment of ant.
The present invention provides termite control microbial inoculum, and active constituent is red silk ribbon attached to an official seal or a medal aspergillus (Aspergillus nomius) Q-1.
The spore concentration of red silk ribbon attached to an official seal or a medal aspergillus Q-1 is 106When, processing termite 20d corrected mortality is 74.32%.The bacterial strain has Prestige plays a significant role in the prevention and treatment of termite.
The beneficial effects of the present invention are:
Red silk ribbon attached to an official seal or a medal aspergillus (Aspergillus nomius) Q-1 that the present invention separates has good control efficiency to termite.
Detailed description of the invention
Fig. 1 is growing state of red silk ribbon attached to an official seal or a medal aspergillus (Aspergillus nomius) Q-1 on PDA;
Fig. 2 is red silk ribbon attached to an official seal or a medal aspergillus (Aspergillus nomius) sporophore of Q-1 and the form of spore.
Specific embodiment
Term as used in the present invention generally has those of ordinary skill in the art usual unless otherwise specified The meaning of understanding.
Combined with specific embodiments below, and referring to the data further detailed description present invention.Following embodiment only be It illustrates the present invention, rather than limits the scope of the invention in any way.
Following example is not construed as limiting the invention in any way for the present invention to be explained further.Under The test method in embodiment is stated, unless otherwise specified, generally conventional method.
The separation and identification of bacterial strain Q-1
The separation of bacterial strain Q-1: in July, 2016 acquires termite from Laoshan.
Collect the worker ant of the termite of natural infection fungi.On superclean bench, whole body is covered into green powdery with tweezers The worker ant of object clips to sterilized PDA culture medium plate top, and lightly fast jitter once, is placed in 26 DEG C of constant incubators Culture, bacterium colony to be grown, picking mycelia see whether, selection and polypide consistent with the fungi on polypide with spore under the microscope Consistent bacterium colony connects in PDA culture medium, sets and carries out purifying culture at 26 DEG C.Strain number after purification is Q-1, and is stored in 4 In DEG C refrigerator.Separation is PDA culture medium with the culture medium saved: 200g potato liquor is filtered to 1000mL, adds 20g grape Sugar, 20g agar.
The identification of bacterial strain: including Morphological Identification and Molecular Identification
Morphological Identification: bacterial strain Q-1 is cultivated on PDA, is white colony, xanthelasma occurs in white colony after a few days Point, entire bacterium colony all turns yellow later.Continue to cultivate, green spot occurs in yellow color colonies center, is gradually diffused as later Entire bacterium colony becomes green (such as Fig. 1), and generates a large amount of conidiums (such as Fig. 2).
Molecular Identification: bacterial strain Q-1 is sent to Shanghai Sheng Gong biotech firm and does Molecular Identification.Fungi primer NS1
(5 ' GTAGTCATATGCTTGTCTC 3 ') and NS6 (5 ' GCATCACAGACCTGTTATTGCCTC 3 ') carry out PCR Amplification, obtains the amplified band (1309bp) of size about 1.3kb, ITS sequencing result:
tgtgaaactgcgaatggctcattaaatcagttatcgtttatttgatagtaccttactacatggatacc tgtggtaattctagagctaatacatgctaaaaacctcgacttcggaaggggtgtatttattagataaaaaaccaat gcccttcggggctccttggtgattcataataacttaacgaatcgcatggccttgcgccggcgatggttcattcaaa tttctgccctatcaactttcgatggtaggatagtggcctaccatggtggcaacgggtaacggggaattagggttcg attccggagagggagcctgagaaacggctaccacatccaaggaaggcagcaggcgcgcaaattacccaatcccgac acggggaggtagtgacaataaatactgatacggggctcttttgggtctcgtaattggaatgagtacaatctaaatc ccttaacgaggaacaattggagggcaagtctggtgccagcagccgcggtaattccagctccaatagcgtatattaa agttgttgcagttaaaaagctcgtagttgaaccttgggtctggctggccggtccgcctcaccgcgagtactggtcc ggctggacctttccttctggggaaccccatggccttcactggctgtggggggaaccaggacttttactgtgaaaaa attagagtgttcaaagcaggcctttgctcgaatacattagcatggaataatagaataggacgtgcggttctatttt gttggtttctaggaccgccgtaatgattaatagggatagtcgggggcgtcagtattcagctgtcagaggtgaaatt cttggatttgctgaagactaactactgcgaaagcattcgccaaggatgttttcattaatcagggaacgaaagttag gggatcgaagacgatcagataccgtcgtagtcttaaccataaactatgccgactagggatcgggcggtgtttctat gatgacccgctcggcaccttacgagaaatcaaagtttttgggttctggggggagtatggtcgcaaggctgaaactt aaagaaattgacggaagggcaccacaaggcgtggagcctgcggcttaatttgactcaacacggggaaactcaccag gtccagacaaaataaggattgacagattgagagctctttcttgatcttttggatggtggtgcatggccgttcttag ttggtggagtgatttgtctgcttaattgcgataacgaacgagacctcggcccttaaatagcccggtccgcgtttgc Gggccgctggcttcttagggggatg identifies that bacterial strain Q-1 is red silk ribbon attached to an official seal or a medal aspergillus (Aspergillus by sequence alignment nomius)。
Toxicity test of the bacterial strain Q-1 to eastern subterranean termite
The preparation of spore liquid: bacterial strain Q-1 is connected in PDA culture medium, 10d is cultivated under 25 DEG C of constant temperature, then with 0.02% Tween-80 sterile water is made into spore liquid, measures spore concentration with blood counting chamber, is then made into 106、105、104With 103A spore The spore liquid of 4 kinds of concentration of son/mL.
Toxicity test of the bacterial strain Q-1 to eastern subterranean termite: being that bacterial strain Q-1 is measured in 60mm culture dish to termite in diameter Virulence.It is to take the bacterium solution of 100 μ L difference spore concentrations uniform in 60mm culture dish that the filter paper that diameter is 55mm, which is placed on diameter, Point drips on filter paper, blank control 0.02%Tween-80,30 worker ants of every ware, and each concentration is repeated 3 times.At 26 ± 1 DEG C It is raised in dark surrounds, 20d is observed continuously, take out dead individuals daily, and record the accumulative of each concentration group and control group in detail The death rate.Must remain environment more wet in experiment culture dish, the test worm death rate control 20% of blank control with It is interior.
Data statistical approach:
The calculation formula of the death rate and corrected mortality:
The death rate (%)=death borer population/for trying total borer population × 100%
Corrected mortality (%)=(the processing death rate-control death rate)/(1- compares the death rate) × 100%
Data are handled using Excel2007, take logarithm as x using concentration and time, corrected mortality is scaled probability value As y, linear regression is carried out, virulence equation is acquired, calculates LC50And relevant parameter.
Toxicity test of the 1 bacterial strain Q-1 of table to eastern subterranean termite
As it can be seen from table 1 spore liquid concentration is 106When a/mL, with the extension of time, the death rate increases, 15 days When, when corrected mortality is more than 50%, 20 days, corrected mortality 74.32%.
The lethal concentration of 50 of the 2 bacterial strain Q-1 of table to eastern subterranean termite
Bacterial strain Q-1's separates again and identifies
The lethal eastern subterranean termite of bacterial strain Q-1 is collected, continues moisturizing, when growing mycelia and spore, its form of microscopy is It is no consistent with bacterial strain Q-1, separated and cultivated by dithering, as the result is shown no matter microscopy or the bacterial strain isolated and bacterial strain Q- 1 is identical.Illustrate that bacterial strain Q-1 is the pathogenic bacteria of termite, can be used for the prevention and treatment of termite.
The above described is only a preferred embodiment of the present invention, being not that the invention has other forms of limitations, appoint What those skilled in the art changed or be modified as possibly also with the technology contents of the disclosure above equivalent variations etc. Imitate embodiment.But without departing from the technical solutions of the present invention, according to the technical essence of the invention to above embodiments institute Any simple modification, equivalent variations and the remodeling made, still fall within the protection scope of technical solution of the present invention.
Sequence table
<110>Qingdao Agricultural University
<120>red silk ribbon attached to an official seal or a medal aspergillus Q-1 and its application in termite control
<130> 2017
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 19
<212> DNA
<213>artificial sequence (Artificial sequence)
<400> 1
gtagtcatat gcttgtctc 19
<210> 2
<211> 24
<212> DNA
<213>artificial sequence (Artificial sequence)
<400> 2
gcatcacaga cctgttattg cctc 24
<210> 3
<211> 1309
<212> DNA
<213>red silk ribbon attached to an official seal or a medal aspergillus (Aspergillus nomiusQ-1)
<400> 3
tgtgaaactg cgaatggctc attaaatcag ttatcgttta tttgatagta ccttactaca 60
tggatacctg tggtaattct agagctaata catgctaaaa acctcgactt cggaaggggt 120
gtatttatta gataaaaaac caatgccctt cggggctcct tggtgattca taataactta 180
acgaatcgca tggccttgcg ccggcgatgg ttcattcaaa tttctgccct atcaactttc 240
gatggtagga tagtggccta ccatggtggc aacgggtaac ggggaattag ggttcgattc 300
cggagaggga gcctgagaaa cggctaccac atccaaggaa ggcagcaggc gcgcaaatta 360
cccaatcccg acacggggag gtagtgacaa taaatactga tacggggctc ttttgggtct 420
cgtaattgga atgagtacaa tctaaatccc ttaacgagga acaattggag ggcaagtctg 480
gtgccagcag ccgcggtaat tccagctcca atagcgtata ttaaagttgt tgcagttaaa 540
aagctcgtag ttgaaccttg ggtctggctg gccggtccgc ctcaccgcga gtactggtcc 600
ggctggacct ttccttctgg ggaaccccat ggccttcact ggctgtgggg ggaaccagga 660
cttttactgt gaaaaaatta gagtgttcaa agcaggcctt tgctcgaata cattagcatg 720
gaataataga ataggacgtg cggttctatt ttgttggttt ctaggaccgc cgtaatgatt 780
aatagggata gtcgggggcg tcagtattca gctgtcagag gtgaaattct tggatttgct 840
gaagactaac tactgcgaaa gcattcgcca aggatgtttt cattaatcag ggaacgaaag 900
ttaggggatc gaagacgatc agataccgtc gtagtcttaa ccataaacta tgccgactag 960
ggatcgggcg gtgtttctat gatgacccgc tcggcacctt acgagaaatc aaagtttttg 1020
ggttctgggg ggagtatggt cgcaaggctg aaacttaaag aaattgacgg aagggcacca 1080
caaggcgtgg agcctgcggc ttaatttgac tcaacacggg gaaactcacc aggtccagac 1140
aaaataagga ttgacagatt gagagctctt tcttgatctt ttggatggtg gtgcatggcc 1200
gttcttagtt ggtggagtga tttgtctgct taattgcgat aacgaacgag acctcggccc 1260
ttaaatagcc cggtccgcgt ttgcgggccg ctggcttctt agggggatg 1309

Claims (3)

1. red silk ribbon attached to an official seal or a medal aspergillus (Aspergillus nomius) Q-1, was preserved on December 7th, 2017: Chinese microorganism strain is protected Administration committee's common micro-organisms center is hidden, deposit number is CGMCC No.14994.
2. red silk ribbon attached to an official seal or a medal aspergillus (Aspergillus nomius) Q-1 described in claim 1 is used to prevent and treat the application in termite.
3. a kind of microbial bacterial agent, which is characterized in that active constituent is red silk ribbon attached to an official seal or a medal aspergillus (Aspergillus nomius) Q-1, institute It states red silk ribbon attached to an official seal or a medal aspergillus (Aspergillus nomius) Q-1 to be preserved on December 7th, 2017: Chinese microorganism strain preservation management Committee's common micro-organisms center, deposit number are CGMCC No.14994.
CN201810001542.9A 2018-01-02 2018-01-02 Red silk ribbon attached to an official seal or a medal aspergillus Q-1 and its application in termite control Active CN107955796B (en)

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CN109971658B (en) * 2019-05-14 2022-05-20 西南林业大学 Aspergillus fungus strain and application thereof
CN112391294B (en) * 2019-08-16 2023-08-08 广西壮族自治区林业科学研究院 Aspergillus peak and application thereof in Huang Yeming control
CN117660202B (en) * 2023-12-11 2024-07-12 云南省农业科学院农业环境资源研究所 Aspergillus peak Aspergillus nomius and application thereof

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* Cited by examiner, † Cited by third party
Title
Resource competition between two fungal parasites in subterranean termites;Thomas Chouvenc等;《Naturwissenschaften》;20121021;第99卷(第11期);第951页左栏 *

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