CN106967613A - Have pale purple purple spore bacteria strain and its application of High pathogenicity to botrytis cinerea - Google Patents
Have pale purple purple spore bacteria strain and its application of High pathogenicity to botrytis cinerea Download PDFInfo
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- CN106967613A CN106967613A CN201710103274.7A CN201710103274A CN106967613A CN 106967613 A CN106967613 A CN 106967613A CN 201710103274 A CN201710103274 A CN 201710103274A CN 106967613 A CN106967613 A CN 106967613A
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION 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
- A01N63/00—Biocides, 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
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Abstract
The present invention relates to a kind of pale purple purple spore bacterium for having High pathogenicity to botrytis cinerea（Purpureocillium lilacinum）Bacterial strain and its application, the bacterial strain is pale purple purple spore bacterium pt362, can be developed into Biological agents is used for biological control to graw mold of tomato, and relatively the chemical agent of preventing and treating botrytis cinerea is compared at present, with efficient, low toxicity, low-residual, it is pollution-free, be not likely to produce drug-fast feature.Bacterial strain of the present invention to botrytis cinerea in addition to having High pathogenicity, and strain culturing is fast, and sporulation quantity is big, and spore germination rate is high, it is easy to prepare, with good market application foreground.
Description
Technical field
The invention belongs to microbial technology field, and in particular to a kind of pale purple purple for having High pathogenicity to botrytis cinerea
Spore bacterium（Purpureocillium lilacinum）Bacterial strain and its application.
Background technology
Gray mold is a kind of global important disease, and it can infect the flowers of 200 various plants, fruit and other green group
Knit, cause plant tissue to rot, lose value.In recent years, gray mold is seriously hindered in the generation of Chinese large area
The raising of yield of vegetables.Wherein solanaceous vegetable is aggrieved most heavy with tomato, and the preventing and treating of graw mold of tomato has turned into protection production development
Key measure, directly decide the yield and economic benefit of tomato.Ash arrhizus bacteria is on tomato mainly with sclerotium in soil
In or survived the winter on invalid body with mycelia and conidium or more summer.The disease is aeroborne disease, mainly by conidium with gas
Stream, irrigation water and farm implements etc. are propagated, and the florescence is the peak period infected.When Second Year spring condition is suitable, sclerotial germination, producing strains
Filament and conidiophore and conidium.Come off, carried out by means of air-flow, rainwater or dewdrop and farming operation after conidium is ripe
Propagate.Output germ tube during sprouting, from host's wound or the organ of aging, withered tissue intrusion, also can directly be invaded by epidermis
Dye causes morbidity, and disease portion produces substantial amounts of conidium and can infected again when moist.Low temperature, high humidity are the necessary bars of morbidity
Part, especially high humidity are the leading factor of Occurrence and development of disease, and temperature is at 15 DEG C or so, and relative humidity is in the starting that 80% is morbidity
Condition.
At present, anti-Zhiduo County for botrytis cinerea uses Chemical control methods, but the frequent of chemical pesticide is used
Cause residues of pesticides, environmental pollution, bio-diversity destruction and botrytis cinerea the problems such as to develop immunity to drugs, and give birth to
Thing agricultural chemicals with it efficiently, low toxicity, low-residual, it is pollution-free, be not likely to produce the resistance to the action of a drug and the characteristic such as raw material is easy to get is by it is believed that be
The preferable surrogate-data technique of following chemical pesticide.Therefore, also gradually cause people's using biological pesticide preventing and treating botrytis cinerea
Pay attention to.The biological control of graw mold of tomato is more with the research of living microorganism or its metabolite, existing result of study table
Bright, many soil, Ye Wei, rhizosphere and interior raw microorganism even have to a certain degree including some phytopathy originals to ash arrhizus bacteria
Inhibitory action.Be currently used in preventing and treating ash arrhizus bacteria biocontrol fungi have Trichoderma viride (Trichodermaviride), breathe out thatch wood
Mould (Trichoderma harzianum), Trichoderma lignorum (Trichoderma lignorum), budding stalk it is short it is mould (Aureobas idium pullulans), light white latent ball yeast (Crypto-coccus albidus), Rhodotorula glutinus (Rhodotoru laglutinis), ball hair shell (Chaetomium globosum), viscous broom it is mould (Gliocladium spp.) etc., bacterium mainly has
Bacillus as (Bacsllus sutizis), bacillus polymyxa (Baeillus polymyxa), bacillus licheniformis
(Bacilluslicheniformis), newborn bacillus (Lactobaeillussp), short body bacillus (Baesllus
Pumilus), the yellow sporangium of thermophilic malt (Xanthomonasmaltophilia), Pseudomonas (PeudomonasSp.), it is glimmering
Light pseudomonas (Peudomonasjluoreseens）Deng.
However, laboratory stage is in the current research for botrytis cinerea biocontrol microorganisms, not yet in agricultural production more
In effective preventing and treating is reached to botrytis cinerea, it would be highly desirable to further separation screening or inducing and acclimating go out the beneficial bacterium that can effectively apply
Strain, and carry out the further investigation of biotic potential.
The content of the invention
, should it is an object of the invention to provide the pale purple purple spore bacteria strain that one plant has High pathogenicity to botrytis cinerea
Bacterial strain is as biological pesticide, with good market application foreground.
To solve problem above, the present invention is achieved through the following technical solutions：
On the basis of the original strain pl36-1 obtained by separated, screening, inventor is further by Agrobacterium tumefaciens mediated
The mutant strain pt362 that T-DNA inserts genetic transformation and obtained, and it is preserved in Chinese Typical Representative culture on November 25th, 2016
Collection（Address：Wuhan University of Wuhan City of Hubei China province, postcode 430072）, deposit number is CCTCC NO：
M2016682。
The invention also discloses the applications of the pale purple purple spore bacteria strain pt362 in prevention and control graw mold of tomato, and in system
The application in the biological pesticide of preventing and treating botrytis cinerea is ready for use on, the conidial suspension and metabolite of bacterial strain are to tomato
Ash arrhizus bacteria has High pathogenicity.
Compared with prior art, the present invention is with following positive beneficial technique effect：
（1）The pale purple purple spore bacteria strain pt362 of the present invention is a kind of primary anti-fungal of nematodiasis, and can be made into Biological agents is used for pair
The biological control of botrytis cinerea, relatively the chemical agent of preventing and treating botrytis cinerea is compared at present, with efficient, low toxicity, low
Residual, it is pollution-free, be not likely to produce drug-fast feature.
（2）The pale purple purple spore bacteria strain pt362 reproductive capacity of the present invention is strong, and culture speed is fast, and sporulation quantity is big, spore germination rate
It is high, it is easy to prepare.
（3）The pale purple purple spore bacteria strain pt362 of present invention spore suspension and metabolite has to botrytis cinerea
Very strong pathogenicity.
Brief description of the drawings
Antagonistic effects of the pale purple purple spore bacteria strain pt362 of Fig. 1 to botrytis cinerea（PDA plate）；
Antagonistic effect of the pale purple purple spore bacteria strain pt362 metabolites of Fig. 2 to botrytis cinerea.
Embodiment
Illustrate the embodiment of the present invention with reference to embodiment, but following examples are used only to describe in detail
The present invention, does not limit the scope of the present invention in any way.Involved instrument and equipment is as without especially in the examples below
Illustrate, be routine instrument device；Involved biochemical reagents are commercially available unless otherwise instructed.
Embodiment 1：Pale purple purple spore bacteria strain pt362 culture, domestication
Pale purple purple spore bacterium original strain pl36-1 teachers Wang Mingzu 1991 Hubei Province out of root-knot nematode body it is isolated（Ginseng
See 1991 and be published in《Plant Pathology》The article of entitled " First Report of Studies of root-knot nematode parasitical fungi of eggs "）, it is stored in
Plant pathology nematode research department of Hua Zhong Agriculture University.Pt362 is by Agrobacterium tumefaciens mediated T-DNA insertion genetic transformations
The mutant strain of acquisition.
The method of transformant genetic stability is with reference to the method once gone in in a big way.Approximately as：Without careless fourth phosphine and carboxin
PDA plate on be inoculated with pale purple purple spore bacterium conversion bacterial strain, 28 DEG C, constant temperature dark culturing is then transferred to 4 DEG C of refrigerators after 3-5 days,
Place 2 days, subsequently 28 DEG C, constant temperature dark culturing 3-5 days is flat to new PDA with card punch picking colony edge mycelium inoculation
Cultivated on plate, so 10 generations of continuous switching, on the PDA plate for inoculating the careless μ g/mL carboxins of fourth phosphine+400 containing 4.5mg/mL,
28 DEG C, constant temperature dark culturing records insertion mutation daughter colony growing state（Once Daxing etc. 2005）.Result of the test shows, passes through
Turn generation 10 times domestication after insertion mutation still can on the PDA plate containing careless fourth phosphine and carboxin normal growth, explanation
Insertion mutation of pale purple purple spore bacterium is during vegetative propagation, and bar and anti-carboxin gene are not lost, with higher something lost
Pass stability.
Embodiment 2：Antagonistic effects of the pale purple purple spore bacteria strain pt362 to botrytis cinerea（PDA plate）
It is prepared by pale purple purple spore bacterium/botrytis cinerea：Full bacterium has been grown on glass culture dish with diameter 0.5cm card punch
Fungus block, is linked into the flat board cooled down, 28 by the pale purple purple spore bacterium of silk/botrytis cinerea colony edge punching with transfer needle
DEG C culture, it is stand-by.
It is prepared by PDA plate：The PDA culture medium melted is cooled to 30 DEG C or so, plate is down flat.Stand 1~2h.
Inoculation：3d botrytis cinerea mycelia edge has been grown on glass culture dish with diameter 0.5cm card punch
Fungus block, is linked into the flat board cooled down (crossing method) by punching with transfer needle.Pale purple purple spore bacterium is inoculated in the middle of flat board,
25 DEG C of cultures, observation, statistics inhibitions.As a result Fig. 1 is seen,（a：Botrytis cinerea；b：Original strain and botrytis cinerea antagonistic effect；c：
Mutant strain pt362 and botrytis cinerea antagonistic effect.）It can be seen that pale purple purple spore bacterium has strong suppression to botrytis cinerea
Effect.
Embodiment 3：Antagonistic effect of the pale purple purple spore bacteria strain pt362 metabolites to ash arrhizus bacteria
It is prepared by pale purple purple spore fermented liquid：Access diameter 0.5cm's in the 250mL triangular flasks equipped with 150mL PDB culture mediums
Pale purple purple spore bacterium fungus block, 28 DEG C, 200r/min shaking table cultures cultivate 6d.Zymotic fluid is taken, 10000r/min centrifugation 15min take
Clear liquid is filtered with biofilter.Resulting liquid is stoste.Stoste is diluted 8 times with sterilized water dilution.
Inoculation：With from leaf age is identical, healthy leaves of the same size, being cleaned with sterilized water, it is standby.Tomato leaf is soaked
Bubble is 20 minutes or so in the zymotic fluid of above-mentioned various concentrations.1 a diameter of 0.5cm ash arrhizus bacteria bacterium is inoculated with the middle of blade
Block, 25 DEG C of moisturizing cultures 3 days.Incidence is observed, the incidence of disease is counted, disease index is calculated.
Disease index (%)=[(control onset area-processing onset area)/control onset area] × 100%
As a result Fig. 2 is seen, wherein left figure is blank control group, middle figure be stoste antagonism group, right figure, can be with to dilute 8 times of antagonism groups
Find out that the concentration of pale purple purple spore bacterium is higher, the fungus block of ash arrhizus bacteria growth is smaller, and the antagonism being subject to is stronger.
Claims (4)
1. a kind of pale purple purple spore bacterium（Purpureocillium lilacinum）Bacterial strain pt362, its deposit number is CCTCC
NO：M2016682.
2. applications of the pale purple purple spore bacteria strain pt362 in graw mold of tomato described in claim 1.
3. the pale purple purple spore bacteria strain pt362 described in claim 1 is preparing the biological pesticide for preventing and treating botrytis cinerea
In application.
4. a kind of Biological agents, including pale purple purple spore bacteria strain pt362 described in claim 1, its conidium or/and its
Metabolite.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107475133A (en) * | 2017-09-26 | 2017-12-15 | 江苏农林职业技术学院 | A kind of pale purple purple spore bacterium and its application in bletilla striata growth is promoted |
| CN110423702A (en) * | 2019-08-05 | 2019-11-08 | 云南大学 | High sporulation quantity purple purple spore bacterium genetic engineering bacterium Δ PlflbC and its construction method and application |
| CN110724640A (en) * | 2018-06-28 | 2020-01-24 | 驻马店市农业科学院 | Biocontrol bacterium for plant root-knot nematodes, preparation and application thereof |
| CN114410518A (en) * | 2022-01-07 | 2022-04-29 | 春华秋实科技集团有限公司 | Compound microbial agent containing streptomyces and application thereof |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107475133A (en) * | 2017-09-26 | 2017-12-15 | 江苏农林职业技术学院 | A kind of pale purple purple spore bacterium and its application in bletilla striata growth is promoted |
| CN107475133B (en) * | 2017-09-26 | 2019-08-30 | 江苏农林职业技术学院 | A kind of pale purple purple spore bacterium and its application in promotion bletilla striata growth |
| CN110724640A (en) * | 2018-06-28 | 2020-01-24 | 驻马店市农业科学院 | Biocontrol bacterium for plant root-knot nematodes, preparation and application thereof |
| CN110724640B (en) * | 2018-06-28 | 2023-06-30 | 驻马店市农业科学院 | Tomato root knot nematode biocontrol bacteria, preparation and application thereof |
| CN110423702A (en) * | 2019-08-05 | 2019-11-08 | 云南大学 | High sporulation quantity purple purple spore bacterium genetic engineering bacterium Δ PlflbC and its construction method and application |
| CN110423702B (en) * | 2019-08-05 | 2022-11-01 | 云南大学 | High-spore-yield purple-spore-bacterium-gene engineering bacterium delta PlflbC and construction method and application thereof |
| CN114410518A (en) * | 2022-01-07 | 2022-04-29 | 春华秋实科技集团有限公司 | Compound microbial agent containing streptomyces and application thereof |
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Application publication date: 20170721 |