CN106167768A - The one pale yellow Cryptococcus of strain and in the application of the preventing and treating postharvest disease of fruits and vegetables such as blue berry - Google Patents

The one pale yellow Cryptococcus of strain and in the application of the preventing and treating postharvest disease of fruits and vegetables such as blue berry Download PDF

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CN106167768A
CN106167768A CN201610557260.8A CN201610557260A CN106167768A CN 106167768 A CN106167768 A CN 106167768A CN 201610557260 A CN201610557260 A CN 201610557260A CN 106167768 A CN106167768 A CN 106167768A
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hmqausz01
cryptococcus
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fruits
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梁晨
赵洪海
郑长英
尉莹莹
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Qingdao Agricultural University
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Abstract

The invention belongs to technical field of plant disease biological control, be specifically related to the separation screening of pale yellow Cryptococcus HMQAUSZ01, fermentation and the biological and ecological methods to prevent plant disease, pests, and erosion determination of activity of the postharvest disease of fruits and vegetables such as a strain preventing and treating blue berry, belong to agricultural biological technical field.Its deposit number is CGMCC No.11984.The invention also discloses the microbial bacterial agent utilizing this bacterial strain to prepare, and the application on the postharvest disease of fruits and vegetables such as blue berry.The present invention provides an efficient microorganism of strain for the preventing and treating postharvest disease of fruits and vegetables such as blue berry, the pale yellow Cryptococcus of the present invention (Cryptococcus flavescens) bacterial strain HMQAUSZ01 has stronger Competition to postharvest disease of fruits and vegetables such as blue berries；Secondly, the present invention pale yellow Cryptococcus (Cryptococcus flavescens) bacterial strain HMQAUSZ01 has preferable preventive effect in the test of the in vitro fruit of postharvest disease of fruits and vegetables such as blue berry, Fructus Vitis viniferae, Fructus Pruni pseudocerasi, Fructus Mali pumilae, Fructus Lycopersici esculenti；Additionally, microbial bacterial agent of the present invention is to people, animal safety, belong to environmentally friendly, there is good development and application prospect.

Description

The one pale yellow Cryptococcus of strain and in the application of the preventing and treating postharvest disease of fruits and vegetables such as blue berry
Technical field
The invention belongs to technical field of plant disease biological control, be specifically related to the postharvest disease of fruits and vegetables such as a strain preventing and treating blue berry The separation screening of pale yellow Cryptococcus HMQAUSZ01, fermentation and biocontrol effect measure, belong to agricultural biological technical field.
Background technology
China is fruit and vegerable big producing countries, and fruit and vegerable often cause a large amount of rotting because of transport and storage improper, cause The phenomenons such as economic benefit reduction, product competitiveness decline and waste, this is also the problem of global concern.Developed country very early will Post-harvest fresh-keeping processing is placed on the top priority of agricultural, it was reported that U.S.'s fruit and vegerable loss rate is 1.7% ~ 5.0%, China then 25 ~ 30%, it is seen then that the preservation technique of China's postharvest fruit and vegetable has the biggest economic potential.
Blue berry belongs to Ericaceae Ericaceae, genus vaccinium Vaccinium, is a kind of to have the new of high economic worth Emerging worldwide small berries fruit tree (Li Yadong 2002), the healthy nutritive value prominent because of it is classified as people by FAO (Food and Agriculture Organization of the United Nation) One of the big health food of class five (Kader & Rovel 1996).At present, blue berry plantation industry has become as domestic planting fruit trees One of industry that industry is the most popular, blue berry market potential is the most gradually developed, to the end of the year 2015, whole nation blue berry cultivated area by Initial 24 hectares are developed into 31210 hectares, and yield is developed into 43244 tons by 2 tons of 2002.
Blue berry belongs to berries fruit, and water content is the highest, and the Blueberry after adopting is the oldest and the most feeble, dehydration occurs, rots Phenomenon, pole not storage tolerance, will cause serious economic loss as processed not in time.This characteristic significantly limit its fresh fruit Shelf life and commodity value.The cause of disease of blue berry storage period disease is caused to have 5 kinds in In The Area of Qingdao at present, wherein Botrytis cinerea and chain Lattice spore is the main pathogenic fungi (Liang Chen etc. 2010).
Along with constantly expanding of blueberry cultivation area and increasing sharply of yield, produce in the urgent need to being suitable for blue berry The real fresh-keeping supporting technology of postharvest storage, to alleviate disparities between supply and demand and the tonnage in market, reduces and loses after adopting, extend supply Phase, thus improve the economic worth of Blueberry and promote the healthy and rapid development (Jiang Aili 2011) of blueberry industry.Traditional Antibacterial and preservation method not only can not solve the problem of rotting well, and bring many potential safety hazards.In view of indigo plant The nutritive value of the certain kind of berries and medical value, the preventing and treating determining blue berry disease should be avoided using chemical agent as far as possible, takes organic The mode of cultivation, develops organic blue berry plant husbandry and must firmly stop the use of chemical agent, therefore for blue berry postharvest disease Preventing and treating should be conceived to develop Biological control means.
Yeast is that it can be at the fruit and vegetable surfaces existence being relatively dried, energy as the great advantage of biological fruit and vegetable preventing and treating Antagonistic Fungi Utilize rapidly nutrition to breed, and affected little (Wisniewski and Wilosn, 1992) and yeast by insecticide Do not produce antibiotics, pathogenic bacteria can be avoided antibiotics to be produced resistance and reduces Biological control and press down disease effect (WiIson and Winsiweksi, 1994), on the other hand can also avoid some antibiotic adverse effect to human body.Therefore new ferment is found Parent resource is the key factor of the postharvest disease of fruits and vegetables biological prevention and control agents such as research and development blue berry.
Summary of the invention
First purpose of the present invention is to provide the persimmon fruit surface isolated one from Chengyang District, Qingdao of Shandong province Strain postharvest disease of fruits and vegetables is had notable preventive and therapeutic effect bio-control yeast-pale yellow Cryptococcus (Cryptococcus flavescens) bacterial strain HMQAUSZ01, enrich the postharvest disease of fruits and vegetables biocontrol fungi microorganism resource such as blue berry, raw for research and development Anti-microbial inoculum lays the foundation.
Second purpose of the present invention is to provide a kind of microbial bacterial agent utilizing above-mentioned bacterial strains HMQAUSZ01 to produce.
The 3rd purpose of the present invention is to provide the preparation method of mentioned microorganism microbial inoculum.
The 4th purpose of the present invention is to provide the mentioned microorganism microbial inoculum sod cultivation to postharvest disease of fruits and vegetables such as blue berries.
The 5th purpose of the present invention is that provide above-mentioned bacterial strains HMQAUSZ01 on the postharvest disease of fruits and vegetables such as blue berry answers With.
The 6th purpose of the present invention is to provide the authentication method of above-mentioned bacterial strains HMQAUSZ01.
Realize technical scheme as described below.
Pale yellow Cryptococcus (Cryptococcus flavescens) HMQAUSZ01 bacterial strain is in preservation on January 7 in 2016 China Committee for Culture Collection of Microorganisms's common micro-organisms center in Microbe Inst., Chinese Academy of Sciences, preservation address: north North Star West Road, Jing Shi Chaoyang District 1 institute 3, deposit number: CGMCC No. 11984.
Utilizing the microbial bacterial agent that above-mentioned pale yellow Cryptococcus HMQAUSZ01 bacterial strain produces, active component is thalline or sends out Ferment stock solution.
The preparation process of mentioned microorganism microbial inoculum is as follows:
(1) actication of culture: by the bacterial strain HMQAUSZ01 of low-temperature preservation 25 DEG C of cultivations on the bean sprouts medium flat board of 10% 48h, obtains the strain of activation；(2) fermentation culture: the bacterial strain HMQAUSZ01 of activation is accessed containing 50mL NYDB culture fluid In 150 mL triangular flasks, making the initial concentration of yeast in fermentation liquid is 2*104Cfu/mL, in 140r/min, after 25 DEG C are shaken training 48h Fermentation liquid is microbial inoculum A；Bacterium solution A is centrifuged at 3,000 rpm 10min, abandons supernatant, collect thalline, and wash bacterium with sterilized water Body twice, and make cell suspending liquid with it, obtain microbial inoculum B；Count with blood counting chamber, adjust the ferment in microbial inoculum A and microbial inoculum B Female bacterium desired concn, 4 DEG C of preservations are stand-by.
A and B viable bacteria body number in mentioned microorganism microbial inoculum should be greater than 1x109cfu/mL。
Bean sprouts medium: 10% bean sprout juice 200mL, agar 20g, glucose 50g, water 800mL, natural pH.
NYDB culture fluid: Carnis Bovis seu Bubali cream 8g, yeast extract 5g, glucose 10g, water 1000mL.
Beneficial effects of the present invention:
1. the pale yellow Cryptococcus HMQAUSZ01 bacterial strain of the present invention has good prevention effect to postharvest disease of fruits and vegetables such as blue berries, Have efficiently, the advantage of has a broad antifungal spectrum；
Pale yellow Cryptococcus HMQAUSZ01 bacterial strain prevention effect the most provided by the present invention is stable, environmentally friendly, on a large scale The fermentation technology produced is simple, low production cost.
The pale yellow Cryptococcus HMQAUSZ01 bacterial strain provided according to the present invention and fermenation raw liquid microbial inoculum can be as single dose or combination systems Agent utilizes or uses, and such preparation can include agriculturally suitably auxiliary agent, solvent, carrier, surfactant or filler.
Detailed description of the invention:
Example below is used for being explained further the present invention, but is construed as limiting the invention never in any form.Following reality Execute the test method in example, if no special instructions, generally conventional method.
The separation screening of embodiment 1 HMQAUSZ01 bacterial strain
Bacterial strain HMQAU SZ01 is the persimmon fruit surface isolated from Chengyang District, Qingdao of Shandong province.In November, 2015 is from mountain Market, Chengyang District, Dong Sheng Qingdao City is bought without disease pest harm, Fructus Jujubae, Fructus Vitis viniferae, Fructus Musae, pears, Fructus Mali pumilae, Fructus Persicae and Fructus Lycopersici esculenti without wound, Fructus Kaki Son is some.Weigh 10g pericarp tissue respectively, join in the conical flask equipped with 90mL bean sprout juice dextrose broth, with Time add the streptomycin of 3000 units, with 120r/min shaken cultivation 48h at 25 DEG C；LmL bacterium solution is drawn, with dense with liquid-transfering gun Degree gradient method is diluted to 10-1～10-7Concentration, by 10-5、10-6、10-7The bacterium solution of concentration takes 0.lmL respectively and has coated bean On the flat board of bud juice glucose agar medium.Every dilution factor is repeated 3 times, and is placed in 25 DEG C of constant incubators cultivation 48h, chooses Take the different single bacterium colony of cultural characteristic and carry out plate line purification.Yeast separation test obtains 40 saccharomycetes, after adopting with blue berry Pathogen rod method is target bacterium, is carried out the screening of antagonistic strain by vitro fruit screening test, and final acquisition one strain is to fruit Real postharvest pathogenic fungi has the bacterial strain HMQAU SZ01 of obvious prevention effect, and preventive effect is 100%.
The biocontrol effect that fruit and vegerable fruit is rotted by embodiment 2 bacterial strain HMQAUSZ01 cell suspending liquid naturally
Microbial inoculum B sterilized water by bacterial strain HMQAUSZ01 is prepared as the treatment fluid of following concentration；(1) 1 × 106cfu/mL;(2) 1 ×107cfu/mL;(3) 1 × 108Cfu/mL, puts in bacteria suspension by blue berry, Fructus Vitis viniferae, Fructus Pruni pseudocerasi, the fruit of Fructus Lycopersici esculenti, after soaking 3min Pull out, air-dry；Being positioned in plastic casing by fruit, overcoat plastic fresh-keeping membrane is to keep humidity (about 95%).Process fruit is stored in 25 DEG C, the rotting rate of periodic logging fruit.Often process 10 fruits, in triplicate.Result of the test shows three kinds of concentration HMQAUSZ01 treatment fluid to fruit naturally rot be respectively provided with obvious inhibition, concentration is the highest, and rear natural occurrence adopted by fruit Rate is the lowest.Wherein concentration is 1 × 108The respective preventive effect of four kinds of fruit nature rotting rates is best by the treatment fluid of cfu/mL, Biocontrol effect to Fructus Lycopersici esculenti is 100%, secondly the preventive effect of blue berry and Fructus Vitis viniferae is respectively 86.96% and 86.11%, the biological and ecological methods to prevent plant disease, pests, and erosion of Fructus Pruni pseudocerasi Effect is minimum, is 61.54%.
Embodiment 3 bacterial strain HMQAUSZ01 cell suspending liquid is to blue berry postharvest pathogen rod method and the life of Botrytis cinerea Anti-effect
Microbial inoculum B sterilized water by bacterial strain HMQAUSZ01 is prepared as the treatment fluid of following concentration: (1) 1 × 105cfu/mL；(2) 1 ×106cfu/mL；(3) 1 × 107cfu/mL；(4) 1 × 108cfu/mL；(5) 1 × 109cfu/mL；Comparison is sterilized water.By nothing The toothpick of bacterium manufactures wound (3mm × 3mm) at the fruit end position of each healthy Blueberry, inoculates above-mentioned place respectively in wound Reason liquid 15 μ L, after 3h dries, inoculates 1 × 10 respectively5 Cfu/mL pathogen spore suspension, after fruit dries, places fruit In plastic casing, overcoat plastic fresh-keeping membrane is to keep humidity.Process fruit and store at 25 DEG C, periodic logging fruit rot rate.Often Process 10 fruits, be repeated 3 times.Result of the test shows along with the increase for the treatment of fluid concentration, and the sickness rate of fruit reduces, wherein 1 ×109Cfu/mL and 1 × 108The sickness rate difference of the treatment fluid of two concentration of cfu/mL is not notable, therefore can use when preventing and treating 1×108Cfu/mL, this concentration for the treatment of to Botrytis cinerea (Botrytis cinerea) and rod method (Alternaria alternara) preventive effect be respectively 87.50% and 84.62%.
Embodiment 4 bacterial strain HMQAUSZ01 cell suspending liquid is to Fructus Mali pumilae postharvest pathogen penicillium expansum, Botrytis cinerea and powder The biocontrol effect of clearance permit end spore
Microbial inoculum B sterilized water by bacterial strain HMQAUSZ01 is prepared as the treatment fluid of following concentration: (1) 1 × 105cfu/mL；(2) 1 ×106cfu/mL；(3) 1 × 107cfu/mL；(4) 1 × 108cfu/mL；(5) 1 × 109cfu/mL；Comparison is sterilized water.By nothing The toothpick of bacterium manufactures wound (3mm × 3mm) at the position, equator of each healthy Apple, inoculates above-mentioned place respectively in wound Reason liquid 15 μ L, after 3h dries, inoculates 1 × 10 respectively5 Cfu/mL pathogen spore suspension, after fruit dries, places fruit In plastic casing, overcoat plastic fresh-keeping membrane is to keep humidity.Process fruit and store at 25 DEG C, periodic measurement lesion diameter.Often locate Manage 3 fruits, be repeated 3 times.Result of the test shows along with the increase for the treatment of fluid concentration, and on fruit, lesion diameter reduces.For three The optimum concentration planting postharvest pathogen penicillium expansum, Botrytis cinerea and trichothecium roseum is respectively 1 × 108Cfu/mL, 1 × 107Cfu/mL and 1 × 109Cfu/mL, corresponding lesion diameter is 0, biocontrol effect 100%.
The different disposal liquid of the embodiment 5 bacterial strain HMQAUSZ01 fermentation liquid biocontrol effect to postharvest diseases of fruit
The bacterial strain HMQAUSZ01 of activation is accessed in the 150 mL triangular flasks containing 50mL NYDB culture fluid, makes ferment in fermentation liquid Female initial concentration is 2*104Cfu/mL, in 140r/min, after 25 DEG C are shaken training 48h, is prepared as following treatment fluid；(1) culture propagation Stock solution: with blood counting chamber count, and with filtrate adjust to concentration be 1 × 108cfu/mL；(2) yeast suspension；Fermentation liquid exists Under 3000r/min, centrifugal 10min, abandons supernatant, washs yeast thalline 2 times with sterilized water, and makes yeast suspension, blood with it Ball count plate count, with sterilized water adjust to concentration be 1 × 108cfu/mL；(3) culture propagation filtrate: culture fluid is at 3000r/ Under min after centrifugal 10min, take supernatant, filter with biofilter (filter membrane 0.22 μm) and get final product；(4) culture propagation is heat-killed Liquid: by culture propagation stock solution autoclaving 20min at 121 DEG C.Comparison is sterilized water.With aseptic toothpick in the equator of Fructus Mali pumilae Position manufactures wound (3mm × 3mm), and at the wound difference above-mentioned four kinds for the treatment of fluid 20 μ L of inoculating strain HMQAUSZ01,3h dries After, inoculate the spore suspension of pathogen (rod method and Botrytis cinerea) respectively, concentration is 1 × 105 Cfu/mL, dries afterwards, Being positioned in plastic casing by fruit, overcoat plastic fresh-keeping membrane is to keep humidity.Process fruit and store at 25 DEG C, after 7 days, measure disease Spot diameter.Often process 3 fruits, be repeated 3 times.Result of the test shows, culture propagation filtrate and the heat-killed liquid of culture propagation are with right According to zero difference, preventive effect is 0, and the preventive effect of culture propagation stock solution and yeast suspension is 100%.
Embodiment 6 bacterial strain HMQAUSZ01 cell suspending liquid is to Fructus Lycopersici esculenti postharvest pathogen rod method and the suppression of Botrytis cinerea Effect
Microbial inoculum B sterilized water by bacterial strain HMQAUSZ01 is prepared as the treatment fluid of following concentration: (1) 1 × 105cfu/mL；(2) 1 ×106cfu/mL；(3) 1 × 107cfu/mL；(4) 1 × 108cfu/mL；(5) 1 × 109cfu/mL；Comparison is sterilized water.By nothing The toothpick of bacterium manufactures wound (3mm × 3mm) at the position, equator of each healthy tamato fruit, inoculates above-mentioned place respectively in wound Reason liquid 15 μ L, after 3h dries, inoculates 1 × 10 respectively5 Cfu/mL pathogen spore suspension, after fruit dries, places fruit In plastic casing, overcoat plastic fresh-keeping membrane is to keep humidity.Process fruit and store at 25 DEG C, periodic measurement lesion diameter.Often locate Manage 3 fruits, be repeated 3 times.Result of the test shows along with the increase for the treatment of fluid concentration, and the lesion diameter of fruit reduces, wherein 1 × 109Cfu/mL and 1 × 108The sickness rate of two concentration treatment fluids of cfu/mL is zero, and difference is not notable, therefore can when preventing and treating Use 1 × 108cfu/mL。
The qualification of embodiment 7 bacterial strain HMQAUSSZ01
(1) morphological feature: on the bean sprout juice flat board of 10% under the conditions of 12h alternation of light and darkness, cultivates 48h for 25 degree, and bacterium colony is rounded, Neat in edge, light yellow, show smooth, thickness is easily provoked, opaque, corrugationless.Microscopy cell ovalize.Train at NYDB Supporting after cultivating 48h in base, bacterium solution is muddy, has precipitation；
(2) physiological feature: strains ferment glucose of the present invention, sucrose, Raffinose, 6-(.alpha.-D-galactosido)-D-glucose., galactose, lactose, trehalose, Maltose, melezitose, methyl-alpha-D-glucose glycosides, cellobiose, Salicin, L-rhamnose, D-xylose, L- Arabinose, D-ribose, ethanol, galactitol, PEARLITOL 25C, D-Sorbitol, inositol, succinate, Portugal (grape) sugar lime；
(3) Molecular Identification: with bacterial strain HMQAUSZ01 genomic DNA as template, utilizes primer NL1/ NL4 nearly to 26S rDNA 5, The D1/D2 region of end expands.Described primer sequence is NL1(5 '-GCA TATCAATAAGCGGAGGAAAAG-') and NL4 (5 '-GGTCCGTGTTTCAAGAC GG-3 ')；Amplification reaction system: 10 × Buffer 2.5 μ L, 5 mM dNTP 2 μ L, 10 μ The each 1 μ L of LM primer NL1 and primer NL4, Taq enzyme (5 U/ μ L) 0.5 μ L, template DNA 2 μ L, moisturizing to 25 μ L.Reaction condition: 94 DEG C of degeneration 1min, 53 DEG C of annealing 1min, 72 DEG C extend 1min20s, 36 circulations.Amplified production is through 1% agarose gel electrophoresis After detection, Sangon Biotech (Shanghai) Co., Ltd. being purified and two-way order-checking, sequencing result is through Sequencher5.0 Software derives contig (Contig) and at NCBI(http after automatically assembling: //www.ncbi.nlm.gov) enter in data base Row BLAST submits to GenBank after analyzing.Suitable sequence, warp is chosen again from GenBank CLUSTALW 2.0 software carries out alignment, and uses adjacent method (neighbor joining with MEGA 5.0 software Analysis, NJ) phylogenetic tree construction, wherein the number of repetition of Bootstrap inspection is 1,000 time.Result shows, should The large subunit sequence of bacterial strain and the pale yellow Cryptococcus from Texas, USACryptococcus flavescensBig sub- The homology of basic sequence (GenBank accession number is FJ743610) reaches 99%.Phylogenetic Analysis result shows, this bacterial strain Large subunit sequence and 4 strainsCryptococcus flavescensThe large subunit sequence of bacterial strain is positioned at same point of phylogenetic tree ?.Sequence analysis data and phylogenetic tree position further demonstrate bacterial strain HMQAUSZ01Cryptococcus flavescens。
Comprehensive above morphological characteristic and 26SrDNA sequence homology comparison analysis result understand HMQAUSZ01 and belong to pale yellow Cryptococcus (Cryptococcus flavescens).
Nucleotides sequence list
<110>Qingdao Agricultural University
<120>one pale yellow Cryptococcus of strain and in the application of the preventing and treating postharvest disease of fruits and vegetables such as blue berry
<160> 1
<210> 1
<211> 641
<212> DNA
<213>pale yellow Cryptococcus (Cryptococcus flavescens)HMQAUSZ01
<400> 1
GCATATCAAT AAGCGGAGGA AAAGAAACTA ACAAGGATTC CCCTAGTAAC GGCGAGTGAA 60
CCGGGAAGAG CTCAAATTTG AAATCTGGCG TGCTCAGTGC GTCCGAGTTG TAATCTATAG120
AGTCGTTTTC CGTGCCGGAC TGTGTCCAAG TCCCTTGGAA CAGGGTATCA AAGAGGGTGA180
TAATCCCGTA CTTGACACAA TGACCGGTGC TCTGTGATAC GTCTTCTACG AGTCGAGTTG240
TTTGGGAATG CAGCTCAAAA TGGGTGGTGA GTTCCATCTA AAGCTAAATA TTGGCGAGAG300
ACCGATAGCG AACAAGTACC GTGAGGGAAA GATGAAAAGC ACTTTGGAAA GAGAGTTAAA360
CAGTACGTGA AATTGTTAAA AGGGAAACGA TTGAAGTCAG TCGTGACTGA GAGGCTCAGC420
CGGTTCTGCC GGTGTATTCC CCTCAGTCGG GTCAACATCA GTTTTGTTCG GTGGATAAGG480
GCGGTTGGAA GGTGGCACCC TCGGGTGTGT TATAGCCAAC TGTCGCATAC ATCGGATGAG540
ACTGAGGAAT GCAGCTCGCC TTTATGGCCG GGGTTCGCCC ACGTTCGAGC TTAGGATGTT600
GACATAATGG CTTTAAACGA CCCGTCTTGA AACACGGACC 640

Claims (8)

1. a pale yellow Cryptococcus of strain (Cryptococcus flavescens) bacterial strain HMQAUSZ01, it is preserved in China Microbiological Culture presevation administration committee common micro-organisms center, deposit number is CGMCC No. 11984.
2. the microbial inoculum prepared by the pale yellow Cryptococcus bacterial strain HMQAUSZ01 described in claim 1.
Microbial inoculum the most according to claim 2, it is characterised in that utilize that above-mentioned pale yellow Cryptococcus HMQAUSZ01 produces 2 Planting microbial bacterial agent (A, B), A active component is HMQAUSZ01 fermenation raw liquid, and B activity composition is yeast suspension；Above-mentioned micro- A and B viable bacteria body number in bacteria agent should be greater than 1x109cfu/mL。
4. the pale yellow Cryptococcus bacterial strain HMQAUSZ01 described in claim 1 is used for preventing and treating the postharvest disease of fruits and vegetables such as blue berry.
5. the microbial inoculum described in Claims 2 or 3 is used for preventing and treating the postharvest disease of fruits and vegetables such as blue berry.
6. the preparation method of the microbial inoculum described in a Claims 2 or 3, it is characterised in that specifically comprise the following steps that
(1) actication of culture: by the bacterial strain HMQAUSZ01 of low-temperature preservation 25 DEG C of cultivations on the bean sprouts medium flat board of 10% 48h, obtains the strain of activation；
(2) fermentation culture: the bacterial strain HMQAUSZ01 of activation is accessed in the 150mL triangular flask containing 50mL NYDB, makes fermentation In liquid, the initial concentration of yeast is 2*104Cfu/mL, in 140r/min, 25 DEG C of fermentation liquids shaken after training 48h are microbial inoculum A；By bacterium solution A It is centrifuged 10min at 3,000 rpm, abandons supernatant, collect thalline, and wash thalline twice with sterilized water, and make cell with it Suspension, obtains microbial inoculum B；
Bean sprouts medium: 10% bean sprout juice 200mL, agar 20g, glucose 50g, water 800mL, natural pH；NYDB culture fluid: Carnis Bovis seu Bubali cream 8g, yeast extract 5g, glucose 10g, water 1000mL.
7. invention also provides one utilizes pale yellow Cryptococcus to carry out postharvest disease of fruits and vegetables prevention and controls；Step is as follows: Concentration is adjusted to 1 × 10 with sterilized water after being activated by pale yellow Cryptococcus HMQAUSZ01, cultivate, be centrifuged and counting6cfu/mL-1 ×109Cfu/mL cell suspending liquid；Fruit is put in bacteria suspension, pull out after soaking 3min, air-dry；Fruit is positioned over plastics In box, overcoat plastic fresh-keeping membrane, to keep humidity (about 95%), processes fruit and stores in 25 DEG C.
8. one kind be used for detecting pale yellow Cryptococcus (Cryptococcus flavescens) method of bacterial strain HMQAUSZ01, its It is characterised by, with the genomic DNA of test strains as masterplate, NL1(5 '-GCATATCAATAAGCGGAGGAAA AG-3 ') NL4 (5 '-GGTCCGTGTTTCAAACGG-3 ') is the large subunit sequence of primer amplification bacterial strain, if gained amplified production is SEQID Nucleotide sequence shown in No:1, is HMQAUSZ01 bacterial strain.
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