CN106635921B - Bacillus amyloliquefaciens Y15 and its application - Google Patents
Bacillus amyloliquefaciens Y15 and its application Download PDFInfo
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Abstract
The invention discloses be directed to all plant disease-proofs, growth-promoting, degeneration-resistant bacillus amyloliquefaciens Y15 and its application.Biocontrol microorganisms Y15 is bacillus amyloliquefaciens (Bacillus amyloliquefaciens), and China Committee for Culture Collection of Microorganisms's common micro-organisms center is preserved on November 22nd, 2016, and culture presevation number is CGMCC NO.13327.Concentration 1 × 10 is made in the bacterium8The microbial inoculum of CFU/mL concentrations above in greenhouse under the conditions of normal management, reaches 50% or more to plurality of plant diseases biocontrol effect after bacillus amyloliquefaciens Y15 microbial inoculum root irrigation；And to the growth-promoting effect of various crop up to 18% or more；In field, to multiple diseases control efficiency 47.05% or more, growth-promoting effect highly significant；And it can significantly propose the resistance of plant.
Description
Technical field
The invention belongs to plant protection arts, are related to bacillus amyloliquefaciens Y15 and application.
Background technique
China is large agricultural country, and China crop (including vegetables, fruit tree, Chinese medicine, cereal crops etc.) has produced the gross area
More than 2,200,000,000 mu, tremendous contribution is made that for world agriculture development.But China is also that pest and disease damage is multiple, takes place frequently and retransmit big
State, pest and disease damage and floods, drought and referred to as three the Nature disaster of China.FAO's estimation, the annual pest and disease damage in China
Caused crop loss is up to 70,000,000,000-900 hundred million yuan, and wherein disease accounts for 33%, and crop disease just becomes limitation China's agricultural increasingly
Industry produces an important factor for sustainable development.
Plant disease mainly includes soil-borne disease and overground part disease.Plant soil-borne diseases usually cause the plant root to be
To the illness of complete stool property, occur and prevalence generally fall into popular disease for many years, such as cause line it is withered, withered, yellow wither, stand it is withered, sudden
, the diseases such as root-rot, green withered, soft corruption, root-knot nematode, cyst nematode, knee.Overground part disease main harm blade, stem, fruit
The grounds position such as real, it is mainly incoming outer outside greenhouse by air-flow and the farming activities of people, as leaf-curl, downy mildew,
Anthracnose, rust, gray mold, virosis etc..These disease pathogens have that host range is wide, resistance is strong, is difficult to eradicate
Feature.
Poor environment includes the bad weathers such as arid, flood, high temperature, cold, can be generated not to the growth of herbaceous crops
Good influence.In the 1990s, area suffered from drought is 27,330,000 hm every year on average for China2, grain because non-irrigated output reduction and total output it
Than having reached 4.7%.And every year because waterlogged disaster loses 62.3 hundred million kg of grain, 46.7 hundred million yuan of average annual economic loss.High temperature is trembled with fear
Cold equal weather can also cause serious harm to crop production.Increase the extraneous poor environment ability of herbaceous crops resistance and receives the country
The great attention of outer researcher, effective method is considerably less at present.
It is in recent years that the generation and induction that disease is controlled using soil beneficial microbe, which improve crop anti-adversity ability,
The hot spot of research, especially these microorganism no pollution to the environment, cause scientists using biological control means control disease,
And the interest of reply poor environment.Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) are a kind of and withered grass
The very high bacterium of bacillus affinity, can produce a series of metabolite in the growth course of its own, these generations
Thanking to product enables bacillus amyloliquefaciens to have the activity or inducing plant resistance for widely inhibiting fungi and bacterium.
Therefore, in recent years a large amount of scholars in the new discovery and new opplication and gene functional research to bacillus amyloliquefaciens and the item that ferments
A large amount of exploration has been done in the optimization of part.It can in different ways or approach influence plant many metabolic processes, increase plant roots
It is long, change plant root species form and build up, promote absorption of the root system to moisture content and nutrient, bacillus amyloliquefaciens can be improved
The disease resistance of plant, resist pathogen infect and anti-adversity ability, enhancing plant reply cold, salinity, the environment such as arid
Stress.
Bacillus amyloliquefaciens are distributed very wide in nature, while can generate inhibition to a variety of fungies and bacterium
Effect, and there is own metabolism product abundant.Therefore, it has broad application prospects in terms of biological control, Ke Yishi
When instead of traditional chemical pesticide.Meanwhile with the raising of molecular studies technology, the gene function of bacillus amyloliquefaciens is more
It mostly is appreciated that, increases applicable feasibility.And the optimization of fermentation condition so that answering for bacillus amyloliquefaciens on a large scale
With also having objective condition.
Summary of the invention
The purpose of the present invention is the deficiencies for existing controlling plant diseases and resistance poor environment technology, provide one kind
Environmentally protective can improve broad-spectrum plant diseases prevention, growth-promoting, degeneration-resistant bacillus amyloliquefaciens Y15.
The purpose of the present invention can be achieved through the following technical solutions:
For the diseases prevention of broad-spectrum plant, growth-promoting, degeneration-resistant bacillus amyloliquefaciens Y15, which is named as Xie Dian
Powder bacillus (Bacillus amyloliquefaciens) is preserved in Chinese microorganism strain guarantor on November 22nd, 2016
Administration committee's common micro-organisms center is hidden, culture presevation number is CGMCC NO.13327.
Bacillus amyloliquefaciens Y15 of the present invention is preparing broad spectrum activity plant disease-proof, growth-promoting, in degeneration-resistant preparation
Using；It is preferred that Chinese medicines such as the horticulture and flowers such as chrysanthemum and dendrobium candidum etc. are all in crops such as water fruits and vegetables, grain and oil crops
Disease on plant promotes it to grow and improve quality, improves the application in the preparation of its resistance.The solution starch gemma
Bacillus Y15 is further preferably in prevention and treatment bacterial fruit blotch of watermelon, tomato yellow leaf curl virus disease, peach leaf-curl, or promotes
Watermelon, tomato, cucumber, rice, water spinach, chrysanthemum, Growth of Dendrobium candidum improve its quality；Improve answering in Drought Resistance of Tomato
With.With the microbial inoculum of the present invention prepared for various plants diseases prevention, growth-promoting, degeneration-resistant bacillus amyloliquefaciens Y15.
The microbial inoculum is preferably prepared by the following method to obtain:
Bacillus amyloliquefaciens Y15 28 DEG C of 180rpm shaken cultivation 12-16h in LB culture solution, then 6000rpm is centrifuged
10min obtains thallus, is diluted thallus with aqua sterilisa and microbial inoculum is made, and viable bacteria total concentration is 1 × 10 in microbial inoculum finished product9-1×
1010CFU/mL。
The preparation method of microbial inoculum of the present invention, it is characterised in that the following steps are included:
Bacillus amyloliquefaciens Y15 28 DEG C of 180rpm shaken cultivation 12-16h in LB culture solution, then 6000rpm is centrifuged
10min obtains thallus, is diluted thallus with aqua sterilisa and microbial inoculum is made, and viable bacteria total concentration is 1 × 10 in microbial inoculum finished product9-1×
1010CFU/mL。
Application of the microbial inoculum of the present invention in herbaceous plant diseases prevention, growth-promoting are degeneration-resistant；It is preferred that making in water fruits and vegetables, grain and oil
The crops such as object, disease on all plants such as Chinese medicines such as the horticulture and flowers such as chrysanthemum and dendrobium candidum promote it to grow and mention
High-quality improves application in the preparation of its resistance.
The microbial inoculum prepared by bacillus amyloliquefaciens Y15 is bent in prevention and treatment bacterial fruit blotch of watermelon, tomato yellowing
Leaf disease viral disease, peach leaf-curl, or promote watermelon, tomato, cucumber, rice, water spinach, chrysanthemum, Growth of Dendrobium candidum, improve it
Quality；Improve the application in Drought Resistance of Tomato.
The microbial inoculum preferably passes through root irrigation and is applied.
Beneficial effect
The present invention, which is utilized, prevents and treats broad spectrum activity plant soil-borne diseases and overground part disease to the microbial bacterial agent of environment nonhazardous
Evil (especially prevention and treatment watermelon fruit blotch), provides a kind of environmentally protective bacillus amyloliquefaciens microbial inoculum, prevents and treats preferably in water
The crops such as fruits and vegetables, grain and oil crop, the soil on all plants such as Chinese medicines such as the horticulture and flowers such as chrysanthemum and dendrobium candidum pass
Disease and overground part disease promote the growth of above-mentioned all kinds of crops and improve quality and to improve its resistance (including drought resisting, anti-
Cold, heat-resisting, waterlogging etc. resist the bad climates such as cold spell in later spring).It is ineffective or have residual to solve other methods such as chemical agent
The problems such as staying protects environment, promotes agricultural sustainable development.
Said preparation is to reaching 52.17% to plurality of plant diseases biocontrol effect after bacillus amyloliquefaciens Y15 root irrigation
More than, it acts on fairly obvious；And plant height to plant and root long have and significantly improve, and there is certain growth-promoting to make plant
With compared with the control group, volume increase surpasses 53.99% to the field-crop handled through Y15.The bacillus amyloliquefaciens microbial inoculum, can room temperature
It saves -1 year 6 months.The plant field disease prevention growth-promoting test result of bacillus amyloliquefaciens Y15 microbial inoculum processing shows to disease
Preventive effect is up to 52.17%, and effect of increasing production is more than 53.99%.
Detailed description of the invention
Fig. 1, bacillus amyloliquefaciens Y15 microbial inoculum handle the biocontrol effect to crop disease
The influence of Fig. 2, the processing of bacillus amyloliquefaciens Y15 microbial inoculum to crop biomass
The influence of Fig. 3, the processing of bacillus amyloliquefaciens Y15 microbial inoculum to crop anti-adversity
Biological deposits
Bacillus amyloliquefaciens Y15 is bacillus amyloliquefaciens Bacillus amyloliquefaciens, in 2016
It was preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on November 22, culture presevation number is CGMCC
NO.13327, address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City institute of microbiology, the Chinese Academy of Sciences.
Specific embodiment
The molecules of 1 bacillus amyloliquefaciens bacterial strain Y15 of embodiment is identified
Y15 is by isolated in the rhizosphere soil of Zhejiang theatre.
After bacterial genomes kit (SBS Genetech company) extracts bacterial gene DNA, with gyrB universal primer gyrB1 (5'-
GAAGTCATCATGACCGTTCTGCAYGCNGGNGGNAARTTYGA-3')；gyrB2R(5'–AGCAGGATACGGATGTGCGA
GCCRTCNACRTCNGCRTCNGTCAT-3' gyrB genetic fragment) is amplified from genomic DNA.Specific amplification system and journey
Sequence (25ul) is as follows: each 1 μ l of primer 1,2 (10 μM)；Mix:12.5μl；Taq:0.5μl.ddH2O:10μl；Template: 1 μ l.94℃
Initial denaturation 5min, 94 DEG C of denaturation 1min, 56 DEG C of annealing 2min, 72 DEG C of extension 2min, 35 circulations, 72 DEG C of extension 10min, 4 DEG C
It saves.Pcr amplification product is detected with 1% agarose gel electrophoresis, 90V, 30min；EB dyes 30min, and detection amplification produces
Object.Amplified production is connected to pMD19-T carrier after purification, and conversion extracts positive colony matter in Escherichia coli DH5 α
Grain, commission TaKaRa company sequencing, is submitted NCBI (National Center for Biotechnology Information)
It is compared.Comparison result shows the bacterial strain and Bacillus amyloliquefaciens similarity is 100%, therefore is accredited as
Bacillus amyloliquefaciens.
Embodiment 2
Bacillus amyloliquefaciens Y15 microbial inoculum the preparation method is as follows:
Bacillus amyloliquefaciens Y15 28 DEG C of 180rpm shaken cultivation 12-16h in LB culture solution, then 6000rpm is centrifuged
10min obtains thallus, is diluted thallus with aqua sterilisa and microbial inoculum is made, and viable bacteria total concentration is 1 × 10 in microbial inoculum finished product9-1×
1010CFU/mL。
The test of 3 bacillus amyloliquefaciens Y15 greenhouse growth-promoting of embodiment
Method: the watermelon that will participate in the experiment, tomato, cucumber seeds 10%H2O2Surface sterilization 10min, sterile water dry after cleaning
It is spare.Each nutritive cube sows 10 seeds.After normal watering management 4 weeks, by the consistent transplantation of seedlings of growing way to 330ml plastic cup
In, by the bacillus amyloliquefaciens Y15 microbial inoculum to have suspended with 1.0 × 10 when transplanting7The concentration pouring root of CFU/mL is applied to crop
Root, every plant of 30mL amount.It is subsequently placed in greenhouse and cultivates, 28 DEG C, 70% relative humidity.Using clear water processing as control, every group
48 young plants are handled, are randomly placed, altogether in triplicate.Watermelon, tomato, cucumber plant plant height, stem thickness, fresh weight, dry weight are measured after 30d
Equal biomass.
To participate in the experiment rice, water spinach seed 10%H2O2Surface sterilization 10min, sterile water dry spare after cleaning.By water
Rice, water spinach seed are with 1.0 × 107The bacillus amyloliquefaciens Y15 microbial inoculum seed soaking 5min of the concentration of CFU/mL.Again in each battalion
Support alms bowl (high 13cm, diameter 9cm) sow 4 seeds, after emergence (7d), by the bacillus amyloliquefaciens Y15 microbial inoculum to have suspended with
1.0×107The concentration pouring root of CFU/mL is applied to crop root, every plant of 30mL amount.Using clear water processing as control.Every group of processing
48 young plants, randomly place, altogether in triplicate.Rice, water spinach plant plant height, stem thickness, fresh weight, dry weight etc. are measured after being inoculated with 30d
Biomass.
Using the chrysanthemum participated in the experiment from the tender tip of the long 12cm of clip on the shoot that perennial root (old root) is sprouted as cutting, from section base portion
It scabbles, cuts the two panels great Ye of base portion, then the leaf on top is repaired into half (reducing moisture evaporation, prevent cutting from wilting)
Cuttage (diameter 9cm, high 13cm) into basin alms bowl.By the bacillus amyloliquefaciens Y15 microbial inoculum to have suspended with 1.0 × 107CFU/mL
Concentration pouring root be applied to crop root, every plant of 30mL amount.Using clear water processing as control.Every group of 48 young plant of processing, puts at random
It sets, altogether in triplicate.The biomass such as plant plant height, stem thickness, fresh weight, dry weight are measured after being inoculated with 30d.
Biomass increases (%)=(processing group plant weights-adjoining tree dry weight)/adjoining tree dry weight × 100%
As shown in table 1,2,3,4,5,6, bacillus amyloliquefaciens Y15 can remarkably promote Solanaceae tomato, Curcurbitaceae watermelon,
Cucumber, rice, leafy vegetable water spinach, the growth of chrysanthemum show the fresh weight dry weight of crop, stem length, stem thickness compared with the control,
Have significant increase, as a result calculate bacillus amyloliquefaciens Y15 to watermelon, Solanaceae tomato, cucumber, rice, leafy vegetable
Water spinach, the biomass increase of chrysanthemum respectively reaches 27.87%, 12.93%, 29.70%, 40.00%, 23.08%,
43.90%.
Influence of the 1 bacillus amyloliquefaciens Y15 of table to watermelon biomass
Note: different lowercase persons indicate in 0.05 horizontal upper significant difference.
Influence of the 2 bacillus amyloliquefaciens Y15 of table to tomato biological amount
Note: different lowercase persons indicate in 0.05 horizontal upper significant difference.
Influence of the 3 bacillus amyloliquefaciens Y15 of table to cucumber biomass
Note: different lowercase persons indicate in 0.05 horizontal upper significant difference.
Influence of the 4 bacillus amyloliquefaciens Y15 of table to rice biological amount
Note: different lowercase persons indicate in 0.05 horizontal upper significant difference.
Influence of the 5 bacillus amyloliquefaciens Y15 of table to water spinach biomass
Note: different lowercase persons indicate in 0.05 horizontal upper significant difference.
Influence of the 6 bacillus amyloliquefaciens Y15 of table to chrysanthemum biomass
Note: different lowercase persons indicate in 0.05 horizontal upper significant difference.
4 bacillus amyloliquefaciens Y15 greenhouse of embodiment is to bacterial disease efficiency test
Method: will participate in the experiment watermelon seed 10%H2O2Surface sterilization 10min, sterile water dry spare after cleaning.
Watermelon seed after vernalization is seeded in equipped in basin Portland, one, every basin keeps soil before sowing in sterile soil
It is more dry.When watermelon grew to for 6 leaf phase, by the bacillus amyloliquefaciens Y15 biocontrol agent to have suspended with 1.0 × 107CFU/
The concentration pouring root of mL is applied to crop root, is subsequently placed in greenhouse and cultivates, and 28 DEG C, it is more than 70% relative humidity, 16/8h light week
Phase.Biocontrol microorganisms to be seeded carried out the inoculation of pathogen, inoculum density 10 after 5 days7CFU/ml, should using hand spray apparatus
Bacterium, which nebulizes, is sprayed on the surface of blade, keeps blade wet and bacterium solution does not flow down just, and the preceding all plant of inoculation do not send out
Disease starts to fall ill and count its disease incidence, and calculate preventive effect after inoculation pathogen 3 days.
Bacterial fruit blotch of watermelon blade grade scale: (Ofir Bahar (2009))
Morbidity statistics (state of an illness statistics):
0 grade: not falling ill；
1 grade: having fragmentary scab on sick leaf, scab accounts for 10% or less leaf area；
: there is a small amount of scab in 2 poles on sick leaf, scab accounts for the 11%~25% of leaf area；
3 grades: Lesion number is medium on sick leaf, and scab accounts for the 26%~50% of leaf area；
4 grades: Lesion number is more on sick leaf, and scab accounts for the 51%~75% of leaf area；
5 grades: there are many scab on sick leaf, scab accounts for the 76%~90% of leaf area；
6 grades: having a large amount of scabs on sick leaf, scab accounts for 90% or more leaf area；
Disease severity %=[Σ (sick grade strain number × represent series)/plant sum × highest represents grade value] × 100%；
Preventive effect %=(control disease incidence-processing diseases prevention rate)/control disease incidence × 100%.
Test result shows that bacillus amyloliquefaciens Y15 can significantly reduce the disease incidence of bacterial fruit blotch of watermelon,
Average preventive effect is up to 50.17%.
Preventive effect of the 7. bacillus amyloliquefaciens Y15 of table to bacterial fruit blotch of watermelon
Note: different lowercase persons indicate in 0.05 horizontal upper significant difference.
The prevention and treatment virosis test of 5 bacillus amyloliquefaciens Y15 microbial inoculum of embodiment processing
Method: by tomato seeds (Shanghai cooperation 903) surface sterilization (70% soaked in absolute ethyl alcohol 1~2min, 0.05% time
Sodium chlorate solution impregnates 10min, and aqua sterilisa rinses 6 times), it is seeded in seedling culture hole plate and is placed in 28 DEG C, the 16/8h photoperiod,
It is cultivated in the greenhouse of 70% relative humidity.(25d) is transplanted when tomato seedling grew to for 3~4 leaf phase.By tomato transplantation of seedlings
Into the basin alms bowl for filling matrix, one plant of every alms bowl, be placed in condition be 28 DEG C, the 16/8h photoperiod, 70% relative humidity greenhouse in training
It educates.Tomato transplants the progress same day, root irrigation biocontrol microorganisms first time, and root irrigation bacterial concentration is about 5 × 107CFU/mL, every plant
Seedling dosage is 30mL, is slowly poured in tomato root, and spray at the same time processing, and bacterial concentration is 5 × 107CFU/mL, according to
Surface active agent tween -20 is added in 0.01% final concentration, to tomato leaf even spraying.Virus is carried out after tomato transplanting 7d to connect
Kind.Inoculation method are as follows: no Bemisia tabaci for carrying TYLCV is raised into 48h on the tomato plant of performance TYLCD symptom, then
Healthy plant to be seeded is moved in aleyrodid greenhouse, every plant tomato 30 or so, is inoculated with after 3d and removes tobacco powder using imidacloprid
Lice, by it is each processing move in insect prevention greenhouse (28 DEG C, the 16/8h photoperiod, 70% relative humidity) (Sade D, Eybishtz A,
Gorovits R,et al.A developmentally regulated lipocalin-like gene is
overexpressed in tomato yellow leaf curl virus-resistant tomato plants upon
Virus inoculation, and its silencing abolishes resistance [J] .Plant Molecular
Biology.2012,80:273-287.).Using clear water processing as blank control, 48 young plants of each processing, 3 repetitions.Virus
It is inoculated with 40d or so, counts the state of an illness.
Tomato yellow leaf curl virus disease disease index grade scale (Friedmann M, Lapidot M, Cohen S, et
al.A novel source of resistance to tomato yellow leaf curl virus exhibiting a
symptomLess reaction to viral infection[J].Journal of the American Society
For Horticultural Science.1998,123:1004-1007.):
0 grade: asymptomatic；
1 grade: yellow that top vane is slight, limb edge slightly crimp, florescence flower variation；
2 grades: top vane moderate yellow, limb edge mild crimp fold, florescence flower variation, fruiting period yield is slight
The underproduction；
3 grades: the serious yellow of blade, curling, fold, florescence flower moderate fall off, the fruiting period yield moderate underproduction；
4 grades: large area blade severe deformities reduce, and plant strain growth is slow, hence it is evident that downgrade, florescence flower severe detachment, as a result
The serious reduction of income of phase yield or total crop failure.
Disease severity %=[∑ (disease plant number × disease series)/(total plant number × highest disease series)] × 100；
Biocontrol effect %=[(control disease severity-processing disease severity)/control disease severity] × 100
Test result shows that bacillus amyloliquefaciens Y15 can significantly reduce the disease incidence of tomato yellow leaf curl virus disease,
Its average preventive effect is up to 47.05%.
Preventive effect of the 8 bacillus amyloliquefaciens Y15 of table to tomato yellow leaf curl virus disease
Note: different lowercase persons indicate in 0.05 horizontal upper significant difference.
The prevention and treatment fruit diseases of 6 bacillus amyloliquefaciens Y15 microbial inoculum of embodiment processing and raising fruit yield test
Method: selection triennial peach when Yu Shangnian autumn basal dressing, is adopted when early spring current year grows young leaves and before blooming
With 1.0 × 107The bacillus amyloliquefaciens Y15 microbial inoculum of CFU/mL concentration is poured in processing group root, and totally 3 times.Every processing selection
100 peach, while clear water control group is set.Peach leaf-curl disease incidence is counted in peach picking time summer current year,
And fruit yield and quality are investigated.
Disease incidence %=[the total plant number of disease plant number/investigation] × 100；
Biocontrol effect %=[(compareing sick disease incidence-processing disease incidence)/compare sick disease incidence] × 100
Test result shows that bacillus amyloliquefaciens Y15 can significantly reduce the disease severity of peach leaf-curl, puts down
Equal preventive effect is up to 57.40%；And yield and quality can be significantly improved.
Preventive effect of the 9 bacillus amyloliquefaciens Y15 of table to peach leaf-curl
Note: different lowercase persons indicate in 0.05 horizontal upper significant difference.
Influence of the 10 bacillus amyloliquefaciens Y15 of table to peach fruit biomass
Note: different lowercase persons indicate in 0.05 horizontal upper significant difference.
The Chinese medicine greenhouse growth promotion test of 7 bacillus amyloliquefaciens Y15 microbial inoculum of embodiment processing
The candidum tissue culturing seedling for having grown about 45d for choosing early-stage preparations is material.In sterile super-clean bench, every
Strain candidum tissue culturing seedling base portion inoculation 1.0 × 107The bacillus amyloliquefaciens Y15 microbial inoculum 1mL of CFU/mL concentration, all experiments
Tissue-cultured seedling carries out (25 ± 1) DEG C constant temperature illumination cultivation, daily continuous illumination 12h, intensity of illumination 2000lx.Routine observation each group
The upgrowth situation for training seedling after 90d, observes number of blade, plant height, stem thickness of tissue-cultured seedling etc., the tissue culture of comparative experiments group and control group
Seedling grows growing way, judges the growth-promoting effect of bacterial strain.
The results show that bacillus amyloliquefaciens Y15 can remarkably promote the growth of dendrobium candidum and improve its effectively at
Divide content.
Influence of the 11 bacillus amyloliquefaciens Y15 of table to Growth of Dendrobium candidum
Note: different lowercase persons indicate in 0.05 horizontal upper significant difference.
The induced resistance test of 8 bacillus amyloliquefaciens Y15 microbial inoculum of embodiment processing
Method: will participate in the experiment tomato seeds 10%H2O2Surface sterilization 10min, sterile water dry spare after cleaning.
Tomato seeds after vernalization are seeded in equipped in basin Portland, one, every basin keeps soil before sowing in sterile soil
It is more dry.When tomato grew to for 6 leaf phase, by the bacillus amyloliquefaciens Y15 biocontrol agent to have suspended with 1.0 × 107CFU/
The concentration pouring root of mL is applied to crop root, is subsequently placed in greenhouse and cultivates, and arid of cutting off the water supply is carried out after 1 month, and drought stress is held
Continue 15d, normally waters after drought stress, observe the restoration ecosystem situation of tomato seedling.Every group of 48 young plant of processing, randomly places,
It is repeated 3 times altogether.It is put in the greenhouse according to randomized complete-block design.
After watering carries out restoration ecosystem 1d, bacillus amyloliquefaciens Y15 processing group plant restore normal growth, and control group
Plant is all dead.
Influence of the 12 bacillus amyloliquefaciens Y15 of table to tomato drought tolerance
Note: different lowercase persons indicate in 0.05 horizontal upper significant difference.
Claims (6)
1. a bacillus amyloliquefaciens Y15, the strain classification are named as bacillus amyloliquefaciens (Bacillus
Amyloliquefaciens), China Committee for Culture Collection of Microorganisms's commonly micro- life is preserved on November 22nd, 2016
Object center, culture presevation number are CGMCC NO.13327.
2. bacillus amyloliquefaciens Y15 described in claim 1 is in prevention and treatment bacterial fruit blotch of watermelon, tomato yellow leaf curl virus
Disease, peach leaf-curl；Or promote watermelon, tomato, cucumber, rice, water spinach, chrysanthemum, Growth of Dendrobium candidum, improve its quality；
Improve the application in Drought Resistance of Tomato.
3. the microbial inoculum prepared with bacillus amyloliquefaciens Y15 described in claim 1.
4. microbial inoculum as claimed in claim 3, it is characterised in that be prepared by the following method to obtain:
By bacillus amyloliquefaciens Y15 described in claim 1 in LB culture solution 28 DEG C of 180rpm shaken cultivation 12-16h, then
6000rpm centrifugation 10min obtain thallus, be diluted with aqua sterilisa and microbial inoculum be made, in microbial inoculum finished product viable bacteria total concentration be 1 ×
109-1×1010CFU/mL。
5. microbial inoculum described in claim 3 is in prevention and treatment bacterial fruit blotch of watermelon, tomato yellow leaf curl virus disease, peach leaf-curl；
Or promote watermelon, tomato, cucumber, rice, water spinach, chrysanthemum, Growth of Dendrobium candidum, improve its quality；Improve Drought Resistance of Tomato
In application.
6. application according to claim 5, which is characterized in that the microbial inoculum passes through root irrigation.
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| CN110384110B (en) * | 2018-04-23 | 2021-08-31 | 拜耳作物科学(中国)有限公司 | Method for preventing and controlling tomato yellow leaf curl disease by using bacillus amyloliquefaciens and application |
| CN111778259B (en) * | 2020-06-11 | 2021-11-23 | 安徽农业大学 | Gene related to iron content of pear leaf and application thereof |
| CN113736685B (en) * | 2021-07-05 | 2023-05-09 | 西北农林科技大学 | Construction method of composite flora |
| CN117757661A (en) * | 2023-11-23 | 2024-03-26 | 齐鲁工业大学(山东省科学院) | Microbial agent for relieving drought stress of plants and application thereof |
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