CN110384247A - Tobacco leaf goes rotten sick biological control method in a kind of Tobacco Leaf Curing - Google Patents
Tobacco leaf goes rotten sick biological control method in a kind of Tobacco Leaf Curing Download PDFInfo
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Abstract
The present invention discloses tobacco leaf in a kind of Tobacco Leaf Curing and goes rotten sick biological control method, is related to technical field of plant disease biological control.The biological control method is sprayed with the microorganism formulation for colloid bacillus cereus (Bacillus mucilaginosus) the bacterial strain YN2011 bacterium solution or the YN2011 containing its bacterial strain that deposit number is CGMCC No.5724 before tobacco flue-curing or impregnates tobacco leaf, then is toasted.Spray tobacco leaf processing to tobacco leaf in baking process go rotten disease relative control effect up to 86.22% or more, impregnate tobacco leaf processing to tobacco leaf in baking process go rotten disease relative control effect up to 97.90% or more, significant effect.The method of the present invention is at low cost, easy to operate, and nontoxic, pollution-free, noresidue, is a kind of environmentally friendly method.
Description
Technical field
The present invention relates to technical field of plant disease biological control, and in particular to one plant of colloid bacillus cereus bacterial strain and its
Preparation controls the mildew and rot disease of tobacco leaf in Tobacco Leaf Curing.
Background technique
The mildew and rot disease of tobacco leaf is a kind of important infectious disease (such as Zeng Tingying baking phase tobacco leaf occurred the tobacco flue-curing phase
The Pathogen identification Chinese tobacco journal for disease of going rotten, the 4th phase of volume 2014,20: 65-68).Cigarette strain is one in entire growth course
It is directly exposed in soil and air, by the threat of various moulds, at later stages because of tobacco, nutriment is rich in blade,
Easily breed various microorganisms.During tobacco flue-curing and in storing process, once condition is suitable for, change of going rotten easily occurs for tobacco leaf
Matter substantially loses utility value, brings heavy economic losses to local tobacco grower and leaf tobacco production.Studies have shown that individual roasting
Room tobacco mildew rate during tobacco flue-curing reaches 50% or more.It is studied according to the Japanese exit or entrance of a clitch and constitution etc., after tobacco mildew, in tobacco leaf
It is rapidly decomposed containing substance, normal configuration and content wreck, and appearance luster is dim, issue strong mould sour stench, inhale
Taste severe exacerbation when going mouldy serious, becomes the graveolent rot of black dirty color completely, loses use value, and going rotten
Mycotoxin is also generated in the process, seriously affects human health, such as cause human allergy and asthma.
Containing various nutriments needed for microorganism growth in tobacco, it is widely present the air of barn, tobacco rod, tobacco leaf table
Face.Tobacco leaf is by mould contamination, because tobacco leaf itself is full of nutrition, under suitable temperature and humidity conditions, will cause tobacco mildew.
Petiole end mildew occurs in baking process for Yunnan Province's report, Hongda tobacco tobacco leaf, and mainly mould causes.Zeng Tingying etc.
The pathogen mildew and rot to Fujian Province's baking phase tobacco leaf identifies that confirming that baking phase tobacco leaf goes rotten for the first time is by Rhizopus oryzae
Infectious disease caused by (Rhizopusoryzae Went et Geerligs).There are many factor for influencing tobacco mildew, in addition to
In surrounding air, baking facility, tobacco leaf surface contain outside a large amount of moulds, and most important chemical factors have temperature, and air is relatively wet
Degree, illumination etc..Some researches show that, the optimum growth temperature of pathogen Rhizopus oryzae of the mildew and rot disease of baking process is 32~40 DEG C,
For relative humidity 75%~85%, the temperature is consistent with tobacco flue-curing changing yellow stage temperature, most has the growth using Rhizopus oryzae and invades
Dye, disease extend rapidly sprawling.
Currently, in relation to few, most of tobacco mildew Prevention Technique collection that tobacco mildew Prevention Technique is reported during baking
During tobacco leaf storage.More common mould inhibitor mainly has sodium benzoate, benzoic acid, sorb in the tobacco leaf production reported at present
Acid, but these mould inhibitors can only inhibit the growth of part mould, and fungistatic effect is smaller.Wang Yongdong etc. using chlorine dioxide and
Sym-closene has effectively prevented the mildew and rot disease of the tobacco leaf during baking, but it is sick to utilize this method prevention and treatment baking process to go rotten,
Tobacco leaf chloride ion content is often exceeded.Currently, lacking good biological prevention and control agent or product come tobacco mildew during preventing and treating baking.
Summary of the invention
It goes rotten diease occurrence object controlling party in order to solve the above technical problems, the present invention provides tobacco leaf in a kind of Tobacco Leaf Curing
Method, this method comprises: using living bacteria count before tobacco flue-curing is 1 × 108Colloid bacillus cereus (the Bacillus of cfu/ml
Mucilaginosus) the 0-100 times of liquid sprinkling of bacterial strain YN2011 or immersion tobacco leaf, then again routinely by tobacco leaf in bamboo pole
On, after short grades surface moisture, routinely it is put into hothouse baking, the colloid bacillus cereus (Bacillus
Mucilaginosus) the deposit number of bacterial strain YN2011 are as follows: CGMCC No.5724.
The biological control method of the mildew and rot disease of tobacco leaf includes: tobacco flue-curing in another kind Tobacco Leaf Curing provided by the invention
Preceding microorganism formulation sprinkling or immersion cigarette with containing colloid bacillus cereus (Bacillus mucilaginosus) bacterial strain YN2011
Leaf after short grades surface moisture, is routinely put into hothouse baking, the glue then again routinely by tobacco leaf on bamboo pole
The deposit number of matter bacillus (Bacillus mucilaginosus) bacterial strain YN2011 are as follows: CGMCC No.5724.
The microorganism formulation is the fermentation of colloid bacillus cereus (Bacillus mucilaginosus) bacterial strain YN2011
Liquid, the living bacteria count of colloid bacillus cereus (Bacillus mucilaginosus) bacterial strain YN2011 is 1 in the fermentation liquid
×106Cfu/ml or more.
The fermentation liquid is prepared by the following method:
(1) colloid bacillus cereus (Bacillus mucilaginosus) bacterial strain YN2011 that -80 DEG C save is connect 1~2
For ring into seed culture medium, 32 DEG C of cultures for 24 hours, obtain colloid bacillus cereus (Bacillus mucilaginosus) bacterial strain of activation
Colloid bacillus cereus (Bacillus mucilaginosus) the bacterial strain YN2011 of activation is inoculated with by YN2011 by 1% inoculum concentration
To seed culture medium, under the conditions of 32 DEG C of cultivation temperature, revolving speed 160rpm, shaking table culture 12h obtains primary seed solution, described
Seed culture medium are as follows: 5~10g/L of sucrose, 0.4~1g/L of yeast extract, 1~2.5g/L of ammonium sulfate, calcium carbonate 1~2g/L, seven
0.5~1g/L of water magnesium sulfate, dipotassium hydrogen phosphate 1~3g/L, pH 7.0~7.5；
(2) 1~3% inoculum concentration is pressed, primary seed solution is transferred to the secondary seed medium in seeding tank, 30~34
DEG C, revolving speed 100rpm, ventilatory capacity 1500L/h, tank pressure cultivate 12h under the conditions of being maintained at 0.04~0.06MPa, obtain second level kind
Sub- liquid；The secondary seed medium is identical as seed culture medium described in step (1)；
(3) 1~3% inoculum concentration is pressed, secondary seed solution is inoculated into the fermentation medium in fermentor, at 35~38 DEG C,
Revolving speed 100rpm, ventilatory capacity 1500L/h, tank pressure cultivate 46~48h, culture to hair under the conditions of being maintained at 0.04~0.06MPa
The You of colloid bacillus cereus (Bacillus mucilaginosus) bacterial strain YN2011 imitates Shuo≤1 × 10 Jun living in zymotic fluid6cfu/
Ml, the fermentation medium are identical as seed culture medium described in step (1).
Above-mentioned biological control method, when impregnating tobacco leaf, entire blade is impregnated, and soaking time is 10 minutes or more.
Above-mentioned biological control method, when spraying tobacco leaf, the sprinkling including petiole end, blade back and blade face, the amount of spraying with
Until blade drips.
The tobacco leaf being organized on bamboo bar is put into 3~4 layers of intensive barn curing of average airflow ascending manner by above-mentioned biological control method
Interior baking.
Compared with prior art, the invention has the following advantages:
1. biological control method provided by the present invention can be such that colloid bacillus cereus bacterial strain YN2011 colonizes in tobacco, from
And achieve the purpose that diseases prevention, disease-resistant.Impregnated or sprayed with colloid bacillus cereus bacterial strain YN2011 bacterium solution or its microorganism formulation to
The tobacco leaf of baking is mildew and rot sick to control tobacco leaf, sprays tobacco leaf, and the relative control effect of the mildew and rot disease of tobacco leaf is reachable in baking process
86.22% or more, tobacco leaf is impregnated, tobacco leaf goes rotten sick relative control effect up to 97.90% or more in Tobacco Leaf Curing.
2. the method for the present invention is a kind of environmentally friendly biological control method, have production cost low, easy to operate, nothing
The advantages that malicious, pollution-free, noresidue.
Specific embodiment
Further illustrate that the present invention, each embodiment are conventional method without specified otherwise below with reference to embodiment.
1 colloid bacillus cereus of embodiment (Bacillus mucilaginosus) bacterial strain YN2011's isolates and purifies, identifies
And preservation
1 colloid bacillus cereus bacterial strain YN2011's isolates and purifies
1.1 material
Pick up from the field soil of plantation tobacco.
The preparation of 1.2 culture mediums
Silicate bacteria solid medium are as follows: sucrose 5.0g/L, sodium phosphate 2.0g/L, epsom salt 0.5g/L, carbonic acid
Calcium 0.1g/L, Iron trichloride hexahydrate 5.0mg/L, agar powder 15g/L, pH 7.0.Above-mentioned silicate bacteria solid medium is melted,
Sterilizes culture dish is poured into respectively, it is cooling, plate is made, for separating and saving bacterium.
1.3 being separately cultured
Aseptically, it weighs and picks up from the field soil 10g soil sample of plantation tobacco and be dissolved in the sterile water of 90mL and sufficiently shake
It swings and gradient dilution is at l0-2、10-3、l0-4Soil supension again, then each dilution respectively takes 100 μ L to be spread evenly across respectively
In above-mentioned silicate bacteria solid medium tablets, 3d is cultivated in 37 DEG C of inversions.It selects colourless, transparent, half glassy and rich in bullet
The bacterium colony of property, is further purified until obtaining pure culture.Finally pure culture bacterial strain obtained is transferred to final concentration of
In the glycerol of 25%w/w and it is sealed in -80 DEG C of refrigerators.
The identification of 2 colloid bacillus cereus bacterial strain YN2011
2.1 Morphological Identification
The above-mentioned silicate bacteria solid medium culture of bacterial strain saved in 1.3 is taken, 32 DEG C are cultivated 36 hours, and shape is carried out
State observation identification, bacterium colony is in the circular protrusions of neat in edge, and colorless and transparent, half is glassy, and surface is smooth wet, high resilience
It is difficult to provoke, it is sticky easily to draw wire.Thallus is rod-shaped.The undergrowth on beef-protein medium.It sees under the microscope
It examines, thallus does not move, and has terminal spore, and abundant pod membrane is generated on nitrogen-free agar.With " common bacteria system identification handbook "
Colloid bacillus cereus category morphological feature described in (east show pearl etc. is write, Science Press, 2001) is almost the same, tentatively sentences
The bacterial strain that breaks belongs to colloid bacillus cereus (Bacillus mucilaginosus), number YN2011, referred to as: bacterial strain
YN2011。
The identification of 2.2 Physiology and biochemistries
Bacterial strain YN2011 Gram's staining, spore staining, Starch Hydrolysis, glucose utilization, citrate utilization, V-P are anti-
It answers, nitrate reduction, gelatin liquefaction etc. are positive.
2.3 Molecular Identification results
According to 16S rDNA universal primer extension increasing sequence, according to sequencing result, the 16S rDNA sequence of bacterial strain YN2011 is passed through
It is compared with ncbi database BLAST, bacterial strain YN2011 sequence is the colloid gemma of YNUCC0001 (AY571332) with strain number
The similarity highest of bacillus Bacillus mucilaginosus, up to 99%, in conjunction with the morphological feature of bacterial strain, physio-biochemical characteristics
And 16S rDNA sequence, bacterial strain YN2011 is accredited as colloid bacillus cereus (Bacillus mucilaginosus), is colloid
Bacillus (Bacillus mucilaginosus) bacterial strain YN2011.
The preservation of 3 colloid bacillus cereus (Bacillus mucilaginosus) bacterial strain YN2011
Colloid bacillus cereus (Bacillus mucilaginosus) the bacterial strain YN2011 was protected on January 12nd, 2012
China Committee for Culture Collection of Microorganisms's common micro-organisms center (referred to as: CGMCC) is ensconced, deposit number is
CGMCCNo.5724, classification naming: colloid bacillus cereus Bacillus mucilaginosus, and in 12 daily test January in 2012
It surveys as survival.The common micro-organisms center address: city, BeiJing, China, North Star West Road 1, Chaoyang District institute 3, postcode: 100101.
The preparation of the microorganism formulation of the present invention of embodiment 2, comprising the following steps:
(1) colloid bacillus cereus (Bacillus mucilaginosus) bacterial strain YN2011 that -80 DEG C save is connect 1~2
For ring into seed culture medium, 32 DEG C of cultures for 24 hours, obtain colloid bacillus cereus (Bacillus mucilaginosus) bacterial strain of activation
Colloid bacillus cereus (Bacillus mucilaginosus) the bacterial strain YN2011 of activation is inoculated with by YN2011 by 1% inoculum concentration
To seed culture medium, under the conditions of 32 DEG C of cultivation temperature, revolving speed 160rpm, shaking table culture 12h obtains primary seed solution；It is described
Seed culture medium composition are as follows: sucrose 8g/L, yeast extract 0.7g/L, ammonium sulfate 1.5g/L, calcium carbonate 1g/L, seven water sulfuric acid
Magnesium 0.8g/L, dipotassium hydrogen phosphate 2g/L, pH 7.5.
(2) 2% inoculum concentration is pressed, primary seed solution is transferred to the secondary seed medium in seeding tank, at 32 DEG C, revolving speed
100rpm, ventilatory capacity 1500L/h, tank pressure cultivate 12h under the conditions of being maintained at 0.05MPa, obtain secondary seed solution；The second level
Seed culture medium is identical as seed culture medium described in step (1).
(3) 3% inoculum concentration is pressed, secondary seed solution is inoculated into the fermentation medium in fermentor, at 32 DEG C, revolving speed
100rpm, ventilatory capacity 1500L/h, tank pressure cultivate 46~48h, culture colloid into fermentation liquid under the conditions of being maintained at 0.05MPa
The You of bacillus (Bacillus mucilaginosus) bacterial strain YN2011 imitates Shuo≤1 × 10 Jun living6When cfu/ml, the fermentation
Liquid is the fermentation liquid of the colloid bacillus cereus (Bacillus mucilaginosus) bacterial strain YN2011, and the fermentation liquid is
For microorganism formulation of the present invention；The fermentation medium is identical as seed culture medium described in step (1).
3 immersion treatment of embodiment waits for the mildew and rot sick method of tobacco leaf in flue-cured tobacco BIOLOGICAL CONTROL Tobacco Leaf Curing
1. test period: in August, 2018 in September, -2018
2. test site: the Qujing City of Yunnan Province Xuanwei City town Ge Yi Yixing flue-cured tobacco cooperative society
3. material to be tested: being cloud and mist 105, the good tobacco leaf of same day harvesting ripe for examination tobacco bred.
4. experimental design: be 4 layers of intensive barn curing of average airflow ascending manner for examination barn, the specification of barn be it is long × wide ×
A height of 8m × 2.7m × 4.1m, circulating fan are No. 7 axial flow blowers, and controller is Liaoning Hai Disheng Machinery Co., Ltd. HDS-6
Type bulk curing barn controller, barn fill smoke 45kgm-3.The 4th layer of barn is the region of mildew and rot disease morbidity most serious, test
It is carried out at the 4th layer.
5. colloid bacillus cereus (Bacillus mucilaginosus) bacterial strain for respectively being prepared embodiment 2 with sterile water
The living bacteria count content of YN2011 is 1.0 × 108The microorganism formulation stoste (hereinafter referred to as: microorganism formulation) of cfu/mL is dilute
10 times of liquid, 50 times of liquid and 100 times of liquid are interpreted into, totally 3 concentration immersion treatments wait for flue-cured tobacco, to pick untreated tobacco leaf directly as sky
White control.When each processing impregnates tobacco leaf with microorganism formulation, entire blade carries out immersion 10min.Each a total of 6 bar of processing,
Every bar compiles 70~80 tobacco leaves, places in 4th layer of point of left and right side of barn, on one side 3 bars, normal with two bars between processing and processing
Untreated tobacco leaf separates, and is roasted by local conventional baking process.Test process are as follows:
CK: the tobacco leaf directly picked.
Processing 1: microorganism formulation × 10, i.e., the dilution for being 10 times with sterile water dilution microorganism formulation, colloid bud
The living bacteria count content of spore bacillus (Bacillus mucilaginosus) bacterial strain YN2011 is 1.0 × 107cfu/mL。
Processing 2: microorganism formulation × 50, i.e., the dilution for being 50 times with sterile water dilution microorganism formulation, colloid bud
The living bacteria count content of spore bacillus (Bacillus mucilaginosus) bacterial strain YN2011 is 2.0 × 106cfu/mL。
Processing 3: microorganism formulation × 100, i.e., the dilution for being 100 times with sterile water dilution microorganism formulation, colloid
The living bacteria count content of bacillus (Bacillus mucilaginosus) bacterial strain YN2011 is 1.0 × 106cfu/mL。
6. investigation method: taking 3 groups in each processing at random, record its incidence.The state of an illness recorded refers to Wang Yongdong
Investigation method (such as Wang Yongdong baking phase tobacco leaf go rotten disease infects derived from prevention and treatment Agriculture of Anhui science, 2017,45 (19)
It 138-142) is classified, is divided into 0,1,2,3,4,5 six rank.0 grade is not fall ill；1 grade is less than 1cm for petiole is mouldy,
Blade is not fallen ill；2 grades are that petiole is mouldy greater than 1cm, and blade is not fallen ill；3 grades are that petiole is mouldy, and the mouldy area of blade is less than
25%；4 grades are that petiole is mouldy, and the mouldy area of blade is up to 25%~50%；5 grades are that petiole is mouldy, and the mouldy area of blade is greater than
50%.
Calculation formula:
Disease index=∑ (the diseased plant numbers at different levels × disease grade value) × 100/ (investigation total strain number × superlative degree value)
Relative control effect (%)=(check plot disease index-treatment region disease index) × 100/ check plot disease index
1 embodiment 3 of table impregnates the control efficiency to flue-cured tobacco to the mildew and rot disease of tobacco leaf in Tobacco Leaf Curing
Note: the English letter in table 1 after number indicates the significant difference under the level of P≤0.05.
7. test result: tobacco leaf passes through baking in 7 days, is counted to its disease occurrence degree and (is shown in Table 1), side of the present invention
The microorganism formulation that method dilutes 10 times reaches 99.22% to the control efficiency of the mildew and rot disease of tobacco leaf in Tobacco Leaf Curing, dilution
50 times of microorganism formulation reaches 99.08% to the go rotten control efficiency of disease of tobacco leaf in Tobacco Leaf Curing, 100 times of dilution it is micro-
Biological agent reaches 97.90% to the control efficiency of the mildew and rot disease of tobacco leaf in Tobacco Leaf Curing.
The sprinkling of embodiment 4 processing is to the mildew and rot sick method of tobacco leaf in flue-cured tobacco BIOLOGICAL CONTROL Tobacco Leaf Curing
Have respectively with colloid bacillus cereus (Bacillus mucilaginosus) bacterial strain YN2011 prepared by embodiment 2
Imitating viable count content is 1.0 × 108The microorganism formulation stoste (hereinafter referred to as: microorganism formulation) of cfu/mL, and with sterile
Water is by aforementioned micro organism preparation diluent at 10 times of liquid, 100 times of liquid, and totally 3 concentration sprinkling processing are to flue-cured tobacco, directly to pick not
Processing tobacco leaf is blank control.When each processing sprays tobacco leaf with microorganism formulation, the spray including petiole end, blade back and blade face
It spills, fountain height is until blade drips.Each a total of 6 bar of processing, every bar compile 70~80 tobacco leaves, left at 4th layer point of barn
The right is placed, on one side 3 bars, and with two bars, normally untreated tobacco leaf is separated between processing and processing, by local conventional baking process
It is roasted.Test process are as follows:
CK: the tobacco leaf directly picked.
Processing 1: microorganism formulation, colloid bacillus cereus (Bacillus mucilaginosus) bacterial strain YN2011 have
Imitating viable count content is 1.0 × 108cfu/mL。
Processing 2: microorganism formulation × 10, i.e., the dilution for being 10 times with sterile water dilution microorganism formulation, colloid bud
The living bacteria count content of spore bacillus (Bacillus mucilaginosus) bacterial strain YN2011 is 1.0 × 107cfu/mL。
Processing 3: microorganism formulation × 100, i.e., the dilution for being 100 times with sterile water dilution microorganism formulation, colloid
The living bacteria count content of bacillus (Bacillus mucilaginosus) bacterial strain YN2011 is 1.0 × 106cfu/mL。
Other processing methods identical with example 3, repeat no more.
Test result: tobacco leaf passes through baking in 7 days, and sick disease occurrence degree mildew and rot to its tobacco leaf is counted and (is shown in Table 2).
The method of the present invention sprays the prevention and treatment effect for going rotten sick to tobacco leaf in Tobacco Leaf Curing to flue-cured tobacco using microorganism formulation stoste
Fruit reaches 95.08%, is sprayed to flue-cured tobacco using 10 times of liquid of microorganism formulation to the mildew and rot disease of tobacco leaf in Tobacco Leaf Curing
Control efficiency reaches 90.07%, is sprayed using 100 times of liquid of microorganism formulation and is gone rotten to flue-cured tobacco to tobacco leaf in Tobacco Leaf Curing
The control efficiency of disease reaches 86.22%.
The sprinkling of 2 embodiment 4 of table is to flue-cured tobacco to the control efficiency of the mildew and rot disease of tobacco leaf in Tobacco Leaf Curing
Note: the English letter in table 2 after number indicates the significant difference under the level of P≤0.05.
Claims (9)
1. the biological control method of the mildew and rot disease of tobacco leaf in a kind of Tobacco Leaf Curing, it is characterised in that: with effective before tobacco flue-curing
Viable count is 1 × 1080-100 times of colloid bacillus cereus (Bacillus mucilaginosus) bacterial strain YN2011 of cfu/ml
Liquid sprinkling or immersion tobacco leaf after short grades surface moisture, are routinely put into roasting then again routinely by tobacco leaf on bamboo pole
Baking room, the deposit number of the colloid bacillus cereus (Bacillus mucilaginosus) bacterial strain YN2011 are as follows: CGMCC
No.5724。
2. the biological control method of the mildew and rot disease of tobacco leaf in a kind of Tobacco Leaf Curing, it is characterised in that: with containing glue before tobacco flue-curing
The microorganism formulation of matter bacillus (Bacillus mucilaginosus) bacterial strain YN2011 sprays or impregnates tobacco leaf, then again
Routinely by tobacco leaf on bamboo pole, after short grades surface moisture, it is routinely put into hothouse baking, the colloid bacillus cereus
The deposit number of (Bacillus mucilaginosus) bacterial strain YN2011 are as follows: CGMCC No.5724.
3. biological control method according to claim 2, which is characterized in that the microorganism formulation is colloid bacillus cereus
The fermentation liquid of (Bacillus mucilaginosus) bacterial strain YN2011, colloid bacillus cereus (Bacillus in the fermentation liquid
Mucilaginosus) living bacteria count of bacterial strain YN2011 is 1 × 106Cfu/ml or more.
4. biological control method according to claim 3, it is characterised in that: the fermentation liquid is prepared by following methods
It arrives:
(1) colloid bacillus cereus (Bacillus mucilaginosus) bacterial strain YN2011 that -80 DEG C save is connect 1~2 ring and arrived
In seed culture medium, 32 DEG C of cultures for 24 hours, obtain colloid bacillus cereus (Bacillus mucilaginosus) bacterial strain of activation
Colloid bacillus cereus (Bacillus mucilaginosus) the bacterial strain YN2011 of activation is inoculated with by YN2011 by 1% inoculum concentration
To seed culture medium, under the conditions of 32 DEG C of cultivation temperature, revolving speed 160rpm, shaking table culture 12h obtains primary seed solution, described
Seed culture medium are as follows: 5~10g/L of sucrose, 0.4~1g/L of yeast extract, 1~2.5g/L of ammonium sulfate, calcium carbonate 1~2g/L, seven
0.5~1g/L of water magnesium sulfate, dipotassium hydrogen phosphate 1~3g/L, pH 7.0~7.5；
(2) 1~3% inoculum concentration is pressed, primary seed solution is transferred to the secondary seed medium in seeding tank, at 30~34 DEG C,
Revolving speed 100rpm, ventilatory capacity 1500L/h, tank pressure cultivate 12h under the conditions of being maintained at 0.04~0.06MPa, obtain secondary seed
Liquid；The secondary seed medium is identical as seed culture medium described in step (1)；
(3) 1~3% inoculum concentration is pressed, secondary seed solution is inoculated into the fermentation medium in fermentor, at 35~38 DEG C, revolving speed
100rpm, ventilatory capacity 1500L/h, tank pressure cultivate 46~48h, culture to fermentation liquid under the conditions of being maintained at 0.04~0.06MPa
The You of middle colloid bacillus cereus (Bacillus mucilaginosus) bacterial strain YN2011 imitates Shuo≤1 × 10 Jun living6Cfu/ml, institute
It is identical as seed culture medium described in step (1) to state fermentation medium.
5. according to claim 1 to biological control method described in any claim in 4, which is characterized in that bamboo bar will be organized in
On tobacco leaf be put into baking in 3~4 layers of intensive barn curing of average airflow ascending manner.
6. according to claim 1 to biological control method described in any claim in 4, which is characterized in that when impregnating tobacco leaf,
Entire blade is impregnated, and soaking time is 10 minutes or more.
7. biological control method according to claim 6, which is characterized in that the tobacco leaf being organized on bamboo bar is put into common gas
Flow baking in 3~4 layers of intensive barn curing of ascending manner.
8. according to claim 1 to biological control method described in any claim in 4, which is characterized in that when sprinkling tobacco leaf,
Sprinkling including petiole end, blade back and blade face, the amount of spraying is until blade drips.
9. biological control method according to claim 8, which is characterized in that the tobacco leaf being organized on bamboo bar is put into common gas
Flow baking in 3~4 layers of intensive barn curing of ascending manner.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110786532A (en) * | 2019-11-20 | 2020-02-14 | 云南省烟草农业科学研究院 | Baking method for reducing TSNAs of different varieties of flue-cured tobaccos by using strains |
| CN111869917A (en) * | 2020-07-31 | 2020-11-03 | 云南省烟草公司曲靖市公司 | Novel method for preventing and treating mildew rot in tobacco leaf baking process |
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