CN108552235A - A kind of bio-bacterial manure and its preparation and application for preventing flue-cured tobacco root rot and improveing soil - Google Patents
A kind of bio-bacterial manure and its preparation and application for preventing flue-cured tobacco root rot and improveing soil Download PDFInfo
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Abstract
The present invention discloses a kind of bio-bacterial manure and its preparation and application of prevention flue-cured tobacco root rot and improvement soil.The bio-bacterial manure is mainly made of following raw material：Bio-bacterial manure seed fermented liquid (Trichoderma harzianum 5~20%, bacillus megaterium 5~20%, bacillus subtilis 5~20%, bacillus pumilus 5~20%, Trichoderma viride 5~20%, lactobacillus acidophilus 5~20%), peanut press pulp；And the two mass ratio is 1：3~8.Trichoderma harzianum, bacillus megaterium, bacillus subtilis, bacillus pumilus, Trichoderma viride have good antagonism and Competition to Pathogens of Root Rot object in the bio-bacterial manure, the vital movement of Pathogens of Root Rot object can effectively be inhibited, inducing plant resistance is resisted simultaneously, improves the anti-root rot ability of flue-cured tobacco；Lactobacillus acidophilus can effectively improve soil, reduce the growth of weeds；Environmentally safe simultaneously, nuisanceless, noresidue, specificity is strong, and activity is high, without side-effects safely to non-target organism.
Description
Technical field
The present invention relates to a kind of prevention flue-cured tobacco root rot and the bio-bacterial manure preparation method for improveing soil, more particularly to one kind
Prevent flue-cured tobacco root rot and improves the bio-bacterial manure and the preparation method and application thereof of soil.
Background technology
Root rot is as one of disease common during flue-cured tobacco grown in field, by a variety of diseases such as rotten mould, sickle-like bacteria, phytophthoras
Original, which infects, to be caused, and is a kind of fungoid soil-borne disease, and since China's vega crop rotation rate is low, it is serious that flue-cured tobacco plants phenomenon year after year, closely
In the past few years in China, main flue-cured tobacco growing district root rot incidence is higher and higher.In the vega that nonirrigated farmland is fallen ill, root rot morbidity year after year
Rate causes large stretch of vega cigarette strain withered or even has no harvest, seriously affected the yield and quality of tobacco, caused huge up to 50% or more
Big economic damage.
Currently, both at home and abroad to the control measure of capsicum epidemic disease mainly improve cultivation technique, using disease-resistant variety, crop rotation,
Spray the means such as chemical agent.Although crop rotation can play the generation for mitigating disease, due to the high-quality plant crude opium in China
Lack and peasant household is difficult to realize to the pursuit of economic interests.The cultivation steps such as high ridge transplanting, light irrigation control water can be to a certain degree
The generation of upper control flue-cured tobacco root rot, but cannot all be inherently eliminated the generation of flue-cured tobacco root rot；It can be with using disease-resistant variety
Mitigate disease generation and prevalence to the maximum extent, but tobacco leaf is as special industrial crops, the inherent quality importance of flue-cured tobacco
More than disease resistance, it is difficult to find can highly resistance root rot inherent quality reaches the new flue-cured tobacco varieties of requirement simultaneously；Chemical bactericide
Major measure that is quick with its, quick and becoming prevention flue-cured tobacco root rot, but chemical pesticide readily largely uses, and can pollute
The biological concentration of water body and pesticide causes high risks to health and ecological environment.The long-term of chemical bactericide makes simultaneously
With so that a variety of cause of diseases such as rotten mould, sickle-like bacteria, phytophthora develop immunity to drugs to it, or even morph, and cause under control effect
Drop, or even do not work to root rot.
The present invention selects have the microorganism of antagonism to be prepared into life using to a variety of cause of diseases such as rotten mould, sickle-like bacteria, phytophthoras
Object bacterial manure carries out biological control, not only can effectively prevent root rot, improves soil, and is not easy to produce resistance and pacifies to environment
Entirely, nuisanceless, noresidue, specificity is strong, and activity is high, without side-effects safely to non-target organism.
Invention content
In order to overcome the disadvantages and deficiencies of the prior art, the primary purpose of the present invention is that providing a kind of prevention flue-cured tobacco root-rot
The bio-bacterial manure of disease and improvement soil.The bio-bacterial manure, which can be reduced, effectively inhibits root rot in flue-cured tobacco grown in field growth course
Generation, improve vega soil, promote the root growth and development of flue-cured tobacco early period, improve the yield and quality of flue-cured tobacco.
Another object of the present invention is to provide the preparations of above-mentioned prevention flue-cured tobacco root rot and the bio-bacterial manure of improvement soil
Method.
It is still another object of the present invention to provide the applications of above-mentioned prevention flue-cured tobacco root rot and the bio-bacterial manure of improvement soil.
The purpose of the invention is achieved by the following technical solution：
A kind of bio-bacterial manure for preventing flue-cured tobacco root rot and improveing soil, is mainly made of following raw material：
Bio-bacterial manure seed fermented liquid, peanut press pulp；Wherein, the mass ratio of bio-bacterial manure seed fermented liquid and peanut press pulp
It is 1：3~8；Preferably 1：4；
The bio-bacterial manure seed fermented liquid includes following percent by volume：Trichoderma harzianum 5~20%, huge gemma
Bacillus 5~20%, bacillus subtilis 5~20%, bacillus pumilus 5~20%, Trichoderma viride 5~20%, acidophilus breast bar
Bacterium 5~20%.
Preferably, the bio-bacterial manure seed fermented liquid includes following percent by volume：Trichoderma harzianum 10~20%,
Bacillus megaterium 15~20%, bacillus subtilis 20%, bacillus pumilus 15~20%, Trichoderma viride 15%, acidophilus
Lactobacillus 10~15%.
The bio-bacterial manure for preventing flue-cured tobacco root rot and improvement soil, is preferably mainly made of raw material：Bio-bacterial manure
Seed fermented liquid (Trichoderma harzianum 20%, bacillus megaterium 15%, bacillus subtilis 20%, bacillus pumilus 15%,
Trichoderma viride 15%, lactobacillus acidophilus 15%), peanut press pulp；Wherein, the mass ratio of bio-bacterial manure seed fermented liquid and peanut press pulp
It is 1：4；
The bio-bacterial manure for preventing flue-cured tobacco root rot and improvement soil, is preferably mainly made of raw material：Bio-bacterial manure
Seed fermented liquid (Trichoderma harzianum 15%, bacillus megaterium 20%, bacillus subtilis 20%, bacillus pumilus 20%,
Trichoderma viride 15%, lactobacillus acidophilus 10%), peanut press pulp；Wherein, the mass ratio of bio-bacterial manure seed fermented liquid and peanut press pulp
It is 1：4；
The bio-bacterial manure for preventing flue-cured tobacco root rot and improvement soil, is most preferably mainly made of raw material：Biological bacteria
Fertile seed fermented liquid (Trichoderma harzianum 10%, bacillus megaterium 20%, bacillus subtilis 20%, bacillus pumilus
20%, Trichoderma viride 15%, lactobacillus acidophilus 15%), peanut press pulp；Wherein, bio-bacterial manure seed fermented liquid and peanut press pulp
Mass ratio is 1：4；
The preparation method for preventing flue-cured tobacco root rot and the bio-bacterial manure for improveing soil, includes the following steps：
A, by the Trichoderma harzianum of gained, bacillus megaterium, bacillus subtilis, bacillus pumilus, Trichoderma viride, thermophilic
The seed fermented liquid of Lactobacillus lactis matches according to a certain percentage, and required bio-bacterial manure seed fermented liquid is made；
B, it is particle of the granular size less than 3mm to crush peanut press pulp, and hot air sterilization obtains the peanut press pulp of complete sterilizing,
By bio-bacterial manure seed fermented liquid and peanut press pulp, according to mass ratio 1：3~8 (preferably 1：4) it is carried out under ratio aseptic condition
Uniformly mixing, is transferred in biochemical cultivation case and further carries out cultivation and fermentation；
C, after the completion of cultivation and fermentation, fermentate is taken out, is placed in 38 DEG C of forced air drying condition or is placed in and be protected from light nature
Under the conditions of air-dry, when to moisture content≤12%, be made described and prevent flue-cured tobacco root rot and improve the bio-bacterial manure of soil.
The effective bacterial content of seed fermented liquid described in step (A) is 5 × 108Cfu/mL or more.
Seed fermented liquid described in step (A), is made by the steps to obtain：
(1), the Trichoderma harzianum access for being stored in sand pipe, being freeze-dried in pipe in a dormant state is trained in Cha Shi solids
Support base in, bacillus megaterium, bacillus subtilis and bacillus pumilus access in beef extract-peptone solid medium,
Trichoderma viride is accessed with lactobacterium acidophilus in MRS solid mediums, each strain is used alone a culture medium and avoids polluting,
28 DEG C of merging carries out culture 48 hours, until slant medium covers with thalline；
(2), inclined-plane thalline oese is inoculated in two rings to manufactured LB (broth bouillon), each strain individually makes
With in a LB (broth bouillon), strain pollution is avoided, in 28 DEG C of constant-temperature table, to be cultivated 48 hours in 110r/min,
Bacterial content is made 5 × 108The bacteria suspension of cfu/mL；
(3), manufactured bacteria suspension is linked into LB (broth bouillon) according to 5% inoculum concentration, is shaken in 28 DEG C of constant temperature
Bed, to be cultivated 48 hours in 110r/min, obtains seed fermented liquid.
The Cha Shi solid mediums are (g/L)：Sodium nitrate 2.8, dipotassium hydrogen phosphate 1.2, potassium chloride 0.5, seven water sulphur
Sour magnesium 0.5, ferrous sulfate 0.02, sucrose 30, agar 20, pH7.5；
The beef extract-peptone solid medium is (g/L)：Beef extract 3.0, peptone 10.0, sodium chloride 5.0, fine jade
Fat 15~25, pH7.5；
The MRS solid mediums are：Peptone 10.0g/L, beef extract 10.0g/L, yeast extract 5.0g/L, citric acid
Hydrogen diammonium 2.0g/L, glucose 20.0g/L, Tween 80 1.0mL/L, sodium acetate 5.0g/L, dipotassium hydrogen phosphate 2.0g/L, seven water sulphur
Sour magnesium 0.58g/L, manganese sulfate monohydrate 0.25g/L, agar 18.0g/L, pH6.4；
The LB (broth bouillon)：Tryptone 10g/L, yeast extract 5g/L, sodium chloride (NaCl) 10g/L,
pH7.4；
The condition of hot air sterilization described in step (B) is that 3 hours of hot air sterilization are carried out in 121 DEG C of autoclaves.
Effective bacterial content is 2 × 10 in bio-bacterial manure described in step (C)8Cfu/g or more.
The bio-bacterial manure can be adapted for the various soil of tobacco leaf planting.
The bio-bacterial manure for preventing flue-cured tobacco root rot and improveing soil is in preventing flue-cured tobacco root rot and improvement soil
Application.
The application process for preventing flue-cured tobacco root rot and the bio-bacterial manure for improveing soil, includes the following steps：
(1) it is turned over to planting vega block, smooth, hack, accomplishes that furrow is straight, ridge body is full, and ridge face is fine crushing；
(2) by bio-bacterial manure and organic fertilizer in mass ratio 1：3~7 (preferably 1：4) ratio is uniformly mixed, and takes double items
Formula fertilizing method, the ditch for being 8~12cm (preferably 9cm) in soil both sides two depth of the reclamation of wasteland for determining plantation tobacco, is sprinkled into
Then ditch is carried out smooth ridging by Special compound fertilizer for tobacco fertilizer as lower layer's fertilizer；Then one is opened up on the position of ridge line
Depth is the ditch of 8~12cm (preferably 9cm), is sprinkled into the mixture of bio-bacterial manure and organic fertilizer as upper layer fertilizer, in this way
Upper layer fertilizer is combined in pairs with lower layer's fertilizer, constitutes double-bar type fertilizing method；
(3) since the humiture of environment, soil are larger to the influential effect of bio-bacterial manure, bio-bacterial manure optimum temperature is 25
DEG C~37 DEG C, it is less than 5 DEG C, is higher than 45 DEG C, application effect is poor；Therefore when base fertilizer applies fertilizer completion, timely ridging epiphragma to wait for
It plants.
The principle of the present invention is：
The Trichoderma harzianum fungus Trichoderma energy parasitic a variety of disease fungus common as biocontrol fungi, increase trichoderma in soil
Bacterium number amount can prevent the generation of certain diseases, and Trichoderma harzianum can secrete generation cell wall degrading enzyme, as cellulase, protease make
The cell wall of pathogen mycelia is locally thinning, resolution, eventually leads to pathogen cell wall rupture, and mycelia fracture, pathogen is dead,
There is an antagonistic ability to the Pathogens Causing Root Rot Disease of various plants, Trichoderma harzianum can not only controlling plant diseases, moreover it is possible to plant be promoted to give birth to
The functions such as long, enhancing resistance and reparation pollution environment.Bacillus subtilis, bacillus pumilus, bacillus megaterium conduct
Common bacillus has be exceedingly fast fertility, stronger adaptive capacity to environment and metabolite diversity, in biological control
It is widely used in research, bacillus colonizes in plant rhizosphere, forms competitive relation with Pathogens Causing Root Rot Disease, while secreting generation
It thanks to object and inhibits Pathogens Causing Root Rot Disease growth, and inducing plant resistance is resisted, and the anti-root rot ability of flue-cured tobacco is improved.Lactic acid bacteria is soil
Beneficial microbe in earth, it is softness that lactobacillus acidophilus can enhance soil, alleviates hardening soil phenomenon, and it is saturating to promote Soil ventilation
Light water permeability, while lactic acid bacteria can generate lactic acid and physiological activator, lactic acid can make the kind skin of weeds soften, it is made to be easy
Moisture content is absorbed, and is influenced and breaking dormancy by physiological activator.
The infecting with flue-cured tobacco disease resistance difference for a variety of cause of diseases such as rotten mould, sickle-like bacteria, phytophthora are pathogenetic of flue-cured tobacco root-rot
This reason.Trichoderma harzianum, bacillus megaterium, bacillus subtilis, bacillus pumilus, Trichoderma viride, lactobacillus acidophilus this
Bio-bacterial manure seed fermented liquid that several beneficial bacteriums are made into according to a certain percentage and peanut press pulp co-fermentation at bio-bacterial manure
A variety of pathogens such as, sickle-like bacteria mould to the corruption of the root rot in soil, phytophthora have good inhibiting effect, can be effectively less
Pathogen quantity in soil, while and improved the soil texture, promote development of flue-cured tobacco, improve flue-cured tobacco disease resistance, subtract
The generation of few flue-cured tobacco root rot, improves the yield and quality of flue-cured tobacco.
The present invention has the following advantages and effects with respect to the prior art：
The present invention bio-bacterial manure in Trichoderma harzianum, bacillus megaterium, bacillus subtilis, bacillus pumilus,
Trichoderma viride has good antagonism and Competition to Pathogens of Root Rot object, can effectively inhibit the life of Pathogens of Root Rot object
Activity, while inducing plant resistance is resisted, and the anti-root rot ability of flue-cured tobacco is improved；Lactobacillus acidophilus can effectively improve soil, subtract
The growth of few weeds；The bio-bacterial manure of the present invention is environmentally safe simultaneously, and nuisanceless, noresidue, specificity is strong, and activity is high, right
Non-target organism is without side-effects safely.
Specific implementation mode
With reference to embodiment, the present invention is described in further detail, and embodiments of the present invention are not limited thereto.
Test method used in following embodiments is conventional method unless otherwise specified；Used material, examination
Agent etc., unless otherwise specified, for the reagent and material commercially obtained.
Trichoderma harzianum used in embodiment (Trichoderma harzianum), is purchased from China General Microbiological strain
Preservation administrative center, CGMCC:5.1213；Bacillus megaterium (Bacillus megaterium) is purchased from Chinese common micro- life
Object culture presevation administrative center, CGMCC:1.9072；Bacillus subtilis (Bacillus subtilis) is purchased from Chinese common
Microbiological Culture Collection administrative center, CGMCC:1.15792；Bacillus pumilus (Bacillus pumilus), is purchased from China
General Microbiological Culture preservation administrative center, CGMCC:1.8714；Trichoderma viride (Trichoderma viride), is purchased from China
General Microbiological Culture preservation administrative center, CGMCC:3.11450；Lactobacillus acidophilus (Lactobacillus
Acidophilus), it is purchased from China General Microbiological culture presevation administrative center, CGMCC:1.12735.
Embodiment 1
A kind of bio-bacterial manure for preventing flue-cured tobacco root rot and improveing soil, each component in the bio-bacterial manure is according to the following ratio
It is made：
Bio-bacterial manure seed fermented liquid (volume percentage：Trichoderma harzianum 20%, bacillus megaterium 15%, withered grass
Bacillus 20%, bacillus pumilus 15%, Trichoderma viride 15%, lactobacillus acidophilus 15%), peanut press pulp；
The preparation method for preventing flue-cured tobacco root rot and the bio-bacterial manure for improveing soil, includes the following steps：
A, the Trichoderma harzianum for being stored in sand pipe, being freeze-dried in pipe in a dormant state is accessed in Cha Shi solid cultures
In base, bacillus megaterium, bacillus subtilis and bacillus pumilus are accessed in beef extract-peptone solid medium, are green
Color trichoderma is accessed with lactic acid bacteria in MRS solid mediums, each strain is used alone a culture medium and avoids polluting, merging 28
Culture 48 hours is carried out in DEG C growth cabinet, until slant medium covers with thalline；
B, inclined-plane thalline oese is inoculated in two rings to manufactured LB (broth bouillon), each strain is used alone
In one LB (broth bouillon), strain pollution is avoided, in 28 DEG C of constant-temperature table, to be cultivated 48 hours in 110r/min, system
At bacterial content 5 × 108The bacteria suspension of cfu/mL；
C, manufactured bacteria suspension is linked into LB (broth bouillon) according to 5% inoculum concentration, in 28 DEG C of constant-temperature table,
To be cultivated 48 hours in 110r/min, seed fermented liquid is obtained.
D, by the different seed fermented liquids of gained according to Trichoderma harzianum 20%, bacillus megaterium 15%, withered grass gemma
Bacillus 20%, bacillus pumilus 15%, Trichoderma viride 15%, lactobacterium acidophilus 15% proportions, required biology is made
Bacterial manure seed fermented liquid；
E, it is particle of the granular size less than 3mm to crush peanut press pulp, and hot air sterilization 3 is carried out in 121 DEG C of autoclaves
Hour, the peanut press pulp of complete sterilizing is obtained, by bio-bacterial manure seed fermented liquid and peanut press pulp, according to mass ratio 1：4 ratio
It is uniformly mixed under aseptic condition, is transferred in biochemical cultivation case and further carries out cultivation and fermentation；
F, after the completion of cultivation and fermentation, fermentate is taken out, is placed in 38 DEG C of air dry oven or is placed in and be protected from light nature item
It is air-dried under part, when to moisture content≤12%, is made described and prevents flue-cured tobacco root rot and improve the bio-bacterial manure of soil.
The application process for preventing flue-cured tobacco root rot and the bio-bacterial manure for improveing soil, includes the following steps：
(1) it is turned over to planting vega block, smooth, hack, accomplishes that furrow is straight, ridge body is full, and ridge face is fine crushing；
(2) by bio-bacterial manure and organic fertilizer in mass ratio 1：4 ratios are uniformly mixed, and take double-bar type fertilizing method,
It determines that the ditch that two depth are 9cm or so is opened up wasteland on the soil both sides of plantation tobacco, is sprinkled into Special compound fertilizer for tobacco fertilizer and (is purchased from
Hubei happiness comes the compound fertilizer of plump industry Co., Ltd, 15-15-15) it is used as lower layer's fertilizer, ditch is then subjected to smooth ridging.
Then the ditch that a depth is 9cm or so is opened up on the position of ridge line, the mixture for being sprinkled into bio-bacterial manure and organic fertilizer is made
For upper layer fertilizer, such upper layer fertilizer is combined in pairs with lower layer's fertilizer, constitutes double-bar type fertilizing method；
(3) since the warm moisture content of environment, soil are larger to the influential effect of bio-bacterial manure, bio-bacterial manure optimum temperature is
25 DEG C~37 DEG C, it is less than 5 DEG C, is higher than 45 DEG C, application effect is poor.Therefore when base fertilizer applies fertilizer completion, timely ridging epiphragma
It waits planting；
Embodiment 2
A kind of bio-bacterial manure for preventing flue-cured tobacco root rot and improveing soil, each component in the bio-bacterial manure is according to the following ratio
It is made：
Bio-bacterial manure seed fermented liquid (volume percentage：Trichoderma harzianum 15%, bacillus megaterium 20%, withered grass
Bacillus 20%, bacillus pumilus 20%, Trichoderma viride 15%, lactobacillus acidophilus 10%), peanut press pulp；
The preparation method for preventing flue-cured tobacco root rot and the bio-bacterial manure for improveing soil, includes the following steps：
A, the Trichoderma harzianum for being stored in sand pipe, being freeze-dried in pipe in a dormant state is accessed in Cha Shi solid cultures
In base, bacillus megaterium, bacillus subtilis and bacillus pumilus are accessed in beef extract-peptone solid medium, are green
Color trichoderma is accessed with lactic acid bacteria in MRS solid mediums, each strain is used alone a culture medium and avoids polluting, merging 28
Culture 48 hours is carried out in DEG C growth cabinet, until slant medium covers with thalline；
B, inclined-plane thalline oese is inoculated in two rings to manufactured LB (broth bouillon), each strain is used alone
In one LB (broth bouillon), strain pollution is avoided, in 28 DEG C of constant-temperature table, to be cultivated 48 hours in 110r/min, system
At bacterial content 5 × 108The bacteria suspension of cfu/mL；
C, manufactured bacteria suspension is linked into LB (broth bouillon) according to 5% inoculum concentration, in 28 DEG C of constant-temperature table,
To be cultivated 48 hours in 110r/min, seed fermented liquid is obtained.
D, by the different seed fermented liquids of gained according to Trichoderma harzianum 15%, bacillus megaterium 20%, withered grass gemma
Bacillus 20%, bacillus pumilus 20%, Trichoderma viride 15%, lactobacterium acidophilus 10% proportions, required biology is made
Bacterial manure seed fermented liquid
E, it is particle of the granular size less than 3mm to crush peanut press pulp, and hot air sterilization 3 is carried out in 121 DEG C of autoclaves
Hour, the peanut press pulp of complete sterilizing is obtained, by bio-bacterial manure seed fermented liquid and peanut press pulp, according to mass ratio 1：4 ratio
It is uniformly mixed under aseptic condition, is transferred in biochemical cultivation case and further carries out cultivation and fermentation；
F, after the completion of cultivation and fermentation, fermentate is taken out, is placed in 38 DEG C of air dry oven or is placed in and be protected from light nature item
It is air-dried under part, when to moisture content≤12%, is made described and prevents flue-cured tobacco root rot and improve the bio-bacterial manure of soil.
The application process for preventing flue-cured tobacco root rot and the bio-bacterial manure for improveing soil, includes the following steps：
(1) it is turned over to planting vega block, smooth, hack, accomplishes that furrow is straight, ridge body is full, and ridge face is fine crushing；
(2) by bio-bacterial manure and organic fertilizer in mass ratio 1：4 ratios are uniformly mixed, and take double-bar type fertilizing method,
It determines that the ditch that two depth are 9cm or so is opened up wasteland on the soil both sides of plantation tobacco, is sprinkled into Special compound fertilizer for tobacco fertilizer and (is purchased from
Hubei happiness comes the compound fertilizer of plump industry Co., Ltd, 15-15-15) it is used as lower layer's fertilizer, ditch is then subjected to smooth ridging.
Then the ditch that a depth is 9cm or so is opened up on the position of ridge line, the mixture for being sprinkled into bio-bacterial manure and organic fertilizer is made
For upper layer fertilizer, such upper layer fertilizer is combined in pairs with lower layer's fertilizer, constitutes double-bar type fertilizing method；
(3) since the warm moisture content of environment, soil are larger to the influential effect of bio-bacterial manure, bio-bacterial manure optimum temperature is
25 DEG C~37 DEG C, it is less than 5 DEG C, is higher than 45 DEG C, application effect is poor.Therefore when base fertilizer applies fertilizer completion, timely ridging epiphragma
It waits planting；
Embodiment 3
A kind of bio-bacterial manure for preventing flue-cured tobacco root rot and improveing soil, each component in the bio-bacterial manure is according to the following ratio
It is made：
Bio-bacterial manure seed fermented liquid (volume percentage：Trichoderma harzianum 10%, bacillus megaterium 20%, withered grass
Bacillus 20%, bacillus pumilus 20%, Trichoderma viride 15%, lactobacillus acidophilus 15%), peanut press pulp；
The preparation method for preventing flue-cured tobacco root rot and the bio-bacterial manure for improveing soil, includes the following steps：
A, the Trichoderma harzianum for being stored in sand pipe, being freeze-dried in pipe in a dormant state is accessed in Cha Shi solid cultures
In base, bacillus megaterium, bacillus subtilis and bacillus pumilus are accessed in beef extract-peptone solid medium, are green
Color trichoderma is accessed with lactic acid bacteria in MRS solid mediums, each strain is used alone a culture medium and avoids polluting, merging 28
Culture 48 hours is carried out in DEG C growth cabinet, until slant medium covers with thalline；
B, inclined-plane thalline oese is inoculated in two rings to manufactured LB (broth bouillon), each strain is used alone
In one LB (broth bouillon), strain pollution is avoided, in 28 DEG C of constant-temperature table, to be cultivated 48 hours in 110r/min, system
At bacterial content 5 × 108The bacteria suspension of cfu/mL；
C, manufactured bacteria suspension is linked into LB (broth bouillon) according to 5% inoculum concentration, in 28 DEG C of constant-temperature table,
To be cultivated 48 hours in 110r/min, seed fermented liquid is obtained.
D, by the different seed fermented liquids of gained according to Trichoderma harzianum 10%, bacillus megaterium 20%, withered grass gemma
Bacillus 20%, bacillus pumilus 20%, Trichoderma viride 15%, lactobacterium acidophilus 15% proportions, required biology is made
Bacterial manure seed fermented liquid
E, it is particle of the granular size less than 3mm to crush peanut press pulp, and hot air sterilization 3 is carried out in 121 DEG C of autoclaves
Hour, the peanut press pulp of complete sterilizing is obtained, by bio-bacterial manure seed fermented liquid and peanut press pulp, according to mass ratio 1：4 ratio
It is uniformly mixed under aseptic condition, is transferred in biochemical cultivation case and further carries out cultivation and fermentation；
F, after the completion of cultivation and fermentation, fermentate is taken out, is placed in 38 DEG C of air dry oven or is placed in and be protected from light nature item
It is air-dried under part, when to moisture content≤12%, is made described and prevents flue-cured tobacco root rot and improve the bio-bacterial manure of soil.
The application process for preventing flue-cured tobacco root rot and the bio-bacterial manure for improveing soil, includes the following steps：
(1) it is turned over to planting vega block, smooth, hack, accomplishes that furrow is straight, ridge body is full, and ridge face is fine crushing；
(2) by bio-bacterial manure and organic fertilizer in mass ratio 1：4 ratios are uniformly mixed, and take double-bar type fertilizing method,
It determines that the ditch that two depth are 9cm or so is opened up wasteland on the soil both sides of plantation tobacco, is sprinkled into Special compound fertilizer for tobacco fertilizer and (is purchased from
Hubei happiness comes the compound fertilizer of plump industry Co., Ltd, 15-15-15) it is used as lower layer's fertilizer, ditch is then subjected to smooth ridging.
Then the ditch that a depth is 9cm or so is opened up on the position of ridge line, the mixture for being sprinkled into bio-bacterial manure and organic fertilizer is made
For upper layer fertilizer, such upper layer fertilizer is combined in pairs with lower layer's fertilizer, constitutes double-bar type fertilizing method；
(3) since the warm moisture content of environment, soil are larger to the influential effect of bio-bacterial manure, bio-bacterial manure optimum temperature is
25 DEG C~37 DEG C, it is less than 5 DEG C, is higher than 45 DEG C, application effect is poor.Therefore when base fertilizer applies fertilizer completion, timely ridging epiphragma
It waits planting；
Embodiment 4
Vega is turned over, is smooth, and hack finishes, and applies fertilizer the stage in base manure, and base manure fertilising is carried out in the method in embodiment 1,
Remaining cultivation step is carried out according to production standard (cured tobacco production technical regulation, DB51/T 1212-2011), 70 days after transplanting,
80 days after transplanting, 90 days investigation root rot incidence and disease index after transplanting, while investigate the economic characters of cured tobacco leaf with
And better-than-average cigarette ratio, this is processing 1.Vega is turned over, is smooth, and hack finishes, and applies fertilizer the stage in base manure, according to production standard
(cured tobacco production technical regulation, DB51/T1212-2011) carries out base manure fertilising, remaining cultivation step is carried out according to production standard,
70 days after transplanting, 80 days after transplanting, 90 days investigation root rot incidence and disease index after transplanting, while investigating cured tobacco leaf
Economic characters and better-than-average cigarette ratio, this for processing 2.Experiment is in 2017 in MeiZhou,GuangDong city Mei County, Fields of Qujing City in Yunnan
Luoping County carries out, and selects the serious field of root rot incidence over the years as experiment field, experimental cultivar cloud and mist 87.
1 Meizhou Mei County of table respectively handles root rot incidence
2 Qujing of Yunnan Luoping of table respectively handles root rot incidence
Known to table 1,2 after the bio-bacterial manure using embodiment 1,70 days Meizhous Mei County, Luoping of Yunnan after flue-cured tobacco transplanting
The incidence of two places is compared with without bio-bacterial manure, and incidence has dropped 8.00,10.80 percentage points respectively, disease index point
1.7,2.4 are not had dropped.After flue-cured tobacco transplanting after 80 days, Meizhou Mei County, Luoping of Yunnan two places incidence with do not have to biological bacteria
Fertilizer is compared, and incidence has dropped 16.40,14.01 percentage points respectively, and disease index has dropped 6.74,6.15 respectively.In flue-cured tobacco
After transplanting after 90 days, Meizhou Mei County, Luoping of Yunnan two places incidence compared with without bio-bacterial manure, incidence has dropped respectively
25.08,22.45 percentage points, disease index has dropped 8.96,8.81 respectively.Illustrate, after using 1 bio-bacterial manure of embodiment
The incidence of flue-cured tobacco root rot can be significantly reduced, the disease index of flue-cured tobacco root rot is reduced, flue-cured tobacco root-rot can be effectively relieved
The generation of disease prevents the further deterioration of disease plant incidence.
3 Meizhou Mei County different disposal cured tobacco leaf economic situation of table
| Processing | First-class cigarette ratio (%) | Upper medium grade cigarette ratio (%) | Yield (kghm-2) | The output value (first hm-2) |
| Bio-bacterial manure | 54.48±1.33 | 95.96±0.99 | 2132.8±5.68 | 54156.7±98.5 |
| Without bio-bacterial manure | 52.85±0.49 | 93.50±0.43 | 1923.3±4.11 | 48368.4±114.6 |
4 Qujing of Yunnan Luoping different disposal cured tobacco leaf economic situation of table
| Processing | First-class cigarette ratio (%) | Upper medium grade cigarette ratio (%) | Yield (kghm-2) | The output value (first hm-2) |
| Bio-bacterial manure | 57.26±1.35 | 96.96±0.89 | 2032.8±5.86 | 58406.7±102.5 |
| Without bio-bacterial manure | 53.34±0.67 | 92.50±0.56 | 1823.3±5.32 | 52248.4±135.6 |
Known to table 3,4 after the bio-bacterial manure using embodiment 1, the better-than-average cigarette ratio of Meizhou Mei County rises 2.46
Percentage point, output increased 209.5kghm-2, the output value improves 5788.3 yuan of hm-2, Qujing of Yunnan Luoping it is better-than-average
Cigarette ratio is 4.46 percentage points above-mentioned, output increased 209.5kghm-2, the output value improves 6158.3 yuan of hm-2, say
It is bright, the better-than-average cigarette ratio of cured tobacco leaf can be significantly significantly improved after using 1 bio-bacterial manure of embodiment, improve the production of flue-cured tobacco
Amount and the output value, effectively improve the income of tobacco grower.
Embodiment 5
Vega is turned over, is smooth, and hack finishes, and applies fertilizer the stage in base manure, and base manure fertilising is carried out in the method in embodiment 2,
Remaining cultivation step according to production standard carry out, 70 days after transplanting, transplanting after 80 days, transplanting after 90 days investigation root rot morbidity
Rate and disease index, while the economic characters of cured tobacco leaf and better-than-average cigarette ratio are investigated, this is processing 1.Vega is turned over, is put down
Whole, hack finishes, and applies fertilizer the stage in base manure, carries out base manure fertilising according to production standard, remaining cultivation step is according to production standard
Carry out, 70 days after transplanting, 80 days after transplanting, 90 days investigation root rot incidence and disease index after transplanting, while investigating roasting
The economic characters of tobacco leaf and better-than-average cigarette ratio afterwards, this is processing 2.Experiment is in 2017 in MeiZhou,GuangDong city Mei County, Yunnan
Qujing Luoping County carries out, and selects the serious field of root rot incidence over the years as experiment field, experimental cultivar cloud and mist 87.
5 Meizhou Mei County of table respectively handles root rot incidence
6 Qujing of Yunnan Luoping of table respectively handles root rot incidence
Known to table 5,6 after the bio-bacterial manure using embodiment 2,70 days Meizhous Mei County, Luoping of Yunnan after flue-cured tobacco transplanting
The incidence of two places is compared with without bio-bacterial manure, and incidence has dropped 6.00,5.81 percentage points respectively, disease index difference
Have dropped 1.23,2.15.After flue-cured tobacco transplanting after 80 days, Meizhou Mei County, Luoping of Yunnan two places incidence with do not have to biological bacteria
Fertilizer is compared, and incidence has dropped 15.66,11.09 percentage points respectively, and disease index has dropped 6.04,4.89 respectively.In flue-cured tobacco
After transplanting after 90 days, Meizhou Mei County, Luoping of Yunnan two places incidence compared with without bio-bacterial manure, incidence has dropped respectively
25.08,18.79 percentage points, disease index has dropped 8.13,8.33 respectively.Illustrate, after using 2 bio-bacterial manure of embodiment
The incidence of flue-cured tobacco root rot can be significantly reduced, the disease index of flue-cured tobacco root rot is reduced, flue-cured tobacco root-rot can be effectively relieved
The generation of disease prevents the further deterioration of disease plant incidence.
7 Meizhou Mei County different disposal cured tobacco leaf economic situation of table
| Processing | First-class cigarette ratio (%) | Upper medium grade cigarette ratio (%) | Yield (kghm-2) | The output value (first hm-2) |
| Bio-bacterial manure | 55.29±1.52 | 95.21±0.87 | 2090.8±6.24 | 53723.7±87.5 |
| Without bio-bacterial manure | 52.85±0.49 | 93.50±0.43 | 1923.3±4.11 | 48368.4±114.6 |
8 Qujing of Yunnan Luoping different disposal cured tobacco leaf economic situation of table
| Processing | First-class cigarette ratio (%) | Upper medium grade cigarette ratio (%) | Yield (kghm-2) | The output value (first hm-2) |
| Bio-bacterial manure | 58.42±1.46 | 97.28±1.13 | 2044.9±6.11 | 58711.7±93.6 |
| Without bio-bacterial manure | 53.34±0.67 | 92.50±0.56 | 1823.3±5.32 | 52248.4±135.6 |
Known to table 7,8 after the bio-bacterial manure using embodiment 2, the better-than-average cigarette ratio of Meizhou Mei County rises 1.71
Percentage point, output increased 167.5kghm-2, the output value improves 5355.3 yuan of hm-2, Qujing of Yunnan Luoping it is better-than-average
Cigarette ratio is 4.78 percentage points above-mentioned, output increased 221.6kghm-2, the output value improves 6463.3 yuan of hm-2, say
It is bright, the better-than-average cigarette ratio of cured tobacco leaf can be significantly significantly improved after using 2 bio-bacterial manure of embodiment, improve the production of flue-cured tobacco
Amount and the output value, effectively improve the income of tobacco grower.
Embodiment 6
Vega is turned over, is smooth, and hack finishes, and applies fertilizer the stage in base manure, and base manure fertilising is carried out in the method in embodiment 3,
Remaining cultivation step according to production standard carry out, 70 days after transplanting, transplanting after 80 days, transplanting after 90 days investigation root rot morbidity
Rate and disease index, while the economic characters of cured tobacco leaf and better-than-average cigarette ratio are investigated, this is processing 1.Vega is turned over, is put down
Whole, hack finishes, and applies fertilizer the stage in base manure, carries out base manure fertilising according to production standard, remaining cultivation step is according to production standard
Carry out, 70 days after transplanting, 80 days after transplanting, 90 days investigation root rot incidence and disease index after transplanting, while investigating roasting
The economic characters of tobacco leaf and better-than-average cigarette ratio afterwards, this is processing 2.Experiment is in 2017 in MeiZhou,GuangDong city Mei County, Yunnan
Qujing Luoping County carries out, experimental cultivar cloud and mist 87.
9 Meizhou Mei County of table respectively handles root rot incidence
10 Qujing of Yunnan Luoping of table respectively handles root rot incidence
Known to table 9,10 after the bio-bacterial manure using embodiment 3,70 days Meizhous Mei County, Luoping of Yunnan after flue-cured tobacco transplanting
The incidence of two places is compared with without bio-bacterial manure, and incidence has dropped 10.13,10.86 percentage points respectively, disease index point
1.53,2.95 are not had dropped.After flue-cured tobacco transplanting after 80 days, Meizhou Mei County, Luoping of Yunnan two places incidence with without biology
Bacterial manure is compared, and incidence has dropped 19.46,17.09 percentage points respectively, and disease index has dropped 7.54,6.69 respectively.Roasting
Cigarette transplanting after after 90 days, Meizhou Mei County, Luoping of Yunnan two places incidence compared with without bio-bacterial manure, incidence declines respectively
28.95,24.79 percentage points, disease index has dropped 9.93,9.53 respectively.Illustrate, is using 3 bio-bacterial manure of embodiment
The incidence of flue-cured tobacco root rot can be significantly reduced afterwards, reduce the disease index of flue-cured tobacco root rot, flue-cured tobacco root can be effectively relieved
The generation of maize ear rot prevents the further deterioration of disease plant incidence.
11 Meizhou Mei County different disposal cured tobacco leaf economic situation of table
| Processing | First-class cigarette ratio (%) | Upper medium grade cigarette ratio (%) | Yield (kghm-2) | The output value (first hm-2) |
| Bio-bacterial manure | 57.28±1.38 | 96.79±0.93 | 2112.8±5.46 | 54451.1±101.3 |
| Without bio-bacterial manure | 52.85±0.49 | 93.50±0.43 | 1923.3±4.11 | 48368.4±114.6 |
12 Qujing of Yunnan Luoping different disposal cured tobacco leaf economic situation of table
| Processing | First-class cigarette ratio (%) | Upper medium grade cigarette ratio (%) | Yield (kghm-2) | The output value (first hm-2) |
| Bio-bacterial manure | 60.42±1.48 | 97.88±1.46 | 2089.7±6.34 | 60612.2±956 |
| Without bio-bacterial manure | 53.34±0.67 | 92.50±0.56 | 1823.3±5.32 | 52248.4±135.6 |
Known to table 11,12 after the bio-bacterial manure of embodiment 3, the better-than-average cigarette ratio of Meizhou Mei County rises 3.29
Percentage point, output increased 189.5kghm-2, the output value improves 6082.7 yuan of hm-2, the better-than-average cigarette of Qujing of Yunnan Luoping
Ratio is 5.38 percentage points above-mentioned, output increased 266.4kghm-2, the output value improves 8363.8hm-2, illustrate,
Using the better-than-average cigarette ratio that can significantly significantly improve cured tobacco leaf after 3 bio-bacterial manure of embodiment, improve the yield of flue-cured tobacco with
The output value effectively improves the income of tobacco grower.
The above embodiment is a preferred embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment
Limitation, it is other it is any without departing from the spirit and principles of the present invention made by changes, modifications, substitutions, combinations, simplifications,
Equivalent substitute mode is should be, is included within the scope of the present invention.
Claims (10)
1. a kind of bio-bacterial manure for preventing flue-cured tobacco root rot and improveing soil, it is characterised in that：Mainly it is made of following raw material：
Bio-bacterial manure seed fermented liquid, peanut press pulp；Wherein, the mass ratio of bio-bacterial manure seed fermented liquid and peanut press pulp is 1：
3~8；
The bio-bacterial manure seed fermented liquid includes following percent by volume：Trichoderma harzianum 5~20%, bacillus megaterium
5~20%, bacillus subtilis 5~20%, bacillus pumilus 5~20%, Trichoderma viride 5~20%, lactobacillus acidophilus 5~
20%.
2. the bio-bacterial manure according to claim 1 for preventing flue-cured tobacco root rot and improveing soil, it is characterised in that：
The mass ratio of the bio-bacterial manure seed fermented liquid and peanut press pulp is 1：4.
3. the bio-bacterial manure according to claim 1 or 2 for preventing flue-cured tobacco root rot and improveing soil, it is characterised in that：
The bio-bacterial manure seed fermented liquid includes following percent by volume：Trichoderma harzianum 10~20%, huge gemma bar
Bacterium 15~20%, bacillus subtilis 20%, bacillus pumilus 15~20%, Trichoderma viride 15%, lactobacillus acidophilus 10~
15%.
4. the bio-bacterial manure according to claim 1 or 2 for preventing flue-cured tobacco root rot and improveing soil, it is characterised in that：
The bio-bacterial manure seed fermented liquid includes following percent by volume：Trichoderma harzianum 20%, bacillus megaterium
15%, bacillus subtilis 20%, bacillus pumilus 15%, Trichoderma viride 15%, lactobacillus acidophilus 15%.
5. the bio-bacterial manure according to claim 1 or 2 for preventing flue-cured tobacco root rot and improveing soil, it is characterised in that：
The bio-bacterial manure seed fermented liquid includes following percent by volume：Trichoderma harzianum 15%, bacillus megaterium
20%, bacillus subtilis 20%, bacillus pumilus 20%, Trichoderma viride 15%, lactobacillus acidophilus 10%.
6. the bio-bacterial manure according to claim 1 or 2 for preventing flue-cured tobacco root rot and improveing soil, it is characterised in that：
The bio-bacterial manure seed fermented liquid includes following percent by volume：Trichoderma harzianum 10%, bacillus megaterium
20%, bacillus subtilis 20%, bacillus pumilus 20%, Trichoderma viride 15%, lactobacillus acidophilus 15%.
7. claim 1~6 any one of them prevents the preparation method of flue-cured tobacco root rot and the bio-bacterial manure for improveing soil,
It is characterized in that including the following steps：
A, by the Trichoderma harzianum of gained, bacillus megaterium, bacillus subtilis, bacillus pumilus, Trichoderma viride, acidophilus breast
The seed fermented liquid of bacillus matches according to a certain percentage, and required bio-bacterial manure seed fermented liquid is made；
B, it is particle of the granular size less than 3mm to crush peanut press pulp, and hot air sterilization obtains the peanut press pulp of complete sterilizing, will give birth to
Object bacterial manure seed fermented liquid and peanut press pulp, according to certain mass than ratio aseptic condition under uniformly mixed, be transferred to
Cultivation and fermentation is further carried out in biochemical cultivation case；
C, after the completion of cultivation and fermentation, fermentate is taken out, is placed in 38 DEG C of forced air drying condition or is placed in and be protected from light natural conditions
It is lower to air-dry, when to moisture content≤12%, it is made described and prevents flue-cured tobacco root rot and improve the bio-bacterial manure of soil.
8. the preparation method according to claim 7 for preventing flue-cured tobacco root rot and the bio-bacterial manure for improveing soil, feature
It is：
The effective bacterial content of seed fermented liquid described in step (A) is 5 × 108Cfu/mL or more；
The condition of hot air sterilization described in step (B) is that 3 hours of hot air sterilization are carried out in 121 DEG C of autoclaves；
Effective bacterial content is 2 × 10 in bio-bacterial manure described in step (C)8Cfu/g or more.
9. claim 1~6 any one of them prevents flue-cured tobacco root rot is preventing flue-cured tobacco root with the bio-bacterial manure for improveing soil
Maize ear rot and the application in improvement soil.
10. claim 1~6 any one of them prevents the application process of flue-cured tobacco root rot and the bio-bacterial manure for improveing soil,
It is characterized by comprising following steps：
(1) it is turned over to planting vega block, smooth, hack, accomplishes that furrow is straight, ridge body is full, and ridge face is fine crushing；
(2) by bio-bacterial manure and organic fertilizer in mass ratio 1：3~7 ratios are uniformly mixed, and double-bar type fertilizing method are taken, true
Surely the ditch that two depth are 8~12cm is opened up wasteland on the soil both sides of plantation tobacco, is sprinkled under the conduct of Special compound fertilizer for tobacco fertilizer
Layer fertilizer, then carries out smooth ridging by ditch；Then the ditch that a depth is 8~12cm is opened up on the position of ridge line, is spread
Enter the mixture of bio-bacterial manure and organic fertilizer as upper layer fertilizer, such upper layer fertilizer is combined in pairs with lower layer's fertilizer, is constituted
Double-bar type fertilizing method；
(3) since the humiture of environment, soil are larger to the influential effect of bio-bacterial manure, bio-bacterial manure optimum temperature is 25 DEG C~
37 DEG C, it is less than 5 DEG C, is higher than 45 DEG C, application effect is poor；Therefore when base fertilizer applies fertilizer completion, timely ridging epiphragma to wait planting.
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