CN107034156A - Lactobacillus plantarum and its application - Google Patents
Lactobacillus plantarum and its application Download PDFInfo
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- CN107034156A CN107034156A CN201710277554.XA CN201710277554A CN107034156A CN 107034156 A CN107034156 A CN 107034156A CN 201710277554 A CN201710277554 A CN 201710277554A CN 107034156 A CN107034156 A CN 107034156A
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- lactobacillus plantarum
- cau
- ensilage
- silage
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- 235000013965 Lactobacillus plantarum Nutrition 0.000 title claims abstract description 72
- 229940072205 lactobacillus plantarum Drugs 0.000 title claims abstract description 72
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- 241000219823 Medicago Species 0.000 claims abstract description 25
- 239000004460 silage Substances 0.000 claims abstract description 25
- 241000219793 Trifolium Species 0.000 claims abstract description 15
- 239000000654 additive Substances 0.000 claims abstract description 10
- 230000000996 additive effect Effects 0.000 claims abstract description 9
- 238000002360 preparation method Methods 0.000 claims description 12
- 241000186660 Lactobacillus Species 0.000 claims description 9
- 230000001580 bacterial effect Effects 0.000 claims description 9
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- 238000003860 storage Methods 0.000 claims description 3
- 244000025254 Cannabis sativa Species 0.000 claims 1
- 238000011081 inoculation Methods 0.000 claims 1
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- KZBUYRJDOAKODT-UHFFFAOYSA-N Chlorine Chemical compound ClCl KZBUYRJDOAKODT-UHFFFAOYSA-N 0.000 description 1
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- 241000194033 Enterococcus Species 0.000 description 1
- 206010016717 Fistula Diseases 0.000 description 1
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- 240000001046 Lactobacillus acidophilus Species 0.000 description 1
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- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
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- 238000000746 purification Methods 0.000 description 1
- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 description 1
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
- C12R2001/25—Lactobacillus plantarum
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Wood Science & Technology (AREA)
- Microbiology (AREA)
- Medicinal Chemistry (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Tropical Medicine & Parasitology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses the Lactobacillus plantarum for belonging to ensilage modulation manufacture field and its application.Lactobacillus plantarum (Lactobacillus plantarum) CAU F1 of the present invention, preserving number is CGMCC No.13610, for preparing ensilage, especially prepares alfalfa silage.Lactobacillus plantarum (Lactobacillus plantarum) CAU F1 of the present invention can quickly reduce the pH value of ensilage, and accelerated fermentation processes improve ensiling speed；The breeding of undesirable microorganism can be suppressed, alfalfa silage quality is improved, preferably save clover nutrition, the problem of clover fermentation is difficult is overcome, increase the Vitro Digestibility of ensilage.The present invention Lactobacillus plantarum (Lactobacillus plantarum) CAU F1 prepare additive silage effect than commercially available lactic bacteria additive more preferably, and cost it is low, safely, be easy to utilize.
Description
Technical field
The invention belongs to ensilage modulation manufacture field, and in particular to Lactobacillus plantarum and its application.
Background technology
Ensilage, which refers to prescind in fresh greenfeed, to be fitted into sealed silage facilities (cellar for storing things, moat, tower, bag etc.), warp
Microbial fermentation effect is crossed, is made a kind of with special aromatic odor, nutritious succulent fodder.Lactic acid bacteria is high-quality ensiling
Amount reproduction under the dominant microflora of feed, anaerobic condition, produces substantial amounts of lactic acid, suppresses the growth of harmful bacteria, so as to be made excellent
Good ensilage.Ensilage fermentation initial stage, the rapid breeding such as leukonid, enterococcus generates a certain amount of lactic acid, pH
Decline, be that other lactic acid bacterias and peptococcus create conditions；Ferment the middle and later periods, Lactobacillus plantarum is dominant, also some other
Lactic acid bacteria, such as L.casei, L.brevis, L.buchneri, L.fermentum, L.acidophilus and L.salivarium
Deng.Lactic acid bacteria plays irreplaceable effect, the sieve of strain excellent because of its unique biological function in ensilage manufacture
Choosing, production increasingly cause the attention of researcher.
Commercially available microbial inoculum produces acid less during alfalfa ensilage, and growth is slow, and the acidity being finally reached is higher, lactic acid producing amount, breast
Sour and acetic acid ratio is not very high.
The content of the invention
The invention aims to overcome in ensilage preparation process in the prior art, lactobacter growth is slow, it is few to produce acid
And cause the problem of alfalfa silage quality is low, propose that one kind can quickly reduce pH value, accelerated fermentation processes effectively improve clover
The Lactobacillus plantarum of Silage Quality.
Therefore, the concrete technical scheme of the present invention is as follows：
A kind of Lactobacillus plantarum (Lactobacillus plantarum), bacterial strain is CAU-F1, and preserving number is CGMCC
No.13610。
Lactobacillus plantarum (Lactobacillus plantarum) CAU-F1 derives from term cattle manure in He Sitan
Just.
The 16S rDNA sequences such as SEQ ID of Lactobacillus plantarum (Lactobacillus plantarum) CAU-F1
Shown in No.1.
The biological characteristics of Lactobacillus plantarum (Lactobacillus plantarum) CAU-F1 is：Gram sun
Property bacillus, glucose homofermentation, acid resistance is strong (can be with normal growth in pH3.0), and fertility wide temperature range (can be resistant to 5 DEG C low
Temperature and 50 DEG C of high temperature).
A kind of silage additive, its active component is Lactobacillus plantarum (Lactobacillus plantarum)
CAU-F1。
The preparation method of the silage additive, comprises the following steps：By described Lactobacillus plantarum
(Lactobacillus plantarum) CAU-F1 is cultivated on MRS culture mediums, obtains silage additive.
Applications of Lactobacillus plantarum (Lactobacillus plantarum) CAU-F1 in ensilage preparation.
In above-mentioned application, the ensilage is alfalfa silage.
A kind of preparation method of ensilage, comprises the following steps：
1) will feed cutting be fermented, be well mixed；
2) to step 1) described Lactobacillus plantarum (Lactobacillus plantarum) CAU-F1 of middle addition；
3) by step 2) in feed vacuum sealing, storage, tunning is ensilage.
Step 1) in treat fermented feed be clover.
Step 1) in feed cutting length to be fermented be 1~2cm.
Step 2) in every gram treat that fermented feed is inoculated with the Lactobacillus plantarum (Lactobacillus described in 5~8log cfu
plantarum)CAU-F1。
Step 3) in holding conditions be：15~25 DEG C of temperature, 30~60d of time.
Beneficial effects of the present invention are：
1st, Lactobacillus plantarum (Lactobacillus plantarum) CAU-F1 of the invention can be used for preparing ensiling feeding
Material, especially prepares alfalfa silage.
2nd, Lactobacillus plantarum (Lactobacillus plantarum) CAU-F1 of the invention can quickly reduce ensilage
PH value, accelerated fermentation processes, improve ensiling speed；The breeding of undesirable microorganism can be suppressed, alfalfa silage quality is improved, preferably
Ground saves clover nutrition, overcomes the problem of clover fermentation is difficult, increases the Vitro Digestibility of ensilage.
3rd, the additives for ensiling prepared with Lactobacillus plantarum (Lactobacillus plantarum) CAU-F1 of the present invention
Silage effect than commercially available lactic bacteria additive more preferably, and cost it is low, safely, be easy to utilize.
Biomaterial preservation is proved
Classification And Nomenclature：Lactobacillus plantarum；Strain number：CAU-F1
Preservation mechanism：China General Microbiological culture presevation administrative center
Preservation mechanism is referred to as：CGMCC
Address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Preservation date：On January 13rd, 2017
Collection is registered on the books numbering：CGMCC No.13610
Embodiment
Following embodiment facilitates a better understanding of the present invention, but is not limited to the present invention.Such as without special in following embodiments
Illustrate, be conventional method.
GFG (business microbial inoculum), purchased from Sichuan Gaofuji Biotechnology Co., Ltd., FG1 (business microbial inoculum), by Japanese livestock products
Grassland Research Institute is provided.Culture medium is from extensive and profound in meaning star bio tech ltd (Beijing) purchase in Beijing.
Material, reagent used etc., unless otherwise specified, are commercially obtained in following embodiments.
Embodiment 1：Lactobacillus plantarum (Lactobacillus plantarum) CAU-F1 separation is identified with bacterial strain
1st, the separation of bacterial strain
Using dilution-plate method separating plant lactobacillus：Term cow manure sample 0.5g in He Sitan is weighed, dress is placed in
In the test tube for having 4.5mL sterile salines, fully shaking is well mixed it, and the dilution factor of this test tube is 10-1, then inhale
Take in 0.5mL to next test tube, sample is diluted to 10 successively-6, the sample 0.1mL for then drawing the dilution factor is applied to
On MRS culture medium flat plates, 30 DEG C of culture 48h of anaerobic culture box, picking form, color on the suitable flat board of colony density are placed in
Etc. different bacterium colonies, continuation, which is rule, cultivates, picking colonies typical single to colonial morphology, progress catalase activity measure,
Gram's staining and microscopy experiment.Every Gram-positive, the bacterial strain of negative catalase can be primarily determined that as breast
Bacillus, isolation and purification culture is continued by it on MRS culture mediums, saves it in the 1mL NB trainings for adding 10% dimethyl sulfoxide (DMSO)
Support in base, preserved in -80 DEG C of ultra low temperature freezers.
MRS culture mediums：Peptone (Proteose peptone NO.3) 10.0g, beef extract (Beef extract)
10.0g, yeast extract (Yeast extract) 5.0g, glucose (Dextrose) 20.0g, tween (Polysorbate
80) 1mL, ammonium citrate (Ammonium citrate) 2.0g, sodium acetate (NaAc) 5.0g, magnesium sulfate (MgSO4·7H2O)
0.1g, manganese sulfate (MnSO4·4H2O) 0.05g, dipotassium hydrogen phosphate (K2HPO4) 2.0g, distilled water (H2O) 1000mL, solid culture
Base adds 20g/L agar (Agar) again, and 121 DEG C, sterilize 15min.
2nd, screening high acid, the lactobacillus of fast growth
Physiology and biochemistry detection is carried out to the lactobacillus separated from cow manure, physiological character, which is determined, to be included：Salt tolerant
(NaCl) concentration, pH range growths, temperature range growth test etc.；Biochemical trait, which is determined, to be included：Catalase test, fermentation
Glucose aerogenesis experiment (homotype or heterofermentation experiment), carbon source through fermentation experiment etc..
Physiological and biochemical test method is as follows：
(1) Gram's staining is with reference to the elegant pearl chief editor's in east《Common bacteria system identification handbook》.
(2) Lactobacillus plantarum growth pH regulations use 2mol/L NaOH and 2mol/L HCl.
(3) Catalase determination is tested：Experimental strain is seeded on MRS culture mediums and in anaerobic culture box culture
48h, picking single bacterium colony, which is coated on, to be added dropwise on the culture dish of 3% (w/w) hydrogenperoxide steam generator, has seen whether bubble generation,
Produce bubble for catalase positive, bubble is not produced is then feminine gender.
(4) glucose fermentation aerogenesis determination test：Experimental strain is seeded on MRS culture mediums and in anaerobic culture box training
48h is supported, picking single bacterium colony is inoculated in the test tube containing 5mLMRS fluid nutrient mediums (pH6.5) that (Du Shi tubules tip upside down on test tube
It is interior), common 37 DEG C of culture 48h of constant incubator are positioned over, observes and records result, produce bubble is designated as the positive, does not produce
Bubble is then feminine gender.
(5) carbon source through fermentation determination test：For Gram-positive, the experimental strain of negative catalase, using containing 8
The biochemical identification suit of carbon source is planted, is compareed as a Maxwell turbidity opacity tube, test method foundation specification progress, 37 DEG C of constant temperature
Cultivate 48h, log.
(6) clover is squeezed the juice cultural method：By the fresh alfalfa of chopping and distilled water according to 1：3 ratio mixing is squeezed the juice and mistake
Filter, by 121 DEG C of sterilization treatment 15min of filtrate, can adapt to clover habitat in this, as Screening of Media and quickly produces the bacterium of acid
Strain.
Physiology and biochemistry method is used, the Lactobacillus plantarum CAU-F1 of high acid, fast growth is filtered out.
As a result show, Lactobacillus plantarum CAU-F1 is inoculated in well-grown in clover juice culture medium, production acid is quick (table 1),
Clover habitat can be well adapted to.Lactobacillus plantarum CAU-F1 is Gram-positive, homo-fermentative bacillus, at 5~50 DEG C
With can all be grown under the conditions of pH3.0~9.0, can preferably acid-fast alkali-proof salt tolerant high temperature resistant it is low temperature resistant (table 2)；It can ferment most of
Carbohydrate (table 3).
The bacterial strain CAU-F1 of table 1 ferments pH and organic acid content (mg/ml) after 24h in clover juice
The Lactobacillus plantarum CAU-F1 of table 2 colonial morphology and physio-biochemical characteristics
Note：-, do not grow；+, normal growth.
The Lactobacillus plantarum CAU-F1 of table 3 carbon source through fermentation experimental result
Note：-, do not grow；W, growing way is faint；+, normal growth.
3rd, the identification of bacterial strain
The Lactobacillus plantarum of screening is seeded in 0.5mL MRS fluid nutrient mediums, 30 DEG C, 180rmp overnight incubations, profit
Strain idenfication is carried out with bacterium solution PCR.
Universal primer of the primer from the 16S rDNA gene magnifications of bacterium：
27F：5’-AGAGTTTGATCCTGGCTCAG-3’
1492R：5’-GGTTACCTTGTTACGACTT-3’
PCR reaction systems (40 μ L)：
Reaction condition：
Amplified production Song Meiji bio tech ltd be sequenced, 16S rDNA sequences as shown in SEQ ID No.1,
Sequence alignment is carried out on NCBI, with reference to the Physiology and biochemistry data of bacterial strain, the final bacterial strain for determining separation belongs to Lactobacillus plantarum
Lactobacillus plantarum, are named as CAU-F1.
Embodiment 2：The preparation of alfalfa silage
The preparation of alfalfa silage comprises the following steps：
(1) clover is dried to moisture 60% or so, and is cut to 1cm~2cm, be well mixed；
(2) into step (1) clover according to 6log cfu g-1Inoculated plant lactobacillus (Lactobacillus
Plantarum) CAU-F1, and be control with no added (Control), GFG (business microbial inoculum), FG1 (business microbial inoculum), each place
Manage 3 repetitions；
(3) by step (2) clover load 30cm × 20cm polyethylene bag silo, per it is packed enter 100g, use vacuum sealing
Machine pumping, sealing, are placed in room temperature storage 60d.
Nutritional quality, fermentation quality, microbiological analysis and External digestion experiment are carried out after alfalfa ensilage.
1st, ensilage nutritional quality and the measure of fermentation quality：
After ensiling, representative sample 10g is weighed, 90mL distilled water is added, is well mixed, leachate is obtained after immersion 12h,
The pH value of leachate is determined with pH meter, with the filtering with microporous membrane that aperture is 0.22 μm, is fitted into 10mL centrifuge tubes, is put into -20
It is DEG C standby, to determine organic acid (lactic acid, acetic acid, propionic acid, butyric acid) and ammoniacal nitrogen (NH3- N) content.Organic acidity test is used
Shimadzu GC-14 type high performance liquid chromatograph (chromatographic columns：KC-811column, Shimadzu, Japan；Detector：SPD-
M10AVP, mobile phase：3mmol L-1Perchloric acid, is accurately weighed in 0.8493g perchloric acid, the volumetric flask of constant volume to 2L, flow velocity
1mL min-1；50 DEG C of column temperature；Detection wavelength 210nm, the μ L of sample size 5), remaining ensiling sample is placed in 65 DEG C of baking ovens and dries 48h,
Determine dry (DM) content.The sample comminution of drying is crossed to be put into valve bag after 1mm sieves and preserved, for determining conventional chemical
Composition and External digestion experiment.The measure of crude protein (CP) content uses Kjeldahl's method；Neutral detergent fiber (NDF), acidity
The measure for washing fiber (ADF) uses Fan Shi fibre analysis methods；Hemicellulose is neutral detergent fiber and acid detergent fiber
Difference；The measure of soluble sugar (WSC) uses anthrone sulphuric acid colorimetry.
Nutritional ingredient after alfalfa ensilage is as shown in table 4.Difference does not show between crude protein and soluble sugar content are respectively handled
Write, but addition Lactobacillus plantarum (Lactobacillus plantarum) CAU-F1 ensilage dry loss amount is minimum,
And neutral detergent fiber, acid detergent fiber and hemicellulose level are slightly below Control, EFG and FG1, therefore, plant is added
The nutritional quality of lactobacillus (Lactobacillus plantarum) CAU-F1 alfalfa silage is slightly higher.
Nutritional quality after the alfalfa ensilage of table 4
Note：With column data difference lowercase letter indication differences significantly (p in table<0.05)；DM, dry weight；FM, fresh weight.
Alfalfa ensilage after fermentation quality as shown in table 5, adds Lactobacillus plantarum (Lactobacillus plantarum)
CAU-F1 alfalfa silage, its pH value is minimum, and ammonia nitrogen content is minimum, and the ratio of lactic acid content, lactic acid and acetic acid will height
In blank control group.Clostridium is the harmful microorganism in ensiling, and it produces butyric acid and ammonia, reduces ensilage quality, and addition is planted
Thing lactobacillus (Lactobacillus plantarum) CAU-F1 alfalfa silage butyric acid composition is very low, considers, and plants
Thing lactobacillus (Lactobacillus plantarum) CAU-F1 can improve alfalfa silage quality.
The alfalfa ensilage after fermentation quality of table 5
Note：With column data difference lowercase letter indication differences significantly (p in table<0.05)；DM, dry weight；FM, fresh weight.
2nd, microbiological analysis：
Content of microorganisms, which is determined, uses spread plate counting method, weighs the 90mL distilled water that 10g ensilings sample adds sterilizing
In, after mixing, then 10 times of gradient dilutions take 10 respectively-1、10-3With 10-5Times each 20 μ L of sample diluting liquid be coated on MRS, she
On red methylene blue, rose bengal medium, detection lactic acid bacteria, Escherichia coli, mould and yeast are respectively used to；By 10-1With 10-2Times
Sample diluting liquid 1mL is after 75 DEG C of water-bath water-bath 15min, and taking 20 μ L to be coated on nutrient agar respectively is used to detect
Bacillus, by these coated 30 DEG C of culture 48h of flat board, wherein MRS culture mediums should be positioned over anaerobic culture box, Qi Tapei
Foster base is positioned over common constant incubator.
Nutrient agar (NB) is also referred to as Nutrient broth bouillons：Peptone 10.0g, powdered beef 5.0g, chlorination
Sodium 5g, distilled water 1L, pH value 7.2 ± 0.2.121 DEG C of high pressure steam sterilization 15min.
Yihong methylene blue medium component (g/L)：Peptone 10g；Beef extract powder 3g；Lactose 10g；Sodium chloride 5g；Eosine
0.4g；Methylenum careuleum 0.065g；Agar 14g.
Rose bengal medium composition (g/L)：Peptone 5g；Dipotassium hydrogen phosphate 1g；Magnesium sulfate 0.5g；Glucose 10g；Chlorine
Mycin 0.1g；Rose-bengal 0.033g；Agar 18.5g.
Microbiological indicator as shown in table 6, adds Lactobacillus plantarum (Lactobacillus plantarum) after alfalfa ensilage
CAU-F1 can significantly reduce the content of the undesirable microorganism such as Escherichia coli and bacillus in alfalfa silage.
Micro organism quantity after the alfalfa ensilage of table 6
Note：With column data difference lowercase letter indication differences significantly (p in table<0.05)；DM, dry weight；FM, fresh weight.
3rd, External digestion experiment：
Rumen fluid is derived from 3 milk cows for being provided with permanent fistula, 1h collections rumen fluid before being raised in morning, and equivalent is mixed after collection
Merge through four layers of filtered through gauze, 39 DEG C of water-bath insulations, be constantly passed through CO2Keep anaerobic environment.0.220g samples are loaded and sent into
In 100mL glass syringes, the buffer solution of rumen fluid and preheating is added, is well mixed, is inoculated in the type microbial fermentations of AGRS- III
Micro aerogenesis automatic recording instrument, 39 DEG C of continuous culture 72h, records gas production.Each processing 3 is parallel, and 3 blank are set in addition.
After fermentation ends, Nylon Bag tap water rinse is limpid to water, is then cleaned with distilled water 2 times, 65 DEG C of drying are calculated to constant weight
Dry digestibility.
As shown in table 7, compared with Control, GFG, FG1, addition plant is newborn for External digestion experiment result after alfalfa ensilage
Bacillus (Lactobacillus plantarum) CAU-F1 can dramatically increase the dry digestibility of alfalfa silage, can
To significantly improve gas production, illustrate addition Lactobacillus plantarum (Lactobacillus plantarum) CAU-F1 to ensilage
Digestion it is beneficial.
The alfalfa silage 72h external digestion aerogenesis dynamics of table 7 and vitro Dry Matter Digestibility
Note：IVDMD, vitro Dry Matter Digestibility；GP, gas production；A, theoretical maximum gas production；C, gas production rate；Lag,
Aerogenesis lag time；AGPR, average gas production rate.
SEQUENCE LISTING
<110>China Agricultural University
<120>Lactobacillus plantarum and its application
<130> 2017
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 1450
<212> DNA
<213>Lactobacillus plantarum（Lactobacillus plantarum）
<400> 1
gtgcttataa tgcaagtcga cgaactctgg tattgattgg tgcttgcatc atgatttaca 60
tttgagtgag tggcgaactg gtgagtaaca cgtgggaaac ctgcccagaa gcgggggata 120
acacctggaa acagatgcta ataccgcata acaacttgga ccgcatggtc cgagtttgaa 180
agatggcttc ggctatcact tttggatggt cccgcggcgt attagctaga tggtgaggta 240
acggctcacc atggcaatga tacgtagccg acctgagagg gtaatcggcc acattgggac 300
tgagacacgg cccaaactcc tacgggaggc agcagtaggg aatcttccac aatggacgaa 360
agtctgatgg agcaacgccg cgtgagtgaa gaagggtttc ggctcgtaaa actctgttgt 420
taaagaagaa catatctgag agtaactgtt caggtattga cggtatttaa ccagaaagcc 480
acggctaact acgtgccagc agccgcggta atacgtaggt ggcaagcgtt gtccggattt 540
attgggcgta aagcgagcgc aggcggtttt ttaagtctga tgtgaaagcc ttcggctcaa 600
ccgaagaagt gcatcggaaa ctgggaaact tgagtgcaga agaggacagt ggaactccat 660
gtgtagcggt gaaatgcgta gatatatgga agaacaccag tggcgaaggc ggctgtctgg 720
tctgtaactg acgctgaggc tcgaaagtat gggtagcaaa caggattaga taccctggta 780
gtccataccg taaacgatga atgctaagtg ttggagggtt tccgcccttc agtgctgcag 840
ctaacgcatt aagcattccg cctggggagt acggccgcaa ggctgaaact caaaggaatt 900
gacgggggcc cgcacaagcg gtggagcatg tggtttaatt cgaagctacg cgaagaacct 960
taccaggtct tgacatacta tgcaaatcta agagattaga cgttcccttc ggggacatgg 1020
atacaggtgg tgcatggttg tcgtcagctc gtgtcgtgag atgttgggtt aagtcccgca 1080
acgagcgcaa cccttattat cagttgccag cattaagttg ggcactctgg tgagactgcc 1140
ggtgacaaac cggaggaagg tggggatgac gtcaaatcat catgcccctt atgacctggg 1200
ctacacacgt gctacaatgg atggtacaac gagttgcgaa ctcgcgagag taagctaatc 1260
tcttaaagcc attctcagtt cggattgtag gctgcaactc gcctacatga agtcggaatc 1320
gctagtaatc gcggatcagc atgccgcggt gaatacgttc ccgggccttg tacacaccgc 1380
ccgtcacacc atgagagttt gtaacaccca aagtcggtgg ggtaaccttt taggaaccag 1440
ccgcctaagg 1450
Claims (10)
1. a kind of Lactobacillus plantarum (Lactobacillus plantarum), bacterial strain is CAU-F1, and preserving number is
CGMCCNo.13610。
2. a kind of silage additive, it is characterised in that the active component of the additive is the plant described in claim 1
Lactobacillus (Lactobacillus plantarum) CAU-F1.
3. the preparation method of the silage additive described in claim 2, it is characterised in that comprise the following steps：By right
It is required that Lactobacillus plantarum (Lactobacillus plantarum) CAU-F1 described in 1 is cultivated on MRS culture mediums, green grass or young crops is obtained
Store feed addictive.
4. Lactobacillus plantarum (Lactobacillus plantarum) CAU-F1 described in claim 1 is prepared in ensilage
In application.
5. application according to claim 4, it is characterised in that the ensilage is alfalfa silage.
6. a kind of preparation method of ensilage, it is characterised in that comprise the following steps：
1) will feed cutting be fermented, be well mixed；
2) to step 1) middle Lactobacillus plantarum (Lactobacillus plantarum) CAU-F1 added described in claim 1；
3) by step 2) in feed vacuum sealing, storage, tunning is ensilage.
7. preparation method according to claim 6, it is characterised in that step 1) in treat that fermented feed is clover.
8. preparation method according to claim 6, it is characterised in that step 1) in feed cutting length to be fermented be 1~
2cm。
9. preparation method according to claim 6, it is characterised in that step 2) in every gram treat fermented feed inoculation 5~
Lactobacillus plantarum (Lactobacillus plantarum) CAU-F1 described in 8logcfu claims 1.
10. preparation method according to claim 6, it is characterised in that step 3) in holding conditions be：Temperature 15~45
DEG C, 30~60d of time.
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| CN107354115A (en) * | 2017-09-04 | 2017-11-17 | 本农国际有限公司 | Lactobacillus plantarum for ensilage fermentation |
| CN108060104A (en) * | 2018-01-29 | 2018-05-22 | 四川农业大学 | A kind of lactobacillus plantarum and its application, screening calibration method |
| CN108265016A (en) * | 2018-01-29 | 2018-07-10 | 四川农业大学 | A kind of lactic bacteria additive and its application, preparation method |
| CN109609410A (en) * | 2019-01-08 | 2019-04-12 | 天津市畜牧兽医研究所 | A kind of preparation method of alfalfa ensilage leavening and alfalfa ensilage |
| CN112410251A (en) * | 2020-11-13 | 2021-02-26 | 河北大学 | Lactobacillus plantarum with quick acid production and high acid production capacity and application thereof |
| CN112746044A (en) * | 2021-01-28 | 2021-05-04 | 郑州大学 | Lactobacillus plantarum mutant strain and application thereof in alfalfa silage |
| CN113969243A (en) * | 2020-07-23 | 2022-01-25 | 中国农业大学 | Lactobacillus plantarum capable of degrading tannin and application thereof |
| CN114058531A (en) * | 2021-03-16 | 2022-02-18 | 中国农业大学 | Bacteriocin-producing lactobacillus plantarum and compound application thereof in silage |
| CN114107085A (en) * | 2021-09-14 | 2022-03-01 | 内蒙古农业大学 | Lactobacillus plantarum ING8 and application thereof |
| CN114231440A (en) * | 2021-11-24 | 2022-03-25 | 内蒙古农业大学 | Lactobacillus plantarum ING1 and application thereof |
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| CN116716228A (en) * | 2023-07-24 | 2023-09-08 | 中国农业大学 | Lactobacillus plantarum EL6 with flavonoid content improving function and application thereof |
| CN117511793A (en) * | 2023-11-06 | 2024-02-06 | 青海省农林科学院 | Low-temperature-resistant lactobacillus plantarum and application thereof in jerusalem artichoke silage |
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| CN108060104A (en) * | 2018-01-29 | 2018-05-22 | 四川农业大学 | A kind of lactobacillus plantarum and its application, screening calibration method |
| CN108265016A (en) * | 2018-01-29 | 2018-07-10 | 四川农业大学 | A kind of lactic bacteria additive and its application, preparation method |
| CN108265016B (en) * | 2018-01-29 | 2019-01-11 | 四川农业大学 | A kind of lactic bacteria additive and its application, preparation method |
| CN108060104B (en) * | 2018-01-29 | 2019-01-29 | 四川农业大学 | A kind of lactobacillus plantarum and its application, screening calibration method |
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| CN113969243A (en) * | 2020-07-23 | 2022-01-25 | 中国农业大学 | Lactobacillus plantarum capable of degrading tannin and application thereof |
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| CN112746044A (en) * | 2021-01-28 | 2021-05-04 | 郑州大学 | Lactobacillus plantarum mutant strain and application thereof in alfalfa silage |
| CN112746044B (en) * | 2021-01-28 | 2022-04-19 | 郑州大学 | Lactobacillus plantarum mutant strain and application thereof in alfalfa silage |
| CN114058531B (en) * | 2021-03-16 | 2023-05-30 | 中国农业大学 | Bacteriocin-producing lactobacillus plantarum and compound application thereof in silage |
| CN114058531A (en) * | 2021-03-16 | 2022-02-18 | 中国农业大学 | Bacteriocin-producing lactobacillus plantarum and compound application thereof in silage |
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