CN102732446A - Brevibacillusreuszeri and application thereof in promoting pine tree growth - Google Patents
Brevibacillusreuszeri and application thereof in promoting pine tree growth Download PDFInfo
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Abstract
The invention discloses a brevibacillusreuszeri which is categorized and named as brevibacillusreuszeri MPt17. The brevibacillusreuszeri is collected in China Center for Type Culture Collection on December 1st, 2011 with a collection number of CCTCC NO:M2011433. The brevibacillusreuszeri MPt17 is advantaged in that: the brevibacillusreuszeri MPt17 can substantially promote the growth of ectomycorrhiza fungus such as pisolithustinctorius and xerocamuschrysenteron. Also, as a result of greenhouse pot culture experiments, the double-inoculated mycorrhiza auxiliary bacterium MPt17 and pisolithustinctorius or xerocamuschrysenteron can substantially promote the growth and development of pinus massoniana seedlings. Therefore, the brevibacillusreuszeri MPt17 provided by the invention assists in providing excellent strain resources for the development of pinus massoniana growth-promoting microbial manure, and has a good application and development prospect.
Description
Technical field
The invention belongs to microbial technology field, be specifically related to a kind of bacillus brevis and the application in promoting the Pinus massoniana Lamb growth thereof.
Background technology
(Ectomycorrhizal Fungi EMF) has been proved as the important monoid in the mycorrhizal fungi plant has been had good short living resistant effect the ectomycorrhiza fungi; The ectomycorrhiza fungi can not cause environmental pollution as the application of microbial fertilizer yet simultaneously, and good environment friendly is arranged.Plant growth-promoting root circle bacterium (Plant Growth Promoting Rhizobacteria; PGPR) be the bacterium monoid that in the rhizosphere microecosystem plant is had optimum effect, PGPR can act on plant separately and plant is shown good short life and resistant effect.The interaction of it and ectomycorrhiza fungi has important researching value; Become the focus that rhizosphere microorganism makes to concern research field mutually in recent years day by day; Big quantity research shows that EMF and PGPR synergy have positive effect to the growth and the dietetic alimentation of host plant.Therefore, deeply probe into the mutual of EMF and PGPR and concern, will help to understand better and utilize beneficial rhizosphere microorganisms,, keep the stable of the agricultural ecosystem and all have great importance improving forest output.
Mycorhiza is assisted bacterium, and (Mycorrhiza bacteria MHB) is one type of special P GPR.The interactively of MHB and EMF is very close, and both synergies have active effect to the growth of plant.Research shows that MHB has the following several kinds of mechanisms of action: 1. MHB has improved the susceptibility that root system infects mycorhiza; 2. MHB has improved the physics microenvironment of rhizosphere soil, thereby helps the growth of plant; 3. MHB plays the instrumentality that root system and mycorrhizal fungi discern each other; 4. MHB can promote the sprouting of fungal spore; 5. MHB can promote the growth of mycorrhizal fungi; 6. MHB promotes plant to nutrient absorbing through acting synergistically with EMF; 7. MHB relies on it to grow surely and competitive edge, changes the soil microorganisms environment, suppresses the growth of pathogenic soil bacterium, can improve the systemic disease resistance of plant simultaneously.Wherein MHB is considered to the most direct, most important effect to the promotion of mycorrhizal fungi growth.Though external existing in a large number about EMF and the mutual research report of doing of MHB, domestic still rarely have report about its research of doing mutually to concern.
Pinus massoniana Lamb (Pinas massoniana) is that the main speed in China south is given birth to one of commerical tree species, is the vanguard tree seed of afforestation, also is the important composition seeds of China's agricultural ecosystem.Its economic worth is high, and purposes is wide, and pine is the important material of using in the industrial and agricultural production.The growth of Pinus massoniana Lamb is very strong to the mycorrhizal fungi dependency; Be typical ectomycorrhiza nutritional type seeds; Poor and barren land plantation Pinus massoniana Lamb; Normal owing to soil depletion causes the Pinus massoniana Lamb poor growth, thus utilize the auxiliary bacterium of mycorhiza to promote the ectomycorrhiza fungi growth, and then promote the growth of Pinus massoniana Lamb to seem most important.At present, screening and the applied research of the auxiliary bacterium of relevant Pinus massoniana Lamb mycorhiza also do not appear in the newspapers.
Summary of the invention
Goal of the invention: the deficiency to existing in the prior art the purpose of this invention is to provide a kind of bacillus brevis (Brevibacillus reuszeri).Another purpose provides the purposes of above-mentioned bacillus brevis.
Technical scheme: in order to realize the foregoing invention purpose, the technical scheme that the present invention adopts is:
A kind of bacillus brevis, its classification called after bacillus brevis MPt17 Brevibacillus reuszeri MPt17 has been preserved in Chinese typical culture collection center, deposit number CCTCC NO:M 2011433, preservation date: on December 1st, 2011.The address of depositary institution: Chinese Wuhan Wuhan University.
Above-mentioned bacillus brevis is promoting the colored beans Lasiosphaera fenzlii of ectomycorrhiza fungi (Pisolithus tinctorius, Pt2) application in the growth.
Above-mentioned bacillus brevis is promoting the red suede lid of ectomycorrhiza fungi suilli fungi (Xerocamus chrysenteron, Xc) application in the growth.
The two application that are seeded in the growth of promotion Pinus massoniana Lamb seedling of above-mentioned bacillus brevis and colored beans Lasiosphaera fenzlii or red suede lid suilli fungi.
Bacillus brevis MPt17; Be that screening obtains in the mycorhiza rhizosphere soil of the colored beans Lasiosphaera fenzlii of the ectomycorrhiza fungi sporophore from 30～40 years living masson pine forests in forest-science academy, Jiangsu Province, can efficiently promote colored beans Lasiosphaera fenzliis (Pt2) of ectomycorrhiza fungi and red suede to cover the growth of suilli fungi (Xc).The main biological property of MPt17 bacterial strain: on the KMB culture medium flat plate, bacterium colony is less, color beige, circular, neat in edge; The thalline rod-short, Gram-positive has gemma.
MPt17 bacterial strain 16SrDNA gene order is seen shown in the SEQ ID No.1.Sequence in survey 16SrDNA sequence and the GenBank DB is carried out the BLAST comparison.The result shows that the similarity of MPt17 bacterial strain and Brevibacillus reuszi is 99%.Combining form, physiological and biochemical property and 16SrDNA sequential analysis are accredited as bacillus brevis (Brevibacillus reuszi).
The potted plant inoculation test of MPt17 microbial inoculum shows that this microbial inoculum and colored beans Lasiosphaera fenzlii or the two inoculations of red suede lid suilli fungi have obviously promoted the growth of Pinus massoniana Lamb seedling.
Beneficial effect: the advantage that bacillus brevis MPt17 of the present invention has comprises: can obviously promote the growth of colored beans Lasiosphaera fenzliis (Pt2) of ectomycorrhiza fungi and red suede lid suilli fungi (Xc); And through greenhouse pot culture test confirmation, MPt17 and colored beans Lasiosphaera fenzlii or the two inoculations of red suede lid suilli fungi can obviously promote growing of Pinus massoniana Lamb seedling.Therefore, bacillus brevis MPt17 of the present invention can provide good strain resource for the short endophytic bacteria fertilizer of exploitation Pinus massoniana Lamb, and has application development prospect preferably.
Description of drawings
Fig. 1 is the as a result figure of MPt17 bacterial strain to colored beans Lasiosphaera fenzliis (Pt2) the mycelial growth influence of external mycorrhizal fungi; Among the figure, left side figure is CK, and right figure is MPt17;
Fig. 2 is the as a result figure of MPt17 bacterial strain born of the same parents extra-metabolite to the influence of colored beans Lasiosphaera fenzlii (Pt2) mycelial growth, and among the figure, left side figure is CK, and right figure is MPt17;
Fig. 3 is the as a result figure of MPt17 bacterial strain born of the same parents extra-metabolite to the influence of colored beans Lasiosphaera fenzlii (Pt2) hypha biomass;
Fig. 4 is MPt17 bacterial strain and the two inoculations of colored beans Lasiosphaera fenzlii (Pt2) figure as a result to Pinus massoniana Lamb seedling growth effect;
Fig. 5 is the as a result figure of MPt17 bacterial strain to the influence of the red suede lid of external mycorrhizal fungi suilli fungi (Xc) mycelial growth; Among the figure, left side figure is CK, and right figure is MPt17;
Fig. 6 is the as a result figure of MPt17 bacterial strain born of the same parents extra-metabolite to the influence of red suede lid suilli fungi (Xc) mycelial growth; Among the figure, left side figure is CK, and right figure is MPt17;
Fig. 7 is the as a result figure of MPt17 bacterial strain born of the same parents extra-metabolite to the influence of red suede lid suilli fungi (Xc) hypha biomass;
Fig. 8 is the as a result figures of the two inoculations of the red suede lid suilli fungi (Xc) of MPt17 bacterial strain and ectomycorrhiza fungi to Pinus massoniana Lamb seedling growth effect.
Embodiment
Below in conjunction with specific embodiment the present invention is done further explanation.
In following examples, the prescription of KMB substratum is: peptone 20g, K
2HPO
41.5g, MgSO
4.7H
2O 1.5g, glycerine 10g, agar 20g, water 1000mL, pH 7.0～7.2.The prescription of PDA substratum is: yam 200g, glucose 20g, agar 20g, water 1000mL, natural pH.
The Pt2 source is: pick up from the wealthy mixed forest of Chuxiong, Yunnan pine.
The Xc source is: available from Northeast Forestry University.
The auxiliary bacterium MPt17 of embodiment 1 mycorhiza answers short the coming into force of the colored beans Lasiosphaera fenzliis of external mycorrhizal fungi (Pt2):
The MPt17 bacterial strain of going bail for and depositing, in the KMB substratum, 28 ℃, behind activation 2～3d, taking a morsel with transfering loop is inoculated in the 250mL triangular flask that 50mL KMB liquid nutrient medium is housed, and 28 ℃, 72h is cultivated in the 180r/min concussion.Bacteria suspension is poured in the centrifuge tube of sterilization, 4 ℃, the centrifugal 5min of 5000r/min removes supernatant, uses saline water rinse thalline twice again, and the quantity according to thalline in the centrifuge tube adds saline water in right amount at last, is thalline suspension liquid (OD behind the mixing
600=1.5～2).
Adopt dried ware antagonism (Dunstan et al, 1998) method to make an experiment.Colored beans Lasiosphaera fenzlii (Pt2) is incubated at the PDA substratum, 25 ℃, cultivate 7-12d, use punch tool at the petridish edge punching of diameter as 7mm, the bacterium piece of getting thickness (about 5mm) basically identical is as material.Colored beans Lasiosphaera fenzlii (Pt2) bacterium piece is inverted in the empty petridish of sterilization; Three of every ware inoculations, triangular shape is placed, and draws 25uL thalline suspension liquid with aseptic pipettor again; Drip on each fungi bacterium piece; Each bacterial isolates is established six repetitions, with drip saline water as contrast, place 25 ℃ of incubators to cultivate.Every glucose solution that adds 25uL at a distance from 6d prevents that the bacterium piece is dry.After treating bacterium piece growth 10～12d, the mycelial growth situation of observing each bacterium piece.Result such as table 1 can find out that with shown in Figure 1 the MPt17 bacterial strain has good promoter action to the growth of colored beans Lasiosphaera fenzliis (Pt2) mycelia of external mycorrhizal fungi, compares with contrast, and mycelia length rate of increase is 48.6%.
Table 1MPt17 is to the influence of colored beans Lasiosphaera fenzlii (Pt2) mycelial growth
Annotate: P＜0.05.
Embodiment 2 mycorhiza are assisted the influence of bacterium MPt17 born of the same parents extra-metabolite to colored beans Lasiosphaera fenzliis (Pt2) mycelial growth of external mycorrhizal fungi:
With (method is with embodiment 1) after the MPt17 activation, with transfering loop take a morsel inoculation be equipped with in the 250mL triangular flask of 50mL KMB liquid nutrient medium, 28 ℃, 72h is cultivated in 180r/min concussion.Bacteria suspension is poured in the centrifuge tube of sterilization, 4 ℃, the centrifugal 5min of 5000r/min draws supernatant with asepsis injector, after biofilter (the filter membrane aperture is 0.22um) filters, gets bacteria-free filtrate.
With 1mL bacteria-free filtrate and 45 ℃, 20mL PDA mixes to fall dull and stereotyped, handles as contrasting with sterilized water.Colored beans Lasiosphaera fenzlii (Pt2) bacterium piece is inoculated in petridish central authorities, and three repetitions of every processing place 25 ℃ of constant temperature culture.Observe its colony radius after cultivating 10～12d.Result such as table 2 can find out that with shown in Figure 2 MPt17 born of the same parents' extra-metabolite has obvious facilitation to the growth of colored beans Lasiosphaera fenzliis (Pt2) mycelia of external mycorrhizal fungi, compares with contrast, and the colony diameter rate of increase is 129.6%.
Table 2MPt17 born of the same parents extra-metabolite is to the influence of colored beans Lasiosphaera fenzlii (Pt2) mycelial growth
Annotate: P＜0.05.
Embodiment 3 mycorhiza are assisted the influence of bacterium MPt17 born of the same parents extra-metabolite to colored beans Lasiosphaera fenzliis (Pt2) living weight of external mycorrhizal fungi:
With cultured colored beans Lasiosphaera fenzlii (Pt2) (the PDA substratum, is cultivated 7-12d by 25 ℃); Lay bacterium cake (d=7mm, ￠=5mm), add 3 of bacterium cakes in every 50mL PDA liquid nutrient medium with the punch tool of sterilization; Add 1mL MPt17 born of the same parents extra-metabolite (for the bacteria-free filtrate of embodiment 2) simultaneously, 25 ℃, the common shake-flask culture 10～12d of 140r/min; Qualitative filter paper filters, and places 60 ℃ of baking ovens to dry to constant weight again, measures with electronic balance then; With the KMB nutrient solution is contrast, and every processing repeats for each 3 times.The result is as shown in Figure 3, can find out, MPt17 born of the same parents' extra-metabolite also has obvious facilitation to the living weight of the colored beans Lasiosphaera fenzliis of external mycorrhizal fungi (Pt2), compares with contrast, and the living weight rate of increase of Pt2 is 124.2%.
The influence that auxiliary bacterium MPt17 of 4 pairs of inoculations of embodiment mycorhiza and the two inoculations of the colored beans Lasiosphaera fenzliis (Pt2) of ectomycorrhiza fungi are grown to the Pinus massoniana Lamb seedling:
With (method is with embodiment 1) after the activation of MPt17 bacterium, taking a morsel with transfering loop is inoculated in the 250mL triangular flask that the 50mLKMB liquid nutrient medium is housed, and 28 ℃, 72h is cultivated in the 180r/min concussion.Bacteria suspension is poured in the centrifuge tube of sterilization, 4 ℃, the centrifugal 5min of 5000r/min removes supernatant, uses saline water rinse thalline twice again, obtains bacterium thalline suspension liquid at last.Adopt photoelectric turbidimetry to measure thalline suspension concentration, and MPt17 inoculation liquid concentration is transferred to 5 * 10
8About cfu/mL.
Cultivate colored beans Lasiosphaera fenzlii (Pt2) on the PDA culture medium flat plate, subsequent use as female bacterial classification.Solid enlarged culturing matrix is vermiculite, Semen Maydis powder and wheat bran; Each mixes with 8: 1: 1 mass ratio; Add the PDA nutritive medium (prescription is with liquid PDA substratum) prepare then, stir once more, get about 400g matrix and pack in the culture bag; With its compacting, and tie sack with packing rope.Culture medium is through 1.01 * 10
6The Pa 120min that sterilizes after the cooling, is inoculated in solid culture matrix with colored beans Lasiosphaera fenzlii (Pt2) under aseptic condition, place 25 ℃ of constant temperature culture, treats that mycelia covers with the solid fungicide that promptly is prepared into the ectomycorrhiza fungi behind the culture medium.
Adopt the bud seedling to cut the root method and transplant the Pinus massoniana Lamb seedling, matrix soil is 10: 1 with the solid fungicide mass ratio of ectomycorrhiza fungi, and the amount of MPt17 microbionation liquid is the 5mL/ strain.Concrete grammar is following: at first, get an amount of aseptic potting media and pack into and cultivate in the cup, cover the solid fungicide of the ectomycorrhiza fungi of about 20g then on the soil layer surface; Secondly; The Pinus massoniana Lamb seedling of cotyledon period is taken out from seedling-growing container lightly; Do not injure root system as far as possible, and the sand of seedling root is rinsed well, transplant to cultivating in the cup after with a small amount of main root of bud seedling intercepting with scalper with clear water; Root system is embedded in as far as possible in the solid fungicide of ectomycorrhiza fungi compacting soil; The 3rd, with the bacterium thalline suspension liquid of aseptic pipette, extract 5mL, to be close to seedlings root it is slowly injected, and then cover one deck matrix soil, the potting media soil of each processing is 200g; The 4th, normal root waters.Only to inoculate MPt17, only to inoculate colored beans Lasiosphaera fenzlii (Pt2) and do not inoculate three kinds of processing of bacterium as contrast, 10 repetitions of every processing.Place 25 ℃ of hot-house cultures, illumination is 12h/d, waters unified management in good time.
MPt17 bacterial strain and the two inoculation Pinus massoniana Lamb of colored beans Lasiosphaera fenzlii (Pt2) seedling growing state after 3 months; Result such as table 3 are with shown in Figure 4; Compare with contrast; Single inoculation (bacterium or fungi) and two inoculation Pinus massoniana Lamb seedling have all played certain promoter action to its growth, and the effect that two inoculation is handled is handled well than single inoculation.Colored beans Lasiosphaera fenzlii (Pt2) height of seedling of single inoculation ectomycorrhiza fungi and stem are slightly compared according to having increased by 32.7% and 17.5%; Auxiliary bacterium MPt17 height of seedling of single inoculation mycorhiza and stem are slightly compared according to having increased by 53.4% and 25.0%; The short fruit of coming into force of the two inoculations of Pt2+MPt17 is best, and height of seedling and stem are slightly compared according to having increased by 78.0% and 46.3%.
Table 3MPt17 and the two inoculations of colored beans Lasiosphaera fenzlii (Pt2) are to the influence of Pinus massoniana Lamb seedling growth
The auxiliary bacterium MPt17 of embodiment 5 mycorhiza answers short the coming into force of the red suede lid suilli fungi of external mycorrhizal fungi (Xc):
After the MPt17 activation with separation and purification (method is with embodiment 1), taking a morsel with transfering loop is inoculated in the 250mL triangular flask that 50mL KMB liquid nutrient medium is housed, and 28 ℃, 72h is cultivated in the 180r/min concussion.Bacteria suspension is poured in the centrifuge tube of sterilization, 4 ℃, the centrifugal 5min of 5000r/min removes supernatant, uses saline water rinse thalline twice again, and the quantity according to thalline in the centrifuge tube adds saline water in right amount at last, is thalline suspension liquid (OD behind the mixing
600=1.5～2).
Adopt dried ware antagonism (Dunstan et al, 1998) method to make an experiment.Red suede lid suilli fungi (Xc) is incubated at the PDA substratum, 25 ℃, cultivate 7-12d, use punch tool at the petridish edge punching of diameter as 7mm, the bacterium piece of getting thickness (about 5mm) basically identical is as material.Mycorrhizal fungi bacterium piece is inverted in the empty petridish of sterilization; Three of every ware inoculations, triangular shape is placed, and draws 25uL thalline suspension liquid with aseptic pipettor again; Drip on each fungi bacterium piece; Each bacterial isolates is established six repetitions, with drip saline water as contrast, place 25 ℃ of incubators to cultivate.Every glucose solution that adds 25uL at a distance from 6d prevents that the bacterium piece is dry.After treating bacterium piece growth 10～12d, the mycelial growth situation of observing each bacterium piece, result such as table 4 are with shown in Figure 5; Can find out; The MPt17 bacterial strain has promoter action preferably to the growth of the red suede lid of external mycorrhizal fungi suilli fungi (Xc) mycelia, compares with contrast, and mycelia length rate of increase is 25.0%.
Table 4MPt17 is to the influence of red suede lid suilli fungi (Xc) mycelial growth
The influence that the auxiliary bacterium MPt17 born of the same parents extra-metabolite of embodiment 6 mycorhiza covers suilli fungi (Xc) mycelial growth to the red suede of external mycorrhizal fungi:
With after the tested bacteria MPt17 activation (the KMB substratum, 28 ℃, 2～3d), with transfering loop take a morsel inoculation be equipped with in the 250mL triangular flask of 50mL KMB liquid nutrient medium, 28 ℃, 72h is cultivated in 180r/min concussion.Bacteria suspension is poured in the centrifuge tube of sterilization, 4 ℃, the centrifugal 5min of 5000r/min draws supernatant with asepsis injector, and biofilter (the filter membrane aperture is 0.22um) promptly gets bacteria-free filtrate after filtering.
With 1mL bacteria-free filtrate and 45 ℃, 20mL PDA mixes to fall dull and stereotyped, handles as contrasting with sterilized water.Red suede lid suilli fungi (Xc) bacterium piece is inoculated in petridish central authorities, and three repetitions of every processing place 25 ℃ of constant temperature culture.Observe its colony radius after cultivating 10～12d; Result such as table 5 can find out that with shown in Figure 6 MPt17 born of the same parents' extra-metabolite has obvious facilitation to the growth of the red suede lid of external mycorrhizal fungi suilli fungi (Xc) mycelia; Compare with contrast, the colony diameter rate of increase is 29.5%.
Table 5MPt17 born of the same parents extra-metabolite is to the influence of red suede lid suilli fungi (Xc) mycelial growth
The influence that embodiment 7 MPt17 born of the same parents' extra-metabolites cover suilli fungi (Xc) living weight to the red suede of external mycorrhizal fungi:
After red suede lid suilli fungi (Xc) cultivated 7～12d, lay bacterium cake (d=7mm, ￠=5mm) with the punch tool of sterilization; Add 3 of bacterium cakes in every 50mL PDA liquid nutrient medium, add 1mL MPt17 born of the same parents extra-metabolite (for the bacteria-free filtrate of embodiment 6) simultaneously, 25 ℃; The common shake-flask culture 10～12d of 140r/min; Qualitative filter paper filters, and places 60 ℃ of baking ovens to dry to constant weight again, measures with electronic balance then.With the KMB nutrient solution is contrast, and every processing repeats for each 3 times, and the result sees Fig. 7; Can find out; MPt17 born of the same parents' extra-metabolite also has obvious facilitation to the living weight of the red suede lid suilli fungi of external mycorrhizal fungi (Xc), compares with contrast, and the living weight rate of increase of red suede lid suilli fungi (Xc) is 34.2%.
The influence that auxiliary bacterium MPt17 of 8 pairs of inoculations of embodiment mycorhiza and the two inoculations of the red suede lid suilli fungi (Xc) of ectomycorrhiza fungi are grown to the Pinus massoniana Lamb seedling:
With (method is with embodiment 1) after the activation of MPt17 bacterium, taking a morsel with transfering loop is inoculated in the 250mL triangular flask that the 50mLKMB liquid nutrient medium is housed, and 28 ℃, 72h is cultivated in the 180r/min concussion.Bacteria suspension is poured in the centrifuge tube of sterilization, 4 ℃, the centrifugal 5min of 5000r/min removes supernatant, uses saline water rinse thalline twice again, obtains bacterium thalline suspension liquid at last.Adopt photoelectric turbidimetry to measure thalline suspension concentration, and MPt17 inoculation liquid concentration is transferred to 5 * 10
8About cfu/mL.
Cultivate red suede lid suilli fungi (Xc) on the PDA culture medium flat plate, subsequent use as female bacterial classification.Solid enlarged culturing matrix is vermiculite, Semen Maydis powder and wheat bran, and each mixes with 8: 1: 1 mass ratio, adds the PDA nutritive medium for preparing then, stirs once more, gets about 400g matrix and packs in the culture bag, with its compacting, and ties sack with packing rope.Culture medium is through 1.01 * 10
6The Pa 120min that sterilizes after the cooling, is inoculated in solid culture matrix with red suede lid suilli fungi (Xc) under aseptic condition, place 25 ℃ of constant temperature culture, treats that mycelia covers with the solid fungicide that promptly is prepared into red suede lid suilli fungi (Xc) behind the culture medium.
Adopt the bud seedling to cut the root method and transplant the Pinus massoniana Lamb seedling, matrix soil is 10: 1 with the solid fungicide mass ratio of red suede lid suilli fungi (Xc), and the amount of MPt17 microbionation liquid is the 5mL/ strain.Concrete grammar is following: at first, get an amount of aseptic potting media and pack into and cultivate in the cup, cover the solid fungicide of the ectomycorrhiza fungi of about 20g then on the soil layer surface; Secondly; The Pinus massoniana Lamb seedling of cotyledon period is taken out from seedling-growing container lightly; Do not injure root system as far as possible, and the sand of seedling root is rinsed well, transplant to cultivating in the cup after with a small amount of main root of bud seedling intercepting with scalper with clear water; Root system is embedded in as far as possible in the solid fungicide of red suede lid suilli fungi (Xc) compacting soil; The 3rd, with the bacterium thalline suspension liquid of aseptic pipette, extract 5mL, to be close to seedlings root it is slowly injected, and then cover one deck matrix soil, the potting media soil of each processing is 200g; The 4th, normal root waters.Only to inoculate the MPt17 bacterium, only to inoculate red suede lid suilli fungi (Xc) and do not inoculate three kinds of processing of bacterium as contrast, 10 repetitions of every processing.Place 25 ℃ of hot-house cultures, illumination is 12h/d, waters unified management in good time.
MPt17 bacterial strain and the two inoculation Pinus massoniana Lamb of red suede lid suilli fungi (Xc) seedling growing state after 3 months; Result such as table 6 are with shown in Figure 8; Compare with contrast; Single inoculation (bacterium or fungi) and two inoculation Pinus massoniana Lamb seedling have all played certain promoter action to its growth, and the effect that two inoculation is handled is handled well than single inoculation.Red suede lid suilli fungi (Xc) height of seedling of single inoculation ectomycorrhiza fungi and stem are slightly compared according to having increased by 39.2% and 23.8%; Single inoculation bacterium MPt17 height of seedling and stem are slightly compared according to having increased by 67.7% and 50.0%; The short fruit of coming into force of the two inoculations of Xc+MPt17 is best, and height of seedling and stem are slightly compared according to having increased by 68.2% and 57.5%.
Table 6MPt17 and the two inoculations of red suede lid suilli fungi (Xc) are to the influence of Pinus massoniana Lamb seedling growth
SEQUENCE?LISTING
< 110>Nanjing Forestry University
< 120>a kind of bacillus brevis and application thereof
<130> 100
<160> 1
<170> PatentIn?version?3.5
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<213> Brevibacillus?reuszeri?MPt17
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ctctagagag?actgccgtcg?acaagacgga?ggaaggcggg?gatgacgtca?aatcatcatg 1140
ccccttatga?cctgggctac?acacgtgcta?caatggttgg?tacaacggga?cgctagcccg 1200
cgagggtatg?ccaatctctt?aaaaccaatc?tcagttcgga?ttgtaggctg?caactcgcct 1260
acatgaagtc?ggaatcgcta?gtaatcgcgg?atcagcatgc?cgcggtgaat?acgttcccgg 1320
gccttgtaca?caccgcccgt?cacaccacgg?gcagcacgcc?aacacgctcg 1370
Claims (4)
1. bacillus brevis, its classification called after bacillus brevis (
Brevibacillus reuszeri) MPt17, be preserved in Chinese typical culture collection center, deposit number CCTCC NO:M 2011433, preservation date: on December 1st, 2011.
The described bacillus brevis of claim 1 promote the colored beans Lasiosphaera fenzlii of ectomycorrhiza fungi (
Pisolithus tinctorius,Pt2) application in the growth.
The described bacillus brevis of claim 1 promote the red suede lid of ectomycorrhiza fungi suilli fungi (
Xerocamus chrysenteron,Xc) application in the growth.
4. the two application that are seeded in the growth of promotion Pinus massoniana Lamb seedling of described bacillus brevis of claim 1 and colored beans Lasiosphaera fenzlii or red suede lid suilli fungi.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108370808A (en) * | 2018-03-27 | 2018-08-07 | 贵州大学 | A kind of Mycorrhizal Fungi of Pinus massoniana seedling method for culturing seedlings |
| CN111363684A (en) * | 2019-09-17 | 2020-07-03 | 南京农业大学 | Composite microbial inoculum for efficiently degrading wood fibers and application thereof in composting |
| CN116622553A (en) * | 2023-04-07 | 2023-08-22 | 四川省食用菌研究所 | Lactarius deliciosus mycorrhiza growth promoting bacterium, and preparation method and application thereof |
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| CN101914474A (en) * | 2010-07-28 | 2010-12-15 | 南京林业大学 | Ectomycorrhizal helper bacteria bacillus pumilus and application thereof |
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| CN101914474A (en) * | 2010-07-28 | 2010-12-15 | 南京林业大学 | Ectomycorrhizal helper bacteria bacillus pumilus and application thereof |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108370808A (en) * | 2018-03-27 | 2018-08-07 | 贵州大学 | A kind of Mycorrhizal Fungi of Pinus massoniana seedling method for culturing seedlings |
| CN111363684A (en) * | 2019-09-17 | 2020-07-03 | 南京农业大学 | Composite microbial inoculum for efficiently degrading wood fibers and application thereof in composting |
| CN116622553A (en) * | 2023-04-07 | 2023-08-22 | 四川省食用菌研究所 | Lactarius deliciosus mycorrhiza growth promoting bacterium, and preparation method and application thereof |
| CN116622553B (en) * | 2023-04-07 | 2023-12-08 | 四川省食用菌研究所 | Lactarius deliciosus mycorrhiza growth promoting bacterium, and preparation method and application thereof |
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