CN106566777A - Trichoderma koningiopsis T5-1 bacterial strain and application thereof in improvement of growth of panax notoginseng and prevention and control of root rot - Google Patents
Trichoderma koningiopsis T5-1 bacterial strain and application thereof in improvement of growth of panax notoginseng and prevention and control of root rot Download PDFInfo
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Abstract
The invention belongs to the technical field of prevention and control of plants diseases and particularly relates to a trichoderma koningiopsis T5-1 bacterial strain and an application thereof in improvement of growth of panax notoginseng and prevention and control of root rot. The bacterial strain is named trichoderma koningiopsis, and is assigned the accession number of CGMCC No. 12779 in China General Microbiological Culture Collection Center. The trichoderma koningiopsis T5-1 bacterial strain has developed mycelia and fluffy and dense aerial mycelia, and is free of generation of pigments. The trichoderma koningiopsis T5-1 bacterial strain can improve growth of panax notoginseng and also can inhibit growth of root rot fungus. The bacterial strain can be hyper-parasitized on the root rot fungus, so that spore production of the root rot fungus is inhibited by means of volatile organic compounds from the bacterial strain in a sealed condition. The bacterial strain lays a good foundation for development and utilization of bio-bacterial fertilizers.
Description
Technical field
The invention belongs to plant disease Control Technology field, and in particular to trichoderma pseudokiningii T5-1 bacterial strains and its promote three
Application in seven growths and root rot prevention and control.
Background technology
The prevention and control of plant disease, the most fast and effectively means are to use chemical bactericide.But it is long-term a large amount of using chemistry
Antibacterial can cause environmental pollution, ecosystem destruction, pathogenic bacteria resistance to drugs and drug-fast generation, and pesticide residues are asked in addition
Topic also seriously restricts crop safety and produces and threaten human health.Current chemical pesticide is still occupied an leading position, but because biocontrol microorganisms have
Have the advantages that using safety, do not develop immunity to drugs, its as it is a kind of green, eco-friendly preventions, especially to soil
Add biocontrol microorganisms in earth and carry out prevention and control soil-borne disease, increasingly paid close attention to by people, and be increasingly becoming the following ecological agriculture and can hold
The important directions of supervention exhibition.Meanwhile, the use of microbial-bacterial fertilizer can also be alleviated because chemical fertilizer is in a large number using the soil for causing
The problems such as hardened, soil physico-chemical property changes.The use premise of microbial-bacterial fertilizer is obtained with efficient bio-active function
Beneficial microbe.Biocontrol microorganisms by with pathogen competitive space, produce antibacterial substance, superparasitism pathogen, induction plant
The direct or indirect prevention and control plant disease of the aspect such as system resistant and promotion plant growing.It is with the development of molecular biology, raw
Maturation of the thing technology to microorganism renovation technique, biocontrol microorganisms are in agricultural production as a kind of pesticide or at least can conduct of substituting
The status for reducing a kind of means of supplementing out economy of chemical pesticide usage amount is more and more firm, while showing huge economic benefit, society
Meeting benefit and ecological benefits, also comply with the target of mankind's environmental protection and sustainable development.
Radix Notoginseng is equal with Radix Ginseng, Radix Panacis Quinquefolii etc. to be belonged to together, enjoys the good reputations such as " southern part of the country SHENCAO ", " Radix Stephaniae Sinicae (Radix Stephaniae Dielsianae) ", is that China is peculiar
Rare Chinese medicine, the effects such as with hemostasis, blood circulation promoting and blood stasis dispelling, blood fat reducing, antithrombotic, especially has special effect to traumatic injury etc..Radix Notoginseng
, growth cycle narrower to the demand of ecological environment more harsh, ecological amplitude is longer, with the increase of Planting Years, disease in soil
Opportunistic pathogen is constantly accumulated, and disease incidence is also continuously increased, and is caused damage up to 10%-40% throughout the year, and serious reaches more than 70%,
The yield and quality of Radix Notoginseng is had a strong impact on, becomes the major obstacle that restriction Radix Notoginseng industry develops in a healthy way.Radix Notoginseng is mainly planted at present
Save in Yunnan Province of China, more than 300,000 mu, worldwide market share is up to more than 85% to cultivated area.With people
Lifting of the raising and society of health perception to cardiovascular diseases drug demand, the market demand of Radix Notoginseng are continuously increased.Thus
The extensive single area plantation that brings, abuse of chemical pesticide etc. so that Radix Notoginseng Pesticide Residue is more projected.Therefore,
Using biological preventions prevention and control notoginseng root rot, the sustainable development tool of quality and healthful Chinese crud drug to lifting Radix Notoginseng product
There is important realistic meaning.
The notoginseng root rot fungal pathogenses reported at present mainly include Fusarium (Fusarium spp.), post spore Pseudomonas
(Cylindrocarpon spp.), Phytophthora (Phytophthora spp.), Phoma (Phoma spp.), Rhizoctonia
(Rhizoctonia spp.) and intend 6 category such as Pestalotia (Pestalotiopsis spp.).
The content of the invention
The purpose of the present invention is to propose to trichoderma pseudokiningii T5-1 bacterial strains and its promote Panax notoginseng Growth and root rot prevention and control
In application.Concrete technical scheme is as follows：
Trichoderma pseudokiningii T5-1 bacterial strains, Classification And Nomenclature are trichoderma pseudokiningii (Trichoderma koningiopsis),
The preserving number of China Microbial Culture Preservation Commission's common micro-organisms center is CGMCC NO.12779.
The trichoderma pseudokiningii T5-1 bacterial strains cultivate 3d in glucose potato agar culture medium PDA, can cover with diameter
For the culture dish of 9cm, mycelia is flourishing, and aerial hyphae is fluffy dense, and non-pigment is produced；The PDA culture medium is consisted of：Rhizoma Solani tuber osi
200g/L, glucose 20g/L, agar 20g/L.
The trichoderma pseudokiningii T5-1 bacterial strains are promoting Panax notoginseng Growth and the application in root rot prevention and control.
Application of the trichoderma pseudokiningii T5-1 bacterial strains in Panax notoginseng Growth is promoted, by using trichoderma pseudokiningii bacterium T5-1
Bacterial strain conidial suspension carries out root irrigation to Radix Notoginseng, promotes Panax notoginseng Growth.
The concentration of the trichoderma pseudokiningii bacterium T5-1 bacterial strain conidial suspensions is 1 × 104～1 × 107Individual mitogenetic spore
Son/mL.
The preparation method of the trichoderma pseudokiningii bacterium T5-1 bacterial strain conidial suspensions, comprises the following steps：
1) trichoderma pseudokiningii T5-1 inoculations are cultivated in PDA culture medium, obtains trichoderma pseudokiningii T5-1 bacterial strain bacterium
Fall；
2) in step 1) culture dish in add 20mL sterilized water, scrape the conidium on surface, entered with 4 layers of sterile gauze
Row is filtered, that is, obtain trichoderma pseudokiningii T5-1 conidial suspensions.
Step 1) in condition of culture be 28 DEG C of dark culturing 6d.
Application of the trichoderma pseudokiningii T5-1 bacterial strains in notoginseng root rot prevention and control, the pathogen of the root rot is destruction post
Spore bacterium (C.destructans), Phytophthora cactorum bacterium (P.cactorum), one kind of Fusarium spp. (F.solani) and more than.
The trichoderma pseudokiningii T5-1 bacterial strains filtrate suppresses pine root fungus growth.
The preparation of the trichoderma pseudokiningii T5-1 bacterial strain filtrates comprises the steps：
1) trichoderma pseudokiningii T5-1 inoculations are cultivated in PDA culture medium, obtains trichoderma pseudokiningii T5-1 bacterial strain bacterium
Fall；
2) from step 1) the trichoderma pseudokiningii T5-1 colony edges that obtain, 20 pieces of mycelia block for taking 5mm is beaten, is seeded to and is contained
In the 500mL triangular flasks of 300mL PDB culture fluid, after shaken cultivation 30d, using vacuum pump sucking filtration, trichoderma pseudokiningii T5- is obtained
1 bacterial strain filtrate；The PDB culture medium is consisted of：Rhizoma Solani tuber osi 200g/L, glucose 20g/L.
Step 1) in condition of culture be 28 DEG C of dark culturing 2d；Step 2) in condition of culture be 25 DEG C, 180rpm/min.
The VOC of the trichoderma pseudokiningii T5-1 bacterial strains suppresses the product of pine root fungus in confined conditions
Spore.
The trichoderma pseudokiningii T5-1 bacterial strains superparasitism pine root fungus.
Beneficial effects of the present invention are：The trichoderma pseudokiningii T5-1 bacterial strains mycelia of screening is flourishing, and aerial hyphae is fluffy dense,
Non-pigment is produced.Trichoderma pseudokiningii T5-1 bacterial strains can promote Panax notoginseng Growth, meanwhile, bacterial strain can suppress pine root fungus growth, weight
Parasitize pine root fungus, the VOC of bacterial strain suppresses the product spore of pine root fungus in confined conditions, reaches promotion
The double effectses of Panax notoginseng Growth and suppression root rot, are that good basis is established in the exploitation and its utilization of bio-bacterial manure.
Biomaterial preservation explanation
Classification And Nomenclature：Trichoderma pseudokiningii (Trichoderma koningiopsis) strain number：T5-1
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 08 08th, 2016
Collection is registered on the books numbering：CGMCC No.12779
Description of the drawings
Fig. 1 is morphological feature of the trichoderma pseudokiningii T5-1 bacterial strains in PDA culture medium, and wherein a is colonial morphology, and b is
Conidiophore form, c are conidium form.
Fig. 2 is inhibition of the trichoderma pseudokiningii T5-1 bacterial strains filtrate to Phytophthora cactorum bacterium D-10, and wherein a is unused T5-1 bacterium
The colonial morphology of the Phytophthora cactorum bacterium D-10 that strain filtrate is processed, b are the suppression after T5-1 bacterial strains filtrate is processed to Phytophthora cactorum bacterium D-10
Effect.
Fig. 3 is inhibition of the trichoderma pseudokiningii T5-1 bacterial strains filtrate to Phytophthora cactorum bacterium D-10, and wherein CK is unused T5-1
Bacterial strain filtrate process, T5-1-Ster-D-10 be T5-1 bacterial strains filtrate after 121 DEG C of high-temperature process 20min to Phytophthora cactorum bacterium D-
10 suppression ratio.
Fig. 4 is the inhibition that trichoderma pseudokiningii T5-1 bacterial strains VOC produces spore to Fusarium spp. F-2, wherein
F-2 is the Sporulation of Fusarium situation that unused T5-1 volatile material is processed, and T5-1-F-2 is to use T5-1 TREATMENT OF VOCs
Sporulation of Fusarium situation afterwards, A and B represent that treatment group has pole significant difference in 1% level with matched group.
Effects of the Fig. 5 for trichoderma pseudokiningii T5-1 superparasitism notoginseng root rot bacterium, wherein a are the weight in the way of spore is adhered to
Parasitic Cylindrocarpon destructans, b are the superparasitism Cylindrocarpon in the way of mycelia winds
Destructans, c are the superparasitism Phtophthora cactorum in the way of forming " hook-shaped " structure.
Specific embodiment
The present invention proposes trichoderma pseudokiningii T5-1 bacterial strains and its application in promotion Panax notoginseng Growth and root rot prevention and control,
With reference to embodiment, the present invention will be further described.
Embodiment 1：The separation and identification of trichoderma pseudokiningii T5-1 bacterial strains
1st, the isolation and purification of bacterial strain
3 years raw sangqi ginseng root tissue samples are obtained from Wenshan Prefecture of Yunnan Province, with tap water by the soil of Radix Notoginseng tissue surface
Etc. rinsing well；Cut the strong intersection piece of tissue of Radix Notoginseng root tissue disease, size about 4mm2；By the piece of tissue for cutting 1% time
Sterilize in sodium chlorate solution 3min, after aseptic water washing 3 times, fully dries on sterilizing filter paper piece；Piece of tissue is positioned over and is added
On PDA plate added with streptomycin sulfate (100ppm), culture 1-3d, routine observation are inverted in 28 DEG C of constant incubators；Treat
After doubtful bacterium colony is grown in culture dish, carry out picking immediately and be transferred on fresh PDA plate being numbered preservation；Using bacterium
Silk end method carries out purification, obtains pure culture as shown in Figure 1.The PDA plate is consisted of：Rhizoma Solani tuber osi 200g/L, Fructus Vitis viniferae
Sugared 20g/L, agar 20g/L.
2nd, the identification of bacterial strain
The genomic DNA of bacterial strain after extraction separation and purification, enters performing PCR amplification using following 3 pairs of primers：
1. forward primer ITS1 (5 '-TCCGTAGGTGAACCTGCGG-3 '), and downstream primer ITS2 (5 '-
GCTGCGTTCTTCATCGATGC-3 '), extension increasing sequence is as shown in SEQ ID No.1.
2. forward primer ITS1 (5 '-TCCGTAGGTGAACCTGCGG-3 '), and downstream primer ITS4 (5 '-
CCTCCGCTTATTGATATGC-3 '), extension increasing sequence is as shown in SEQ ID No.2.
3. forward primer tef1 (5 '-GTGAGCGTGGTATCACCATCG-3 '), and downstream primer tef1 (5 '-
GCCATCCTTGGAGACCAGC-3 '), extension increasing sequence is as shown in SEQ ID No.3.
By above-mentioned extension increasing sequence trichoderma data base TrichoBLAST (http://www.isth.info/tools/ blast/preblast.php) in compare, Classification And Nomenclature be trichoderma pseudokiningii (Trichoderma koningiopsis),
Strain Designation is T5-1.
Embodiment 2：Growth-promoting functions of the trichoderma pseudokiningii T5-1 bacterial strains conidial suspension to Radix Notoginseng
By trichoderma pseudokiningii T5-1 inoculations in the PDA culture medium 28 DEG C of dark culturing 6d, obtain trichoderma pseudokiningii T5-
1 bacterial strain bacterium colony；20mL sterilized water is added in culture dish, the conidium on surface is scraped, is filtered with 4 layers of sterile gauze,
Trichoderma pseudokiningii T5-1 conidial suspensions are obtained, concentration is 1 × 107Individual conidium/mL.
The annual Radix Notoginseng plant in field and biennial Radix Notoginseng plant, are provided by Yunnan Province Miao Xiang Radix Notoginseng Technology Park.Using with
The EXPERIMENTAL DESIGN of machine district's groups, the plot area/Radix Notoginseng strain number of setting is：About 100 plants of annual Radix Notoginseng 0.5m × 0.5m/；2 years
About 30 plants of raw sangqi ginseng 1.5m × 0.5m/；The spore suspension concentration of the trichoderma pseudokiningii T5-1 bacterial strains of 3 gradient dilutions is set
(A dilutes 10 times；B dilutes 100 times；C, dilutes 1000 times) regular root irrigation is carried out, each concentration arranges 3 repetitions, each
Cell pouring root amount is 500mL, carries out a root irrigation every 7d, carries out 4 pouring roots altogether, and matched group is substituted with tap water.
According to five point sampling, annual Radix Notoginseng 20d after treatment is sampled during biennial Radix Notoginseng 45d.1 year
Raw sangqi ginseng each cell samples 10 plants (totally 30 plants of 3 repetitions)；Biennial Radix Notoginseng each cell samples 5 plants of (3 repetitions totally 15
Strain).The measurement of middle period length, middle leaf width, plant height, overground part fresh weight, underground part fresh weight is carried out to the individual plant of samples taken, to each
All samples in repeating carry out overground part dry weight, the measurement of underground part dry weight gross weight.By above measurement index, use
DPS softwares carry out significant difference analysis.As a result such as Tables 1 and 2, as a result show, trichoderma pseudokiningii T5-1 bacterial strains have to Radix Notoginseng
Growth-promoting functions.
Growth-promoting effect of the 1 trichoderma pseudokiningii T5-1 bacterial strains of table to annual Radix Notoginseng
Note：* is noted after data, variant significance (P compared with the control is illustrated<0.05).
Growth-promoting effect of the 2 trichoderma pseudokiningii T5-1 bacterial strains of table to biennial Radix Notoginseng
Note：* is noted after data, variant significance (P compared with the control is illustrated<0.05).
Embodiment 3：Inhibitory action of the trichoderma pseudokiningii T5-1 bacterial strains filtrate to Phytophthora cactorum.
By trichoderma pseudokiningii T5-1 inoculations in the PDA culture medium 28 DEG C of dark culturing 2d, obtain trichoderma pseudokiningii T5-
1 bacterial strain bacterium colony；20 pieces of mycelia block for taking 5mm is beaten, is seeded in the 500mL triangular flasks of the culture fluid of PDB containing 300mL, 25 DEG C,
Under the conditions of 180rpm/min after shaken cultivation 30d, using vacuum pump sucking filtration, trichoderma pseudokiningii T5-1 bacterial strain filtrates are obtained；Filtrate
In 121 DEG C of heat treated 20min, and with PDA culture medium according to volume ratio 1:4 are sufficiently mixed, and are down flat plate, per plate about 20mL；With
The card punch of 5mm, takes the Phytophthora cactorum colony edge mycelia block of activation 4d, is positioned over 9cm flat boards central authorities, after culture 10d, adopts ten
Word interior extrapolation method measures the colony diameter of Phytophthora cactorum bacterium, calculates suppression ratio.As a result such as Fig. 2 and Fig. 3, the filter of trichoderma pseudokiningii T5-1 bacterial strains
Liquid is obvious to the inhibition of Phytophthora cactorum bacterium.
Embodiment 4：Inhibitory action of the trichoderma pseudokiningii T5-1 bacterial strains VOC to Sporulation of Fusarium
Using the method for make-up culture.Trichoderma pseudokiningii T5-1 bacterial strains and Fusarium spp. are activated respectively, 3 days to be grown
Afterwards, beaten in the colony edge of trichoderma pseudokiningii T5-1 bacterial strains and Fusarium spp. with 5mm card punch respectively and take fresh mycelia block；Health will be intended
Family name's Trichoderma spp. T5-1 bacterial strain mycelia blocks are placed on the central authorities of the 9cm culture dishs containing 20mLPDA, and it is new that Fusarium spp. mycelia block is placed on another
The fresh central authorities of the 9cm culture dishs containing 20mLPDA；By two ware bottoms mutual make-up, 4 layers of parcel are carried out with sealed membrane and is processed, each
Process 3 repetitions；The one side of the block of mycelia containing Fusarium spp. upper, the one side of the bacterial strain mycelia blocks of T5-1 containing trichoderma pseudokiningii under, with
Two sides all connects the process of Fusarium spp. as blank, carries out culture 7d in 28 DEG C of constant incubators；Beat to take with 5mm card punch and receive
Trichoderma pseudokiningii T5-1 bacterial strains VOC process after Fusarium spp. mycelia block and be transferred to fresh PDA culture medium
On carry out restoration ecosystem 5d after, near vaccination, midpoint, 3 positions of colony edge of vaccination and colony edge line, use
5mm card punch is beaten and takes mycelia block, and being placed in carries out the statistics of sporulation quantity after shaking up in the PCR pipe containing 1mL aquesterilisa.As a result as schemed
4, trichoderma pseudokiningii T5-1 bacterial strains VOC produces spore to Fusarium spp. F-2 and produces inhibition.
Embodiment 5：The effect of trichoderma pseudokiningii T5-1 superparasitism notoginseng root rot bacterium
Using the method for opposite culture.By trichoderma pseudokiningii T5-1, destroy post spore bacterium (C.destructans) and Phytophthora cactorum
Bacterium (P.cactorum) is activated, and after 3 days to be grown, is beaten with 5mm card punch respectively and is taken mycelia block；By trichoderma pseudokiningii T5-1
Mycelia block is placed on the 9cm culture dishs side containing 20mL PDA (away from edge about 2cm or so), destroys post spore bacterium with Phytophthora cactorum bacterium
Mycelia block is placed on culture dish opposite side (away from edge about 2cm or so), and culture is inverted in 28 DEG C of constant incubators.Respectively in culture
During 4d and 8d, mycelia intersection is cut with sterilizing blade, size about 4mm × 4mm is scanned electron microscope observation.
As a result such as Fig. 5, trichoderma pseudokiningii T5-1 can the superparasitism Cylindrocarpon destructans in the way of spore is adhered to；With
The mode superparasitism Cylindrocarpon destructans of mycelia winding；The superparasitism in the way of forming " hook-shaped " structure
Phtophthora cactorum。
SEQUENCE LISTING
<110>China Agricultural University
<120>Trichoderma pseudokiningii T5-1 bacterial strains and its application in promotion Panax notoginseng Growth and root rot prevention and control
<130> 2016
<160> 3
<170> PatentIn version 3.3
<210> 1
<211> 235
<212> DNA
<213>Trichoderma pseudokiningii（Trichoderma koningiopsis）
<400> 1
agccgaatcg ttgatgagtt ttgattcatt ttgaattttt gctcagagct gtaagaaata 60
acgtccgcga ggggactaca gaaagagttt ggttggtccc tccggcgggc gcctggttcc 120
ggggctgcga cgcacccggg gcgtgacccc gccgaggcaa cagtttggta tggttcacat 180
tgggtttggg agttgtaaac tcggtaatga tccctccgca ggttcaccta cggaa 235
<210> 2
<211> 568
<212> DNA
<213>Trichoderma pseudokiningii（Trichoderma koningiopsis）
<400> 2
gaacagcttt cctacctgat cgaggtcaca tttcagaaag ttgggtgttt tacggacgtg 60
gacgcgccgc gctcccggtg cgagttgtgc aaactactgc gcaggagagg ctgcggcgag 120
accgccactg tatttcgggg ccgggatccc gtcttagggg ttcccgatcc ccaacgccga 180
ccccccggag gggttcgagg gttgaaatga cgctcggaca ggcatgcccg ccagaatact 240
ggcgggcgca atgtgcgttc aaagattcga tgattcactg aattctgcaa ttcacattac 300
ttatcgcatt tcgctgcgtt cttcatcgat gccagaacca agagatccgt tgttgaaagt 360
tttgattcat tttgaatttt tgctcagagc tgtaagaaat aacgtccgcg aggggactac 420
agaaagagtt tggttggtcc ctccggcggg cgcctggttc cggggctgcg acgcacccgg 480
ggcgtgaccc cgccgaggca acagtttggt atggttcaca ttgggtttgg gagttgtaaa 540
ctcggtaatg atccctccgc aggtacgc 568
<210> 3
<211> 217
<212> DNA
<213>Trichoderma pseudokiningii（Trichoderma koningiopsis）
<400> 3
tgtccctgcg tcgagactcc agtactatgt caccgtcatt ggtatgttat tcctggctct 60
tgacatgtcg aaatcatcat tctaacgtgc caatacagac gctcccggcc accgtgattt 120
catcaagaac atgatcactg gtacctccca ggctgactgc gctatcctga ttatcgctgc 180
cggtactggt gagttcgagg ctggtcccaa ggatggc 217
Claims (14)
1. trichoderma pseudokiningii T5-1 bacterial strains, it is characterised in that the strain classification is named as trichoderma pseudokiningii (Trichoderma
Koningiopsis), the preserving number in China Microbial Culture Preservation Commission's common micro-organisms center is CGMCC
NO.12779。
2. trichoderma pseudokiningii T5-1 bacterial strains according to claim 1, it is characterised in that the bacterial strain is in glucose Rhizoma Solani tuber osi
3d is cultivated on agar culture medium PDA, the culture dish of a diameter of 9cm can be covered with, mycelia is flourishing, and aerial hyphae is fluffy dense, colourless
Element is produced；The PDA culture medium is consisted of：Rhizoma Solani tuber osi 200g/L, glucose 20g/L, agar 20g/L.
3. trichoderma pseudokiningii T5-1 bacterial strains are promoting Panax notoginseng Growth and the application in root rot prevention and control.
4. application of the trichoderma pseudokiningii T5-1 bacterial strains according to claim 3 in Panax notoginseng Growth is promoted, it is characterised in that
Root irrigation is carried out to Radix Notoginseng using trichoderma pseudokiningii bacterium T5-1 bacterial strains conidial suspension, promotes Panax notoginseng Growth.
5. application according to claim 4, it is characterised in that the trichoderma pseudokiningii bacterium T5-1 bacterial strain sectional growing spore suspensions
The concentration of liquid is 1 × 104～1 × 107Individual conidium/mL.
6. application according to claim 4, it is characterised in that the trichoderma pseudokiningii bacterium T5-1 bacterial strain sectional growing spore suspensions
The preparation method of liquid, comprises the following steps：
1) trichoderma pseudokiningii T5-1 inoculations are cultivated in PDA culture medium, obtains trichoderma pseudokiningii T5-1 bacterial strain bacterium colonies；
2) in step 1) culture dish in add 20mL sterilized water, scrape the conidium on surface, carried out with 4 layers of sterile gauze
Filter, that is, obtain trichoderma pseudokiningii T5-1 conidial suspensions.
7. application according to claim 6, it is characterised in that step 1) in condition of culture be 28 DEG C of dark culturing 6d.
8. application of the trichoderma pseudokiningii T5-1 bacterial strains according to claim 3 in notoginseng root rot prevention and control, its feature exist
In the pathogen of the root rot is destruction post spore bacterium (Cylindrocarpon destructans), Phytophthora cactorum bacterium
(Phtophthora cactorum), one kind of Fusarium spp. (Fusarium solani) and more than.
9. application of the trichoderma pseudokiningii T5-1 bacterial strains according to claim 3 in notoginseng root rot prevention and control, its feature exist
In trichoderma pseudokiningii T5-1 bacterial strains filtrate suppresses pine root fungus growth.
10. application according to claim 9, it is characterised in that the preparation bag of the trichoderma pseudokiningii T5-1 bacterial strain filtrates
Include following steps：
1) trichoderma pseudokiningii T5-1 inoculations are cultivated in PDA culture medium, obtains trichoderma pseudokiningii T5-1 bacterial strain bacterium colonies；
2) from step 1) the trichoderma pseudokiningii T5-1 colony edges that obtain, 20 pieces of mycelia block for taking 5mm is beaten, is seeded to containing 300mL
In the 500mL triangular flasks of PDB culture fluid, after shaken cultivation 30d, using vacuum pump sucking filtration, trichoderma pseudokiningii T5-1 bacterial strains are obtained
Filtrate；The PDB culture medium is consisted of：Rhizoma Solani tuber osi 200g/L, glucose 20g/L.
11. applications according to claim 10, it is characterised in that step 1) in condition of culture be 28 DEG C of dark culturing 2d；
Step 2) in condition of culture be 25 DEG C, 180rpm/min.
Application of the 12. trichoderma pseudokiningii T5-1 bacterial strains according to claim 3 in notoginseng root rot prevention and control, its feature exist
In the VOC of trichoderma pseudokiningii T5-1 bacterial strains suppresses the product spore of pine root fungus in confined conditions.
Application of the 13. trichoderma pseudokiningii T5-1 bacterial strains according to claim 3 in notoginseng root rot prevention and control, its feature exist
In trichoderma pseudokiningii T5-1 bacterial strain superparasitism pine root fungus.
14. applications according to claim 13, it is characterised in that trichoderma pseudokiningii T5-1 bacterial strains are in the way of spore adhesion
Superparasitism C.destructans, the superparasitism C.destructans in the way of mycelia winds, to form the side of " hook-shaped " structure
Formula superparasitism P.cactorum.
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Application publication date: 20170419 Assignee: Jiangsu Tenglong Biological Pharmaceutical Co.,Ltd. Assignor: YUNNAN CI & CP BIOTECH ENGINEERING Co.,Ltd. Contract record no.: X2021530000004 Denomination of invention: Trichoderma pseudokoningii t5-1 strain and its application in promoting the growth of Panax notoginseng and controlling root rot Granted publication date: 20190813 License type: Common License Record date: 20210323 |
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