CN105969672B - A kind of mixing endogenetic fungus that acacia confusa height of seedling and ground diameter can be promoted to increase - Google Patents
A kind of mixing endogenetic fungus that acacia confusa height of seedling and ground diameter can be promoted to increase Download PDFInfo
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- CN105969672B CN105969672B CN201610452322.9A CN201610452322A CN105969672B CN 105969672 B CN105969672 B CN 105969672B CN 201610452322 A CN201610452322 A CN 201610452322A CN 105969672 B CN105969672 B CN 105969672B
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- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
- C12R2001/885—Trichoderma
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G17/00—Cultivation of hops, vines, fruit trees, or like trees
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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Abstract
The invention discloses a kind of mixing endogenetic fungus that acacia confusa height of seedling and ground diameter can be promoted to increase, the mixing endogenetic fungus by acacia confusa endogenetic fungus trichoderma (Trichoderma sp.) A bacterial strain and Fusarium (Fusarium sp.) E bacterial strain composition, preservation is registered in China Committee for Culture Collection of Microorganisms's common micro-organisms center on January 27th, 2016, wherein the deposit number of A bacterial strain is CGMCC No.11911, and the deposit number of E bacterial strain is CGMCC No.11909.The present invention, which mixes contained bacterial strain in endogenetic fungus, to be isolated and purified from acacia confusa plant, which is prepared into using bacterium solution, has the function of that acacia confusa height of seedling and ground diameter is promoted to increase, to promote plant strain growth.
Description
Technical field
The present invention relates to a kind of mixing endogenetic fungus that acacia confusa height of seedling and ground diameter can be promoted to increase and its applications.
Background technique
The research of endophyte of plant starts from the end of the 19th century, and Vogl divides from rye grass Lolium temulentum L. seed
Separate out first plant of endophyte.But the endophyte really started in numerous studies plant originates in last century the eighties, mainly
It conducts a research in the vegetation of Temperate Region in China, subtropical zone and torrid areas.
The distribution of plant endogenesis epiphyte is wide, type is more, and forefathers are studies have shown that can be isolated in most plants
Endogenetic fungus, it can be seen that endogenetic fungus is prevalent in plant.At least belong to more than 290 at more than 80 in worldwide
Endophyte is had found in the gramineae farm crop of kind.The endo-mycorrhiza separated from plant at present can according to the unique function that it has
To be divided into: nitrogen-fixing bacteria consolidate plant growth-promoting bacterias, the biocontrol microorganisms with disease and insect resistance such as potassium bacterium, solid phosphorus bacterium and have promotion plant pair
The resistance bacterium of the repair ability of poor environment.
Through consulting literatures material, the influence research grown at present in relation to endogenetic fungus to acacia confusa is also blank.
Summary of the invention
The purpose of the present invention is to provide a kind of mixing endogenetic fungus that acacia confusa height of seedling and ground diameter can be promoted to increase,
Bacterial strain uses therefor is isolated and purified from acacia confusa plant, and manufactured application bacteria liquid has promotes acacia confusa height of seedling
And the effect that ground diameter increases.
To achieve the above object, the present invention adopts the following technical scheme:
A kind of mixing endogenetic fungus that acacia confusa height of seedling and ground diameter can be promoted to increase, by acacia confusa endogenetic fungus wood
It is mould belong to (Trichoderma sp.) A bacterial strain and Fusarium (Fusarium sp.) E bacterial strain composition；The trichoderma
(Trichoderma sp.) A bacterial strain and Fusarium (Fusarium sp.) E bacterial strain on January 27th, 2016 China it is micro-
Biological inoculum preservation administration committee's common micro-organisms center registers preservation, and (address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Number), wherein the deposit number of A bacterial strain is CGMCC No. 11911, and the deposit number of E bacterial strain is CGMCC No. 11909.
The separation of acacia confusa endogenetic fungus used, purification process include:
A, material: being divided into three parts for the root for collecting different year, branch stem, leaf, cleaned up with tap water, point
It is attached in valve bag, is saved in 4 DEG C of refrigerators；
B, sterilize: 75% alcohol rinses 30s → aseptic water washing 3-4 times → 15% sodium hypochlorite and rinses (root 10min, stem
5min, leaf 2min) → aseptic water washing 4-5 times；Water after sample last time is embathed is contained in the small beaker of sterilizing, flat
Whether lining out is used as control, thorough with the disinfection of sample survey, generates behind several days if any bacterium colony, then shows sample surfaces
Disinfection is not thorough, and need to continue to adjust sterilization method；
C, the selection of inoculation position: blade: leaf tip, leaf central part and leaf base 3 processing；Branch stem: upper, middle and lower is divided into 3
Position, wherein the 3rd position is branch stem junction；Root: it is divided into 2 parts of the tip of a root and foundation；
D, it is inoculated with: the explant after disinfection being cut with vaccinating lancet, is laid in media surface, in 28 DEG C of constant incubators
Bacterium colony upgrowth situation is observed after interior culture 5-7 days；Rapidly, the bacterial strain that can be first generated separate pure for some bacterial strain breedings
Change；The bacterial strain of slow growth and negligible amounts can extend observing time, purify after its growth is stablized；
Solid culture: using improvement Martin's solid medium, be formulated for peptone 5.0g/L, yeast extract powder 2.0g/L,
Glucose 20.0g/L, dipotassium hydrogen phosphate 1.0g/L, magnesium sulfate 0.5g/L, agar 14.0g/L, it is other be sterile water, pH value 6.4
± 0.2, cultivation temperature is 28 DEG C, and training method is plate culture, incubation time 48h；
E, different types of endogenetic fungus access plating medium that separation is bred is purified, by 2-3 point
The above-mentioned of single culture is respectively obtained after connecing purifying, switchingTrichoderma sp.WithFusarium sp.Function stem.
The mixing endogenetic fungus that gained can promote acacia confusa height of seedling and ground diameter to increase can be prepared into using bacterium solution, be used for platform
The rhizosphere soil of gulf yearning between lovers nursery stock, which pours, grants nursery stock and is directly inoculated with.
The preparation method using bacterium solution be respectively by trichoderma (Trichoderma sp.) A bacterial strain and Fusarium
(Fusarium sp.) E bacterial strain access fluid nutrient medium, shaking table shaken cultivation, cultivation temperature is 28 DEG C, 48 ~ 72h of incubation time,
Bacterial concentration is calculated using blood counting chamber, bacterium solution is then diluted to 5.5 × 10 with ultrapure water6Cfu/mL, then by volume
Two kinds of bacterium solutions are mixed to prepare by 1:1；The fluid nutrient medium is improvement Martin's culture medium, wherein peptone 5.0g/L, yeast leaching
Out powder 2.0g/L, glucose 20.0g/L, dipotassium hydrogen phosphate 1.0g/L, magnesium sulfate 0.5g/L, it is other for sterile water, pH value 6.4 ±
0.2。
This is that gained application bacterium solution is poured to the rhizosphere for imposing on acacia confusa by every plant of 100mL using the application method of bacterium solution.
Bacterial strain uses therefor of the present invention is isolated and purified from acacia confusa plant, is had and is promoted acacia confusa height of seedling
And the effect that ground diameter increases, to promote plant strain growth.
Mixing endogenetic fungus preparation is inoculated in acacia confusa seedling by pouring the method applied using bacterium solution, measurement connects
The height of seedling of acacia confusa, ground diameter at kind 4 months, 6 months.The results show that acacia confusa endogenetic fungus AE processing plant with it is right
The height of seedling growth rate of height of seedling growth rate, ground diameter growth rate significant difference according to plant, strains A E processing is 1.5 times of bacterial strains of control
The ground diameter growth rate of AE processing is 2.3 times of control, it was demonstrated that the growth of the mixing endogenetic fungus AE to acacia confusa height of seedling, ground diameter
With apparent facilitation.
Specific embodiment
A kind of mixing endogenetic fungus that acacia confusa height of seedling and ground diameter can be promoted to increase, by acacia confusa endogenetic fungus wood
It is mould belong to (Trichoderma sp.) A bacterial strain and Fusarium (Fusarium sp.) E bacterial strain composition；The trichoderma
(Trichoderma sp.) A bacterial strain and Fusarium (Fusarium sp.) E bacterial strain on January 27th, 2016 China it is micro-
Biological inoculum preservation administration committee's common micro-organisms center registers preservation, wherein the deposit number of A bacterial strain is CGMCC No.
The deposit number of 11911, E bacterial strains is CGMCC No. 11909.
The mixing endogenetic fungus that acacia confusa height of seedling and ground diameter can be promoted to increase can be prepared into using bacterium solution, be used for platform
The rhizosphere soil of gulf yearning between lovers nursery stock, which pours, grants nursery stock and is directly inoculated with.
The preparation method using bacterium solution be respectively by trichoderma (Trichoderma sp.) A bacterial strain and Fusarium
(Fusarium sp.) E bacterial strain access fluid nutrient medium, shaking table shaken cultivation, cultivation temperature is 28 DEG C, 48 ~ 72h of incubation time,
Bacterial concentration is calculated using blood counting chamber, bacterium solution is then diluted to 5.5 × 10 with ultrapure water6Cfu/mL, then by volume
Two kinds of bacterium solutions are mixed to prepare by 1:1；The fluid nutrient medium is improvement Martin's culture medium, wherein peptone 5.0g/L, yeast leaching
Out powder 2.0g/L, glucose 20.0g/L, dipotassium hydrogen phosphate 1.0g/L, magnesium sulfate 0.5g/L, it is other for sterile water, pH value 6.4 ±
0.2。
In order to make content of the present invention easily facilitate understanding, With reference to embodiment to of the present invention
Technical solution is described further, but the present invention is not limited only to this.
Rhizosphere soil, which pours, grants nursery stock and is directly inoculated with application
This test uses earth culture pot experiment, and test material therefor is the Taiwan phase that Municipal Forestry Bureau, Zhangzhou City, Fujian Province provides
Think one year seedling.Select the consistent acacia confusa seedling planting of growing way in diameter 15cm, high 10cm plastic tub in.Soil used
It is the stringent yellow soil Jing Guo fumigation.It is weighed by mixing, every basin is put into 3kg soil.By restorative life in one month
After length, it is 5.5 × 10 that continuous three days, which apply 100mL concentration in acacia confusa rhizosphere,6The bacterium solution AE of cfu/mL, it is another water-soluble with distilling
Liquid, which pours, to be applied as blank control CK.
Strains tested: being respectively connected to the fluid nutrient medium of 40mL by bacterium solution preparation, and culture medium is improvement Martin's culture medium,
Middle peptone 5.0g/L, yeast extract powder 2.0g/L, glucose 20.0g/L, dipotassium hydrogen phosphate 1.0g/L, magnesium sulfate 0.5g/L,
Other is sterile water, pH value 6.4 ± 0.2；The culture for passing through 72h in constant-temperature shaking incubator presses ten with sterile saline
Times dilution method dilutes bacterium solution, calculates bacterial concentration using blood counting chamber, bacterium solution is then diluted to 5.5 with ultrapure water ×
106Cfu/mL, then two kinds of bacterium solutions are mixed to prepare by 1:1 by volume.
When being inoculated with 4 months, 6 months, height of seedling is measured using steel ruler, electronic digital indicator measures ground diameter, the results are shown in Table 1.
Influence of 1 different disposal of table to acacia confusa seedling ground diameter and height of seedling
As shown in table 1, the seedling height of seedling growth rate of inoculating strain AE is 47.432%, and control is only 26.438%, the side of passing through
Difference analysis can obtain, and treated that acacia confusa seedling height of seedling growth rate reaches extremely significant level (sig compared with the control group by strains A E
=0.000, the level of signifiance 0.01).The seedling ground diameter growth rate of inoculating strain AE is 35.466%, and control is only 15.190%, with
Control group otherness reaches extremely significant horizontal (sig=0.001, the level of signifiance 0.01).
It can be seen that mixing endogenetic fungus provided by the invention can remarkably promote the growth of acacia confusa height of seedling and ground diameter,
To promote plant strain growth.
The foregoing is merely presently preferred embodiments of the present invention, all equivalent changes done according to scope of the present invention patent with
Modification, is all covered by the present invention.
Claims (3)
1. a kind of mixing endogenetic fungus that acacia confusa height of seedling and ground diameter can be promoted to increase, it is characterised in that: mixing Nei Shengzhen
Bacterium by acacia confusa endogenetic fungus trichoderma (Trichoderma sp.) A bacterial strain and Fusarium (Fusarium sp.) E bacterial strain
Composition；The trichoderma (Trichoderma sp.) A bacterial strain and Fusarium (Fusarium sp.) E bacterial strain is in 2016 1
The moon 27 registered preservation in China Committee for Culture Collection of Microorganisms's common micro-organisms center, wherein the deposit number of A bacterial strain
For CGMCC No. 11911, the deposit number of E bacterial strain is CGMCC No. 11909.
2. the application for the mixing endogenetic fungus that one kind can promote acacia confusa height of seedling and ground diameter to increase as described in claim 1,
Be characterized in that: the rhizosphere soil for being prepared into using bacterium solution, for acacia confusa nursery stock by the mixing endogenetic fungus, which pours, grants seedling
Wood is directly inoculated with.
3. the application for the mixing endogenetic fungus that acacia confusa height of seedling and ground diameter can be promoted to increase according to claim 2, special
Sign is: the preparation method using bacterium solution be respectively by trichoderma (Trichoderma sp.) A bacterial strain and Fusarium
(Fusarium sp.) E bacterial strain access fluid nutrient medium, shaking table shaken cultivation, cultivation temperature is 28 DEG C, 48 ~ 72h of incubation time,
Bacterial concentration is calculated using blood counting chamber, bacterium solution is then diluted to 5.5 × 10 with ultrapure water6Cfu/mL, then by volume
Two kinds of bacterium solutions are mixed to prepare by 1:1；
The fluid nutrient medium is improvement Martin's culture medium, wherein peptone 5.0g/L, yeast extract powder 2.0g/L, glucose
20.0g/L, dipotassium hydrogen phosphate 1.0g/L, magnesium sulfate 0.5g/L, it is other be sterile water, pH value 6.4 ± 0.2.
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| CN104762219A (en) * | 2015-04-14 | 2015-07-08 | 福建农林大学 | Endophytic fungus for promoting biomass growth of aleurites montana in low-phosphorus environment |
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| CN104762219A (en) * | 2015-04-14 | 2015-07-08 | 福建农林大学 | Endophytic fungus for promoting biomass growth of aleurites montana in low-phosphorus environment |
Non-Patent Citations (3)
| Title |
|---|
| Identification and Characterization of Halophilic, Endophytic Fungi from Acacia ampliceps;S. Piriyaprin等;《Thai Journal of Agricultural Science》;20131231;第46卷(第3期);149-156 * |
| 相思树种根瘤菌的生物学特性;吕成群等;《微生物学通报》;20031231;第30卷(第4期);1-5 * |
| 石灰岩地区台湾相思根瘤内生细菌的初步研究;石前;《万方数据知识服务平台》;20101008;全文 * |
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