CN104818217B - One plant can promote the endogenetic fungus that aleurite montana biomass increases - Google Patents
One plant can promote the endogenetic fungus that aleurite montana biomass increases Download PDFInfo
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- CN104818217B CN104818217B CN201510174387.7A CN201510174387A CN104818217B CN 104818217 B CN104818217 B CN 104818217B CN 201510174387 A CN201510174387 A CN 201510174387A CN 104818217 B CN104818217 B CN 104818217B
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Abstract
The present invention relates to one plant of endogenetic fungus that aleurite montana biomass can be promoted to increase.The endogenetic fungus is disk stey（Pestalotia sp.）ZJ2, registers preservation, preserving number is CGMCC No. 10327 on January 30th, 2015 in China Committee for Culture Collection of Microorganisms's common micro-organisms center.Aleurite montana endogenetic fungus ZJ2 is inoculated in aleurite montana earth culture seedling, the biomass when height of seedling of aleurite montana when measuring just inoculation July and during 60 days Septembers of inoculation, ground diameter and inoculation 60 days.Plant handled by aleurite montana endogenetic fungus ZJ2 and the height of seedling growth rate, ground diameter growth rate and biomass significant difference compareed.
Description
Technical field
The present invention relates to one plant of endogenetic fungus that aleurite montana biomass can be promoted to increase.
Background technology
The research of endophyte of plant starts from the end of the 19th century, and Vogl divides from rye grass Lolium temulentum L. seeds
Separate out first plant of endophyte.But the endophyte really started in numerous studies plant originates in last century the eighties, mainly
Conduct a research in the vegetation of Temperate Region in China, subtropical zone and torrid areas.
The distribution of plant endogenesis epiphyte is wide, species is more, and forefathers' research shows to separate out of most plants
To endogenetic fungus, it can be seen that endogenetic fungus is prevalent in plant.At least belong to 290 at more than 80 in worldwide
Endophyte is found that in a variety of gramineae farm crops.The unique function that separated endo-mycorrhiza has according to it from plant at present
It can be divided into：Nitrogen-fixing bacteria, consolidate the plant growth-promoting bacterias such as potassium bacterium, solid phosphorus bacterium, the biocontrol microorganisms with disease and insect resistance and with promotion plant
To the resistance bacterium of the repair ability of poor environment.Consulting literatures material, the influence research grown in relation to endogenetic fungus to aleurite montana
Also there are blank.
The content of the invention
It is an object of the invention to provide one plant of endogenetic fungus that aleurite montana biomass can be promoted to increase, the fungi are more for disk
Hair spore（Pestalotia sp.）ZJ2, it is commonly micro- in China Committee for Culture Collection of Microorganisms on January 30th, 2015
Bio-Centers register preservation, and address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, and preserving number is CGMCC No. 10327.
To achieve the above object, the present invention adopts the following technical scheme that：
Separation, the purifying of the bacterium include：
A, material：The root for collecting different year, branch stem, leaf are divided into three parts, cleaned up with tap water, point
It is attached in valve bag, is preserved in 4 DEG C of refrigerators；
B, sterilize：The sodium hypochlorite rinsing of 75% alcohol rinsing 30s → aseptic water washing 3-4 times → 15%（Root 7min, stem
5min, leaf 3min）→ 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, is generated behind some 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：3 leaf tip, leaf central part and leaf base 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：Explant after disinfection is cut with vaccinating lancet, is laid in media surface, in 28 DEG C of constant temperature incubations
Culture observes colony growth situation after 5-7 days in case.Some bacterial strain breedings are rapid, and the bacterial strain that can be first produced is separated
Purifying；Slow-growing and negligible amounts bacterial strains can extend observing time, be purified after its growth stabilization；
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 tablet culture；
E, the different types of endogenetic fungus access plating medium bred separation is purified, by 2-3 point
The above-mentioned of single culture is obtained after connecing purifying, switchingPestalotiaFunction stem；
A kind of preparation method of above-mentioned endogenetic fungus application bacterium solution that aleurite montana biomass can be promoted to increase, being will be above-mentioned
Bacterial strain access fluid nutrient medium, shaking table shaken cultivation, cultivation temperature be 28 DEG C, 48~72h of incubation time, utilizes blood count
Plate calculates bacterial concentration, and it is that finished product is spare that bacterium solution is diluted to 1.0-9.0 × 106cfu/ml with ultra-pure water；Liquid Culture
Base：To improve Martin's culture medium, wherein peptone 5.0g/L, yeast extract powder 2.0g/L, glucose 20.0g/L, phosphoric acid hydrogen two
Potassium 1.0g/L, magnesium sulfate 0.5g/L, it is other for sterile water, pH value 6.4 ± 0.2.
A kind of application process of above-mentioned endogenetic fungus application bacterium solution that aleurite montana biomass can be promoted to increase, is to apply to be somebody's turn to do
The bacterium solution of bacterial strain, which is poured, imposes on Seedling rhizosphere soil.
A kind of application process of above-mentioned endogenetic fungus application bacterium solution that aleurite montana biomass can be promoted to increase, is with 5.5
× 106cfu/ml bacterium solutions, are poured in the rhizosphere of every plant of aleurite montana by every plant of 100ml in forest land.
Bacterial strain of the present invention isolates and purifies to obtain from aleurite montana plant, through being inoculated in aleurite montana earth culture seedling,
According to indices Comprehensive Evaluation, further confirm that it has the function that plant strain growth to promote aleurite montana biomass to increase, seek
Find and the beneficial function endogenetic fungus for promoting biomass to increase can be applied to produce.
The method applied is poured by bacterium solution and is inoculated in aleurite montana seedling, measures when being just inoculated with July and be inoculated with thousand during 60 days Septembers
The biomass when height of seedling of year paulownia, ground diameter and inoculation 60 days.Plant handled by aleurite montana endogenetic fungus ZJ2 and the seedling compareed
High growth rate, ground diameter growth rate and biomass significant difference.Height of seedling compares display, height of seedling of the bacterial strain ZJ2 processing compared with control group
Growth-promoting works well, its height of seedling growth rate is 5.75 times of control group.Ground diameter compares display, and the ground diameter growth rate of bacterial strain ZJ2 is
1.98 times of control.Increment compares display, and the increment of bacterial strain ZJ2 is 1.98 times of control.Show bacterial strain ZJ2 to aleurite montana
Biomass increase there is obvious facilitation.
Brief description of the drawings
Fig. 1 is influence of the different disposal to aleurite montana Seedling Biomass.
Embodiment
Rhizosphere soil, which pours, grants nursery stock and is directly inoculated with application
Test material is that aleurite montana seedling is the aleurite montana one year seedling that Jianyang Municipal Forestry Bureau provides.This experiment uses earth culture
Pot experiment, selects the consistent nursery stock of growing way to be colonized in diameter 15cm in May, 2014, in the plastic tub of high 10cm.Put per basin
Enter the 3kg yellow soils of equal mass, in order to ensure that later stage nursery stock connects the accuracy of bacterium, 10 ~ 15 drop first are instilled into per basin soil
Aldehyde (formalin) thimerosal, and cover basin mouth, disinfecting time 24h with plastic film.Treat that formaldehyde infiltration has sterilized in soil
Entirely, plastic film is removed, remaining formaldehyde is evaporated, in order to avoid influence the transplanting success of seedling.Nursery stock is implanted into, by one
Month restorative growth, in the bacterium solution 100ml that aleurite montana seedling rhizosphere applies that bacterial strain concentration is 5.5 × 106cfu/ml, continuous three
It, which is poured, applies, and blank control is used as by the use of distilled water solution processing.
It is prepared by bacterium solution：By the fluid nutrient medium of strains tested access 50mL, culture medium is improvement Martin's culture medium, wherein egg
It is white 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
For sterile water, pH value 6.4 ± 0.2, the culture of 72h is passed through in constant-temperature shaking incubator.It is dilute by ten times with sterile saline
Interpretation of the law dilutes bacterium solution, and calculating bacterial concentration using blood counting chamber is configured to 5.5 × 106cfu/mL.Inoculation carries out after 60 days
The measure of the indexs such as height of seedling, ground diameter, biomass, detection method are as follows：
（1）The measure of height of seedling ground diameter
Steel ruler measures height of seedling, electronic digital indicator measure ground diameter.
（2）Biomass estimation
The different parts of plant are respectively put into envelope and put signs on, are put into 105 DEG C of drying half an hour of baking oven（Passivation
Enzyme）, dry at 60 DEG C, after 12h, placed into after weighing in baking oven, replication is untill constant weight.Claim to obtain sample
Weight（W）, it is sample dry weight, the biomass of plant is represented with this, is accurate to 0.01g.
As a result with analysis
Influence of the different disposal to aleurite montana seedling height of seedling and ground diameter
As shown in table 1, the seedling height of seedling growth rate of inoculating strain ZJ2 is 44.38%, and control is only 7.72%, thus,
The seedling height of seedling growth rate of inoculating strain ZJ2 adds 475.2% than control.The seedling ground diameter growth rate of inoculating strain ZJ2 is
55.10%, with comparing display, the seedling ground diameter growth rate of inoculating strain ZJ2 adds 98.2% than control.
Influence of 1 different disposal of table to aleurite montana seedling height of seedling and ground diameter
Note：Data are the means standard deviation of three repetitions in table.
Influence of the different disposal to aleurite montana Seedling Biomass
Biomass is the important indicator for reflecting nursery stock production power level, and there is very close for its growth and development with nursery stock
Relation.Biomass the results show such as Fig. 1, can be obtained by variance analysis, bacterial strain ZJ2 processing after aleurite montana Seedling Biomass with
Control, which is compared, reaches the level of signifiance（Sig=0.003, the level of signifiance 0.05）, bacterial strain ZJ2 adds 98.2% than control.
In conclusion plant and height of seedling growth rate, ground diameter growth rate and growth handled by aleurite montana blade fungi ZJ2
Significant difference is measured, significant facilitation is played compared to growth of the control to aleurite montana nursery stock.Illustrate that the endogenetic fungus can
Aleurite montana nursery stock is set preferably to carry out photosynthesis, so that promote the growth of height of seedling, ground diameter, biomass increase.
Present invention discover that one plant can promote the endogenetic fungus that aleurite montana biomass increases, by this plant of aleurite montana endogenetic fungus bacterium
Strain is inoculated in aleurite montana seedling using the method for pouring root.By the measure to the height of seedling of plant, ground diameter and biomass, draw and connect
Seedling height growth rate, ground diameter growth rate and the increment of the processing of the kind bacterial strain largely higher than control, show it for promoting
Growth into plant height of seedling, ground diameter and biomass has very big function, so as to further confirm that obtaining this plant of endogenetic fungus can promote
Into the growth of aleurite montana biomass.
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, should all belong to the covering scope of the present invention.
Claims (4)
1. one plant can promote the endogenetic fungus that aleurite montana biomass increases, it is characterised in that：The fungi is disk stey
（Pestalotia sp.）ZJ2, on January 30th, 2015 in China Committee for Culture Collection of Microorganisms's common micro-organisms
Preservation is registered at center, and preserving number is CGMCC No. 10327.
2. the application bacterium solution of a kind of one plant as claimed in claim 1 endogenetic fungus that aleurite montana biomass can be promoted to increase, its
It is characterized in that：The preparation method using bacterium solution, by the disk stey（Pestalotia sp.）ZJ2 bacterial strains access liquid
Body culture medium, shaking table shaken cultivation, cultivation temperature are 28 DEG C, 48~72h of incubation time, and it is dense to calculate bacterium solution using blood counting chamber
Degree, 1.0 × 10 are diluted to by bacterium solution with ultra-pure water6-9.0×106Cfu/ml, it is spare.
3. the application bacterium solution of the one plant according to claim 2 endogenetic fungus that aleurite montana biomass can be promoted to increase, it is special
Sign is：The fluid nutrient medium is to improve Martin's culture medium, wherein peptone 5.0g/L, yeast extract powder 2.0g/L, grape
Sugared 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.
A kind of 4. purposes as claimed in claim 2 for applying bacterium solution, it is characterised in that：The application bacterium solution is used for aleurite montana seedling
Wooden rhizosphere soil, which pours, grants nursery stock and is directly inoculated with.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1420180A (en) * | 2002-08-23 | 2003-05-28 | 厦门大学 | Method for early diagnosis and detection of pine nematode disease |
| CN103173364A (en) * | 2013-03-05 | 2013-06-26 | 福建农林大学 | Endophytic fungus promoting casuarina equisetifolia biomass growth |
| CN104004666A (en) * | 2014-06-05 | 2014-08-27 | 福建农林大学 | Endophytic fungi capable of promoting China fir growth |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1420180A (en) * | 2002-08-23 | 2003-05-28 | 厦门大学 | Method for early diagnosis and detection of pine nematode disease |
| CN103173364A (en) * | 2013-03-05 | 2013-06-26 | 福建农林大学 | Endophytic fungus promoting casuarina equisetifolia biomass growth |
| CN104004666A (en) * | 2014-06-05 | 2014-08-27 | 福建农林大学 | Endophytic fungi capable of promoting China fir growth |
Non-Patent Citations (1)
| Title |
|---|
| "拟盘多毛孢属真菌的新种";陈育新 等;《菌物系统》;20020331;第21卷(第3期);第316-323页 * |
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