CN106010984A - Endophytic fungus promoting acacia confusa biomass growth under low-phosphorous environment - Google Patents
Endophytic fungus promoting acacia confusa biomass growth under low-phosphorous environment Download PDFInfo
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- CN106010984A CN106010984A CN201610454595.7A CN201610454595A CN106010984A CN 106010984 A CN106010984 A CN 106010984A CN 201610454595 A CN201610454595 A CN 201610454595A CN 106010984 A CN106010984 A CN 106010984A
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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
- A01G9/00—Cultivation in receptacles, forcing-frames or greenhouses; Edging for beds, lawn or the like
- A01G9/02—Receptacles, e.g. flower-pots or boxes; Glasses for cultivating flowers
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Abstract
The invention relates to an endophytic fungus promoting acacia confusa biomass growth under a low-phosphorous environment. The endophytic fungus is Penicillium sp. and is registered and preserved in Center of general microbiology of China Committee for Culture Collection of Microorganisms on January 27, 2016, and the preservation number is CGMCC No.11913. Acacia confusa normal plantlets are inoculated with an acacia confusa endophytic fungus I, by measuring the heights and the biomass of plants, it is concluded that the heights and the biomass of the treated plants inoculated with the endophytic fungus I under the low-phosphorous environment are much higher than those of contrasts, it shows that the endophytic fungus I has a very large effect on promoting the plant biomass growth under the low-phosphorous environment, and therefore it is further proved that the endophytic fungus can promote the acacia confusa biomass growth under the low-phosphorous environment.
Description
Technical field
The present invention relates to the endogenetic fungus that a strain promotes acacia confusa Biomass to increase under low-phosphorous environment.
Background technology
The research of endophyte of plant started from for 19 end of the centurys, and Vogl is from rye grass Lolium temulentum
L., seed is isolated the first strain endophyte.But really start the endophyte in numerous studies plant and originate in the eighties in last century, 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, kind is many, and forefathers' research shows in most plants can isolated endogenetic fungus, it can be seen that endogenetic fungus is prevalent in plant.At least in more than 80 gramineae farm crop belonging to kind more than 290, it is found that endophyte in worldwide.The endo-mycorrhiza separated from plant at present can be divided into according to its unique function having: the plant growth-promoting bacterias such as nitrogen-fixing bacteria, solid potassium bacterium, solid phosphorus bacterium, has the biocontrol microorganisms of disease and insect resistance and has the promotion plant resistance bacterium to the repair ability of poor environment.
The influence research grown acacia confusa about endogenetic fungus there is also blank.
Summary of the invention
It is an object of the invention to provide the endogenetic fungus that a strain promotes acacia confusa Biomass to increase under low-phosphorous environment, this fungus be Penicillium (Penicillium sp.), register preservation on January 27th, 2016 at China Committee for Culture Collection of Microorganisms's common micro-organisms center, address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, and preserving number is CGMCC No. 11913.
For achieving the above object, the present invention adopts the following technical scheme that
The separation of this bacterium, purification include:
A, material: by collecting the root of different year, branch stem, leaf are divided into three parts, clean up with tap water, are dispensed in valve bag, in 4 DEG C of Refrigerator stores；
B, sterilization: sodium hypochlorite rinsing (root 10min, stem 5min, leaf 2min) → aseptic water washing of 75% ethanol rinsing 30s → aseptic water washing 3-4 time → 15% 4-5 time.Water after sample embathes for the last time is contained in the small beaker of sterilizing, and at flat lining out as comparison, the sterilization with sample survey is the most thorough, generates if any bacterium colony behind some skies, then show that sample surfaces sterilization is not thorough, need to continue to adjust sterilization method；
C, the selection of inoculation position: blade: leaf tip, leaf central part and leaf base 3 process；Branch stem: upper, middle and lower is divided into 3 positions, and wherein the 3rd position is branch stem junction；Root: be divided into the tip of a root and 2 parts of foundation；
D, inoculation: cut by the outer implant vaccinating lancet after sterilization, be laid in media surface, observe colony growth situation in 28 DEG C of constant incubators after cultivating 57 days.Rapidly, the bacterial strain that can be first produced carries out isolated and purified the breeding of some bacterial strains；The bacterial strain of poor growth and negligible amounts can extend observing time, until purification after its growth is stable；
Solid culture: use improvement Martin's solid medium, formula is 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, other be sterilized water, pH value 6.4 ± 0.2, cultivation temperature is 28 DEG C, training method is that flat board is cultivated, and incubation time is 48h；
E, different types of endogenetic fungus separation bred access plating medium and are purified, and respectively obtain the above-mentioned of single culture after 2-3 point connects purification, switchingPenicillium sp.Function stem；
The preparation method of above-mentioned a kind of endogenetic fungus application bacterium solution promoting acacia confusa Biomass to increase under low-phosphorous environment, it is that above-mentioned bacterial strain is accessed fluid medium, shaking table shaken cultivation, cultivation temperature is 28 DEG C, incubation time 48～72h, utilize blood counting chamber to calculate bacterial concentration, bacterium solution ultra-pure water is diluted to 1.0-9.0 × 106It is standby that cfu/ml is finished product；Fluid medium: for 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, other be sterilized water, pH value 6.4 ± 0.2.
The application process of above-mentioned a kind of endogenetic fungus application bacterium solution promoting acacia confusa Biomass to increase under low-phosphorous environment, is to apply the bacterium solution of this bacterial strain to water to impose on acacia confusa Seedling rhizosphere soil or nursery stock direct inoculation.
The application process of above-mentioned a kind of endogenetic fungus application bacterium solution promoting acacia confusa Biomass to increase under low-phosphorous environment, is with 5.5 × 106Cfu/ml bacterium solution, is watered in the rhizosphere of every strain acacia confusa in forest land by every strain 100ml.
The bacterial strain that the present invention relates to is isolated and purified from acacia confusa plant to be obtained, through being inoculated in acacia confusa earth culture Seedling, according to indices Comprehensive Evaluation, it is further characterized by it and under low-phosphorous environment, plant strain growth is had the effect promoting that acacia confusa Biomass increases, search out promoting that the plant useful function endogenetic fungus that Biomass increases under low-phosphorous environment can be applicable to produce.
Watered the method executed by bacterium solution and be inoculated in acacia confusa seedling, and carry out low-phosphorus stress test after inoculating 15 days, measure its height of seedling when coercing 0d, when coercing 90d, measure its height of seedling and Biomass thereof.According to the indices Comprehensive Evaluation recorded, it is further characterized by it and plant strain growth is had the effect alleviating low-phosphorus stress, search out promoting that the function endogenetic fungus that biomass is useful can be applicable to produce.By the mensuration to plant height of seedling and Biomass, show that height of seedling rate of increase and the Biomass of the process inoculating this bacterial strain are higher than comparison the most largely, show that it has the biggest function to promotion biomass growth under low-phosphorous environment, thus be further characterized by obtaining this strain endogenetic fungus and can promote that under low-phosphorous environment acacia confusa biomass grows.
Accompanying drawing explanation
Fig. 1 is the different disposal impact on acacia confusa seedling height of seedling；
Fig. 2 is the different disposal impact on acacia confusa Seedling Biomass.
Detailed description of the invention
One, rhizosphere soil waters and grants the application of nursery stock direct inoculation
This test uses earth culture pot experiment, the acacia confusa one year seedling that test material therefor provides for Municipal Forestry Bureau of Zhangzhou City of Fujian Province.Select the acacia confusa seedling planting that growing way is consistent in diameter 15cm, the plastic tub of high 10cm.Soil used is strictly through the yellow soil of fumigation.Weighing through mixing, the 3kg soil of equivalent put into by every basin.After the restorative growth of month, continuous three days apply concentration in acacia confusa rhizosphere is 5.5 × 106The 100mL bacterium solution of cfu/ml, with distilled water solution as blank.
Prepared by bacterium solution: strains tested accesses the fluid medium of 40mL, culture 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, other be sterilized water, pH value 6.4 ± 0.2, through the cultivation of 72h in constant-temperature shaking incubator.Press decimal dilution method with physiological saline solution bacterium solution to be diluted, utilize blood counting chamber to calculate bacterial concentration and be configured to 5.5 × 106cfu/mL。
Two, low-phosphorus stress EXPERIMENTAL DESIGN
The substrate of pot experiment soil is yellow soil.In this soil, each Nutrients is shown in Table 1 after measured.Carry out low-phosphorus stress test process after inoculating 15 days, the most mainly allow inoculating strain invade plant.
Table 1 matrix soil nutrient situation
Unit: mg/g
4 phosphorus of this EXPERIMENTAL DESIGN process levels, each level 3 repetition (table 2).Phosphate fertilizer uses KH2PO4, regular nitrogen fertilizer application, potash fertilizer and other trace element, until results.Low-phosphorus stress was carried out on May 4th, 2015.Keep the skin wet for acacia confusa Seedling in time according to weather conditions.
Table 2 low-phosphorus stress EXPERIMENTAL DESIGN
Unit: mg/kg
Measure the height of seedling of plant time when coercing 0d, 90d, after 90d, measure its Biomass.Detection method is as follows:
(1) mensuration of height of seedling
Steel ruler measures height of seedling
(2) mensuration of Biomass
Being respectively put in envelope by the different parts of plant and put signs on, putting into 105 DEG C of baking oven and dry half an hour (inactive enzyme), dry, after 12h, place in baking oven after weighing at 60 DEG C, replication is till constant weight.Claim to obtain the weight (W) of sample, be sample dry weight, the Biomass representing plant with this, it is accurate to 0.01g.
(3) data process
Data compilation uses Excel2003 with mapping, and the statistical analysis of data uses SPSS20.0.
Three, result and analysis
(1) different disposal impact on acacia confusa height of seedling under different low-phosphorus stress
Four different low-phosphorus stress conditions are respectively as follows: severe water stress, moderate is coerced, mild stress, normal condition.As it is shown in figure 1, under four different low-phosphorus stress degree, the ratio comparison big 113.48%, 130.17%, 73.11%, 40.24% respectively of the height of seedling rate of increase of the seedling of inoculating strain I, it follows that bacterial strain I may advantageously facilitate the growth of plant height of seedling.
(2) different disposal impact on acacia confusa Biomass under different low-phosphorus stress
Biomass is the important indicator of reflection nursery stock production force level, and it also exists very close relationship with the growth promoter of nursery stock.As in figure 2 it is shown, under four different low-phosphorus stress degree, the ratio comparison big 92.60%, 103.20%, 10.38%, 12.82% respectively of the Biomass of the seedling of inoculating strain I, thus can obtain, bacterial strain I may advantageously facilitate the growth of biomass.
Present invention discover that a strain can promote the endogenetic fungus that acacia confusa Seedling Biomass increases under low-phosphorous environment, use the method watering root to be inoculated in acacia confusa seedling this strain acacia confusa endogenetic fungal bacterial strain.By to the height of seedling of plant, the mensuration of Biomass, show that the inoculation height of seedling of process of this bacterial strain, Biomass increase the most largely higher than comparison, show that it has the biggest function for the growth of promotion biomass, thus be further characterized by obtaining this strain endogenetic fungus and can promote the growth of acacia confusa Biomass under low-phosphorous environment.
The foregoing is only presently preferred embodiments of the present invention, all impartial changes done according to scope of the present invention patent and modification, all should belong to the covering scope of the present invention.
Claims (3)
1. a strain promotes the endogenetic fungus that acacia confusa Biomass increases under low-phosphorous environment, it is characterised in that: this fungus be Penicillium (Penicillium sp.), register preservation on January 27th, 2016 at China Committee for Culture Collection of Microorganisms's common micro-organisms center, preserving number is CGMCC No. 11913.
2. the application bacterium solution of the endogenetic fungus that a strain as claimed in claim 1 promotes acacia confusa Biomass to increase under low-phosphorous environment, it is characterised in that: the preparation method of described application bacterium solution, by described Penicillium (Penicillium sp.) bacterial strain access fluid medium, shaking table shaken cultivation, cultivation temperature is 28 DEG C, incubation time 48～72h, utilizes blood counting chamber to calculate bacterial concentration, bacterium solution ultra-pure water is diluted to 1.0-9.0 × 106Cfu/ml, standby；Fluid medium: for 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, other be sterilized water, pH value 6.4 ± 0.2.
3. the purposes of an application bacterium solution as claimed in claim 2, it is characterised in that: described application bacterium solution is watered for acacia confusa Seedling rhizosphere soil and is executed or nursery stock direct inoculation.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110257258A (en) * | 2019-07-01 | 2019-09-20 | 福建农林大学 | A kind of endogenetic fungus that Schima superba phosphorus can be promoted to absorb |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101974438A (en) * | 2010-11-18 | 2011-02-16 | 福建农林大学 | Eucalyptus endophyte and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101974438A (en) * | 2010-11-18 | 2011-02-16 | 福建农林大学 | Eucalyptus endophyte and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| CHIU-CHUNG YOUNG: "effects of phosphorus-solubilizing bacteria and vesicular-arbuscular mycorrhizal fungi on the growth of tree species in subtropical-tropical soils", 《SOIL SCIENCE AND PLANT NUTRITION》 * |
| 南宏伟等: "相思树根瘤菌优良抗逆性菌株的筛选", 《西北林学院学报》 * |
| 谢安强等: "内生真菌对低磷胁迫下尾巨桉生理及土壤特性的影响", 《西南林业大学学报》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110257258A (en) * | 2019-07-01 | 2019-09-20 | 福建农林大学 | A kind of endogenetic fungus that Schima superba phosphorus can be promoted to absorb |
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