CN106010984B - One plant of endogenetic fungus for promoting acacia confusa biomass to increase under low-phosphorous environment - Google Patents

One plant of endogenetic fungus for promoting acacia confusa biomass to increase under low-phosphorous environment Download PDF

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CN106010984B
CN106010984B CN201610454595.7A CN201610454595A CN106010984B CN 106010984 B CN106010984 B CN 106010984B CN 201610454595 A CN201610454595 A CN 201610454595A CN 106010984 B CN106010984 B CN 106010984B
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biomass
plant
endogenetic fungus
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acacia confusa
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CN106010984A (en
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谢安强
陈灿
林晗
吴承祯
洪滔
周艳芬
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Fujian Agriculture and Forestry University
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    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • A01G9/02—Receptacles, e.g. flower-pots or boxes; Glasses for cultivating flowers
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Abstract

The endogenetic fungus that the present invention relates to one plant to promote acacia confusa biomass to increase under low-phosphorous environment.The endogenetic fungus be Penicillium (Penicillium sp.), preservation is registered in China Committee for Culture Collection of Microorganisms's common micro-organisms center on January 27th, 2016, deposit number is CGMCC No.11913.Acacia confusa endogenetic fungus I is inoculated in acacia confusa earth culture seedling, pass through the measurement to plant height of seedling, biomass, show that the height of seedling, the biomass that are inoculated with processing plant of the bacterial strain under low-phosphorous environment are largely higher than control substantially, show that it increases with very big function promotion biomass under low-phosphorous environment, to further confirm that obtaining this plant of endogenetic fungus can promote acacia confusa biomass to grow under low-phosphorous environment.

Description

One plant of endogenetic fungus for promoting acacia confusa biomass to increase under low-phosphorous environment
Technical field
The endogenetic fungus that the present invention relates to one plant to promote acacia confusa biomass to increase under low-phosphorous environment.
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 out of most plants research shows that can separate 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 had found in a variety of gramineae farm crops.The unique function that the endo-mycorrhiza separated from plant at present has according to it Can 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 To the resistance bacterium of the repair ability of poor environment.
In relation to endogenetic fungus, to the influence research that acacia confusa is grown, there is also blank.
Summary of the invention
The endogenetic fungus that the purpose of the present invention is to provide one plant to promote acacia confusa biomass to increase under low-phosphorous environment, The fungi be Penicillium (Penicillium sp.), on January 27th, 2016 in Chinese microorganism strain preservation conservator Meeting common micro-organisms center registers preservation, and address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, and deposit number is CGMCC No. 11913。
To achieve the above object, the present invention adopts the following technical scheme:
The separation of the bacterium, purifying 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.In the small beaker that water after sample last time is embathed is contained in 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 temperature incubations Bacterium colony upgrowth situation is observed after culture 5-7 days in case.Rapidly, the bacterial strain that can be first generated is separated for some bacterial strain breedings Purifying；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, switchingPenicillium sp.Function stem；
A kind of preparation of above-mentioned endogenetic fungus application bacterium solution for promoting acacia confusa biomass to increase under low-phosphorous environment Method is that above-mentioned bacterial strain is accessed to fluid nutrient medium, shaking table shaken cultivation, and cultivation temperature is 28 DEG C, incubation time 48~ 72h calculates bacterial concentration using blood counting chamber, bacterium solution is diluted to 1.0-9.0 × 10 with ultrapure water6Cfu/ml is finished product It is spare；Fluid nutrient medium: to improve 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.
A kind of application of above-mentioned endogenetic fungus application bacterium solution for promoting acacia confusa biomass to increase under low-phosphorous environment Method is to pour to impose on acacia confusa Seedling rhizosphere soil or nursery stock is directly inoculated with using the bacterium solution of the bacterial strain.
A kind of application of above-mentioned endogenetic fungus application bacterium solution for promoting acacia confusa biomass to increase under low-phosphorous environment Method is with 5.5 × 106Cfu/ml bacterium solution is poured in forest land in the rhizosphere of every plant of acacia confusa by every plant of 100ml.
Bacterial strain of the present invention is isolated and purified from acacia confusa plant, and acacia confusa earth culture is inoculated in Seedling further confirms that it has plant strain growth under low-phosphorous environment and promotes acacia confusa raw according to indices Comprehensive Evaluation The effect that object amount increases, the promotion plant beneficial function endogenetic fungus that biomass increases under low-phosphorous environment can be answered by searching out For producing.
The method applied is poured by bacterium solution and is inoculated in acacia confusa seedling, and carries out low-phosphorus stress test after inoculation 15 days, Its height of seedling is measured when coercing 0d, measures its height of seedling and its biomass when coercing 90d.It is commented according to the indices synthesis measured Sentence, further confirm that it has the function of plant strain growth to alleviate low-phosphorus stress, searches out beneficial to promotion biomass Function endogenetic fungus can be applied to produce.By the measurement to plant height of seedling and biomass, the processing for being inoculated with the bacterial strain is obtained Height of seedling growth rate and biomass are largely higher than control substantially, show that it increases promotion biomass under low-phosphorous environment It is long that there is very big function, to further confirm that obtaining this plant of endogenetic fungus can promote acacia confusa plant raw under low-phosphorous environment Object amount growth.
Detailed description of the invention
Fig. 1 is influence of the different disposal to acacia confusa seedling height of seedling；
Fig. 2 is influence of the different disposal to acacia confusa Seedling Biomass.
Specific embodiment
One, 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, the plastic tub of high 10cm.Soil used It is the stringent yellow soil Jing Guo fumigation.It is weighed by mixing, every basin is put into the 3kg soil of equivalent.It is extensive by one month After renaturation growth, it is 5.5 × 10 that continuous three days, which apply concentration in acacia confusa rhizosphere,6The 100mL bacterium solution of cfu/ml, uses distilled water Solution is as blank control.
Bacterium solution preparation: by the fluid nutrient medium of strains tested access 40mL, 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, calculates bacterial concentration using blood counting chamber and is configured to 5.5 × 106cfu/mL。
Two, low-phosphorus stress experimental design
The matrix of pot experiment soil is yellow soil.Each Nutrients are shown in Table 1 in the soil after measured.Inoculation 15 days laggard Row low-phosphorus stress test process, this period mainly allow inoculating strain to invade plant.
1 matrix soil nutrient situation of table
Unit: mg/g
The 4 phosphorus processing of this experimental design are horizontal, each level 3 repetitions (table 2).Phosphate fertilizer uses KH2PO4, periodically apply nitrogen Fertilizer, potash fertilizer and other microelements, until harvest.Low-phosphorus stress was carried out on May 4th, 2015.It is in time according to weather conditions Acacia confusa seedling keeps the skin wet.
2 low-phosphorus stress experimental design of table
Unit: mg/kg
The height of seedling of plant is measured when coercing 0d, when 90d, measures its biomass after 90d.Detection method is as follows:
(1) measurement of height of seedling
Steel ruler measures height of seedling
(2) measurement of biomass
The different parts of plant are respectively put into envelope and are put signs on, 105 DEG C of drying half an hour (passivation of baking oven are put into Enzyme), it dries, after 12h, is placed into baking oven after weighing, replication is until constant weight at 60 DEG C.Claim to obtain sample Weight (W), as sample dry weight, the biomass of plant is indicated with this, is accurate to 0.01g.
(3) data processing
Data preparation and mapping use Excel2003, and the statistical analysis of data uses SPSS20.0.
Three, result and analysis
(1) influence of the different disposal to acacia confusa height of seedling under different low-phosphorus stress
Four different low-phosphorus stress conditions are respectively as follows: severe water stress, moderate stress, mild stress, normal condition.Such as Fig. 1 It is shown, under four different low-phosphorus stress degree, the height of seedling growth rate of the seedling of inoculating strain I respectively than compareing big 113.48%, 130.17%, 73.11%, 40.24%, it follows that bacterial strain I may advantageously facilitate the growth of plant height of seedling.
(2) influence of the different disposal to acacia confusa biomass under different low-phosphorus stress
Biomass is the important indicator for reflecting nursery stock production power level, and there is very close for the growth and development of it and nursery stock Relationship.As shown in Fig. 2, the biomass of the seedling of inoculating strain I is bigger than compareing respectively under four different low-phosphorus stress degree 92.60%, 103.20%, 10.38%, 12.82%, thus, bacterial strain I may advantageously facilitate the growth of biomass.
Present invention discover that one plant of endogenetic fungus that acacia confusa seedling biomass can be promoted to increase under low-phosphorous environment, by the strain Acacia confusa endogenetic fungal bacterial strain is inoculated in acacia confusa seedling using the method for pouring root.Pass through height of seedling, the biomass to plant Measurement, show that the height of seedling of processing for being inoculated with the bacterial strain, biomass increase and be largely higher than control, show it for promoting The growth of biomass has very big function, to further confirm that obtaining this plant of endogenetic fungus can promote under low-phosphorous environment The growth of acacia confusa 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, is all covered by the present invention.

Claims (1)

1. the purposes using bacterium solution of one plant of endogenetic fungus for promoting acacia confusa biomass to increase under low-phosphorous environment, feature Be: the application bacterium solution is poured for acacia confusa Seedling rhizosphere soil and is applied or nursery stock is directly inoculated with；The endogenetic fungus is blueness It is mould belong to (Penicillium) I, on January 27th, 2016 in China Committee for Culture Collection of Microorganisms's common micro-organisms Preservation is registered at center, and deposit number is CGMCC No. 11913；The preparation method using bacterium solution, by the Penicillium (Penicillium) I 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 diluted to 1.0 × 10 with ultrapure water6-9.0×106Cfu/ml, it is spare；Liquid Body culture medium: to improve Martin's culture medium, wherein peptone 5.0g/L, yeast extract powder 2.0g/L, glucose 20.0g/L, phosphorus Sour hydrogen dipotassium 1.0g/L, magnesium sulfate 0.5g/L, it is other be sterile water, pH value 6.4 ± 0.2.
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CN110257258B (en) * 2019-07-01 2020-11-27 福建农林大学 Endophytic fungus capable of promoting phosphorus absorption of schima superba

Citations (1)

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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
CN101974438A (en) * 2010-11-18 2011-02-16 福建农林大学 Eucalyptus endophyte and application thereof

Non-Patent Citations (3)

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effects of phosphorus-solubilizing bacteria and vesicular-arbuscular mycorrhizal fungi on the growth of tree species in subtropical-tropical soils;Chiu-Chung Young;《soil science and plant nutrition》;19901231;全文 *
内生真菌对低磷胁迫下尾巨桉生理及土壤特性的影响;谢安强等;《西南林业大学学报》;20130630;全文 *
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