CN110200018A - It is a kind of promote plant establishment best DSE connect bacterium amount - Google Patents
It is a kind of promote plant establishment best DSE connect bacterium amount Download PDFInfo
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- CN110200018A CN110200018A CN201910505631.1A CN201910505631A CN110200018A CN 110200018 A CN110200018 A CN 110200018A CN 201910505631 A CN201910505631 A CN 201910505631A CN 110200018 A CN110200018 A CN 110200018A
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- plant
- dark color
- endogenetic fungus
- dse
- organ
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K17/00—Soil-conditioning materials or soil-stabilising materials
- C09K17/14—Soil-conditioning materials or soil-stabilising materials containing organic compounds only
Abstract
The invention discloses a kind of best DSE for promoting plant establishment to connect bacterium amount.It is (0.468-1.404) mg/mL that the best DSE disclosed by the invention for promoting plant establishment, which connects dry mycelial weight in bacterium amount, it is that having containing dark color for (0.468-1.404) mg/mL impregnates plant tissue or organ every the liquid of endogenetic fungus using dry mycelial weight, it realizes and promotes plant establishment, there are root long, radical and root diameter after the culture solution of endogenetic fungus handles plant organ to dramatically increase using dark color, and having with dark color every the concentration dilution of endogenetic fungus is 20% (i.e. dry mycelial weight is 0.468mg/mL) best.The present invention, which connects bacterium amount processing plant tissue or organ using the best DSE of plant establishment, can assist plant establishment, promote plant growth, there is positive ecological significance for improving land reclamation efficiency, also provide a kind of microbial technique support for land reclamation in mining area and ecological reconstruction.
Description
Technical field
The present invention relates in field of biotechnology, a kind of best DSE promoting plant establishment connects bacterium amount.
Background technique
Dark color has and has the biology similar with mycorrhizal fungi every endogenetic fungus (Dark Septate Endophytes, DSE)
Function is learned, the epidermis in health plant root, the cortex even intracellular or space between cells of vascular tissue is mainly colonized, is formed altogether
Raw body, without causing pathological changes of plant.DSE ecologicaI distribution is extensive, and colonizing in different niches plant shows them seldom or do not have
There is host specificity, and the physiology that can be established mutual reciprocity and mutual benefit with plant and mutually adjust is whole, but respectively has its form
Feature.Studies have shown that DSE plays even more important effect in stressful environmental, plant is expanded by the growth of mycelia
Root absorbing area improves the Nutrient Absorption ability except primitive root system absorption region.
Summary of the invention
The technical problem to be solved by the present invention is to how promote plant establishment.
In order to solve the above technical problems, present invention firstly provides a kind of method for promoting plant establishment, the method packet
It includes: having using dark color and handle plant tissue or organ every endogenetic fungus, realize and promote plant establishment.
In the above method, have using dark color and handle plant tissue or organ every endogenetic fungus can include: using containing described
Dark color, which has, impregnates the plant tissue or organ every the liquid of endogenetic fungus.
In the above method, having containing the dark color every dry mycelial weight in the liquid of endogenetic fungus can be (0.468-1.404)
mg/mL.Have containing the dark color every the concretely 0.468mg/mL or 1.404mg/mL of dry mycelial weight in the liquid of endogenetic fungus.
Simultaneously drying to constant weight to have the liquid collection mycelia every endogenetic fungus containing the dark color using filter paper filtering for the dry mycelial weight
Obtained dry mycelial weight.
In the above method, it is described have containing the dark color can have every endogenetic fungus for the dark color every the liquid of endogenetic fungus
Culture solution or the culture solution dilution.
The culture solution can be to have using dark color described in MMN culture medium culture every endogenetic fungus gained culture solution.
The dilution can dilute the culture solution using water and obtain.
The MMN culture medium can be by by CaCl20.05g, MgSO40.15g, NaCl 0.025g, FeCl30.01g,
KH2PO40.5g, Vitamin B1 (thiamine) 0.0001g, (NH4)2HPO40.25g, Glucose (glucose) 10g,
After Citric acid (citric acid) 0.2g and Malt extract (malt extract) 10g mixing, distilled water 1000ml is added,
Sterilizing obtains.
In the above method, the plant tissue or organ are impregnated every the liquid of endogenetic fungus using having containing the dark color
When, the half of the plant tissue or organ can be in air.
It during processing, can be by adding the MMN culture medium so that the half of the plant tissue or organ is in
In air.
In the above method, the time of the processing is 15 days.
The condition of the processing can are as follows: carries out in the dark within first 3 days, temperature is 22 DEG C, relative air humidity 70%；Place
Reason is after three days, illumination cultivation, intensity of illumination 1500-2000Lux, and the photoperiod is 16h illumination/8h dark, under illumination and dark
Cultivation temperature is respectively 25 and 22 DEG C, relative air humidity 70%.
It is described that plant establishment is promoted to may be embodied in the elongation for promoting plant roots, radical increase and/or root in the above method
In overstriking.
In the above method, the plant organ can be vegetable seeds.
In the above method, it can be common in China Committee for Culture Collection of Microorganisms that the dark color, which has every endogenetic fungus,
The deposit number at microorganism center is CGMCC No.17464.
In the above method, the plant can be a1) or a2) or a3):
A1) monocotyledon or dicotyledon；
A2) gramineae plant；
A3) corn.
The plant tissue or organ can be sterile tissue or organ.
The processing can carry out in gnotobasis.
Following any applications of the method for promoting plant establishment, also belong to protection scope of the present invention:
X1) plant production；
X2) planting；
X3) promote plant growth；
X4) promote land reclamation.
The plant can be a1) or a2) or a3):
A1) monocotyledon or dicotyledon；
A2) gramineae plant；
A3) corn.
In one embodiment of the invention, the corn be in glutinous No.1.
In the method for promotion plant establishment of the invention, is had using dark color and handle plant organ every the culture solution of endogenetic fungus
Root long, radical and root diameter afterwards, which is all remarkably higher than, does not have the control group every endogenetic fungus processing using dark color, using deeply
Color has when endogenetic fungus handles plant organ, and having with dark color every the concentration of endogenetic fungus is 20% best.Promotion of the invention
The method of plant establishment can assist plant establishment, promote plant growth, have positive life for improving land reclamation efficiency
State meaning also provides a kind of microbial technique support for land reclamation in mining area and ecological reconstruction.
Biomaterial preservation explanation
The classification naming of biomaterial: Darksidea zeta
The strain number of biomaterial: needle A1-3
Depositary institution's title of biomaterial: China Committee for Culture Collection of Microorganisms's common micro-organisms center
Depositary institution's abbreviation of biomaterial: CGMCC
The depositary institution address of biomaterial: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Chinese Academy of Sciences microorganism
Research institute, postcode: 100101
The preservation date of biomaterial: on April 8th, 2019
The collection of biomaterial is registered on the books number: CGMCC No.17464
Detailed description of the invention
Fig. 1 is the influence that the DSE culture solution of various concentration is soaked seed to corn root.CK indicates to soak seed using sterile water, MMN
CK indicates to soak seed using sterile MMN fluid nutrient medium, and 20%, 40% and 60% respectively indicates and utilizes 20%DSE culture solution, 40%
DSE culture solution and the seed soaking of 60%DSE culture solution；With there were significant differences between the data in figure, indicating different lowercases, phase is indicated
With between the data of lowercase without significant difference.
Specific embodiment
The present invention is further described in detail With reference to embodiment, and the embodiment provided is only for explaining
The bright present invention, the range being not intended to be limiting of the invention.Experimental method in following embodiments is unless otherwise specified
Conventional method.Material as used in the following examples, reagent, instrument etc., are commercially available unless otherwise specified.
Quantitative test in following embodiment, is respectively provided with three repeated experiments, and results are averaged.
In following embodiments, the experimental method for connecing bacterium unless otherwise specified, is completed in aseptic operating platform.DSE and jade
Rice seed co-cultures in growth cabinet (the intelligent growth cabinet of RXZ).
Influence of the DSE culture solution to maize rooting of embodiment 1, various concentration
Dark color has every endogenetic fungus (DSE) (Darksidea zeta) (its entitled needle A1-3): by inventor from Inner Mongol
Obtained strains are isolated and purified in the Stipa capillata root system of ancient Xilinguole League, autonomous region Xilinhot City nortel Shengli Opencast Mine, according to as follows
Method, which is accredited as dark color, to be had every endogenetic fungus: detecting the ITS sequence of the bacterial strain, sequence is sequence 1 in sequence table.
The bacterial strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms on April 8th, 2019
The heart, deposit number are CGMCC No.17464.
Corn variety be in glutinous No.1 (authentication number: 0103006-2000, farming development in science and technology Co., Ltd in Beijing).
One, the culture of DSE
PDA culture medium: after potato leaching powder 3.0g, glucose 20.0g and agar 14.0g are mixed, distilled water is added
Dissolution is boiled in 1000ml, heating, packing, and 121 DEG C of high pressure sterilization 15min are spare.
DSE is inoculated into PDA culture medium, it is dark at 28 DEG C to be inverted culture 15 days.
Two, the preparation of DSE culture solution
MMN fluid nutrient medium: by CaCl20.05g, MgSO40.15g, NaCl 0.025g, FeCl30.01g, KH2PO4
0.5g, Vitamin B1 (thiamine) 0.0001g, (NH4)2HPO40.25g, Glucose (glucose) 10g, Citric acid
After (citric acid) 0.2g and Malt extract (malt extract) 10g mixing, distilled water 1000ml, packing, 121 DEG C of height are added
Sterilizing 15min is pressed, it is spare.
After step 1 culture, the bacteria cake of diameter 6mm is taken, is inoculated into MMN fluid nutrient medium, is trained in 25 DEG C of concussions
It supports, shaking speed first day is 180r/min, start within second day to be changed to 160r/min, after cultivating 15 days, obtains DSE culture solution,
(i.e. DSE culture solution is filtered through filter paper, and the mycelia after collecting mycelia drying to constant weight is dry for DSE dry mycelial weight in 1mL DSE culture solution
Weight) it is 2.34mg, it is spare.
Three, influence of the DSE culture solution of various concentration to plant establishment effect
It is diluted with sterile water DSE culture solution resulting to step 2, obtains the culture solution of three concentration, i.e., 20%
DSE culture solution (20%DSE culture solution: take DSE culture solution 20ml described in step 2, be diluted to 100ml with sterile water), 40%
DSE culture solution (40%DSE culture solution: taking DSE culture solution 40ml described in step 2, be diluted to 100ml with sterile water) and 60%
DSE culture solution (60%DSE culture solution: takes DSE culture solution 60ml described in step 2, is diluted to 100ml with sterile water), for use.
Sterile triangular flask (100ml) is taken, is divided into five groups, every group 3, is put into glutinous No.1 corn in 5 in each triangular flask
Seed.The DSE culture solution of three concentration achieved above is separately added into three groups of triangular flasks, a kind of every group of DSE training of concentration
Nutrient solution guarantees every bottle of culture solution 5ml (i.e. liquid there be not seed just half).Remaining one group is used sterile water as control, separately
One group using sterile MMN fluid nutrient medium as control.All triangular flasks are sealed with sterile PVA film.Primary nothing was added every 5 days
After bacterium MMN nutrient solution co-cultures 15 days, Maize Roots situation is analyzed using CI600 software statistics.Culture first 3 days in dark
Middle progress, temperature are 22 DEG C, relative air humidity 70%；Culture three days after, illumination cultivation, intensity of illumination be 1800Lux (
In incubator, illumination is within the scope of 1500-2000Lux, average out to 1800Lux), the photoperiod is 16h illumination/8h dark, light
According to being respectively 25 and 22 DEG C with cultivation temperature under dark, relative air humidity 70%.
Various concentration DSE culture solution seed soaking growth-promoting root influence result as shown in figure 1 and table 1, additional amount 20%DSE
The corn of culture solution is averaged the total root long of single plant and average root diameter is all remarkably higher than the corresponding indexs of other each group plant roots,
The corn of the 20% and 60%DSE culture solution single plant radical that is averaged is all remarkably higher than other each groups.Show that 20%DSE culture solution has
Help that corn seed is promoted to take root.
Influence of the DSE culture solution seed soaking of table 1, various concentration to corn root
In table 1, total root long indicates the average total root long of single plant, and radical indicates average single plant radical；In the data of same row, mark
Have between the data of different lowercases that there were significant differences, indicates between the data of identical lowercase without significant difference.
<110>China Mining Univ. (Beijing), Beijing synbiotic ecological environment engineering technology Co., Ltd
<120>a kind of best DSE for promoting plant establishment connects bacterium amount
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 590
<212> DNA
<213>dark color has every endogenetic fungus (Darksidea zeta)
<400> 1
cttccgtaag gtgacctgcg gaaggatcat tacctggcct tgggccgctc gcgggagcta 60
gtcgcttgcg acgacgctac cgagggcgct tagcccttga ctatcacctt gactacgtgc 120
accttttgtt gtttcctcgg caggtcacct gccgccagga accctctaaa ccttttgcaa 180
tagcatccaa acttctgaaa acaaaccaaa ttatttacaa cttttaacaa tggatctctt 240
ggttctggca tcgatgaaga acgcagcgaa atgcgataag tagtgtgaat tgcagaattc 300
agtgaatcat cgaatctttg aacgcacatt gcgccccatg gtattccgtg gggcatgcct 360
gttcgagcgt catttacccc ctcaagctcc gcttggtgtt gggcgtctgt cccgcttcgc 420
gcgcggactc gccccaaagg tattggcagc ggtcgtgcca gcttctcgcg cagcacattg 480
cgcttctcga ggcaccggcg ggcccgcgtc catcaagctc acccccccag tttgacctcg 540
gatcaggtag ggatacccgc tgaacttaag catatcaata agcggaggaa 590
Claims (9)
1. promoting the method for plant establishment, comprising: have using dark color and handle plant tissue or organ every endogenetic fungus, realize and promote
Plant establishment.
2. according to the method described in claim 1, handling plant tissue or device every endogenetic fungus it is characterized by: having using dark color
Official includes: to utilize the liquid immersion plant tissue or organ having containing the dark color every endogenetic fungus.
3. according to the method described in claim 2, it is characterized by: having containing the dark color every mycelia in the liquid of endogenetic fungus
Dry weight is (0.468-1.404) mg/mL.
4. according to the method in claim 2 or 3, it is characterised in that: the liquid having containing the dark color every endogenetic fungus
Body is the dilution that the dark color has culture solution or the culture solution every endogenetic fungus.
5. according to the method any in claim 2-4, it is characterised in that: have using containing the dark color every endogenetic fungus
Liquid when impregnating the plant tissue or organ, the half of the plant tissue or organ is in air.
6. any method in -5 according to claim 1, it is characterised in that: the time of the processing is 15 days；
And/or the condition of the processing are as follows: carry out in the dark for first 3 days, temperature is 22 DEG C, relative air humidity 70%；Place
Reason is after three days, illumination cultivation, intensity of illumination 1500-2000Lux, and the photoperiod is 16h illumination/8h dark, under illumination and dark
Cultivation temperature is respectively 25 and 22 DEG C, relative air humidity 70%.
7. any method in -6 according to claim 1, it is characterised in that: the promotions plant establishment is embodied in promotion plant
In elongation, radical increase and/or the overstriking of root of object root；
And/or the plant organ is vegetable seeds；
And/or it is in China Committee for Culture Collection of Microorganisms's common micro-organisms center that the dark color, which has every endogenetic fungus,
Deposit number be CGMCC No.17464.
8. any method in claim 1-7, it is characterised in that: the plant is a1) or a2) or a3):
A1) monocotyledon or dicotyledon；
A2) gramineae plant；
A3) corn.
9. following any applications of any the method in claim 1-8:
X1) plant production；
X2) planting；
X3) promote plant growth；
X4) promote land reclamation.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111304101A (en) * | 2020-04-17 | 2020-06-19 | 中国矿业大学(北京) | DSE (Deuteroxylin-beta) dry bacterium agent and application thereof in promoting plant growth |
| CN111394256A (en) * | 2020-03-20 | 2020-07-10 | 中国矿业大学(北京) | Efficient large-scale production and transportation linkage production method for deep-color endophytic fungus liquid |
| CN111394259A (en) * | 2020-04-17 | 2020-07-10 | 中国矿业大学(北京) | Preparation method of DSE dry microbial inoculum capable of promoting plant growth and easy to store and transport |
| CN111471597A (en) * | 2020-04-17 | 2020-07-31 | 中国矿业大学(北京) | Preparation and application method of DSE (Deuteroxylin-N-acetylneuraminidase) fungicide and symbiotic effect sensitive period monitoring |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111394256A (en) * | 2020-03-20 | 2020-07-10 | 中国矿业大学(北京) | Efficient large-scale production and transportation linkage production method for deep-color endophytic fungus liquid |
| CN111394256B (en) * | 2020-03-20 | 2021-10-15 | 中国矿业大学(北京) | Efficient large-scale production and transportation linkage production method for deep-color endophytic fungus liquid |
| CN111304101A (en) * | 2020-04-17 | 2020-06-19 | 中国矿业大学(北京) | DSE (Deuteroxylin-beta) dry bacterium agent and application thereof in promoting plant growth |
| CN111394259A (en) * | 2020-04-17 | 2020-07-10 | 中国矿业大学(北京) | Preparation method of DSE dry microbial inoculum capable of promoting plant growth and easy to store and transport |
| CN111471597A (en) * | 2020-04-17 | 2020-07-31 | 中国矿业大学(北京) | Preparation and application method of DSE (Deuteroxylin-N-acetylneuraminidase) fungicide and symbiotic effect sensitive period monitoring |
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