CN108753638A

CN108753638A - The salicylic rice field Azotica of one plant of production and its application in rice field

Info

Publication number: CN108753638A
Application number: CN201810397659.3A
Authority: CN
Inventors: 贺鸿志; 包江桥; 周伊薇; 卢玉真; 陈桂葵; 黎华寿; 秦俊豪
Original assignee: South China Agricultural University
Current assignee: South China Agricultural University
Priority date: 2018-04-28
Filing date: 2018-04-28
Publication date: 2018-11-06
Anticipated expiration: 2038-04-28
Also published as: CN108753638B

Abstract

The invention discloses the salicylic rice field Azotica of one plant of production and its applications in rice field.Nostoc (Nostoc piscinale) SCAU-003 is given birth in the entitled pond of the bacterial strain, and deposit number is CCTCC NO：M 2018194, the deposit date is on April 11st, 2018, depositary institution was the China typical culture collection center positioned at Wuhan, China Wuhan University.The bacterial strain can generate that the plant hormones such as high concentration salicylic acid, growth is fast, fixed nitrogen carbon sequestration capacity is strong, can be used for new applying mode and used as bio-feritlizer in rice terrace, can be instead of the chemical nitrogen fertilizer of half, while increase lodging resistance in rice and disease resistance ability.

Description

The salicylic rice field Azotica of one plant of production and its application in rice field

Technical field

The present invention relates to microorganism field and microbial manure crop field application field, more particularly to the one plant salicylic rice of production Field Azotica and its application in rice field.

Background technology

After nineteen thirty-nine India scientist reports fertilized the soil with cyanobacteria for the first time, many countries have carried out correlative study in succession, grind Studying carefully proof inoculation cyanobacteria in rice terrace can increase production.Starting Li Shanghao the fifties in last century, part provinces and regions carry out in south China Support the experiment of algae in rice field, the results showed that increasing production of rice is up to 10~30%.Azotica is put in rice field in a suitable place to breed not only can provide nitrogen for crop Element and organic matter improve yield, reduce fertilizer amount, improve Phosphate solubility, reduce water and soil pollution, also have secretion plant Hormone promotes the beneficial ecological functions such as plant growth.

Plant hormone be plant cell receive specific environment signal induction generate, in extremely low concentration be adjusted physiology it is anti- The active material answered, they are all a little simple small molecular organic compounds, are divided into six major class, i.e., auxin, the basic element of cell division, Abscisic acid, gibberellin, ethylene and rape element sterol etc..Plant hormone has important adjusting control to make the growth and development of plant With.Wherein, heteroauxin and salicylic acid etc. can improve the resistance of crop while promoting plant growth, and salicylic acid exists Play an important roll in terms of induction crop resistance.And both at home and abroad, some researches show that Azoticas can generate including auxin With the various plants hormone including the basic element of cell division etc., but salicylic report is not generated about rice field Azotica.? Under the current double overall background subtracted of national requirements fertilizer and pesticide, if can screen, growth is rapid, fixed nitrogen carbon sequestration capacity is strong and can produce It is important that plant hormone induces the Azotica of plant disease-resistant to have it for agricultural production as bio-feritlizer and biological pesticide Meaning.

Invention content

The primary purpose of the present invention is that the shortcomings that overcoming the prior art and deficiency, it is solid to provide one plant of salicylic rice field of production Nitroblue algae.

Another object of the present invention also resides in the application for providing the above-mentioned salicylic rice field Azotica of production in rice field.

The purpose of the invention is achieved by the following technical solution：One plant of salicylic rice field Azotica of production, entitled pond Raw nostoc (Nostoc piscinale) SCAU-003, deposit number are CCTCC NO：M2018194, the deposit date is 2018 On April 11, in, depositary institution are the China typical culture collection center positioned at Wuhan, China Wuhan University.

The salicylic rice field Azotica of the production can be used as bio-feritlizer and be applied in rice field, preferably comprise as Lower step：The salicylic rice field Azotica of production is put into rice field on suitable opportunity.

The suitable opportunity is when the salicylic rice field Azotica of production is put into rice field, and environmental condition has Conducive to the salicylic rice field Azotica survival of production；Preferably temperature be 25~30 DEG C, it is sunny, put after algae 5 days Inside the environmental condition or temperature of heavy rain are not less than 15 DEG C of environmental condition.

State of the described salicylic rice field Azotica of production when being put into rice field be preferably high density and floating at Film.

Preferably culture obtains the salicylic rice field Azotica of the production as follows：

(1) aseptically by the salicylic rice field Azotica culture of production to exponential phase, algae is obtained Kind liquid；

(2) algae seedling solution that step (1) obtains is inoculated into the culture medium of sterilizing, is cultivated under natural environment, obtain production water The rice field Azotica algae solution of poplar acid.

Culture described in step (1) is preferably 14 in 25~30 DEG C, 2500~3500LX of illumination, brightness time~ 18h：6~10h is cultivated；It is more preferably 16h in 28 DEG C, illumination 3000lx, brightness time:8h is cultivated.

The culture medium used in culture described in step (1) is preferably BG11₀Fluid nutrient medium；

BG11₀The composition of fluid nutrient medium is as follows：K₂HPO₄.3H₂O 0.04g/L、MgSO₄.7H₂O 0.075g/L、 CaCl₂.2H₂O 0.036g/L, citric acid 0.006g/L, ferric citrate 0.006g/L, EDTA 0.001g/L, Na₂CO₃ 0.02g/L, trace element A₅1mL；

Micro- A₅Composition it is as follows：H₃BO₃ 2.860g/L、NaMoO₄.2H₂O 0.021g/L、ZnSO₄.7H₂O 0.222g/L、CuSO₄.5H₂O 0.079g/L、MnCl₂.4H₂O 1.810g/L、NiSO₄.6H₂O 0.479g/L。

The addition of algae seedling solution is preferably pressed the 0.5% of culture volume and is calculated in inoculation described in step (2).

Culture medium described in step (2) is preferably the BG11 fluid nutrient mediums (BG11 without sodium nitrate₀)。

Natural environment described in step (2) is preferably peasant household's Around the house vacant lot.

Container is preferably open-top receptacle used in culture described in step (2)；More preferably plastic tub.

The step of culture described in step (2) is preferably：Upper mosquito net or window screen cloth are covered in the opening of culture vessel, Prevent mosquito from laying eggs；It is placed in the place culture of direct sunlight daytime, moves at night indoor；Wait for that algae grows to form algae film.

The present invention has the following advantages and effects with respect to the prior art：

The present invention obtains generating the plant hormones such as high concentration salicylic acid, grows fast, fixed nitrogen carbon sequestration by isolating and purifying screening The strong rice field Azotica of ability, can be used for new applying mode and is used as bio-feritlizer in rice terrace, can replace one Half chemical nitrogen fertilizer, while increasing lodging resistance in rice and disease resistance ability.

Description of the drawings

Fig. 1 is rice-Azotica cultivation results figure that foster-banded sclerotial blight seedling stage microchamber is inoculated with altogether；Wherein, figure A is to burn The photo figure cultivated in cup；Figure B is the rice seedlings that different disposal culture obtains；From left to right it is followed successively by blank control group, SCAU- 003 group, SCAU-010 groups, SCAU-012 groups and FACHB119 groups.

Fig. 2 is the Azotica figure under light microscope；Wherein, figure A is the photo figure for amplifying 300 times, and figure B is amplification The photo figure of 1500 times (oil mirror).

Fig. 3 is the BLAST comparison result figures of Azotica 16srDNA gene orders.

Fig. 4 is the BLAST comparison result figures of Azotica rbcLX gene orders.

Fig. 5 is the growth performance testing result figure of SCAU-003；Wherein, figure A is growth curve chart, and figure B is than growth speed Rate calculates figure.

Specific implementation mode

Present invention will now be described in further detail with reference to the embodiments and the accompanying drawings, but embodiments of the present invention are unlimited In this.

Embodiment 1：Rice field Azotica algae strain SCAU-003 is isolated and purified and is identified

(1) algae isolation and purification

Surface layer soil sample：Pick up from Agricultural University Of South China Zengcheng Linxi base rice terrace.

The separation and purifying of algae：Take pedotheque that 20mL BG11 are added by 1g₀In fluid nutrient medium, in illumination cultivation 28 DEG C of temperature, illumination 3000lx, brightness time are 16h in case:8h precultures 7 days.Then clear liquid of the upper layer containing algae is taken, is coated on 9cm contains BG11₀On the plate of solid medium.After growing macroscopic bacterium colony, carefully chooses and draw tablet again, microscopy, This process is repeated 3 times above to obtain the Azotica of monoculture.Most afterwards through ultraviolet light irradiation and Multiple Classes of Antibiotics joint at Reason obtains axenic culture.Specific method is：20 microlitres of the algae solution that OD680 is adjusted to 0.05 or so is carefully coated on quartz colorimetric utensil On inner wall, irradiate 20-30 minutes in the UV lamp.With 40 microlitres of BG11 after processing₀Culture medium applies after rinsing frustule It is distributed in and contains BG11₀On the agar solid plate of culture medium.After growing naked eyes visible colonies, chooses and be placed in BG11₀Fluid nutrient medium It is middle to cultivate to exponential phase.Pass through the BG11 of the penicillin containing 10mg/L successively₀Fluid nutrient medium is handled 3 days, and centrifuging and taking algae is thin Born of the same parents use BG11₀After fluid nutrient medium rinses suspension, then with 1mg/L gentamicins and 1mg/L kanamycins by penicillin processing side After method is handled successively, it is finally suspended in BG11₀Culture medium obtains sterile frustule.

BG11₀The formula of culture medium is as follows：

1 BG11 of table₀Culture medium drug inventory

2 trace element A of table₅Inventory

(2) Azotica-rice supports disease resistance assessment altogether

It selects three plants of isolated sterile algaes to be detected, is respectively designated as SCAU-003, SCAU-010, SCAU- 012.Simultaneously as a contrast with the Azotica algae strain FACHB119 purchased from Chinese Academy of Sciences's fresh water algae library (Hubei Wuhan).

According to document (Xu Guojuan etc., rice in China science, 2015,29 (1)：97-105) method is improved, and is established The rice algae co-cultivation of rice seedling Semi-solid cell culture-algae Liquid Culture, sheath blight fungus microchamber inoculation technique.It is as follows： By 3 rice paddy seeds (Huang Huazhan is purchased from the grand embroidery done with golden or silver thread coiled evenly seed-grain industry Co., Ltd in Shenzhen) after sprouting, it is placed in 1/2MS containing 400mL In the 1L open glass beakers of semisolid culturemedium, in the controlled environment chamber in 29 DEG C of temperature, humidity 85%, illumination 3000lx, light The dark time is 16h:8h is cultivated.When young rice seedlings growth to 21 heart of leaf, 40mL is added in BG11₀It is grown into culture medium The Azotica algae solution (being respectively SCAU-003, SCAU-010, SCAU-012 and FACHB119) of exponential phase, is cultivated 7 days Afterwards, Rhizoctonia solani Kuhn (the Guangdong Microbes Inst strain library in Chinese Guangdong Guangzhou, number are placed in rice leaf sheath GIM3.512) bacteria cake.It is sleeved on glass beaker with the 2.5L sky plastic barrels for cutting off bottom, is made with forming opposite closed environment at this time Banded sclerotial blight is fallen ill.Using the processing cup for being not added with algae as blank control, each three repetitions are handled and compareed.Algae is had evaluated to support altogether with rice To the resistance of rice sheath blight disease, 3 and Fig. 1 are the results are shown in Table.The Plant Height of Rice and the state of an illness handled from SCAU-003 known to table 3 and Fig. 1 Inhibiting rate is above blank control group and FACHB119 processing groups.Therefore, selection SCAU-003 is identified.

3 algae rice of table supports-sheath blight fungus microchamber inoculation technique assessment result altogether

(3) identification of algae

The algae plant shape state of SCAU-003 is as shown in Fig. 2, be the Filamentous Azotica with heterocyst.

Algae strain carries out sequence analysis using 16SrDNA methods：With 16S pair of primers 27F (AGAGTTTGATCMTGGCTCAG) PCR amplification is carried out with 1492R (TACGGYTACCTTGTTACGACTT), 16srDNA sequences (as follows) are determined by sequencing, Blast comparative analysis is carried out by Gene bank again, the results are shown in Figure 3, shows the algae and known Nostoc sp.PCC 9426 close source relationships are nearest, illustrate that the algae belongs to Nostoc.

16srDNA sequences：

CCTTGGCGGTTGCTTACCATGCAGTCGAACGGAATCTTAGGATTTAGTGGCGGACGGGTGAGTAACGCGTGAGAATC TAGCTTCAGGTCGGGGATAACTACTGGAAACGGTGGCTAATACCGGATGTGCCGAAAGGTGAAAGGCTTGCTGCCTG AAGATGAGCTCGCGTCTGATTAGCTAGTTGGTGTGGTAAGAGCGCACCAAGGCGACGATCAGTAGCTGGTCTGAGAG GATGATCAGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATTTTCCGCAATGGG CGAAAGCCTGACGGAGCAATACCGCGTGAGGGAGGAAGGCTCTTGGGTTGTAAACCTCTTTTCTCAAGGAATAAGTT CTGAAGGTACTTGAGGAATAAGCATCGGCTAACTCCGTGCCAGCAGCCGCGGTAATACGGAGGATGCAAGCGTTATC CGGAATGATTGGGCGTAAAGCGTCCGCAGGTGGCTGTGTAAGTCTGCTGTTAAAGAGTCTAGCTTAACTAGATAAAA GCAGTGGAAACTACATAGCTAGAGTACGTTCGGGGTAGAGGGAATTCCTGGTGTAGCGGTGAAATGCGTAGAGATCA GGAAGAACACCGGTGGCGAAGGCGCTCTACTAGGCCGTAACTGACACTGAGGGACGAAAGCTAGGGGAGCGAATGGG ATTAGATACCCCAGTAGTCCTAGCCGTAAACGATGGATACTAGGCGTTGCGAGTATCGACCCTCGCAGTGCCGGAGC CAACGCGTTAAGTATCCCGCCTGGGGAGTACGCACGCAAGTGTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAG CGGTGGAGTATGTGGTTTAATTCGATGCAACGCGAAGAACCTTACCAAGACTTGACATGTCGCGAATCCTCTTGAAA GGGAGGAGTGCCTTCGGGAGCGCGAACACAGGTGGTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAA GTCCCGCAACGAGCGCAACCCTCGTTTTTAGTTGCCAGCATTAAGTTGGGCACTCTAGAGAGACTGCCGGTGACAAA CCGGAGGAAGGTGGGGATGACGTCAAGTCAGCATGCCCCTTACGTCTTGGGCTACACACGTACTACAATGCTCCGAA CAGAGGGCAGCGAGCTAGCGATAGCAAGCAAATCCCGGAAATCGGAGCTCAGTTCAGATCGAAGGCTGCAACTCGCC TTCGTGAAGGAGGAATCGCTAGTAATTGCAGGTCAGCATACTGCAGTGAATTCGTTCCCGGGCCTTGTACACACCGC CCGTCACACCATGGAAGCTGGCAACGCCCGAAGTCATTACCCCAACTTTCGAGAGGGGGATGCCTAAGGCA。

In order to further identify algae strain, after extracting the algae genomic DNA, with two pairs of primers：CX(5'- GGCGCAGGTAAGAAAGGGTTTCGTA-3 ') and CW (5 '-CGTAGCTTCCGGTGGTATCCACGT-3 ') to Azotica spy Allogene rbcLX expand and (as follows) is sequenced.It is as shown in Figure 4 according to BLAST analysis results, it is seen then that the algae with Known Nostoc piscinale CENA21 homologys are high (99%).I.e. the algae is accredited as Nostoc piscinale CENA21。

RbcLX gene orders：

GGTGATGACTCCGTACTACAATTTGGTGGTGGTACACTCGGACACCCCTGGGGTAATGCTCCTGGTGCAACCGCTAA CCGTGTAGCTTTGGAAGCGTGTATCCAAGCTCGTAACGAAGGCCGCAACTTGGCGCGTGAAGGTAACGATGTTATTC GTGAAGCTGCTAAGTGGTCTCCTGAACTAGCTGCTGCTTGCGAACTGTGGAAAGAAATCAAGTTCGAGTTTGAGGCA ATGGATACCGTCTGATCATCAAGTAAAAGGTAAAAAGTTAAAAGGTAAAAGAAAATCTTTTTACTTATTACTTTTTA CTTTTAACTTAAATTTGGGCTGGGTCAAGCATGAATCTCAAGCAAATTGCGAAAGATACAGCCAAAACTCTCCAAAG CTACCTAACATATCAGGCGCTAAGGATTGTACTGGCACAGCTAGGCGAAACGAATCCACCGTTAGCTGTTTGGCTGC ATAACTTTTCTGCCGGAAAAGTTCAGGATGGAGAAGCTTATATTGAACAACTTTTCCAACACAAACCTGACTTAGCA CAGCGAATTATGACAGTTAGAGAACATATAGCCGAAGAAGTGGCAGAGTTCTTACCAGAAATGGTTCGCATTGGTAT TAAGCAAGCCAATATGGAACAGCGTCGCCAACACTTAGAACGCATCACGCAGCTGAGTTTAACTAACCCCAGTCCTG AATCAGAACAACAGCAATTTTCCGATCCTGACTGGGATAACTTAGCCAGTTAGGAAACATCCAAGTCGTCACTCAAT AGCAAACCTTTATTATTAGCTATGC。

It is named as pond life nostoc (Nostoc piscinale) SCAU-003, deposit number is CCTCC NO： M2018194, the deposit date is on April 11st, 2018, depositary institution was the Chinese Typical Representative culture positioned at Wuhan, China Wuhan University Object collection.

Embodiment 2：SCAU-003 growths, fixed nitrogen, carbon sequestration and biomass capability evaluation

With above-mentioned BG11₀After fluid nutrient medium inoculation grows into the algae SCAU-003 of exponential phase, in illumination cultivation In case, 28 DEG C of temperature, illumination 3000lx, brightness time are 16h:8h is cultivated, growth curve such as Fig. 5 of algae strain SCAU-003 It is shown, OD₆₈₀Value is increased to the 0.65 of the 13rd day from 0.08 when inoculation, increases above 5 times.Result of calculation shows that the algae has Higher specific growth rate (μ=0.163d^-1)。

It with same method culture algae, samples on day 4, algae solution is taken to measure fixed nitrogen by Acetylene Reduction-gas chromatography Enzymatic activity, measurement result show that the nitrogenase activity of the algae is 0.309 ± 0.032 μm of ol C₂H₂·h^-1·(μg Chla)^-1.? It samples within 15th day, collects frustule, TOC instrumental method frustule total carbons and total nitrogen are used after freeze-drying.Measure frustule total carbon and Total nitrogen content is respectively 1.457 ± 0.599% and 0.037 ± 0.017%.

In addition, in BG11₀Culture medium, 28 DEG C of temperature, illumination 3000lx, 10L basin open wide culture.The algae weight in wet base can reach 15.384 ± 2.251g/L, dry weight is up to 0.413 ± 0.047g/L.Illustrate that the algae has stronger Biomass Accumulation ability and fixed nitrogen Carbon sequestration capacity can provide organic matter and nitrogen source for Rice Production.

Embodiment 3：SCAU-003 generates the plant hormones capability evaluations such as salicylic acid

SCAU-003 algaes are in BG11₀20d, the freeze-drying of centrifuging and taking frustule are cultivated in culture medium.Supernatant is filtered with 0.22 micron Membrane filtration.The 1ml Bieleski buffer solutions of containing the internal standard (5mg/L DHZ) are added after taking frustule to grind broken wall in liquid nitrogen (60% methanol, 25% chloroform, 10% formic acid and 5% water), (frequency 20kHz, power 200W, work/intermittently are 30s/ to ultrasound 30s) 3min is fully to extract.Supernatant is collected by centrifugation, precipitation is surpassed again with solution of the 0.3ml containing 2% formic acid and 50% methanol Sound 3min extractions, are collected by centrifugation supernatant, and the supernatant of extraction merges with first time, after 40 DEG C of decompression rotary evaporated to dryness, use Methanol aqueous solution (the first alcohol and water by volume 1 of pH=3:1 proportioning, is adjusted to 3) be settled to 1mL with 1mol/L hydrochloric acid pH, with Standby UPLC-MS-MS is measured.Above-mentioned culture solution Bieleski buffer extractions, after taking 40 DEG C of rotary evaporations of organic phase to close do, With the 1 of pH=3:It is spare that 1 methanol aqueous solution is settled to 1mL.

Chromatographic condition：Instrument is UPLC-MS-MS (Waters), and chromatographic column is Acquity UPLC BEH C18 columns (2.1 × 50mm, 1.7 μm, Waters).Mobile phase:0.1% formic acid water (A)/0.1% formic acid acetonitrile (B) system for it is positive and negative from Sub- mode determination.Flow velocity 0.3mL/min.10 μ L of sample size.40 DEG C of column temperature, 25 DEG C of sample temperature.Condition of gradient elution is:0~ 2min, 90%~75%A；2~3min, 75%~40%A；3~3.5min, 40%~90%A；3.5~5min, 90%A.Matter Spectral condition：Capillary voltage 1.50kV；Desolventizing air-flow N2, flow velocity 800L/h；400 DEG C of desolventizing temperature；Taper hole air-flow N2, stream Fast 150L/h；150 DEG C of ion source temperature；Collide chamber pressure 3.1 × 10^-3mbar；Collision gas is argon gas, the following institute of mass spectrometry parameters Show.

4 mass spectrometry parameters of table

Note：(1) it is quota ion

Phytohormone concentration in frustule and culture solution is shown in Table 5, and frustule can generate more highly concentrated from the experimental results Auxin substance IBA, cytokinin-like substance TZR and the salicylic acid SA of degree.The salicylic yield of frustule is μ g/g It is horizontal.And salicylic acid can induce crop to improve resistance, research both at home and abroad has been found that it in anti-high and low-temp, drought-resistant, anti- Play an important roll in terms of lodging and disease and insect resistance.

Each hormone concentration in 5 frustule of table and culture solution

Embodiment 4：Azotica rice field crop field cell puts experiment in a suitable place to breed

(1) experimental design

Carry out Azotica fixed nitrogen on Jiangmen city Xinhui District farm December in July, 2017-to substitute chemical nitrogen fertilizer and carry High paddy rice anti contravariance experiment.Experiment sets 3 processing (adding same NPK fertilizer as base manure)：

Processing one：Conventional fertilizer application compares CK (top dressing of full dose)

Processing two：Azotica SCAU-003 is applied, the same CK of other fertilizer (add by algae in addition to nitrogenous fertilizer is not applied in top dressing： Respectively 3-5 days after intertillage, tillering stage and when Feng Hangqian 5-8 days be added in three times.

The expansion cultural method of Azotica SCAU-003 is as follows：

1) under the aseptic condition of laboratory, BG11 is used₀Liquid Culture is based on 28 DEG C of temperature, illumination 3000lx, brightness time For 16h:8h is cultivated, and 1L levels are amplified to from 20mL cultures.

2) by 100mL BG11₀Culture medium concentrated nutrient solution (100 times) tap water or the well water for boiling 10min are dilute It releases 100 times and arrives 10L, be placed in big plastic tub and be added algae seedling solution 50mL, basin overlying mosquito net or window screen cloth cover surrounding long-tail Clamping prevents mosquito from laying eggs, and is placed in the place culture of direct sunlight, moves at night indoor.Take the high density of culture 10-14 days And 15~20L/ mus of the algae of floating film forming, it is discharged into rice field under conditions of not destroying algae film as possible.The each cell of field experiment For 5m*5m (long * wide), three repetitions are each handled.

(2) test result

Since the upper buoyancy of algae strain is strong, it is easy to form algae film, resistance is very strong outdoors, than better suited temperature and light It can rapid mass propagation according under the conditions of.After the success of crop field, it is numerous to realize fast-growth expansion.Typhoon three times is being undergone, algae rice is applied Field lodging is substantially reduced compared with control, and the later stage, it is aggrieved that large area all occur in experiment contrast and periphery rice field when bacterial leaf-blight is broken out, But influence very little to applying algae rice field.Last yield realizes growth, is shown in Table 6, actual measurement yield amplification is more than 17%.

6 algae of table handles the influence to rice yield

(3) conclusion

Illustrate in Rice Production plus algae can substitute additional nitrogenous fertilizer, dramatically increase the resistance of rice, increases yield.

The above embodiment is a preferred embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment Limitation, it is other it is any without departing from the spirit and principles of the present invention made by changes, modifications, substitutions, combinations, simplifications, Equivalent substitute mode is should be, is included within the scope of the present invention.

Sequence table

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tggttcgcat tggtattaag caagccaata tggaacagcg tcgccaacac ttagaacgca 660

tcacgcagct gagtttaact aaccccagtc ctgaatcaga acaacagcaa ttttccgatc 720

ctgactggga taacttagcc agttaggaaa catccaagtc gtcactcaat agcaaacctt 780

tattattagc tatgc 795

Claims

1. one plant of salicylic rice field Azotica of production, it is characterised in that：Entitled pond life nostoc (Nostoc Piscinale) SCAU-003, deposit number are CCTCC NO：M 2018194, the deposit date is on April 11st, 2018, preservations Unit is the China typical culture collection center positioned at Wuhan, China Wuhan University.

2. the application of the salicylic rice field Azotica of production described in claim 1, it is characterised in that：The production is salicylic Rice field Azotica is applied as bio-feritlizer in rice field.

3. the application of the salicylic rice field Azotica of production according to claim 2, it is characterised in that comprise the following steps： The salicylic rice field Azotica of production is put into rice field on suitable opportunity.

4. the application of the salicylic rice field Azotica of production according to claim 3, it is characterised in that comprise the following steps： The suitable opportunity be temperature be 25~30 DEG C, it is sunny, put after algae the not environmental condition of heavy rain, or temperature in 5 days Environmental condition of the degree less than 15 DEG C.

5. the application of the salicylic rice field Azotica of production according to claim 3, it is characterised in that：The production bigcatkin willow State of the rice field Azotica of acid when being put into rice field is high density and floating film forming.

6. producing the application of salicylic rice field Azotica according to claim 3~5 any one of them, it is characterised in that：Institute The salicylic rice field Azotica of production stated is cultivated as follows to be obtained：

(1) aseptically by the salicylic rice field Azotica culture of production to exponential phase, algae seedling solution is obtained；

(2) algae seedling solution that step (1) obtains is inoculated into the culture medium of sterilizing, is cultivated under natural environment, obtain production salicylic acid Rice field Azotica algae solution.

7. the application of the salicylic rice field Azotica of production according to claim 6, it is characterised in that：

Culture described in step (1) be in 25~30 DEG C, 2500~3500LX of illumination, brightness time be 14~18h：6~10h It is cultivated；

The culture medium used in culture described in step (1) is BG11₀Fluid nutrient medium；

BG11₀The composition of fluid nutrient medium is as follows：K₂HPO₄.3H₂O 0.04g/L、MgSO₄.7H₂O 0.075g/L、CaCl₂.2H₂O 0.036g/L, citric acid 0.006g/L, ferric citrate 0.006g/L, EDTA 0.001g/L, Na₂CO₃0.02g/L, micro member Plain A₅1mL；

Micro- A₅Composition it is as follows：H₃BO₃ 2.860g/L、NaMoO₄.2H₂O 0.021g/L、ZnSO₄.7H₂O 0.222g/ L、CuSO₄.5H₂O 0.079g/L、MnCl₂.4H₂O 1.810g/L、NiSO₄.6H₂O 0.479g/L。

8. the application of the salicylic rice field Azotica of production according to claim 7, it is characterised in that：

Culture described in step (1) be in 28 DEG C, illumination 3000lx, brightness time be 16h:8h is cultivated.

9. the application of the salicylic rice field Azotica of production according to claim 6, it is characterised in that：

The addition of algae seedling solution is calculated by the 0.5% of culture volume in inoculation described in step (2)；

Culture medium described in step (2) is BG11₀Fluid nutrient medium；

Natural environment described in step (2) is peasant household's Around the house vacant lot；

Container used in culture described in step (2) is open-top receptacle；

The step of culture described in step (2) is：In the upper mosquito net of opening covering or window screen cloth of culture vessel, mosquito is prevented Oviposition；It is placed in the place culture of direct sunlight daytime, moves at night indoor；Wait for that algae grows to form algae film.