CN102628027A - Method for rapidly activating and abundantly producing anabaena azotica ley dry powder - Google Patents
Method for rapidly activating and abundantly producing anabaena azotica ley dry powder Download PDFInfo
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- CN102628027A CN102628027A CN2012101016879A CN201210101687A CN102628027A CN 102628027 A CN102628027 A CN 102628027A CN 2012101016879 A CN2012101016879 A CN 2012101016879A CN 201210101687 A CN201210101687 A CN 201210101687A CN 102628027 A CN102628027 A CN 102628027A
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Abstract
The invention discloses a method for rapidly activating anabaena azotica ley dry powder. The method comprises the following steps of: a, preparing a basic culture solution from a certain amount of dipotassium phosphate, ferric citrate, molybdic acid and calcium carbonate; b, dissolving anabaena azotica ley dry powder into the basic culture solution to obtain an anabaena azotica ley solution; and c, putting the anabaena azotica ley solution into the basic culture solution, adding a certain amount of potassium nitrate, adjusting the pH to 7-9, and culturing for 2-3 days to realize rapid activation of anabaena azotica ley. A method for abundantly producing anabaena azotica ley activated with the method comprises the following steps of: transferring the activated anabaena azotica ley into an opaque open container filled with a basic culture solution of glucose or cane sugar; and culturing for 5-7 days to obtain abundantly-produced anabaena azotica ley. Due to the adoption of the method, the germination proportion of dry algae powder is greatly increased under an effective activating condition, the duration of germination is shortened, and the stress tolerance of the anabaena azotica ley is enhanced due to the use of the glucose or cane sugar.
Description
Technical field
The present invention relates to a kind of biotechnology, be specifically related to a kind of quick active and mass-produced method thereof of fixed nitrogen anabena dry algae powder.
Background technology
Fixed nitrogen anabena (Anabaena azotica) provides (numbering: FACHB-686) by Inst. of Hydrobiology, Chinese Academy of Sciences algae kind storehouse; And at " Jianying Shen, Antonio DiTommaso, Mingquan Shen; Wei Lu, and Zhengming Li; Molecular basis for differential metabolic responses to monosulfuron in three nitrogen-fixing cyanobacteria; Weed Science; 2009; 57:178-188 ", " Jianying Shen, Jing Jiang, Peizhong Zheng; Effects of Light and Monosulfuron on growth and Photosynthetic Pigments of Anabaena flos-aquae Breb; Water Resource and Protection; 2009,1:408-413 ", " Zheng Peizhong, Shen Jianying; Organic solvent-acetone is to fixed nitrogen blue algae growth Study on Effect, Shanghai Agricultural journal, 2010.4:33-38 " etc. in open.
The fixed nitrogen anabena is the nomadic nitrogen in the atmosphere fixedly, is a kind of of fixed nitrogen blue-green algae.Its growth and breeding is rapid, and amount of nitrogen fixation reaches 20-55kg/hm
2, its frond can be absorbed by crop root after sinking to soil, and distinctive algae somatotropin can promote that crop grows vigorously, and makes crop yield 10-30%, so this is a kind of good fertilizer.And the production of agrochemicals fertilizer and agricultural chemicals, use and can bring a large amount of greenhouse gas emissions, therefore develop agro-ecology fixed nitrogen technology, reduce applying quantity of chemical fertilizer, be the important channel of current development low-carbon (LC) agricultural.
The purposes of fixed nitrogen anabena mainly contains: extract natural pigment, phycobiliprotein and polysaccharide, biological nitrogen fixation and other aspects etc.The dry algae powder that is prepared into often has germination rate shortcoming on the low side, so the fertilizer efficiency function in rice field fails to fully play.
So far both at home and abroad seldom to the correlative study of fixed nitrogen anabena dry algae powder; Aspect the activation of dry algae powder; The attachment energy that rarely seen Huang has fragrant grade in " the dried algae kind of fixed nitrogen blue-green algae prepares and effect in the rice field ", to mention wheat husk or rice husk improves sprouts effect, and exploration in other respects is rare report then.
Summary of the invention
The object of the invention is exactly the above-mentioned deficiency that exists to prior art, and a kind of quick active and mass-produced method thereof of fixed nitrogen anabena dry algae powder is provided.
To achieve these goals, the present invention has adopted following technical scheme: a kind of quick active method of fixed nitrogen anabena dry algae powder may further comprise the steps:
A, get quantitative potassium hydrogenphosphate, ironic citrate, molybdic acid and dissolution of calcium carbonate in water; Be prepared into basic culture solution, and make the final concentration of each component in the basic culture solution be: potassium hydrogenphosphate 75mg/L, ironic citrate 10mg/L, molybdic acid 10mg/L, lime carbonate 100mg/L;
B, fixed nitrogen anabena dry algae powder is dissolved in the basic culture solution and smashes with making the algae machine, be prepared into fixed nitrogen anabena solution;
C, fixed nitrogen anabena solution is placed basic culture solution, add quantitative saltpetre, adjusting pH is 7-9, cultivated 2-3 days the quick active of realization fixed nitrogen anabena down at half state that open wide.
The quantitative saltpetre of described interpolation is meant, makes the concentration of saltpetre reach 380-500mg/L; Described adjusting pH is that 7-9 passes through sodium hydroxide and hydrochloric acid is regulated; Described under the state that partly opens wide, the cultivation is with petridish timing stir culture.
Described timing stir culture is meant and is stirring once at the same time every day.
Described stirring is meant breaks up cell even by the liquid-transfering gun of triangle rod of the bacterium of going out or 5ml.
The mass production method of aforesaid method activated fixed nitrogen anabena is transferred in the opaque open container of the basic culture solution that is added with glucose or sucrose through activating gained fixed nitrogen anabena, cultivates 5-7 days, obtains the fixed nitrogen anabena of volume production.
The concentration of each composition is in the described basic culture solution that is added with glucose or sucrose: potassium hydrogenphosphate 75mg/L, ironic citrate 10mg/L, molybdic acid 10mg/L, lime carbonate 100mg/L, glucose 130-200mg/L or sucrose 70-130mg/L, surplus is a water.
The present invention is directed to and widely popularize the low-carbon (LC) agricultural at present, reduce using of chemical fertilizer and agricultural chemicals, improve the consumption of fertilizer and the policy of fertilizer efficiency; With the germination rate drawback on the low side of fixed nitrogen anabena dry algae powder, and the fixed nitrogen anabena defective that is not easy to gather in the crops, providing a kind of is raw material with fixed nitrogen anabena dry algae powder; Adding physico chemical factor efficiently activates; Make its quick growth and breeding, and stimulate, by plastic film through appropriate condition; The fixed nitrogen anabena of the high vigor that mass production resistance is stronger is a kind of technology of production microbial-bacterial fertilizer of safe ready.
The invention has the advantages that: effectively activation condition has improved the sprouting ratio of dry algae powder greatly, has shortened sprout time, and the use of glucose or sucrose has strengthened the resistance of fixed nitrogen anabena; The technology that the efficient Activiation method of fixed nitrogen anabena dry algae powder and mass production method thereof relate to is simple; Fertilizer is used safe and efficient, is easy to promotion and implementation; Can not cause new pollution after the enforcement to environment, food safety; So the present invention has great theoretical value and using value.
Description of drawings
Fig. 1 be among the present invention under the activation of different saltpetre concentration, the fixed nitrogen anabena is at 7 days content of cultured continuously.
Fig. 2 be among the present invention under the activation of different pH, the fixed nitrogen anabena is at 7 days content of cultured continuously.
Fig. 3 be among the present invention under the activation of different opening-wide states, the fixed nitrogen anabena is at 7 days content of cultured continuously.
Fig. 4 be among the present invention under the effect of different sugar, the fixed nitrogen anabena is at 7 days content of cultured continuously.
Fig. 5 is two types of cells of fixed nitrogen anabena among the present invention.
Fig. 6-the 8th, the transmission electron microscope picture of fixed nitrogen anabena among the present invention.
Embodiment
Below in conjunction with accompanying drawing and specific embodiment the present invention is elaborated.The component of used basic culture solution and concentration are among the embodiment: potassium hydrogenphosphate 75mg/L, ironic citrate 10mg/L, molybdic acid 10mg/L, lime carbonate 100mg/L.
Take by weighing the fixed nitrogen anabena dry algae powder of 0.01g; Be dissolved in the small beaker that contains the 10mL basic culture solution; The algae machine of beating with the sterilization of 75% alcohol is smashed 3 times, and each 10s transfers to algae liquid in the little petridish of 90 * 15mm again; Be poured in the petridish with basic culture solution flushing beaker, last petridish is added basic culture solution to 35mL again.Add saltpetre, make its concentration reach 380-500mg/L, pH is transferred to 7, be placed on light intensity at last and be 3000 ± 200Lx, temperature and be in 30 ± 2 ℃ the sterilisable chamber and cultivate with sodium hydroxide and hydrochloric acid.General cultivate that the fixed nitrogen anabena begins by a large amount of activation after 2-3 days; Transfer to activated fixed nitrogen anabena in the opaque open container this moment, replenishes an amount of basic culture solution, takes by weighing an amount of sucrose and be added in the container; Make its concentration reach 70-130mg/L; Cultivated again 5-7 days, and poured out unnecessary basic culture solution, results fixed nitrogen anabena.
Take by weighing the fixed nitrogen anabena dry algae powder of 0.01g; Be dissolved in the small beaker that contains the 10mL basic culture solution; The algae machine of beating with the sterilization of 75% alcohol is smashed 3 times, and each 10s transfers to algae liquid in the little petridish of 90 * 15mm again; Be poured in the petridish with basic culture solution flushing beaker, last petridish is added basic culture solution to 35mL again.Add saltpetre, make its concentration reach 380-500mg/L, pH is transferred to 9, be placed on light intensity at last and be 3000 ± 200Lx, temperature and be in 30 ± 2 ℃ the sterilisable chamber and cultivate with sodium hydroxide and hydrochloric acid.General cultivate that the fixed nitrogen anabena begins by a large amount of activation after 2-3 days; Transfer to activated fixed nitrogen anabena in the opaque open container this moment, replenishes an amount of basic culture solution, takes by weighing an amount of sucrose and be added in the container; Make its concentration reach 70-130mg/L; Cultivated again 5-7 days, and poured out unnecessary basic culture solution, results fixed nitrogen anabena.
Take by weighing the fixed nitrogen anabena dry algae powder of 0.01g; Be dissolved in the small beaker that contains the 10mL basic culture solution; The algae machine of beating with the sterilization of 75% alcohol is smashed 3 times, and each 10s transfers to algae liquid in the little petridish of 90 * 15mm again; Be poured in the petridish with basic culture solution flushing beaker, last petridish is added basic culture solution to 35mL again.Add saltpetre, make its concentration reach 380-500mg/L, pH is transferred to 9, be placed on light intensity at last and be 3000 ± 200Lx, temperature and be in 30 ± 2 ℃ the sterilisable chamber and cultivate with sodium hydroxide and hydrochloric acid.General cultivate that the fixed nitrogen anabena begins by a large amount of activation after 2-3 days; Transfer to activated fixed nitrogen anabena in the opaque open container this moment, replenishes an amount of basic culture solution, takes by weighing an amount of glucose and be added in the container; Make its concentration reach 130-200mg/L; Cultivated again 5-7 days, and poured out unnecessary basic culture solution, results fixed nitrogen anabena.
Above embodiment is not a limitation of the present invention; Can in scope thereof, prepare the basic culture solution of big volume during practical implementation by a certain percentage; As adding 13-20mg glucose or 7-13mg sucrose in the 100mL basic culture solution, add 0.03g fixed nitrogen anabena dry algae powder; Add 100-200mg glucose or 70-150mg sucrose in the 1L basic culture solution, add 0.30g fixed nitrogen anabena dry algae powder etc.
Fig. 1 be among the present invention under the activation of different saltpetre concentration, the fixed nitrogen anabena is at 7 days content of cultured continuously.
Fig. 2 be among the present invention under the activation of different pH, the fixed nitrogen anabena is at 7 days content of cultured continuously.
Fig. 3 be among the present invention under the activation of different opening-wide states, the fixed nitrogen anabena is at 7 days content of cultured continuously.
Fig. 4 be among the present invention under the effect of different sugar, the fixed nitrogen anabena is at 7 days content of cultured continuously.
Fig. 5 is two types of cells of fixed nitrogen anabena among the present invention.
Fig. 6-the 8th, the transmission electron microscope picture of fixed nitrogen anabena among the present invention.
Claims (6)
1. the quick active method of a fixed nitrogen anabena dry algae powder is characterized in that, may further comprise the steps:
A, get quantitative potassium hydrogenphosphate, ironic citrate, molybdic acid and dissolution of calcium carbonate in water; Be prepared into basic culture solution, and make the final concentration of each component in the basic culture solution be: potassium hydrogenphosphate 75mg/L, ironic citrate 10mg/L, molybdic acid 10mg/L, lime carbonate 100mg/L;
B, fixed nitrogen anabena dry algae powder is dissolved in the basic culture solution and smashes with making the algae machine, be prepared into fixed nitrogen anabena solution;
C, fixed nitrogen anabena solution is placed basic culture solution, add quantitative saltpetre, adjusting pH is 7-9, cultivated 2-3 days the quick active of realization fixed nitrogen anabena down at half state that open wide.
2. the quick active method of fixed nitrogen anabena dry algae powder according to claim 1 is characterized in that: the quantitative saltpetre of described interpolation is meant, makes the concentration of saltpetre reach 380-500mg/L; Described adjusting pH is that 7-9 passes through sodium hydroxide and hydrochloric acid is regulated; Described under the state that partly opens wide, the cultivation is with petridish timing stir culture.
3. the quick active method of fixed nitrogen anabena dry algae powder according to claim 2 is characterized in that: described timing stir culture is meant and is stirring once at the same time every day.
4. the quick active method of fixed nitrogen anabena dry algae powder according to claim 3 is characterized in that: described stirring is meant breaks up cell even by the liquid-transfering gun of triangle rod of the bacterium of going out or 5ml.
5. according to the mass production method of the said method activated of claim 1 fixed nitrogen anabena; It is characterized in that: transfer in the opaque open container of the basic culture solution that is added with glucose or sucrose through activating gained fixed nitrogen anabena; Cultivated 5-7 days, and obtained the fixed nitrogen anabena of volume production.
6. the mass production method of fixed nitrogen anabena according to claim 5; It is characterized in that: the concentration of each composition is in the described basic culture solution that is added with glucose or sucrose: potassium hydrogenphosphate 75mg/L, ironic citrate 10mg/L, molybdic acid 10mg/L, lime carbonate 100mg/L, glucose 130-200mg/L or sucrose 70-130mg/L, surplus is a water.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102618480A (en) * | 2012-04-10 | 2012-08-01 | 上海交通大学 | Preparation method of dry algae powder of anabaena azollae |
| CN104604605A (en) * | 2015-01-08 | 2015-05-13 | 上海交通大学 | Composite control method for paddy field weeds |
| CN106479949A (en) * | 2016-09-08 | 2017-03-08 | 上海交通大学 | Activity In Anabaena Azotica cell is by the restorative procedure of ALS inhibitor class herbicide injury |
| CN107852884A (en) * | 2017-12-04 | 2018-03-30 | 中国科学院水生生物研究所 | A kind of method for improveing acidified soil using Azotica and lifting soil fertility |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102242078A (en) * | 2011-02-25 | 2011-11-16 | 上海交通大学 | Induction technology of anabaena flosaquae chlamydospore and preparation method of dry algae powder thereof |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102242078A (en) * | 2011-02-25 | 2011-11-16 | 上海交通大学 | Induction technology of anabaena flosaquae chlamydospore and preparation method of dry algae powder thereof |
Non-Patent Citations (2)
| Title |
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| 喻国策等: "有机碳化合物对鱼腥藻7120生长的影响", 《水生生物学报》, no. 03, 20 May 2003 (2003-05-20) * |
| 黄有馨等: "固氮蓝藻干藻种制备及其在稻田中的应用效果", 《微生物学报》, 31 December 1983 (1983-12-31) * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102618480A (en) * | 2012-04-10 | 2012-08-01 | 上海交通大学 | Preparation method of dry algae powder of anabaena azollae |
| CN102618480B (en) * | 2012-04-10 | 2013-05-08 | 上海交通大学 | Preparation method of dry algae powder of anabaena azollae |
| CN104604605A (en) * | 2015-01-08 | 2015-05-13 | 上海交通大学 | Composite control method for paddy field weeds |
| CN104604605B (en) * | 2015-01-08 | 2017-01-25 | 上海交通大学 | Composite control method for paddy field weeds |
| CN106479949A (en) * | 2016-09-08 | 2017-03-08 | 上海交通大学 | Activity In Anabaena Azotica cell is by the restorative procedure of ALS inhibitor class herbicide injury |
| CN106479949B (en) * | 2016-09-08 | 2019-12-10 | 上海交通大学 | Method for repairing anabaena azotica cells damaged by ALS inhibitor herbicides |
| CN107852884A (en) * | 2017-12-04 | 2018-03-30 | 中国科学院水生生物研究所 | A kind of method for improveing acidified soil using Azotica and lifting soil fertility |
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