CN102242078A - Induction technology of anabaena flosaquae chlamydospore and preparation method of dry algae powder thereof - Google Patents
Induction technology of anabaena flosaquae chlamydospore and preparation method of dry algae powder thereof Download PDFInfo
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- CN102242078A CN102242078A CN 201110046706 CN201110046706A CN102242078A CN 102242078 A CN102242078 A CN 102242078A CN 201110046706 CN201110046706 CN 201110046706 CN 201110046706 A CN201110046706 A CN 201110046706A CN 102242078 A CN102242078 A CN 102242078A
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- wawter bloom
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Abstract
The invention relates to an induction method of anabaena flosaquae chlamydospore in the field of biotechnologies. The induction method comprises the following steps of: culturing the anabaena flosaquae for 3-4 days under the conditions of adding soil extract, cane sugar or glucose to a liquid culture medium containing dipotassium phosphate, ferric citrate, molybdic acid and calcium carbonate and regulating pH to be 6-7 to realize induction of the chlamydospore; and laying the anabaena flosaquae on a porcelain disc or a big culture vessel of 150*15mm, placing the porcelain disc or the big culture vessel into an oven and baking or placing the porcelain disc or the big culture vessel into a disinfection chamber of 30 DEG C and drying for 3-5 days to obtain dry algae powder. In the invention, the chlamydospore content is greatly improved because of effective and reasonable conditions, the dry algae powder can be obtained more efficiently through a simple drying mode, the method for preparing the dry algae powder with bloom cyanobacteria as a raw material is simple in process, the application of fertilizer is safe and efficient, the method is easy for popularization and implementation, and after the fertilizer is utilized, new pollution on environment and food safety is not brought about, therefore, the induction method of the anabaena flosaquae chlamydospore has great theoretical value and application value.
Description
Technical field
What the present invention relates to is a kind of optimization production method of biological technical field, specifically is the inductive technology of a kind of wawter bloom anabena chlamydospore and the preparation method of dry algae powder thereof.
Background technology
Wawter bloom anabena (Anabaena flos-aquae) provides (numbering: FACHB-245) by Inst. of Hydrobiology, Chinese Academy of Sciences algae kind storehouse, be recorded in " Jianying Shen, Antomo DiTommaso, Mingquan Shen, Wei Lu, and Zhengming Li; Molecular basis for differential metabolic responses to monosulfuron inthree nitrogen-fixing cyanobacteria, Weed Science, 2009,57:178-188 ", " Jianying Shen, JingJiang, Peizhong Zheng; Effects of Light and Monosulfuron on growth and PhotosyntheticPigments 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 wawter bloom 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 10-51kg/ha, and 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 wawter bloom anabena mainly contains: extract natural pigment, phycobiliprotein and polysaccharide, algae toxin study and utilization, biological nitrogen fixation and other aspects etc.Though purposes is many, but, takies volume and cause being difficult to transportation greatly, use and be inconvenient to cause and big area promote, thereby be not easy to utilize because wawter bloom anabena water content height causes going mouldy easily.For this blue-green algae of a large amount of and convenient application, can induce chlamydospore to generate in a large number by applying some condition, the algae liquid drying of wawter bloom anabena, be prepared into dry algae powder again, so that long-distance transportation and long-time the preservation.Under field conditions (factors), chlamydospore is high more a lot of than the survival rate of vegetative cell, uses easier sprouting behind the rice field.The known chlamydospore that contains trace in the existing bloom blue algae, chlamydospore is not only low temperature resistant, at high temperature also is difficult for dead.Naturally occurring chlamydospore is because of denier, so germination rate is not high after being applied to the rice field, fertilizer efficiency is not obvious.
Both at home and abroad the heterocyst of bloom blue algae is possessed some special knowledge so far, the inductive research of bloom blue algae chlamydospore is not then almost had.Publication number is the patent documentation of CN 1380901A, discloses the method for the mycelial chlamydospore of mass production Trichoderma herzianum SK-55.Trichoderma herzianum SK-55 belongs to fungi, and the wawter bloom anabena then belongs to bacterium, and Shang Weijian is open about the mass-produced technology of wawter bloom anabena chlamydospore.
The drying means of blue-green algae has a lot: spread drying on sheet glass out, be placed on drying in the big porcelain dish, also have directly drying in big or small culture dish, but these drying meanss are difficult to all the exsiccant blue algae collecting is stored.Simple and easy drying means of the present invention is that the wawter bloom anabena that removes nutrient solution is transferred in the big culture dish or porcelain dish that are coated with plastic film of the bacterium of going out, and blue-green algae can automatically peel off with plastic film when to be dried, is convenient to very much collection.
Summary of the invention
The present invention is directed to the prior art above shortcomings, the inductive technology of a kind of wawter bloom anabena chlamydospore and the preparation method of dry algae powder thereof are provided, at wideling 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, water content height with bloom blue algae, it is big to take volume, be not easy to transport the drawback of preservation, and the wawter bloom anabena defective that is not easy to gather in the crops, the invention provides a kind of is raw material with the bloom blue algae, adds physico chemical factor and induces chlamydospore to form in a large number and prepare the technology that the highly effective and safe of dry algae powder is produced fertilizer easily by plastic film.
The present invention is achieved by the following technical solutions:
The present invention relates to a kind of induction method of wawter bloom anabena chlamydospore, the liquid nutrient medium that use contains dipotassium hydrogen phosphate, ironic citrate, molybdic acid and lime carbonate is transferred to 6-7 at pH, sucrose or glucose are transferred under the condition of 80-100mg/L and were cultivated the wawter bloom anabena 3-4 days, realize inducing of wawter bloom anabena chlamydospore.
Described wawter bloom anabena is meant: with the wawter bloom anabena dry algae powder of water content about 10% airtight be stored in 4 ℃ the refrigerator two week the back take out, transfer in the small beaker that is added with nutrient solution and smash with making the algae machine.
The component of described nutrient solution is: 75g/L dipotassium hydrogen phosphate, 10g/L ironic citrate, 10g/L molybdic acid, 100g/L lime carbonate, surplus are the soil extract of 50g/L.
The described cultivation under pH is transferred to the environment of 6-7 is meant: with the wawter bloom anabena is that 280-300mg/L is seeded in the liquid nutrient medium with the inoculum size, is 3000 ± 200Lx in light intensity, and temperature is 30 ± 2 ℃, and pH is the environment of the 6-7 stir culture of regularly uncapping down.
Described timing is meant: stir once in section at the same time every day.
Described stirring is meant: cell is broken up even by the triangle rod of the bacterium of going out or glass stick or liquid-transfering gun.
The present invention relates to the preparation method of the dry algae powder of above-mentioned wawter bloom anabena chlamydospore, by with in the described wawter bloom anabena chlamydospore tiling porcelain dish, place 35-40 ℃ oven for drying or 30 ℃ the dry 3-5 of sterilisable chamber days to obtain dry algae powder.
Described tiling is meant: the height flat that adopts thickness to be no more than 1cm is layered on the porcelain dish bottom and it is evenly scattered.
Described porcelain dish is meant: the ceramic whiteware dish that is covered with the plastic film of the bacterium of going out.
The invention has the advantages that: effectively physico chemical factor has improved the content of chlamydospore in a large number; Easy drying mode can be gathered in the crops dry algae powder more efficiently; Be that the technology that relates to of the method for feedstock production dry algae powder is simple with the bloom blue algae; 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 is the inducing and the schematic diagram of dry algae powder preparation of wawter bloom anabena chlamydospore among the present invention
Fig. 2 be among the embodiment under the inductive condition of different pH, wawter bloom anabena chlamydospore is at 7 days content of cultured continuously.
Fig. 3 be among the embodiment under the inductive condition of different nutrient solutions, wawter bloom anabena chlamydospore is at 7 days content of cultured continuously.
Fig. 4 be among the embodiment under the inductive condition of different storage temperatures, wawter bloom anabena chlamydospore is at 7 days content of cultured continuously.
Fig. 5 is three class cells of wawter bloom anabena among the embodiment.
Fig. 6 is the transmission electron microscope picture of wawter bloom anabena chlamydospore among the embodiment.
Embodiment
Below embodiments of the invention are elaborated, present embodiment is being to implement under the prerequisite with the technical solution of the present invention, provided detailed embodiment and concrete operating process, but protection scope of the present invention is not limited to following embodiment.
Main raw: sodium hydroxide, hydrochloric acid, sucrose, nutrient solution, dry algae powder, plastic film, big porcelain dish.
As shown in Figure 1, concrete scheme is the wawter bloom dry algae powder that takes by weighing about 0.01g, be dissolved in the small beaker that contains the 10mL nutrient solution, the algae machine of beating with the sterilization of 75% alcohol is smashed 3 times, each 10s, algae liquid is transferred in the little culture dish of 90 * 15mm again, be poured in the culture dish with nutrient solution flushing beaker again, last culture dish is added nutrient solution to 35mL.With sodium hydroxide and hydrochloric acid pH is transferred to 6, adds 3mg sucrose then in culture dish, be placed on light intensity at last and be 3000 ± 200Lx, temperature and be in 30 ± 2 ℃ the sterilisable chamber and cultivate.General cultivate that chlamydospore content will reach peak value after 3 days, pour out unnecessary nutrient solution this moment, keep the wawter bloom anabena.Blue-green algae is transferred in the big porcelain dish that is coated with plastic film of the bacterium of going out, tiling thickness is no more than 1cm, and it is evenly scattered, and places 30 ℃ illumination chamber again, general after 3-5 days the wawter bloom anabena will peel off very easy the results automatically with film.
Microscopic count: cultivating the 3rd day, microscopic count is carried out in sampling, learns that as calculated total cellular score is 6.15 * 105/mL, and the chlamydospore number is 1.03 * 10
5Individual/mL, account for 16.75% of total cellular score.
As shown in Figure 5, be three class cells of wawter bloom anabena among the embodiment; Correspondence is the transmission electron microscope picture of wawter bloom anabena chlamydospore as shown in Figure 6.
Main raw: sodium hydroxide, hydrochloric acid, glucose, nutrient solution, dry algae powder, plastic film, big porcelain dish.
Concrete scheme is the wawter bloom dry algae powder that takes by weighing about 0.01g, be dissolved in the small beaker that contains the 10mL nutrient solution, the algae machine of beating with the sterilization of 75% alcohol is smashed 3 times, each 10s, again algae liquid is transferred in the little culture dish of 90 * 15mm, be poured in the culture dish with nutrient solution flushing beaker, last culture dish is added nutrient solution to 35mL again.With sodium hydroxide and hydrochloric acid pH is transferred to 6, adds 3mg glucose then in culture dish, be placed on light intensity at last and be 3000 ± 200Lx, temperature and be in 30 ± 2 ℃ the sterilisable chamber and cultivate.General cultivate that chlamydospore content will reach peak value after 3 days, pour out unnecessary nutrient solution this moment, keep the wawter bloom anabena.Blue-green algae is transferred in the big porcelain dish that is coated with plastic film of the bacterium of going out, tiling thickness is no more than 1cm, and it is evenly scattered, and places 30 ℃ illumination chamber again, general after 3-5 days the wawter bloom anabena will peel off very easy the results automatically with film.
Microscopic count: cultivating the 3rd day, microscopic count is carried out in sampling, learns that as calculated total cellular score is 1.02 * 10
6Individual/mL, the chlamydospore number is 1.13 * 10
5Individual/mL, account for 11.08% of total cellular score.
Main raw: sodium hydroxide, hydrochloric acid, glucose, nutrient solution, dry algae powder, plastic film, big culture dish, baking oven.
Concrete scheme is the wawter bloom dry algae powder that takes by weighing about 0.01g, be dissolved in the small beaker that contains the 10mL nutrient solution, the algae machine of beating with the sterilization of 75% alcohol is smashed 3 times, each 10s, again algae liquid is transferred in the little culture dish of 90 * 15mm, be poured in the culture dish with nutrient solution flushing beaker, last culture dish is added nutrient solution to 35mL again.With sodium hydroxide and hydrochloric acid pH is transferred to 7, adds 3mg glucose then in culture dish, be placed on light intensity at last and be 3000 ± 200Lx, temperature and be in 30 ± 2 ℃ the sterilisable chamber and cultivate.General cultivate that chlamydospore content will reach peak value after 3 days, pour out unnecessary nutrient solution this moment, keep the wawter bloom anabena.Again blue-green algae is transferred in the big culture dish of the 150 * 15mm that is coated with plastic film of the bacterium of going out, tiling thickness is no more than 1cm, and it is evenly scattered, and places 40 ℃ baking oven dry, the wawter bloom anabena will be peeled off with film automatically behind the general 12h, the very easy results.
Microscopic count: cultivating the 3rd day, microscopic count is carried out in sampling, learns that as calculated total cellular score is 8.23 * 10
5Individual/mL, the chlamydospore number is 1.53 * 10
5Individual/mL, account for 18.59% of total cellular score.
Main raw: sodium hydroxide, hydrochloric acid, sucrose, nutrient solution, dry algae powder, plastic film, big culture dish, baking oven.
Concrete scheme is the wawter bloom dry algae powder that takes by weighing about 0.01g, be dissolved in the small beaker that contains the 10mL nutrient solution, the algae machine of beating with the sterilization of 75% alcohol is smashed 3 times, each 10s, again algae liquid is transferred in the little culture dish of 90 * 15mm, be poured in the culture dish with nutrient solution flushing beaker, last culture dish is added nutrient solution to 35mL again.With sodium hydroxide and hydrochloric acid pH is transferred to 7, adds 3mg sucrose then in culture dish, be placed on light intensity at last and be 3000 ± 200Lx, temperature and be in 30 ± 2 ℃ the sterilisable chamber and cultivate.General cultivate that chlamydospore content will reach peak value after 3 days, pour out unnecessary nutrient solution this moment, keep the wawter bloom anabena.Again blue-green algae is transferred in the big culture dish of the 150 * 15mm that is coated with plastic film of the bacterium of going out, tiling thickness is no more than 1cm, and it is evenly scattered, and places 40 ℃ baking oven dry, the wawter bloom anabena will be peeled off with film automatically behind the general 12h, the very easy results.
Microscopic count: cultivating the 3rd day, microscopic count is carried out in sampling, learns that as calculated total cellular score is 5.83 * 10
5Individual/mL, the chlamydospore number is 1.15 * 10
5Individual/mL, account for 19.73% of total cellular score.
Above embodiment is not a limitation of the present invention, can prepare the nutrient solution of large volume when specifically implementing in the claim restricted portion by a certain percentage, as adding 9mg glucose or sucrose in the 100mL nutrient solution, adds 0.03g wawter bloom dry algae powder; Can be with the big porcelain dish that is coated with plastic film of the bacterium of going out when shifting wawter bloom.As Fig. 2-shown in Figure 4, under different pH, different nutrient solution, different storage temperature inductive condition, wawter bloom anabena chlamydospore at 7 days content of cultured continuously relatively.
Claims (9)
1. the induction method of a wawter bloom anabena chlamydospore, it is characterized in that, use contains the liquid nutrient medium of dipotassium hydrogen phosphate, ironic citrate, molybdic acid and lime carbonate on the basis of adding soil extract, sucrose or glucose, pH is transferred to the environment of 6-7 and cultivated the wawter bloom anabena 3-4 days down, realizes inducing of chlamydospore.
2. the induction method of wawter bloom anabena chlamydospore according to claim 1, it is characterized in that, described wawter bloom anabena is meant: with the wawter bloom anabena dry algae powder of water content about 10% airtight be stored in 4 ℃ the refrigerator two week the back take out, transfer in the small beaker that is added with nutrient solution and smash with making the algae machine.
3. the induction method of wawter bloom anabena chlamydospore according to claim 2, it is characterized in that, the component of described nutrient solution is: 75g/L dipotassium hydrogen phosphate, 10g/L ironic citrate, 10g/L molybdic acid, 100g/L lime carbonate, surplus are the 50g/L soil extraction.
4. the induction method of wawter bloom anabena chlamydospore according to claim 1 is characterized in that, cultivates under the described environment of being transferred to 6-7 at pH to be meant: by adding sodium hydroxide and hydrochloric acid, and the environment that pH the is adjusted to 6-7 stir culture of regularly uncapping down.
5. the induction method of wawter bloom anabena chlamydospore according to claim 4 is characterized in that, described timing is meant: stir once in section at the same time every day.
6. the induction method of wawter bloom anabena chlamydospore according to claim 4 is characterized in that, described stirring is meant: cell is broken up even by the triangle rod of the bacterium of going out or glass stick or liquid-transfering gun.
7. preparation method who prepares the dry algae powder of wawter bloom anabena chlamydospore according to the described method of above-mentioned arbitrary claim, it is characterized in that, by with in the described wawter bloom anabena chlamydospore tiling porcelain dish, place 35-40 ℃ oven for drying or 30 ℃ the dry 3-5 of sterilisable chamber days to obtain dry algae powder.
8. the preparation method of dry algae powder according to claim 7 is characterized in that, described tiling is meant: the height flat that adopts thickness to be no more than 1cm is layered on the porcelain dish bottom and it is evenly scattered.
9. the preparation method of dry algae powder according to claim 7 is characterized in that, described porcelain dish is meant: the porcelain dish that is coated with the white of plastic film.
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Cited By (3)
| 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 |
| CN102628027A (en) * | 2012-04-09 | 2012-08-08 | 上海交通大学 | Method for rapidly activating and abundantly producing anabaena azotica ley dry powder |
| CN107047972A (en) * | 2017-04-24 | 2017-08-18 | 苏州维淼生物工程有限公司 | A kind of functional feed for adding bloom blue algae extract |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1492037A (en) * | 2003-09-02 | 2004-04-28 | 中国科学院水生生物研究所 | Process for preparing water bloom blue-green algae harvest on lake |
-
2011
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1492037A (en) * | 2003-09-02 | 2004-04-28 | 中国科学院水生生物研究所 | Process for preparing water bloom blue-green algae harvest on lake |
Non-Patent Citations (4)
| Title |
|---|
| 《Archiv fur Hydrobiologie》 20050531 Kim,B.H.等 Relationship between akinete germination and vegetative population of Anabaena flos-aquae(Nostocales,Cyanobacteria)in Seokchon reservoir(Seoul,Korea) 第49-64页 1-9 第163卷, * |
| 《Hydrobiologia》 20000131 Peter D.Baker等 Environmental influnences on akinete germination of Anabaena circinalis and implications for management of cyanobacterial blooms 第65-73页 1-9 第427卷, 第1期 * |
| 《Journal of phycology》 19970831 Renhui Li等 Akinete formation in planktonic Anabaena spp.(cyanobacteria)by treatment with low temperature 第576-584页 1-9 第33卷, 第4期 * |
| 《热带海洋学报》 20060831 张燕英等 海洋固氮蓝藻Calothrixsp.与Lyngbyasp.固氮生理的研究 第46-50页 1-9 第25卷, 第4期 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102628027A (en) * | 2012-04-09 | 2012-08-08 | 上海交通大学 | Method for rapidly activating and abundantly producing anabaena azotica ley dry powder |
| CN102628027B (en) * | 2012-04-09 | 2013-07-10 | 上海交通大学 | Method for rapidly activating and abundantly producing anabaena azotica ley dry powder |
| 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 |
| CN107047972A (en) * | 2017-04-24 | 2017-08-18 | 苏州维淼生物工程有限公司 | A kind of functional feed for adding bloom blue algae extract |
| CN107047972B (en) * | 2017-04-24 | 2020-12-11 | 苏州维淼生物工程有限公司 | Functional feed added with water bloom blue algae extract |
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