CN113637601B - Preparation method of composite microbial agent capable of improving yield of konjak - Google Patents
Preparation method of composite microbial agent capable of improving yield of konjak Download PDFInfo
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- 229920002752 Konjac Polymers 0.000 title claims abstract description 72
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- 230000000813 microbial effect Effects 0.000 title claims abstract description 42
- 239000002131 composite material Substances 0.000 title claims abstract description 21
- 238000002360 preparation method Methods 0.000 title claims abstract description 10
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 35
- 239000002689 soil Substances 0.000 claims abstract description 24
- 238000000034 method Methods 0.000 claims abstract description 18
- 241000193744 Bacillus amyloliquefaciens Species 0.000 claims abstract description 16
- 241000194108 Bacillus licheniformis Species 0.000 claims abstract description 16
- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 16
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 16
- 150000001875 compounds Chemical class 0.000 claims abstract description 14
- 241000194103 Bacillus pumilus Species 0.000 claims abstract description 13
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- 238000002386 leaching Methods 0.000 claims description 7
- 238000002156 mixing Methods 0.000 claims description 7
- 235000010469 Glycine max Nutrition 0.000 claims description 6
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- 230000001737 promoting effect Effects 0.000 abstract description 3
- 239000000126 substance Substances 0.000 abstract description 3
- 239000000575 pesticide Substances 0.000 abstract description 2
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- LUEWUZLMQUOBSB-FSKGGBMCSA-N (2s,3s,4s,5s,6r)-2-[(2r,3s,4r,5r,6s)-6-[(2r,3s,4r,5s,6s)-4,5-dihydroxy-2-(hydroxymethyl)-6-[(2r,4r,5s,6r)-4,5,6-trihydroxy-2-(hydroxymethyl)oxan-3-yl]oxyoxan-3-yl]oxy-4,5-dihydroxy-2-(hydroxymethyl)oxan-3-yl]oxy-6-(hydroxymethyl)oxane-3,4,5-triol Chemical compound O[C@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@@H](O[C@@H]2[C@H](O[C@@H](OC3[C@H](O[C@@H](O)[C@@H](O)[C@H]3O)CO)[C@@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O LUEWUZLMQUOBSB-FSKGGBMCSA-N 0.000 description 2
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- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- FZWLAAWBMGSTSO-UHFFFAOYSA-N Thiazole Chemical compound C1=CSC=N1 FZWLAAWBMGSTSO-UHFFFAOYSA-N 0.000 description 1
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C21/00—Methods of fertilising, sowing or planting
- A01C21/005—Following a specific plan, e.g. pattern
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
- A01G22/25—Root crops, e.g. potatoes, yams, beet or wasabi
-
- 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
- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
- A01N63/22—Bacillus
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- Tropical Medicine & Parasitology (AREA)
- General Engineering & Computer Science (AREA)
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- Agronomy & Crop Science (AREA)
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Abstract
The invention belongs to the technical field of application of microbial technology in fertilizers, and discloses a composite microbial agent capable of improving konjak yield and a preparation method thereof, wherein the composite microbial agent is prepared by combining 5 bacillus such as bacillus subtilis, bacillus licheniformis, bacillus amyloliquefaciens, bacillus pumilus and bacillus cereus according to a specific proportion. The effective viable count in the composite microbial agent is more than or equal to 1.0 hundred million/g (cfu). The compound microbial agent capable of improving the yield of konjak is beneficial to promoting the growth and development of konjak, improving the agronomic characters of konjak, has better control effect on soft rot and southern blight of konjak, has optimal control effect on soft rot, and obviously reduces the incidence rate and dead plant rate of soft rot. In addition, the method can regulate and control the micro-ecological environment of the soil, improve the number of beneficial microorganisms in the soil and reduce the number of pathogenic bacteria in the soil, thereby reducing the occurrence of konjak diseases and reducing the dosage of chemical fertilizers and pesticides.
Description
Technical Field
The invention belongs to the technical field of application of microbial technology in fertilizers, and particularly relates to a compound microbial agent capable of improving yield of konjak and a preparation method thereof.
Background
At present: konjak belongs to perennial host herbaceous plants of konjak of Araceae, and is the only special cash crop which can provide a large amount of glucomannan. The konjak glucomannan is a high molecular water-soluble dietary fiber, is widely applied to a plurality of fields of agriculture, food, medicine, health care, chemical industry, textile and the like, and has great commercial value. With the development of modern planting industry to intensive agriculture mode, continuous cropping obstacles such as poor konjak growth, disease aggravation, yield reduction and the like caused by long-term continuous cropping become serious. Especially soft rot accompanied by continuous cropping, especially seed-borne and soil-borne bacterial diseases which are most common and serious in damage in the konjak growing process, and even cause harvest in a large area, the yield is seriously affected, and the peasant is seriously lost.
Through the above analysis, the problems and defects existing in the prior art are as follows: problems and defects existing in the prior art are as follows: the development of modern planting industry to intensive agriculture mode causes poor konjak growth, disease aggravation and yield reduction due to long-term continuous cropping.
The difficulty of solving the problems and the defects is as follows: at present, chemical agents are mainly adopted to prevent and control diseases of konjak in the planting process, and commonly used chemical agents mainly comprise organic copper bactericides, inorganic copper bactericides, thiazole bactericides and the like, but the long-term use of the chemical agents easily causes serious problems such as environmental pollution, ecological damage and the like. In recent years, the prevention and control of konjak diseases by using biocontrol microbial agents has become a hot spot for research and application due to the advantages of low cost, high efficiency, no residue, environmental friendliness and the like. However, at present, more biocontrol microorganisms are mainly actinomycetes, and the actinomycetes are mainly used for preventing and controlling soft rot of konjak, but the actinomycetes can kill pathogenic bacteria and inhibit beneficial microorganisms in soil, so that the microbial ecological environment of the planting soil is affected to a certain extent. Therefore, the development of the compound microbial agent which can solve the problems of disease prevention and control and continuous cropping obstacle of konjak planting land, improve the micro-ecological environment of soil and promote the improvement of konjak yield and improve konjak yield is a difficult problem of biological prevention and control of the crops.
The meaning of solving the problems and the defects is as follows: (1) The 5 bacillus adopted by the invention belong to the same genus, have close relationship, and cannot cause antagonistic effect, so that the characteristics of single strain can be furthest exerted in the soil and the harmonic position.
(2) The compound microbial agent capable of improving the yield of konjak can secrete various products such as bacteriocin, protease, cellulase and the like, wherein the bacteriocin can inhibit pathogenic fungi, and other enzymes can decompose organic matters and mineral nutrition in soil so as to supply beneficial microorganisms and growth and development of crops in the soil, finally improve the living environment of the soil microorganisms, reduce continuous cropping obstacles and improve the yield and quality of konjak.
(3) In order to improve the adaptability of the strain, the invention adopts the soil leaching solution prepared from the soil of the konjak planting area in Yunnan Fuyuan to ferment and cultivate the strain so as to improve the tolerance of the strain and enable the prepared microbial inoculum to be more suitable for the soil environment for planting konjak.
Disclosure of Invention
Aiming at the problems existing in the prior art, the invention provides a compound microbial agent capable of improving the yield of konjak and a preparation method thereof.
The invention is realized in such a way that the composite microbial agent capable of improving the yield of konjak consists of 5 microorganisms of bacillus subtilis, bacillus licheniformis, bacillus amyloliquefaciens, bacillus pumilus and bacillus cereus, and after the bacillus subtilis, the bacillus licheniformis, the bacillus amyloliquefaciens and the bacillus pumilus are mixed according to the ratio of 1:1:1:1,then mixed with bacillus cereus cultured independently according to the ratio of 1:1. The strain is prepared from bacillus subtilis (purchased from a Chinese microbial strain resource library, strain number: ACCC 10719), bacillus licheniformis (purchased from a Chinese microbial strain resource library, strain number: ATCC 11946), bacillus amyloliquefaciens (purchased from a Chinese microbial strain resource library, strain number: ATCC 23350), and bacillus pumilus (purchased from a Chinese microbial strain resource library, strain number:BAA-1434 TM) and bacillus cereus (purchased from the chinese microorganism strain resource pool, strain number: ATCC 9259) and 5 bacillus bacteria according to a specific proportion. The effective viable count in the composite microbial agent is more than or equal to 1.0 hundred million/g (cfu).
Furthermore, the effective viable count of the composite microbial agent capable of improving the yield of konjak is more than or equal to 1.0 hundred million/g (cfu).
Another object of the present invention is to provide a method for preparing the composite microbial agent capable of improving the yield of konjak, comprising the steps of:
firstly, carrying out mixed fermentation culture on bacillus subtilis, bacillus licheniformis, bacillus amyloliquefaciens and bacillus pumilus by adopting an improved tryptone soybean liquid culture medium;
step two, solely inoculating bacillus cereus into a modified beef extract peptone liquid culture medium for fermentation culture;
and thirdly, mixing the two fermentation liquids after fermentation is finished, wherein the mixed bacterial liquid is the composite microbial agent stock solution capable of improving the yield of konjak.
Further, the tryptone soybean liquid culture medium is composed of tryptone 12.0g/L, soytone 3.0g/L, sodium chloride 3.0g/L, soil leaching liquid 5.0ml/L and H 2 O 1000ml，pH 7.2-7.5。
Further, the fermentation time of the first step is 48 hours.
Further, the beef extract peptone liquid cultureThe base is 2.5g/L of beef extract, 12.5g/L of peptone, 2.5g/L of sodium chloride, 5.0ml/L of soil leaching liquid and H 2 O 1000ml，pH7.5-7.8。
Further, the fermentation time of the second step is 36h.
Further, after the fermentation is finished, the two fermentation liquids are mixed according to the ratio of 1:1. The parameters are determined by a plurality of experiments, and the effective viable count and the field application effect of the compound microbial inoculum prepared according to the parameters are stable.
Another object of the present invention is to provide a method for improving the yield of konjak, which comprises diluting the compound microorganism bacterium agent capable of improving the yield of konjak by 1:500 times and irrigating roots.
Further, the method for improving the yield of the konjak is respectively applied in three periods of konjak planting process, namely a seeding period, a seedling emergence period and a tuber fast-growing period, and the single dosage per mu is 1000ml.
By combining all the technical schemes, the invention has the advantages and positive effects that: the compound microbial agent capable of improving the yield of konjak is beneficial to promoting the growth and development of konjak, improving the agronomic characters of konjak, has better control effect on soft rot and southern blight of konjak, has optimal control effect on soft rot, and obviously reduces the incidence rate and dead plant rate of soft rot. In addition, the microbial inoculum can be used for regulating and controlling the micro-ecological environment of the soil, improving the quantity of beneficial microorganisms in the soil and reducing the quantity of pathogenic bacteria in the soil, thereby reducing the occurrence of konjak diseases, reducing the dosage of chemical fertilizers and pesticides and achieving the effect of increasing the yield.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are needed in the embodiments of the present invention will be briefly described below, and it is obvious that the drawings described below are only some embodiments of the present invention, and other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
Fig. 1 is a flowchart of a preparation method of a composite microbial agent capable of improving the yield of konjak provided by the embodiment of the invention.
Detailed Description
The present invention will be described in further detail with reference to the following examples in order to make the objects, technical solutions and advantages of the present invention more apparent. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the scope of the invention.
Aiming at the problems existing in the prior art, the invention provides a method for improving the yield of konjak by utilizing a compound microbial agent capable of improving the yield of konjak and application thereof, and the invention is described in detail below with reference to the accompanying drawings.
The composite microbial agent capable of improving the yield of konjak provided by the invention consists of bacillus subtilis, bacillus licheniformis, bacillus amyloliquefaciens and bacillus pumilus. The bacillus subtilis, the bacillus licheniformis and the bacillus amyloliquefaciens are mixed according to equal proportion and then are mixed with the bacillus cereus which is cultured independently according to the ratio of 1:1.
The present invention will be described in further detail with reference to the following examples in order to make the objects, technical solutions and advantages of the present invention more apparent. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the scope of the invention.
Aiming at the problems existing in the prior art, the invention provides a method for improving the yield of konjak by utilizing a compound microbial agent capable of improving the yield of konjak and application thereof, and the invention is described in detail below with reference to the accompanying drawings.
The composite microbial agent capable of improving the yield of konjak provided by the invention consists of bacillus subtilis, bacillus licheniformis, bacillus amyloliquefaciens and bacillus pumilus. The bacillus subtilis, the bacillus licheniformis and the bacillus amyloliquefaciens are mixed according to equal proportion and then are mixed with the bacillus cereus which is cultured independently according to the ratio of 1:1.
As shown in FIG. 1, the preparation method of the compound microbial agent capable of improving the yield of konjak provided by the invention comprises the following steps:
s101: bacillus subtilis, bacillus licheniformis, bacillus amyloliquefaciens and Bacillus pumilus are prepared from modified tryptone soybean liquid medium (tryptone 12.0g/L, soytone 3.0g/L, sodium chloride 3.0g/L, soil leaching solution 5.0ml/L, H) 2 O1000 ml, pH 7.2-7.5) for 48h;
s102: bacillus cereus was inoculated separately into modified beef extract peptone liquid medium (beef extract 2.5g/L, peptone 12.5g/L, sodium chloride 2.5g/L, soil extract 5.0ml/L, H) 2 O1000 ml, ph 7.5-7.8) for 36h;
s103: and after fermentation, mixing the two fermentation liquids according to the proportion of 1:1, wherein the mixed bacterial liquid is the composite microbial agent stock solution capable of improving the yield of konjak.
The preparation method of the composite microbial agent capable of improving the yield of konjak provided by the invention can be implemented by other steps by a person skilled in the art, and the preparation method of the composite microbial agent capable of improving the yield of konjak provided by the invention in fig. 1 is only one specific embodiment.
The technical scheme of the invention is further described below with reference to specific embodiments.
The invention relates to a method for preparing a strain number (ACCC 10719) by utilizing bacillus subtilis purchased from a Chinese microbial strain resource library; bacillus licheniformis, strain number (ATCC 11946), bacillus amyloliquefaciens, strain number (ATCC 23350); bacillus pumilus, strain number [ ]BAA-1434 TM ) The method comprises the steps of carrying out a first treatment on the surface of the Bacillus cereus, strain number (ATCC 9259). Firstly, adopting modified tryptone soybean liquid culture medium (tryptone 12.0g/L, soytone 3.0g/L, sodium chloride 3.0g/L, soil leaching liquid 5.0ml/L and H) for bacillus subtilis, bacillus licheniformis, bacillus amyloliquefaciens and bacillus pumilus 2 O1000 ml, pH 7.2-7.5) for 48h. ThenBacillus cereus was inoculated separately into modified beef extract peptone liquid medium (beef extract 2.5g/L, peptone 12.5g/L, sodium chloride 2.5g/L, soil extract 5.0ml/L, H) 2 O1000 ml, pH 7.5-7.8) for 36h. After fermentation, mixing the two fermentation liquids according to the proportion of 1:1, wherein the mixed bacterial liquid is the composite microbial agent stock solution capable of improving the konjak yield, and the effective viable count in the microbial agent is more than or equal to 1.0 hundred million/g (cfu). The application method comprises the steps of diluting the microbial inoculum according to the ratio of 1:500, and then irrigating the roots, wherein the microbial inoculum is applied in three periods of konjak planting process, namely a seeding period, a seedling emergence period and a tuber fast-growing period, and the single dosage per mu is 1000ml. After the microbial inoculum is applied, the growth and development of konjak are promoted, the agronomic characters of konjak are improved, the morbidity and death rate of konjak soft rot are obviously reduced, the yield is increased by 28.64%, and the effect is obvious.
The technical effects of the present invention will be described in detail with reference to experiments.
The application test of the composite microbial agent capable of improving the yield of konjak is carried out in the New Minvillage of Chua family community in the Qin mountain area of Kunming city, and the konjak variety is Lijiang flower konjak. The test result shows that the compound microbial agent capable of improving the yield of konjak has obvious growth promoting effect in the growth and development period of konjak, and the prevention effect on soft rot and southern blight of konjak reaches 28.54% and 64.32%, both reach extremely obvious level. The yield of the konjak single plant is increased compared with the control, the yield per mu is increased by 28.64% compared with the control, and the yield increasing effect is obvious.
The foregoing is merely illustrative of specific embodiments of the present invention, and the scope of the invention is not limited thereto, but any modifications, equivalents, improvements and alternatives falling within the spirit and principles of the present invention will be apparent to those skilled in the art within the scope of the present invention.
Claims (3)
1. The preparation method of the compound microbial agent capable of improving the yield of konjak is characterized by comprising the following steps of:
firstly, carrying out mixed fermentation culture on bacillus subtilis, bacillus licheniformis, bacillus amyloliquefaciens and bacillus pumilus by adopting an improved tryptone soybean liquid culture medium;
step two, solely inoculating bacillus cereus into a modified beef extract peptone liquid culture medium for fermentation culture;
thirdly, mixing the two fermentation liquids after fermentation is finished, wherein the mixed bacterial liquid is the composite microbial agent stock solution capable of improving the yield of konjak;
the improved tryptone soybean liquid culture medium is tryptone 12.0g/L, soytone 3.0g/L, sodium chloride 3.0g/L, soil leaching liquid 5.0ml/L and H 2 O 1000ml，pH 7.2-7.5；
The fermentation time of the first step is 48 hours;
the modified beef extract peptone liquid culture medium is beef extract 2.5g/L, peptone 12.5g/L, sodium chloride 2.5g/L, soil leaching liquid 5.0ml/L and H 2 O 1000ml，pH7.5-7.8；
The fermentation time of the second step is 36h;
mixing the two fermentation liquids according to the proportion of 1:1 after the fermentation is finished;
the composite microbial agent capable of improving the yield of konjak consists of bacillus subtilis, bacillus licheniformis, bacillus amyloliquefaciens and bacillus pumilus; mixing bacillus subtilis, bacillus licheniformis and bacillus amyloliquefaciens according to equal proportion, and then mixing with bacillus cereus which is independently cultured according to the ratio of 1:1;
the strain number of the bacillus subtilis is as follows: ACCC10719, bacillus licheniformis, strain number: ATCC11946, bacillus amyloliquefaciens, strain No.: ATCC 23350, bacillus pumilus, strain number:BAA-1434 TM ；
the effective viable count of the composite microbial agent capable of improving the yield of konjak is more than or equal to 1.0 hundred million cfu/g.
2. A method for improving the yield of konjak is characterized in that the method for improving the yield of konjak uses the compound microorganism bacterium agent capable of improving the yield of konjak according to claim 1 to carry out root irrigation after dilution according to 1:500 times.
3. The method for improving the yield of konjak according to claim 2, wherein the method for improving the yield of konjak is applied at three periods of konjak planting process, namely a seeding period, a seedling emergence period and a tuber fast-growing period, respectively, and the single dosage per mu is 1000ml.
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| CN102899266A (en) * | 2012-07-12 | 2013-01-30 | 云南农业大学 | Konjak endophytic bacteria Pantoea agglomerans bacterial strain1-7 and application |
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