CN114455986A - Carya illinoensis bacterial fertilizer and application thereof - Google Patents
Carya illinoensis bacterial fertilizer and application thereof Download PDFInfo
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- CN114455986A CN114455986A CN202210140809.9A CN202210140809A CN114455986A CN 114455986 A CN114455986 A CN 114455986A CN 202210140809 A CN202210140809 A CN 202210140809A CN 114455986 A CN114455986 A CN 114455986A
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- 241001453450 Carya illinoinensis Species 0.000 title claims abstract description 26
- 235000009025 Carya illinoensis Nutrition 0.000 title claims abstract description 25
- 230000001580 bacterial effect Effects 0.000 title claims abstract description 20
- 239000003337 fertilizer Substances 0.000 title claims abstract description 6
- 238000000855 fermentation Methods 0.000 claims abstract description 42
- 230000004151 fermentation Effects 0.000 claims abstract description 42
- 235000013399 edible fruits Nutrition 0.000 claims abstract description 33
- 239000000843 powder Substances 0.000 claims abstract description 13
- 238000005507 spraying Methods 0.000 claims abstract description 10
- 241000235058 Komagataella pastoris Species 0.000 claims abstract description 8
- 241000233866 Fungi Species 0.000 claims abstract description 5
- 241000521553 Pichia fermentans Species 0.000 claims abstract description 4
- 241001523965 Xylaria Species 0.000 claims abstract description 4
- 239000003085 diluting agent Substances 0.000 claims abstract description 3
- 241000894006 Bacteria Species 0.000 claims description 21
- 239000000706 filtrate Substances 0.000 claims description 9
- 239000007788 liquid Substances 0.000 claims description 9
- 210000003608 fece Anatomy 0.000 claims description 8
- 239000010871 livestock manure Substances 0.000 claims description 8
- 239000007921 spray Substances 0.000 claims description 6
- 239000000758 substrate Substances 0.000 claims description 3
- 238000001914 filtration Methods 0.000 claims description 2
- 238000002156 mixing Methods 0.000 claims description 2
- 230000005526 G1 to G0 transition Effects 0.000 claims 1
- 240000005979 Hordeum vulgare Species 0.000 claims 1
- 235000007340 Hordeum vulgare Nutrition 0.000 claims 1
- 244000061456 Solanum tuberosum Species 0.000 claims 1
- 235000002595 Solanum tuberosum Nutrition 0.000 claims 1
- 238000011081 inoculation Methods 0.000 claims 1
- 238000004321 preservation Methods 0.000 claims 1
- 239000007787 solid Substances 0.000 claims 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 11
- 201000010099 disease Diseases 0.000 abstract description 10
- 230000000694 effects Effects 0.000 abstract description 8
- 230000017260 vegetative to reproductive phase transition of meristem Effects 0.000 abstract description 8
- 230000010152 pollination Effects 0.000 abstract description 7
- 238000000034 method Methods 0.000 abstract description 6
- 241001523964 Xylariaceae Species 0.000 abstract description 5
- 206010027146 Melanoderma Diseases 0.000 abstract description 3
- 238000001035 drying Methods 0.000 abstract description 2
- 241000235648 Pichia Species 0.000 abstract 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 abstract 1
- 241000196324 Embryophyta Species 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 239000011550 stock solution Substances 0.000 description 4
- 241000238631 Hexapoda Species 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 241000607479 Yersinia pestis Species 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000007865 diluting Methods 0.000 description 2
- 238000001694 spray drying Methods 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 241000193738 Bacillus anthracis Species 0.000 description 1
- 241000723418 Carya Species 0.000 description 1
- AUNGANRZJHBGPY-UHFFFAOYSA-N D-Lyxoflavin Natural products OCC(O)C(O)C(O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O AUNGANRZJHBGPY-UHFFFAOYSA-N 0.000 description 1
- 241000758791 Juglandaceae Species 0.000 description 1
- 229920002774 Maltodextrin Polymers 0.000 description 1
- 239000005913 Maltodextrin Substances 0.000 description 1
- GXCLVBGFBYZDAG-UHFFFAOYSA-N N-[2-(1H-indol-3-yl)ethyl]-N-methylprop-2-en-1-amine Chemical compound CN(CCC1=CNC2=C1C=CC=C2)CC=C GXCLVBGFBYZDAG-UHFFFAOYSA-N 0.000 description 1
- AUNGANRZJHBGPY-SCRDCRAPSA-N Riboflavin Chemical compound OC[C@@H](O)[C@@H](O)[C@@H](O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O AUNGANRZJHBGPY-SCRDCRAPSA-N 0.000 description 1
- 229930003451 Vitamin B1 Natural products 0.000 description 1
- 229930003471 Vitamin B2 Natural products 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 235000019606 astringent taste Nutrition 0.000 description 1
- 229940065181 bacillus anthracis Drugs 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 239000013043 chemical agent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 208000029078 coronary artery disease Diseases 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 238000009396 hybridization Methods 0.000 description 1
- 230000009027 insemination Effects 0.000 description 1
- 238000009533 lab test Methods 0.000 description 1
- 229940035034 maltodextrin Drugs 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 235000021281 monounsaturated fatty acids Nutrition 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 239000000575 pesticide Substances 0.000 description 1
- 238000005498 polishing Methods 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000003223 protective agent Substances 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000004080 punching Methods 0.000 description 1
- 229960002477 riboflavin Drugs 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 229960003495 thiamine Drugs 0.000 description 1
- DPJRMOMPQZCRJU-UHFFFAOYSA-M thiamine hydrochloride Chemical compound Cl.[Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N DPJRMOMPQZCRJU-UHFFFAOYSA-M 0.000 description 1
- 235000010374 vitamin B1 Nutrition 0.000 description 1
- 239000011691 vitamin B1 Substances 0.000 description 1
- 235000019164 vitamin B2 Nutrition 0.000 description 1
- 239000011716 vitamin B2 Substances 0.000 description 1
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Classifications
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
- C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
- C05G3/60—Biocides or preservatives, e.g. disinfectants, pesticides or herbicides; Pest repellants or attractants
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G5/00—Fertilisers characterised by their form
- C05G5/20—Liquid fertilisers
- C05G5/23—Solutions
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Pest Control & Pesticides (AREA)
- Organic Chemistry (AREA)
- Soil Sciences (AREA)
- Plant Pathology (AREA)
- Environmental Sciences (AREA)
- Biochemistry (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
From fungi of the family xylariaceae (Xylaria radix) sj18 strain and fermented Pichia yeast (Pichia pastoris) ((R))Pichia fermentans) 15B1 strain mixed fermentation broth or dry powder obtained by concentrating and drying the fermentation broth to form the apocarya bacterial fertilizer. Spraying 10 times of diluent of fermentation liquor for 1-3 times within three months from the time when male flowers of the carya illinoensis begin to loose pollen to the time when female flowers bear fruits, wherein the spraying amount of each tree is 1-2 liters, so that the carya illinoensis pollen activity-maintaining and prolonging capacity is achieved, the flowering asynchronism is effectively improved, and the pollination rate is increased; meanwhile, the method inhibits the generation of black disease spots in the forming process of the carya illinoensis, and has the effects on the phenomena of low yield of carya illinoensis woodland, serious flower and fruit dropping, poor fruit quality caused by the black spot phenomenon of fruits and the like in ChinaHas remarkable improvement effect.
Description
Technical Field
The invention relates to the technical field of biology, in particular to xylariaceae fungi (A)Xylaria radix) sj18 Strain and Pichia pastoris (A), (B)Pichia fermentans) Application of a mixed fermentation product of 15B1 strain in pecan fructification.
Background
Carya illinoensis is one of the most economically valuable species of Carya of the Juglandaceae family, the origin of the United states. In the original producing area, the apocarya has more than 1000 varieties, large fruits, thin shells, high kernel yield (50-70%), easy kernel extraction and high yield (1500-2250 kg/hectare, 100-150 kg/mu). The nuts are delicious, have no astringent taste, are rich in nutrition (containing 72% of oil, 11% of protein and 13% of carbohydrate), have high amino acid content, and are rich in vitamin B1 and B2. The core oil contains 73% of monounsaturated fatty acid, has a calorific value of 32 kcal/kg, can reduce the incidence rate of coronary heart disease after being eaten, and is an ideal health-care product and food additive. The crushed fruit shell is a high-grade polishing material with the price equivalent to that of the kernel. The full bearing period of the apocarya is generally more than 50 years. The introduction of the apocarya in China has been over 100 years old, and although the apocarya grows well for a long time, the yield difference of different places is large, and particularly the yield of Jiangzhe and Zhejiang is unstable and most of the apocarya is low;
the carya illinoensis is a wind-borne flower cross-crossing plant with the same male and female plants, and is divided into a male-type variety and a female-type variety according to the sequence of the male and female florescence, and the cross among different varieties has the problem of flowering asynchronism, so that the insemination is insufficient, the fruit setting rate is low, and the fruit is thin and flat, so that the yield and the fruit quality are seriously influenced; on the other hand, due to different climatic factors such as temperature, humidity and the like, the staggered time of male and female florescence is unequal, so that a plurality of reasonable variety proportions are configured in some areas, but the florescence is possibly missed in other areas, unreasonable is achieved, and the yield and quality are influenced by the missed florescence; in addition, due to the difference of micro-ecological environments in different regions, some new regional pests occur, and the yield and the fruit quality are influenced. For example, in recent years, it has been found that the quality of pecan fruits is deteriorated due to the presence of black spots in the pecan fruits, and the pecan fruits are detected as anthracnose. At present, the method mainly improves the yield and controls the plant diseases and insect pests by using chemical agents, pesticides and the like in production, most of the method is used in the flowering phase and the fruit phase, plays a role in controlling the plant diseases, but also influences the hybrid pollination, reduces the fruit quality and increases the potential safety hazard of food. Therefore, a safe and effective biological regulation and control method is urgently needed to be developed in production, so that the pollination efficiency is improved, the yield is increased, and the fruit quality is improved while the fruit diseases can be effectively controlled. The applicant proposes two inventions of a xylariaceae fungus and an application thereof (invention 201710795984.0) and a fermented pichia pastoris and an application thereof (invention 202110032614.8) in the earlier stage, the two fungi related to the two inventions are verified to have the capabilities of resisting and expelling germs and insects in laboratory experiments, and the fermented pichia pastoris 15B1 strain can fully utilize the fermentation product of the sj18 strain to further ferment, simultaneously has the capability of keeping and prolonging the pollen activity of the carya illinoensis, and can effectively improve the problems of flowering asynchronism and the like. The bacterial manure which can be sprayed in the flowering phase and the fruit phase of the carya illinoensis is obtained by utilizing the mixed fermentation of the two fungi, the pollen activity of the carya illinoensis can be obviously improved, the pollination rate is increased, and the yield is improved; meanwhile, the generation of disease spots of the carya illinoensis is inhibited, and the quality of fruits is obviously improved.
Disclosure of Invention
Aiming at the problems in the prior art, the invention aims to provide a mixed fermentation bacteria liquid of a xylariaceae fungus sj18 and a fermentation pichia pastoris 15B1, which is used for improving the pollination rate of apocarya pollen, improving the yield, reducing the formation of fruit disease spots and improving the quality of fruits;
the strain is xylariaceae fungus (CGMCC No. 12780)Xylaria radix) The strain sj18 and the fermented pichia pastoris (CGMCC No. 19317)Pichia fermentans) Strain 15B 1;
the mixed fermentation liquid can obviously maintain and prolong the pollen activity of the apocarya and increase the pollination rate, wherein the laboratory achievement of the 15B1 strain for maintaining and recovering the pollen activity of the apocarya is patented (a fermentation pichia pastoris and application thereof, ZL 202110032614.8);
the fermentation liquor of the mixed fermentation is used as an application agent for biological germ resistance and insect expelling, can reduce fruit disease spots, promotes fruit growth and plays a role of bacterial manure. The strain is disease-resistant plant endophytic fungi, and the fermentation process and the fermentation product of the strain are environment-friendly, pollution-free and residue-free. Because the growth advantages of the two bacteria are complementary, the mixed fermentation liquor can obviously promote pollination efficiency, improve setting percentage, reduce fruit diseases and improve fruit quality in production;
the application method and the dosage of the bacterial fertilizer are as follows:
diluting the mixed fermentation stock solution according to the ratio of the fermentation stock solution to water of 1:10 within 1-3 months at the early stage of flowering and fruiting of carya illinoensis fruit trees, and then dissolving 1-2g of each tree in 1-2L of water to spray the solution of each tree with the spray-dried dry powder of the fermentation stock solution, wherein the spray-dried powder is 0.1-0.2L of the stock solution. Spraying for 1-3 times within 1-3 months of the initial stage of flowering and fruiting, with interval of 1 month.
Drawings
FIG. 1: and (3) a viable bacteria detection plate for the bacterial manure fermentation liquid dry powder.
Detailed Description
The present invention is further illustrated by the following examples. The following examples are not intended to limit the invention.
Example 1: mixed fermentation of sj18 strain and 15B1 strain and filtrate collection
Punching 2-3 pieces on PDA plate inoculated with sj18 with 1cm puncher to obtain 2-3 pieces of bacterial blocks, inoculating the bacterial blocks into 1L shake flask filled with 500mL PDB culture medium, shaking at 200rpm, culturing at 32 deg.C for 4-5 days to obtain initial sj18 strain;
meanwhile, selecting 15B1 monoclonal on a 15B1 strain PDA plate, inoculating the monoclonal into a 50mL tube filled with 20mL PDB culture medium, shaking at 200rpm, culturing at 32 ℃ for 48h to obtain initial 15B1 strain;
drying a small amount of sj18 and 15B1 bacterial liquids, determining the proportion of the mixed fermentation bacterial liquids to be approximately sj18:15B1 to be 10-100, and mixing the two bacteria and inoculating the mixture into a 50L stirring type fermentation tank containing 25L culture solution for fermentation. Fermentation conditions are as follows: fermenting with 10% malt as fermentation substrate at stirring speed of 200rpm and fermentation temperature of 32 deg.C, controlling pH at 5.5-6.0, feeding air pressure of 0.1MPa, and air inflow of 25L/min. After fermenting for 6 days, filtering the fermentation liquor by a 200-mesh and 300-mesh filter screen to obtain hypha and fermentation substrate, collecting the filtrate, and properly diluting the filtrate to spray the carya illinoensis plants.
Example 2: analysis of bacterial content of different preparations of mixed fermentation filtrate of sj18 bacteria and 15B1 bacteria
3L of a fermentation product of a mixed bacterium liquid of sj18 bacteria and 15B1 bacteria is centrifugally concentrated to 300mL, 300g of maltodextrin is added to serve as a protective agent, and spray drying is carried out by adopting a YAMATO ADL311 fog dryer, wherein the spray parameters are inlet temperature 115 ℃ and outlet temperature 50 ℃, so that 13.7g of dry powder is obtained. Dissolving 0.01g of dry powder in 1mL of sterile waterThen, 100. mu.L of the solution was applied onto a 9 cm-sized culture medium plate, and the number of viable bacteria contained in the spray-dried powder was calculated to be about 1500 on average by calculating the amount of bacteria in the plate, so that the number of viable bacteria contained in the plate was about 1.5X 106Per gram. The dry powder obtained after spray drying is a state convenient for storing and transporting active bacteria;
FIG. 1 is a viable bacteria detection plate for bacterial manure fermentation liquid dry powder.
Example 3: effect of spraying mixed fermentation filtrate of sj18 strain and 15B1 strain in carya illinoensis woodland
Spray test method of 15B1 bacteria: selecting variety 'Jinhua No. 1' (Hangzhou Jiande Carya illinoensis garden) in Hangzhou Jiande Carya illinoensis gardenCarya illinoinensis) 12 individuals are grouped and paired according to the yield of the last year, the yield is similar and is divided into the same group, and the groups are marked by numbers of-1 and-2, and spraying and non-spraying are respectively arranged. When spraying, 20g of dry powder of mixed fermentation filtrate of sj18 bacteria and 15B1 bacteria is dissolved in 20L of water, and spraying is carried out once in 5 months and 10 days, 6 months and 10 days and 7 months and 10 days respectively. Statistics of the yield of the current year is shown in the table I, and the yield of the carya illinoensis is obviously increased after bacterial manure spraying. Some hybridization combinations that were previously difficult to obtain also show consequences. In addition, the quality of the fruits is also obviously improved, 100 fruits which are not sprayed with the bacterial manure are randomly peeled, 8% of the fruits have black spots, the black spots are identified as the black spot disease caused by the bacillus anthracis through molecules, and the 100 fruits which are sprayed with the bacterial manure are also randomly peeled, and no black spot appears;
TABLE-results of 3 sprays of mixed fermentation filtrates from sj18 and 15B1 bacteria control
Forest research results show that the mixed fermentation filtrate diluent of the sj18 strain and the 15B1 strain is intermittently sprayed for 3 times within 3 months of the flowering and initial fruiting of the carya illinoensis, and the effects of improving the fruiting rate, reducing fruit diseases and improving the fruit quality are obvious.
Claims (5)
1. A Carya illinoensis bacterial fertilizer is prepared from Carnikoaceae fungi with preservation number of CGMCC No.12780Bacteria (A), (B)Xylaria radix) sj18 bacterial strain and fermented pichia pastoris (CGMCC No. 19317)Pichia fermentans) 15B1 strain is prepared by mixing fermented liquid or dry powder of fermented liquid.
2. The proportional mixed fermentation of claim 1, wherein barley or potato is used as fermentation substrate, and the ratio of inoculation of bacteria is sj18:15B1 is greater than 10 and less than 100.
3. The fermentation broth of claim 1, wherein after the fermentation has reached a stationary phase, a filtrate obtained by filtering the mycelia and containing viable bacteria is taken as a component of the bacterial manure.
4. The dry, concentrated fermentation broth powder of claim 1, wherein the fermentation broth is lyophilized at low temperature or spray dried to produce a viable bacteria-containing solid.
5. The application of the carya illinoensis bacterial fertilizer as claimed in claim 1, characterized in that within three months from the time when male flowers of carya illinoensis start to loose powder to the time when female flowers bear fruit, 10 times of diluent of fermentation liquor is sprayed for 1-3 times, and the spraying amount of each tree is 1-2 liters.
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| CN202210140809.9A CN114455986A (en) | 2022-02-16 | 2022-02-16 | Carya illinoensis bacterial fertilizer and application thereof |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108117998A (en) * | 2017-09-06 | 2018-06-05 | 浙江省林业科学研究院 | A kind of Xylariaceae fungi and its application |
| CN112438277A (en) * | 2021-01-12 | 2021-03-05 | 浙江省林业科学研究院 | Xylariaceae fungus sj18 and application thereof in prevention and treatment of pine wood nematodes |
| CN112574896A (en) * | 2021-01-12 | 2021-03-30 | 浙江省林业科学研究院 | Fermented pichia pastoris and application thereof |
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- 2022-02-16 CN CN202210140809.9A patent/CN114455986A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108117998A (en) * | 2017-09-06 | 2018-06-05 | 浙江省林业科学研究院 | A kind of Xylariaceae fungi and its application |
| CN112438277A (en) * | 2021-01-12 | 2021-03-05 | 浙江省林业科学研究院 | Xylariaceae fungus sj18 and application thereof in prevention and treatment of pine wood nematodes |
| CN112574896A (en) * | 2021-01-12 | 2021-03-30 | 浙江省林业科学研究院 | Fermented pichia pastoris and application thereof |
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