CN116158447B - Biological resistance inducer for corn sprouts and application thereof - Google Patents
Biological resistance inducer for corn sprouts and application thereof Download PDFInfo
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- CN116158447B CN116158447B CN202310143542.3A CN202310143542A CN116158447B CN 116158447 B CN116158447 B CN 116158447B CN 202310143542 A CN202310143542 A CN 202310143542A CN 116158447 B CN116158447 B CN 116158447B
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- 239000000411 inducer Substances 0.000 title claims abstract description 27
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- 238000000034 method Methods 0.000 claims abstract description 18
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Classifications
-
- 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
- A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
- A01N63/38—Trichoderma
-
- 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
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/08—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing solids as carriers or diluents
- A01N25/10—Macromolecular compounds
-
- 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
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/02—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms
- A01N43/04—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom
- A01N43/14—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings
- A01N43/16—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings with oxygen as the ring hetero atom
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P21/00—Plant growth regulators
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Zoology (AREA)
- Environmental Sciences (AREA)
- Wood Science & Technology (AREA)
- Pest Control & Pesticides (AREA)
- Plant Pathology (AREA)
- Dentistry (AREA)
- Agronomy & Crop Science (AREA)
- Microbiology (AREA)
- Virology (AREA)
- Biotechnology (AREA)
- Mycology (AREA)
- Toxicology (AREA)
- Botany (AREA)
- Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The application discloses a corn sprout biological resistance inducer and application thereof; the biological attractant comprises trichoderma atroviride, luteolin, a carrier and a cross-linking agent, and is prepared by embedding according to a certain proportion. After the corn seeds are treated by induction, the stress resistance of corn sprouts is further improved. The biological attractant can effectively protect trichoderma atroviride and luteolin from being damaged by severe natural environments such as insolation, flooding and the like, improve the survival rate and the biological activity of the biocontrol bacteria in soil, and provide a protective environment for the active ingredients to play a role. Meanwhile, after the biological resistance inducer induces corn seeds, various substances related to plant stress resistance are produced, so that the growth capacity of the corn seeds can be improved, and the biological resistance inducer has a promoting effect on improving the yield and quality of corn. The method is simple, and the product is green and environment-friendly and has no pollution and high application and popularization value.
Description
Technical Field
The invention belongs to the technical field of plant protection, relates to a corn sprout biological attractant, and in particular relates to a corn sprout biological attractant and application thereof.
Background
Corn is an important cereal crop in China, the total yield of the corn is second to that of rice, and the corn is an important cereal, feed and industrial raw crop in China. At present, main diseases seriously affecting the production of corns are corn big spot, small spot, tumor black powder, stem basal rot and the like. The traditional pesticides and fertilizers have increasingly serious negative effects on agricultural systems in the use process, and the partial or full utilization of biocontrol agents or biological methods has positive effects on solving the problems.
Plants form multiple levels of defense mechanisms in the long-term interaction process with multiple pathogens in nature, and the damage of the external bad environment to the plants is resisted. The defense system of plants can be divided into three layers: constitutive defenses, innate immunity and systemic immunity. The constitutive defenses are formed in the early stage of plant evolution and form a first defense line for plants against pathogen infection, and can block infection of most of pathogen. Therefore, the stress resistance of the corn seedlings to soil-borne diseases, salt stress, drought and waterlogging and the like is improved by inducing resistance, the survival of the seedlings and the growth of plants are facilitated, the use of chemical pesticides and fertilizers is reduced, and the corn seedlings have no pollution to the atmosphere and soil and do not hurt natural enemies.
The corn seeds are treated to induce the seedlings to generate defending reaction, strengthen the capability of the seedlings for resisting pathogen infection, accelerate the growth and development of crops, increase the crop yield and improve the crop quality. The history of using the seed treating agent in China is long, warm soup soaking and medicament soaking are carried out in ancient times, and records are carried out in the book of agriculture, shengshen, in the West Han years. The function of the seed coating agent is mainly realized by the active ingredient, and the function of the inactive ingredient is realized by promoting the active ingredient to better function. The areas of China are wide, the climate conditions, soil conditions, environments and the like in different areas are greatly different, the use effect of the biological resistance inducer is easily influenced by the field environment or cannot be shown in a short period, for example, the biological seed coating agent taking pseudomonas fluorescens as a main component has high requirements on temperature and humidity, and the activity of strains is different under different temperature and humidity conditions.
The biological immobilization technique is a technique of immobilizing cells in a limited spatial region by physical or chemical means and maintaining the original biological activity thereof. The biological immobilization technology has the advantages of good stability, simple operation, easy realization and the like, can avoid the influence of adverse factors in the external environment on cells to a certain extent, and is beneficial to the growth and propagation of the cells. The biological immobilization method can be classified into an adsorption method, a crosslinking method, an embedding method, a complex immobilization method, etc., according to the type of the carrier and the form of the carrier combined with the living beings. The embedding method is to embed the organism in the gel grid of the high polymer or in the high polymer semipermeable membrane, and the small molecular substrate and the product can freely diffuse through the high polymer or the semipermeable membrane, so that the influence on the organism is small.
Disclosure of Invention
The application aims at providing a corn sprout biological attractant and application thereof; the biological resistance inducer has stable effect and long lasting period, is not influenced by soil environment, temperature and humidity, ph and the like, can induce corn seedlings to generate stress-resistant substances, and improves the stress resistance of crops to soil-borne diseases, salt stress, drought and waterlogging and the like by selecting proper biocontrol bacteria, plant active ingredients and biological fixation embedding technology.
In order to achieve the above purpose, the present invention provides the following technical solutions:
in a first aspect, the present invention provides a corn sprout biostimulant comprising a microorganism, a plant active ingredient, a carrier and a cross-linking agent; the microorganism is Trichoderma atroviride sp.CJCDSJ-F-KY 10039CCTCC No. M20221135, the plant active ingredient is luteolin, and the carrier is sodium alginate, chitosan and carrageenan. By adopting the technical scheme, the microorganism and the plant active ingredient are taken as the effective ingredients together, so that the problems of slow action of the microorganism and short acting duration of the plant active ingredient can be solved; in addition, the carrier adopts three substances, so that the limitation caused by singly using one carrier can be overcome. Sodium alginate is used as a carrier and has the advantages of no toxicity to microorganisms, good mass transfer property, low price, good operation and the like, and has the defects of wide application, low mechanical strength, easiness in being decomposed by microorganisms in soil and the like. The chitosan and the carrageenan have stable performance determined by molecular structures, can improve the strength of embedded particles, are easy to fix, have high survival rate and reproduction rate of microorganisms, and can provide carbon sources for the reproduction and growth of microorganisms.
As one embodiment, the crosslinker is a calcium chloride solution.
In a second aspect, the invention also relates to a preparation method of the corn sprout biological elicitor, which comprises the following steps:
s1, inoculating Trichoderma atroviride strain CJCSJ-F-KY 10039 into PDA culture medium, culturing in a 27-30deg.C illumination incubator for 3-5d, preparing spore suspension, inoculating with 1×10 concentration in PD 5 -10 7 Culturing Trichoderma spores at 27-30deg.C and 150-300rpm for 4-6 days to obtain bacterial liquid, and centrifuging at 9000-12000rpm for 15-25 min to obtain thallus; in some examples, after 4d incubation in an illumination incubator at 28 ℃, a spore suspension was prepared and inoculated at a concentration of 1×10 in PD 6 Culturing Trichoderma spores per ml at 28 ℃ and 200rpm for 5 days to obtain bacterial liquid, and centrifuging at 10000rpm for 20 minutes to obtain thalli;
s2, taking sodium alginate, chitosan and carrageenan as carriers, and embedding the thallus-plant active ingredients into the carriers after sterilization operation to obtain a thallus-plant active ingredient-carrier mixture;
s3, dripping the thallus-plant active ingredient-carrier mixture into a sterile cross-linking agent through a physical extrusion mode to prepare a spherical object, washing the spherical object with sterile physiological saline for 8-15 times to obtain trichoderma atroviride CJCSJ-F-KY 10039-luteolin embedded beads, drying the beads at 26-30 ℃ for 20-30h and storing the beads at 0-4 ℃. In some examples, the embedded beads were washed 10 times with sterile physiological saline to yield Trichoderma atroviride CJCSJ-F-KY 10039, dried at 27℃for 24h, and stored at 4 ℃. By adopting the technical scheme, calcium ions in the fixative calcium chloride are chelated with carrier molecules to form water-insoluble gel so as to fix cells.
As one embodiment, the carrier is a carrier solution with a mass concentration of 1-3% formed by dissolving sodium alginate, chitosan and carrageenan in sterile water; the dosage ratio of the carrier solution to the thallus-plant active ingredient is 5-20 ml/1 g; the mass ratio of sodium alginate, chitosan and carrageenan in the carrier is 1-4:2-5:1-10. In some embodiments, a carrier solution is formed with a mass concentration of 2%; the dosage ratio of the carrier solution to the thalli is 10ml to 1g; the mass ratio of sodium alginate, chitosan and carrageenan in the carrier is 1:3:2.
As one embodiment, the effective viable count in the bacterial liquid is 1×10 9 More than one/ml。
As one embodiment, the ratio of the bacterial cells to the plant active ingredient is 10 (1-7).
In step S2, the carrier is subjected to a sterilization operation under steam sterilization at 121 ℃ and a pressure of 0.12MPa for 20min.
As one embodiment, the crosslinking agent is a calcium chloride solution having a mass concentration of 0.5 to 2%; the dosage ratio of the calcium chloride solution to the thalli is 20ml to 1g. In some embodiments, the crosslinker is a 1% calcium chloride solution by mass.
As one embodiment, the physical extrusion mode is to uniformly drop the microbial inoculum-plant active ingredient-carrier mixture into the cross-linking agent by using a liquid-transferring gun, wherein the volume of each drop is 0.02ml.
As one embodiment, the embedded beads have a cure time of 15 to 20 hours. Preferably, the embedded beads have a curing time of 18 hours.
In a third aspect, the present invention relates to the use of a corn sprout biological elicitor for inducing the production of stress-resistant substances in corn sprouts. Corn seeds are induced to colonize by a biostimulant.
In the invention, trichoderma atroviride sp.CJCDSJ-F-KY 10039 is preserved in China Center for Type Culture Collection (CCTCC) for 7 months and 19 days in 2022, the preservation address is the university of Wuhan in China, and the preservation number is: cctccc No. m 20221135.
Compared with the prior art, the invention has the following beneficial effects:
1. the specific biological resistance inducer is adopted to induce corn seeds, so that corn sprouts generate stress-resistant substances, the stress resistance of crops to soil-borne diseases, salt stress, drought and waterlogging and the like is improved, the yield and the quality are improved, and the biological resistance inducer is safe and nontoxic, thereby reducing the use of chemical pesticides and chemical fertilizers, and meeting the development requirements of green agriculture and carbon emission reduction.
2. The biological attractant adopts a composition of microorganisms and plant active ingredients, so that the problems of slow action of the microorganisms and short acting duration of the plant active ingredients can be avoided.
3. The invention avoids the defect that after the biological resistance inducer is applied into the soil, the bacterial cells are directly exposed in the open and changeable soil environment to cause massive death of the bacterial cells, and dominant bacterial groups are not easy to form and the bacterial agents are difficult to exert the effective effects.
4. The invention screens and combines the embedded carriers, overcomes the defects of weak mechanical strength, low microorganism survival rate and the like caused by singly using one carrier, and better plays a role in protecting microorganism active ingredients after being applied into the soil and resisting impact and the like in natural environment.
5. The specific biological resistance inducer can induce corn sprouts to generate various stress-resistant substances such as malondialdehyde, calcium ions, jasmonic acid and salicylic acid to be increased, so that the adaptation of crops to drought, saline-alkali and other stress environments is improved, and the product has the advantages of simple preparation method, convenience in use, safety and reliability and better application potential and value.
Detailed Description
The present invention will be described in detail with reference to examples. The following examples will assist those skilled in the art in further understanding the present invention, but are not intended to limit the invention in any way. It should be noted that several modifications and improvements can be made by those skilled in the art without departing from the inventive concept. These are all within the scope of the present invention. The raw materials used in the present application are all commercially available.
Examples
1. Preparation of bioelectric elicitor
(1) Preparation of cells
Trichoderma atroviride strain CJCSJ-F-KY 10039 was inoculated into PDA medium, cultured in an illumination incubator at 28℃for 4d, spore suspension was prepared, and inoculated in PD at a concentration of 1X 10 6 Culturing Trichoderma spores at 28 deg.C and 200rpm for 5 days to obtain bacterial liquid with effective viable count of 1×10 9 After centrifugation at 10000rpm for 20 minutes per ml, cells were obtained.
(2) Preparation of thallus-plant active ingredient-carrier mixture
Preparing a mixed carrier of sodium alginate, chitosan and carrageenan. Preparing 10ml of carrier solutions Z1, Z2, Z3, Z4 and Z5 of sodium alginate, chitosan, carrageenan and polyvinyl alcohol according to different mass proportions, wherein the mass concentration of the carrier is 2%, the specific compositions are shown in table 1, the sodium alginate, the chitosan, the carrageenan and the polyvinyl alcohol are all chemically pure, and a certain amount of the sodium alginate, the chitosan, the carrageenan and the polyvinyl alcohol are respectively dissolved in hot water at 80 ℃ before use and cooled to about 35 ℃ for constant temperature for standby; steam sterilizing at 121deg.C and 0.12MPa for 20min, adding different thallus-plant active ingredient compositions shown in Table 2, and mixing under shaking on a constant temperature magnetic stirrer.
TABLE 1 Carrier composition in different proportions
| Carrier body | Sodium alginate | Chitosan | Carrageenan gum | Polyvinyl alcohol |
| Z1 | 1 | 3 | 2 | 0 |
| Z2 | 0 | 3 | 2 | 0 |
| Z3 | 1 | 0 | 2 | 0 |
| Z4 | 1 | 3 | 0 | 0 |
| Z5 | 0 | 3 | 2 | 1 |
TABLE 2 different thallus-plant active ingredient compositions
| Active ingredient | Trichoderma viride | Luteolin |
| A1 | 1 | 0.1 |
| A2 | 1 | 0.3 |
| A3 | 1 | 0.7 |
| A4 | 0 | 0.3 |
| A5 | 1 | 0 |
| A6 | 0 | 0 |
(3) Cross-linking embedding fixation
Dripping the thallus-plant active ingredient-carrier mixture into 20mL containing 1% CaCl at a certain speed with a pipette 2 In the crosslinking solution of (2), the volume of each drop is 0.02ml, crosslinking is carried out for 18 hours, particles are filtered out, the solution is washed for 10 times by sterile normal saline, and the trichoderma atroviride CJCSJ-F-KY 10039 embedded particles, namely the resistance inducer, are respectively obtained, dried at 27 ℃ for 24 hours and stored at 4 ℃.
2. Performance test
The activity, mechanical strength, etc. of the biostimulant directly affect the actual effect of the biostimulant.
(1) Determination of the mechanical Strength of the elicitor particles
The bioaugmented agents were prepared using the active ingredient A1 as an example, and the bioaugmented agents Y1, Y2, Y3, Y4 and Y5 were prepared with the carriers in the respective proportions shown in table 1. And respectively taking 5 biological antigen-inducing agent particles Y1, Y2, Y3, Y4 and Y5 with the same size, putting the particles on a glass plate in a square shape, putting a glass dish on the glass plate, slowly adding broken codes into the glass dish until the particles are completely deformed, and recording the glass dish which enables the particles to be completely deformed and the total weight of the added codes as 1/4 of the weight of each embedded particle, thereby representing the mechanical strength of the pellets. Each treatment was repeated 3 times and the results averaged.
TABLE 3 determination of mechanical Strength of Carrier biological resistance inducer particles in different proportions
(2) Biological attractant particle viable count determination
1g of fresh embedded particles are weighed, the volume is fixed to 10mL in 0.2mol/L sodium citrate solution respectively, the particles are fully oscillated by a vortex oscillator until the particles are fully dissolved, the bacterial cells are fully dissolved out, and the bacterial cells are cultivated for 3d at 28 ℃ by a dilution plate method, and the bacterial colonies are counted. Each treatment was repeated 3 times and the results averaged. The various indices are calculated as follows.
Particle viable count (cfu/g) = (viable count of a certain dilution x bacterial liquid amount)/particle weight
TABLE 4 determination of the viable count of Carrier biological resistance inducer particles in different proportions
As can be seen from tables 3 and 4, the mechanical strength of the bioelectric antigen Y1 was maximized to 50.9g/g, and the viable count of the particles was maximized to 13X 10 8 cfu/g, which shows that the mass ratio of the carrier Z1 to the sodium alginate, chitosan and carrageenan is 1:3:2, the bacteria-plant active substance-carrier is crosslinked, embedded and immobilized, so that the biological attractant has the best performance, can resist the interference of different external natural environments, and is beneficial to the delivery of the active substances. The carrier adopts three substances, and can overcome the limitation caused by the single use of one carrier. Sodium alginate is used as a carrier and has the advantages of no toxicity to microorganisms, good mass transfer property, low price, good operation and the like, and has the defects of wide application, low mechanical strength, easiness in being decomposed by microorganisms in soil and the like. Chitosan and carrageenan are determined by molecular structureThe chitosan has stable performance, can improve the strength of embedded particles, is easy to fix, has high survival rate and reproduction rate of microorganisms, and can provide carbon sources for reproduction and growth of microorganisms.
3. Induced stress test
The corn seeds are firstly subjected to surface sterilization by 75% ethanol for 10min and placed in an ultra clean bench for air drying. The carrier Z1 is selected, and the biological resistance inducer is prepared according to the composition of different thallus-plant active ingredients in the table 2, wherein the biological resistance inducer comprises Y1, Y6, Y7, Y8, Y9 and Y10 respectively in mass ratio of 10:1, pouring the mixture into a beaker, shaking the beaker, mixing uniformly and then performing colonization. The blank is corn seed that was not induced.
(1) Germination rate investigation
Samples were placed in a dark incubator at 28℃and the germination count was counted daily.
Germination percentage% = number of sprouts/total number of corn × 100%
(2) Effect on maize seedling growth
The induced seeds were sown in pots, 10 per pot, and each treatment was repeated three times, with the uninduced seeds as controls. The germination rate was calculated after 7 days, and root length, root number, plant height, fresh weight and dry weight were measured after 14 days, see table 5.
TABLE 5 determination of different indicators
As can be seen from Table 5, the germination percentage, root length, root number, plant height, fresh weight and dry weight of the corn seeds treated with the bioaugmenters Y1, Y6 and Y7 were all higher than those of the corn seeds which were single active substances, without active substances or were not treated with the bioaugmenters. It is demonstrated that under the same carrier components, the biological elicitor prepared by the active ingredient composition of the invention is favorable for germination of corn seeds and absorption of nutrient substances, and has good growth vigor.
(3) Determination of stress resistance index of corn sprouts
And respectively measuring indexes related to plant stress resistance, such as malondialdehyde, calcium ions, jasmonic acid, salicylic acid and the like in the seed sprouts 7 days after the treatment.
TABLE 6 determination of stress resistance index for different treated seeds
Malondialdehyde is an organic substance produced by peroxidation of membrane lipids of tissues or organs of plant organs due to senescence or damage in adverse circumstances. The starch in the plant body can be converted into glucose, and the freezing point of the sugar solution is relatively low, so that the body fluid of the plant is not easy to solidify in a low-temperature environment so as to adapt to a cold environment. The plant stress resistance can be judged by measuring the content of the plant malondialdehyde.
Calcium ion is a macroelement necessary for plant growth and development, and it is simultaneously involved in regulating stress-tolerance of plants to environmental stress as an important messenger. An increase in calcium ion content indicates an increase in the ability of plants to regulate under environmental stress.
Jasmonic acid and its derivatives are lipid-based plant hormones that regulate various processes in plants, from growth and photosynthesis to reproductive development. In particular, jasmonic acid is critical to plant defense against herbivores and plants' responses to harsh environmental conditions and other types of abiotic and biological challenges.
Salicylic acid is one of the ubiquitous endogenous signal molecules in plants, has been identified as a phytohormone, and has important physiological functions. Research shows that the salicylic acid has obvious effects on disease resistance, low temperature resistance, drought resistance, salt resistance and the like of plants, fruit ripening, product preservation, seed germination and the like.
As can be seen from Table 6, the content of malondialdehyde, calcium ions, jasmonic acid and salicylic acid substances in the corn seed sprouts treated by the biological inducers Y1, Y6 and Y7 is higher than that of the corn seed sprouts treated by the biological inducers Y8, Y9 and Y10 and the corn seed sprouts treated by the biological inducers Y7. The corn sprouts treated by the biological resistance inducer prepared by the active ingredient composition of the invention contain higher plant stress-resistance regulating substances under the same carrier components, and can be better adapted to growth in a stress environment, thereby improving the yield and quality of corn. As a biological inducer, the microorganism and the plant active ingredient are used together as the effective ingredients, so that the problems of slow action of the microorganism and short acting duration of the plant active ingredient can be solved.
The foregoing describes specific embodiments of the present invention. It is to be understood that the invention is not limited to the particular embodiments described above, and that various changes and modifications may be made by one skilled in the art within the scope of the claims without affecting the spirit of the invention.
Claims (8)
1. A corn sprout biological resistance inducer, which is characterized by comprising microorganisms, plant active ingredients, a carrier and a cross-linking agent; the microorganism is Trichoderma atroviride sp.CJCDSJ-F-KY 10039CCTCC No. M20221135, the plant active ingredient is luteolin, and the ratio of thallus to plant active ingredient is 10 (1-7); the carrier is sodium alginate, chitosan and carrageenan; dissolving sodium alginate, chitosan and carrageenan in sterile water to form a carrier solution with the mass concentration of 1-3%, wherein the dosage ratio of the carrier solution to the microorganism-plant active ingredient is 5-20 ml/1 g; the mass ratio of sodium alginate, chitosan and carrageenan in the carrier is 1-4:2-5:1-10.
2. The corn sprout biological inducer of claim 1 wherein the cross-linking agent is a calcium chloride solution.
3. A method of preparing a corn sprout biological attractant according to claim 1, comprising the steps of:
s1, inoculating Trichoderma atroviride strain CJCSJ-F-KY 10039 into PDA culture medium, culturing in a 27-30deg.C illumination incubator for 3-5d, preparing spore suspension, inoculating with 1×10 concentration in PD 5 -10 7 Culturing Trichoderma spores at 27-30deg.C and 150-300rpm for 4-6 days to obtain bacterial liquid, and centrifuging at 9000-12000rpm for 15-25 min to obtain thallus;
s2, dissolving sodium alginate, chitosan and carrageenan in sterile water to form a carrier solution with the mass concentration of 1-3%; embedding the thallus-plant active ingredient into a carrier after sterilization operation to obtain a thallus-plant active ingredient-carrier mixture;
s3, dripping the thallus-plant active ingredient-carrier mixture into a sterile cross-linking agent through a physical extrusion mode to prepare a spherical object, washing the spherical object with sterile physiological saline for 8-15 times to obtain trichoderma atroviride CJCSJ-F-KY 10039-luteolin embedded beads, drying the beads at 26-30 ℃ for 20-30h and storing the beads at 0-4 ℃.
4. The method for preparing a biological attractant for maize seedlings according to claim 3, wherein the effective viable count in the bacterial liquid is 1×10 9 More than one per ml.
5. The method for preparing a biological attractant for maize seedlings according to claim 3, wherein in the step S2, the carrier is subjected to a sterilization operation under the sterilization condition of 121 ℃ and the pressure of 0.12MPa for 20min.
6. The method for preparing a corn sprout biological inducer according to claim 3, wherein the cross-linking agent is a calcium chloride solution with a mass concentration of 0.5-2%; the dosage ratio of the calcium chloride solution to the thalli is 20ml to 1g.
7. The method for preparing a corn sprout biological inducer according to claim 3, wherein the physical extrusion method is to uniformly drop the mixture of the microbial inoculum and the plant active ingredient and the carrier into the cross-linking agent by a pipette, and the volume of each drop is 0.02ml; the curing time of the embedded beads is 15-20h.
8. Use of the corn sprout biological inducer according to claim 1 or the corn sprout biological inducer according to claim 3 for inducing corn sprout to produce stress-resistant substance.
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