CN112501028A - Efficient microbial agent suitable for dendrobium wilt and preparation method thereof - Google Patents
Efficient microbial agent suitable for dendrobium wilt and preparation method thereof Download PDFInfo
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Abstract
The invention provides a high-efficiency microbial agent suitable for dendrobium wilt and a preparation method thereof, and relates to the technical field of dendrobium planting. The mixture of the mosaicdou ductal vesiculomycete agent and the trichoderma harzianum agent is used as a microbial agent for preventing and treating dendrobium wilt. The high-efficiency microbial agent suitable for the dendrobium wilt can promote the growth of the root system of the dendrobium seedling, and the growth promoting effect of the bursa of Moxifragans microbial agent and the trichoderma harzianum microbial agent is better than that of the bursa of Moxifragans microbial agent or the trichoderma harzianum microbial agent which is inoculated independently, so that the synergistic effect of the bursa of Moxifragans and the trichoderma harzianum in the aspect of preventing and treating the dendrobium wilt is proved.
Description
Technical Field
The invention relates to the technical field of dendrobium planting, in particular to a high-efficiency microbial agent suitable for dendrobium wilt, and a preparation method and application thereof.
Background
Dendrobe (Dendrobium spp.) is a larger genus of epiphytic plants in Orchidaceae, about 1000 species are found in the world at present, 74 and 2 varieties exist in China, and common Dendrobium officinale, Dendrobium devonianum, Dendrobium moniliforme, Dendrobium huoshanense and the like exist, wherein the Dendrobium officinale and the Dendrobium nobile are both recorded by the pharmacopoeia of the national calendar, have the effects of nourishing yin, promoting the production of body fluid and the like, and are important Chinese medicinal material resources. The dendrobium is an epiphytic plant, a fleshy root, a typical mycorrhizal plant which is grown on a bark, a trunk or in a stone crack in a wild way and is rich in endophytic fungi. The dendrobium officinale has long planting period, is generally harvested about two years after being transplanted, has long harvesting period, can be harvested circularly, and can generally harvest about five years, so the management of the dendrobium officinale after being transplanted is particularly important; in the cultivation process of the dendrobium officinale, blight often occurs to cause large-area yield reduction and even total harvest of the dendrobium officinale.
The dendrobium wilt disease is mainly to damage the root of the plant, pathogenic bacteria infect the root system of the dendrobium to cause the plant to wilt, and the root system is dead. The epidermis of the stem at the base of the plant becomes rough and occasionally has cracks, white or pink mildew can be seen when the humidity is high, and some rootstock bases can also see water-soaked scabs. When the root of the rhizome is transversely or longitudinally cut, the vascular bundle has brown or black brown ring changes.
The dendrobium wilt is prevented and treated by chemical agents in the existing dendrobium planting process. However, there are disadvantages as follows:
1. the prevention and treatment effect is poor; 2. high chemical residue and soil pollution.
Therefore, a microbial agent capable of preventing and treating dendrobium wilt is needed.
Disclosure of Invention
In view of the problems, the invention provides the efficient microbial agent suitable for the dendrobium wilt and the preparation method thereof, and the strains are not antagonistic and are synergistic with each other, so that the aim of the invention is to be beneficial to preventing and treating the dendrobium wilt and promoting the growth of dendrobium.
In order to achieve the purpose, the invention provides a preparation method of a high-efficiency microbial agent suitable for dendrobium wilt, the trachelospermum moxidense is planted and propagated through corns in advance, a mixture of spores, extra-root hyphae and root soil infecting corn root segments of the trachelospermum moxidense is used as a trachelospermum moxidense microbial agent, and each gram of the trachelospermum moxidense microbial agent contains 25-35 spores;
activating trichoderma harzianum, and inoculating the activated trichoderma harzianum to a trichoderma harzianum liquid seed culture medium for culture to obtain trichoderma harzianum fermented seed liquid; wherein, the spore concentration of the Trichoderma harzianum strain liquid is adjusted to 1 x 106Per ml;
weighing 51% of straw, 25.5% of wheat bran, 1.5% of rice bran, 21.5% of shell powder and 0.5% of (NH)4)2SO4Mixing to obtain fermented material, adding water to make water content of the fermented material reach 75%, stirring, packaging into polypropylene fungus bags, sterilizing at 121 deg.C for 20min, 150g per bag to obtain sterilized fermented material;
adding 5ml of Trichoderma harzianum fermentation seed liquid into the sterilized fermentation material, and fermenting at room temperature for 12 days; after culturing for 12 days, drying the fermentation product, separating the fermentation material from conidia by screening, collecting spore powder and storing at 4 ℃ to obtain a trichoderma harzianum microbial inoculum;
and mixing the trichoderma harzianum microbial inoculum with the moscilla bursa mycorrhiza microbial inoculum to obtain the high-efficiency microbial inoculum suitable for the dendrobium wilt.
Further, preferably, in the microbial agent, the ratio of the trichoderma harzianum microbial agent to the pipefish mosaicensis microbial agent in parts by weight on a dry basis is 3-5: 1.
the invention also protects a high-efficiency microbial agent suitable for dendrobium wilt, and the effective viable count of Moxidou trachelospermum and Trichoderma harzianum in the microbial agent is not less than 1 x 1011CFU/g。
The invention also protects the application of the high-efficiency microbial agent suitable for the dendrobium wilt in the dendrobium wilt.
The invention has the following beneficial effects:
the high-efficiency microbial agent suitable for the dendrobium wilt provided by the invention takes the mixture of the bursa of mossease fungal agent and the trichoderma harzianum fungal agent as the microbial agent, so that the probability of the dendrobium to get the wilt is effectively reduced.
The survival rate of dendrobium seedlings can be improved and the growth of the seedlings can be accelerated by endophytic fungi such as bursa of mochi fungi, trichoderma harzianum and the like, the problem that the symbiotic relationship is unbalanced due to overlarge bacterial content inoculated by a bacterial liquid is avoided after a microbial inoculum is prepared, and the dendrobium seedlings cannot be polluted due to the residue of a culture medium.
Meanwhile, the mixed fungicide obviously promotes the growth of the root system of the dendrobium seedling, and the growth promoting effect of the mosaicdou ductus vesiculosus fungicide and the trichoderma harzianum fungicide is superior to that of the mosaicdou ductus vesiculosus fungicide or the trichoderma harzianum fungicide which is inoculated independently by utilizing the synergistic effect of the mosaicdou ductus vesiculosus fungicide and the trichoderma harzianum fungicide, so that the synergistic effect of the mosaicdou ductus vesiculosus fungicide and the trichoderma harzianum fungicide in the aspect of preventing and treating dendrobium wilt is proved;
the microbial inoculum obtained by using the solid fermentation mode to obtain the trichoderma harzianum has high viable bacteria content and simple operation, and further improves the control effect of the dendrobium wilt.
Drawings
FIG. 1 is a graph showing the determination of the effective viable count of each microbial agent in an example of the highly effective microbial agent suitable for dendrobe blight of the present invention;
FIG. 2 is a graph showing the effect of each microbial agent on the total root length in an example of the highly effective microbial agent suitable for dendrobe blight of the present invention;
FIG. 3 is a graph showing the effect of each microbial agent on the total surface area of roots in an example of a highly effective microbial agent suitable for dendrobe wilt disease of the present invention;
FIG. 4 is a graph showing the effect of each microbial agent on the average diameter of roots in examples of the highly effective microbial agent suitable for dendrobe wilt of the present invention;
FIG. 5 is a graph showing the effect of each microbial agent on the total root volume of an example of a highly effective microbial agent suitable for dendrobe wilt disease of the present invention.
Detailed Description
It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention. The examples do not show specific techniques or conditions, the techniques or conditions are described in the literature in the field, or the specifications of the products, and the reagents or apparatuses used are not shown in the manufacturers, and the conventional products can be purchased from normal distributors.
In the prior art, Plant growth-promoting rhizobacteria (PGPR) are typical Plant root symbiotic microorganisms, and Plant growth-promoting rhizobacteria (PGPR) are soil microorganisms that have a combined action of stimulating Plant growth and inhibiting Plant pathogenic bacteria and the like in Plant rhizosphere life; the PGPR has the biological control effect on harmful pathogenic microorganisms and non-parasitic rhizosphere harmful microorganisms in soil, has the promotion effect on the absorption and utilization of mineral nutrition by plants, and can produce metabolites beneficial to the growth of the plants, thereby promoting the growth and development of the plants.
Arbuscular Mycorrhizal (AM) fungi are beneficial soil microorganisms that can infect most higher plants in the terrestrial ecosystem and form a "mycorrhizal" symbiotic structure, and are one of the important functional flora of the ecosystem. AM fungi, Trichoderma and plant root growth-promoting bacteria (PGPR) are typical plant root symbiotic microorganisms. The mosaicina paucicostata is one of mycorrhizal fungi.
Trichoderma (Trichoderma spp.) belongs to fungi kingdom, binuclear mycoderm, deuteromycotina, hyphomycetes, Moniliales and Moniliaceae, and is favored to grow on substrates rich in corrosive substances such as wood chips and barks, is a fungus with strong adaptability, high propagation speed and high biocontrol value, and has prevention and treatment effects on various plant diseases. Part of trichoderma can be colonized on the dendrobium tissue, and trichoderma harzianum, trichoderma atroviride, trichoderma viride and the like can generate symbiotic relation with plants.
By utilizing the synergistic effect of the branched fungi and the trichoderma, the blight control effect of the trichoderma moschatum (fusanloformis mossea) fungicide and the trichoderma harzianum fungicide is better than that of the trichoderma moschatum fungicide or the trichoderma harzianum fungicide which is inoculated independently, so that the synergistic effect of the trichoderma moschatum and the trichoderma harzianum is proved to exist in the aspect of dendrobium wilt.
Among them, Trichoderma harzianum (Trichoderma) and Tugassum mossambica (Fusneliformis mossea) are commercially available.
Preparation example 1
The bursa of Moxifrage is planted and propagated by corns in advance, the mixture of spores, extra-root hyphae and root soil of infected corn root segments of the bursa of Moxifrage is used as a bursa of Moxifrage microbial inoculum, and each gram of the bursa of Moxifrage microbial inoculum contains 25-35 spores. Specifically, the propagating step comprises:
takes the spores, hyphae and soil samples of infected plant root segments of the Muscosculus tussississimus as an initial inoculant and takes corn or Sudan grass as host plants.
Use the mixture of farmland soil and fertilizer as expanding numerous matrix, dry farmland soil and fertilizer respectively and cross 2mm sieve, sieve the back, with farmland soil and fertilizer according to volume ratio 2: 1, uniformly mixing, and sterilizing before or after mixing the farmland soil and the organic fertilizer to obtain the propagation expanding matrix. Wherein the sterilization method of the propagation matrix comprises steam sterilization at 121 ℃, sterilization for 1-2 hours, taking out and cooling to room temperature.
Adding water into the propagation matrix, wherein the adding amount of the water is 15-20% of the total mass of the propagation matrix and the initial inoculant; after the host plant is sowed and in the growth process, the host plant is managed conventionally to ensure the normal growth of the host plant, the host plant is cultured for 3-4 months, the overground part of the host plant is cut off, the cut root section is uniformly mixed with a substrate, and the air-dried substrate containing the root section of the host plant, the spores of the mycosphaerella pusilla and the hyphae outside the root is the single mycosphaerella pusilla fungicide.
Activating trichoderma harzianum, and inoculating the activated trichoderma harzianum to a trichoderma harzianum liquid seed culture medium for culture to obtain trichoderma harzianum fermented seed liquid; wherein, the spore concentration of the Trichoderma harzianum strain liquid is adjusted to 1 x 106Per ml;
weighing 51% of straw, 25.5% of wheat bran, 1.5% of rice bran, 21.5% of shell powder and 0.5% of (NH)4)2SO4Mixing to obtain fermented material, adding water to make water content of the fermented material reach 75%, stirring, packaging into polypropylene fungus bags, sterilizing at 121 deg.C for 20min, 150g per bag to obtain sterilized fermented material;
adding 5ml of Trichoderma harzianum fermentation seed liquid into the sterilized fermentation material, and fermenting at room temperature for 12 days; after culturing for 12 days, drying the fermentation product, separating the fermentation material from conidia by screening, collecting spore powder and storing at 4 ℃ to obtain a trichoderma harzianum microbial inoculum;
mixing the trichoderma harzianum microbial inoculum and the pipefish mossease microbial inoculum according to the ratio of 3:1, and using attapulgite as a carrier to obtain a microbial inoculum I suitable for the dendrobium wilt.
Preparation example 2
The preparation method is the same as that of preparation example 1, wherein the mosaicina paulownia microbial inoculum and the trichoderma harzianum microbial inoculum are mixed according to the mass ratio of 1: 5 to form mixed bacterial powder, namely the microbial agent II suitable for the dendrobium wilt. Wherein, the carrier of the microbial agent is attapulgite.
Preparation example 3
The preparation method is the same as that of preparation example 1, wherein the mosaicina paulownia microbial inoculum and the trichoderma harzianum microbial inoculum are mixed according to the mass ratio of 1: 4 to form mixed bacterial powder, namely the microbial agent III suitable for the dendrobium wilt. Wherein, the carrier of the microbial agent is attapulgite.
Preparation example 4
The bursa of Moxifrage is planted and propagated by corns in advance, the mixture of spores, extra-root hyphae and root soil of infected corn root segments of the bursa of Moxifrage is used as a bursa of Moxifrage microbial inoculum, and each gram of the bursa of Moxifrage microbial inoculum contains 25-35 spores. Specifically, the propagating step comprises:
takes the spores, hyphae and soil samples of infected plant root segments of the Muscosculus tussississimus as an initial inoculant and takes corn or Sudan grass as host plants.
Use the mixture of farmland soil and fertilizer as expanding numerous matrix, dry farmland soil and fertilizer respectively and cross 2mm sieve, sieve the back, with farmland soil and fertilizer according to volume ratio 2: 1, uniformly mixing, and sterilizing before or after mixing the farmland soil and the organic fertilizer to obtain the propagation expanding matrix. Wherein the sterilization method of the propagation matrix comprises steam sterilization at 121 ℃, sterilization for 1-2 hours, taking out and cooling to room temperature.
Adding water into the propagation matrix, wherein the adding amount of the water is 15-20% of the total mass of the propagation matrix and the initial inoculant; after the host plant is sowed and in the growth process, the host plant is managed conventionally to ensure the normal growth of the host plant, the host plant is cultured for 3-4 months, the overground part of the host plant is cut off, the cut root section is uniformly mixed with a substrate, and the air-dried substrate containing the root section of the host plant, the spores of the mycosphaerella pusilla and the hyphae outside the root is the single mycosphaerella pusilla fungicide.
And mixing the Moxidoutongguan mildew agent and the purchased Trichoderma harzianum wettable powder according to the mass ratio of 1:3 to form mixed bacterial powder, namely the microbial agent IV for the dendrobium wilt.
Effect example 1
And obtaining the four equivalent microbial agents, namely a microbial agent I, a microbial agent II, a microbial agent III and a microbial agent IV, wherein the sample numbers are 1, 2, 3 and 4, and counting the viable bacteria of the 4 complex microbial agents. And (3) repeating each microbial inoculum, and measuring the number of viable conidia in the microbial inoculum by adopting a dilution coating plate method, wherein the detection culture medium is a PDA (potato dextrose agar) culture medium. PDA culture medium: 200g of potato, 20g of glucose, 15g of agar and 1000ml of distilled water. The measurement results are shown in FIG. 1, and FIG. 1 shows the measurement of the effective viable cell count of each microbial agent.
The results of observing the figure 1 show that the effective viable count of the microbial agents I, II and III cultured by solid state fermentation is obviously increased compared with the microbial agent IV containing the Trichoderma harzianum wettable powder.
Example 1
The fertilizer efficiency test is carried out on dendrobium in the Cistus area of Puer city in 2019 from 4 months to 2019 and 8 months.
The test is carried out by setting 5 treatments, namely a treatment group (microbial agent is used for irrigating roots) and a control group (common pesticide is used for irrigating roots), wherein the area of each cell is 150m2Each treatment was repeated 3 times at a planting density of 45 plants/m2。
Treatment groups 1-4: for four microbial agents: numbering a first microbial agent, a second microbial agent, a third microbial agent and a fourth microbial agent to form a treatment group 1, a treatment group 2, a treatment group 3 and a treatment group 4;
irrigating roots of the dendrobium with high-content microbial inoculum 1-4 (adding 100g of high-content microbial inoculum into 50 kg/mu of water) in the middle growth stage;
control group 5: the soil fertilization is carried out according to the conventional fertilization of farmers (common pesticides are irrigated to roots), and the management measures are carried out according to the conventional requirements.
Sample collection method
In each processing cell, the area is randomly selected to be 1m2As a sample collection area.
Test results and analysis
The root system is the active absorption organ and synthesis organ of the plant, and the growth condition of the root is directly related to the nutrition level of the upper part of the plant. The shape of the root system of the dendrobium seedling is analyzed, and the result is as follows.
The influence of different treatments on the total root length, the total root surface area, the average root diameter and the total root volume of the dendrobium seedlings is shown in fig. 2-5, the growth promoting effect of different microbial inoculum treatments on dendrobium is shown in fig. 2-5, wherein fig. 2 shows the influence of each microbial inoculum on the total root length; FIG. 3 shows a graph of the effect of various microbial agents on the total surface area of roots; FIG. 4 shows a graph of the effect of various microbial agents on the mean diameter of roots; FIG. 5 shows a graph of the effect of each microbial agent on the total volume of roots.
As can be seen from fig. 2-5, the total root length, the total root surface area, the average root diameter and the total root volume of the dendrobium nobile seedlings treated by the microbial inoculum are all significantly higher than those of the control group 5, which indicates that the microbial inoculum is beneficial to promoting the growth of the dendrobium nobile root system; the treatment groups 1-3 are obviously higher than the treatment group 4, which shows that the growth promoting effect of the mixed microbial inoculum of the bursa of Moxifrage microbial inoculum and the trichoderma harzianum microbial inoculum cultured by solid state fermentation is higher than that of the microbial inoculum containing trichoderma harzianum wettable powder.
The healing effects of different treatments on dendrobium wilt disease are shown in table 1. 1-3 microbial agents I, II and III are used for irrigating the dendrobium plants, and the morbidity of dendrobium wilt is lower than 9%; the plants of dendrobium stem irrigated with common pesticide have 55 diseases, and the disease incidence rate is 40.7%. In addition, the morbidity of dendrobium wilt of the compound microbial inoculant in the treatment groups 1-3 is lower than 8.2%; the incidence rate of dendrobium wilt disease of the treatment group 4 (namely the treatment group contains the Trichoderma harzianum wettable powder) is higher than 8.8 percent, so that the growth promoting effect of the mixed microbial inoculum of the bursa of mosaicensis microbial inoculum and the Trichoderma harzianum microbial inoculum cultured by solid state fermentation is higher than that of the microbial inoculum containing the Trichoderma harzianum wettable powder.
TABLE 1 Effect of different treatments on Dendrobium wilt
Through the comprehensive comparison, the high-content microbial agent for dendrobium wilt disclosed by the invention is used for irrigating roots with the microbial agent in the growth period of dendrobium on the basis of local conventional fertilization, so that the effect of promoting the growth and development of dendrobium is remarkable, wherein the incidence rate of dendrobium wilt is reduced by 30%, and the high-content microbial agent has a certain popularization value.
Although the invention has been described in detail hereinabove by way of general description, specific embodiments and experiments, it will be apparent to those skilled in the art that many modifications and improvements can be made thereto based on the invention. Accordingly, such modifications and improvements do not depart from the spirit of the invention and are intended to be included within the scope of the invention.
Claims (4)
1. A preparation method of a high-efficiency microbial agent suitable for dendrobium wilt, which is characterized in that,
the method comprises the following steps of planting and breeding the trachelospermum moxidense through corns in advance, taking a mixture of spores, extra-root hyphae and root soil of infected corn root segments of the trachelospermum moxidense as a trachelospermum moxidense microbial inoculum, wherein each gram of the trachelospermum moxidense microbial inoculum contains 25-35 spores;
activating trichoderma harzianum, and inoculating the activated trichoderma harzianum to a trichoderma harzianum liquid seed culture medium for culture to obtain trichoderma harzianum fermented seed liquid; wherein, the spore concentration of the Trichoderma harzianum strain liquid is adjusted to 1 x 106Per ml;
weighing 51% of straw, 25.5% of wheat bran, 1.5% of rice bran, 21.5% of shell powder and 0.5% of (NH)4)2SO4Mixing to obtain fermented material, adding water to make water content of the fermented material reach 75%, stirring, packaging into polypropylene fungus bags, sterilizing at 121 deg.C for 20min, 150g per bag to obtain sterilized fermented material;
adding 5ml of Trichoderma harzianum fermentation seed liquid into the sterilized fermentation material, and fermenting at room temperature for 12 days; after culturing for 12 days, drying the fermentation product, separating the fermentation material from conidia by screening, collecting spore powder and storing at 4 ℃ to obtain a trichoderma harzianum microbial inoculum;
and mixing the trichoderma harzianum microbial inoculum with the moscilla bursa mycorrhiza microbial inoculum to obtain the high-efficiency microbial inoculum suitable for the dendrobium wilt.
2. The preparation method of the efficient microbial agent suitable for the dendrobium wilt according to claim 1, wherein in the microbial agent, the proportion of the trichoderma harzianum microbial agent to the pipewort mossease microbial agent in parts by weight on a dry basis is 3-5: 1.
3. the efficient microbial agent suitable for dendrobium wilt is characterized in that the number of effective viable bacteria of Moxidou Tunica and Trichoderma harzianum in the microbial agent is not less than 1 x 1011CFU/g。
4. The use of the highly effective microbial inoculant for controlling blight of dendrobium nobile lindl according to claim 3 in controlling blight of dendrobium nobile lindl.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116235867A (en) * | 2022-11-30 | 2023-06-09 | 乐施康农业科技有限公司 | Pathogen for preventing and treating cotton wilt and preparation method thereof |
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