阅读说明：本技术 一种复配生防菌剂及其制备方法与应用 (A kind of compounding biocontrol agent and the preparation method and application thereof ) 是由 王傲雪 王慧 刘佳音 张瑶 陈秀玲 冯明芳 仇有文 于 2019-05-29 设计创作，主要内容包括：本发明涉及一种复配生防菌剂及其制备方法与应用,属于微生物技术领域。为解决单一菌种防效不稳定,抑菌谱范围窄的问题,本发明提供了一种复配生防菌剂,包括一定体积比的哈茨木霉菌悬液和粉红粘帚菌菌悬液,所述哈茨木霉菌悬液和粉红粘帚菌菌悬液的活菌数均为10<Sup>6</Sup>cfu/mL。本发明提供的复配生防菌剂抑菌谱广、拮抗作用强、持效期长、生产成本低、发酵工艺简单,将其应用于农业生物防治方面,可防治灰霉病菌、茎腐病菌、立枯病菌、炭疽病菌、镰刀菌和枯萎病菌等真菌引起的作物病害；其对农作物种子及幼苗具有促生作用,可促进种子萌发、胚根和幼苗生长,还具有防腐保鲜的作用,将其应用于果蔬防腐保鲜能够降低腐烂率和失重。(The present invention relates to a kind of compounding biocontrol agents and the preparation method and application thereof, belong to microorganisms technical field.It is unstable to solve single culture preventive effect, the problem of antimicrobial spectrum narrow range, the present invention provides a kind of compounding biocontrol agents, Trichoderma harzianum bacteria suspension and Parasitism broom bacterium bacteria suspension including certain volume ratio, and the viable count of the Trichoderma harzianum bacteria suspension and Parasitism broom bacterium bacteria suspension is 10 6 cfu/mL.Compounding biocontrol agent antimicrobial spectrum provided by the invention is wide, antagonism is strong, the lasting period is long, production cost is low, zymotechnique is simple, in terms of being applied to Strategies of Agricultural Bio-control, the fungus-caused crop disease such as ash arrhizus bacteria, Pathogen, rhizoctonia solani, anthrax bacteria, sickle-like bacteria and wilt can be prevented and treated；It has growth-promoting functions to crop seeds and seedling, can promote seed sprouting, radicle and growth of seedling, also has the function of anti-corrosive fresh-keeping, being applied to antiseptic preservation of fruits and vegetables can reduce rotting rate and weightlessness.)

1. a kind of compounding biocontrol agent, which is characterized in that Trichoderma harzianum bacteria suspension and Parasitism broom bacterium including certain volume ratio The viable count of bacteria suspension, the Trichoderma harzianum bacteria suspension and Parasitism broom bacterium bacteria suspension is 10⁶cfu/mL。

2. a kind of compounding biocontrol agent according to claim 1, which is characterized in that the Trichoderma harzianum bacteria suspension and Parasitism The volume ratio of broom bacterium bacteria suspension is 0.5~1.5:1.

3. a kind of preparation method for compounding biocontrol agent, which comprises the steps of:

Step 1: taking wheat bran, rice chaff, wheat according to certain mass than preparing solid fermentation culture medium, steam pressure sterilizing；

Step 2: Trichoderma harzianum fermentation seed liquid or Parasitism broom bacterium fermentation seed liquid to be inoculated in the sterilizing of step 1 preparation In solid fermentation culture medium, room temperature is fermented 15 days, keeps primary to solid fermentation culture medium horizontal oscillations daily；

It tiles Step 3: step 2 is completed fermentation gained fermentation material into sterile tray, is covered on fermentation material with aseptic filter paper On, the fermentation material in supporting plate is stirred daily, continues 10 days, is completely dried to the fermentation material in pallet, to fermentation material into Row crushes, sieving obtains trichoderma harzianum powder or Parasitism broom bacterium bacterium powder, takes trichoderma harzianum powder or Parasitism broom bacterium bacterium respectively Powder and sterile water mixed preparing contain the Trichoderma harzianum bacteria suspension or Parasitism broom bacterium bacteria suspension of certain viable count, by certain volume It obtains compounding biocontrol agent than mixing Trichoderma harzianum bacteria suspension with Parasitism broom bacterium bacteria suspension.

4. a kind of preparation method for compounding biocontrol agent according to claim 3, which is characterized in that wheat bran described in step 1, Rice chaff, wheat mass ratio be 1.67:1:7.23；The initial water content of the solid fermentation culture medium is 75%；The steam The temperature of pressure sterilization is 121 DEG C, pressure 20psig, sterilization time 30min, repeats sterilizing 3 times；Thatch is breathed out described in step 2 The inoculum concentration of trichoderma fermentation seed liquid or Parasitism broom bacterium fermentation seed liquid is 3mL/40g.

5. according to a kind of preparation method for compounding biocontrol agent of claim 3 or 4, which is characterized in that sent out described in step 3 It is 200 mesh bacterium powder that ferment object, which crushes gained bacterium powder, and viable count contained by the Trichoderma harzianum bacteria suspension or Parasitism broom bacterium bacteria suspension is 10⁶The volume ratio of cfu/mL, the Trichoderma harzianum bacteria suspension and Parasitism broom bacterium bacteria suspension is 0.5~1.5:1.

6. a kind of application of compounding biocontrol agent as claimed in claim 1 or 2 in terms of Strategies of Agricultural Bio-control.

7. application of a kind of compounding biocontrol agent in terms of Strategies of Agricultural Bio-control according to claim 6, which is characterized in that packet Include the prevention and treatment to ash arrhizus bacteria, Pathogen, rhizoctonia solani, anthrax bacteria, sickle-like bacteria and wilt.

8. application of the compounding biocontrol agent according to claim 6 or 7 in terms of Strategies of Agricultural Bio-control, which is characterized in that Including the application in terms of improving agro-ecology disease resistance.

9. a kind of application of compounding biocontrol agent as claimed in claim 1 or 2 in terms of crop seeds and seedling growth-promoting.

10. a kind of application of compounding biocontrol agent a kind of as claimed in claim 1 or 2 in terms of fruit and vegetable anti-corrosive fresh-keeping.

Technical field

The invention belongs to microorganisms technical fields more particularly to a kind of compounding biocontrol agent and the preparation method and application thereof.

Background technique

Gray mold is prevented and treated in agricultural production at present still takes the Preventing Countermeasures based on chemical prevention.Common fungicide is main Including benzimidazole class, dicarboximide class, N- carbanilate class, anilino-pyrimidine and protective fungicide.This A little chemical bactericide mechanisms of action are different, and wherein benzimidazole is by the tubulin phase with graw mold of tomato germ In conjunction with, its function is destroyed, thus inhibit the separation and elongation of germ mycelia, the mitosis and morphogenesis of block cell；And The mechanism of action of dicarboximide fungicide is currently still not clear, it has been reported that it may be by the cell function of interference germ Can, influence membrane permeability makes cell rupture and realizes.Although above-mentioned chemical mechanism control efficiency is considerable, due to gray mold germ fauna Genetically there is heterocaryosis, there are diversity in population is also easy to produce resistant strain under the action of medicament natural selection, Gradually decline so as to cause the using effect of medicament, even forfeiture curative effect.Though thus the short-term preventive effect of chemical prevention is obvious, long Phase application chemical pesticide inherently makes Quality of Tomato Fruit decline, pathogen develop drug resistance, seriously pollutes environment, jeopardizes people and animals Health, or even destroy Agro-ecological System.

But the biocontrol agent majority for graw mold of tomato of current research is made of based on single culture, actual Phenomena such as unstable preventive effect, antimicrobial spectrum narrow range are frequently present of in production application.

Summary of the invention

To solve the problem of the unstable antimicrobial spectrum narrow range of single culture preventive effect, the present invention provides a kind of compounding biological and ecological methods to prevent plant disease, pests, and erosions Microbial inoculum and the preparation method and application thereof.

Technical solution of the present invention:

A kind of compounding biocontrol agent, Trichoderma harzianum bacteria suspension and Parasitism broom bacterium bacteria suspension including certain volume ratio, institute The viable count for stating Trichoderma harzianum bacteria suspension and Parasitism broom bacterium bacteria suspension is 10⁶cfu/mL。

Further, the volume ratio of the Trichoderma harzianum bacteria suspension and Parasitism broom bacterium bacteria suspension is 0.5~1.5:1.

A kind of preparation method compounding biocontrol agent, includes the following steps:

Step 1: taking wheat bran, rice chaff, wheat according to certain mass than preparing solid fermentation culture medium, steam pressure sterilizing；

Step 2: Trichoderma harzianum fermentation seed liquid or Parasitism broom bacterium fermentation seed liquid have been inoculated in step 1 preparation It sterilizes in solid fermentation culture medium, room temperature is fermented 15 days, keeps primary to solid fermentation culture medium horizontal oscillations daily；

It tiles Step 3: step 2 is completed fermentation gained fermentation material into sterile tray, is covered on hair with aseptic filter paper On ferment object, the fermentation material in supporting plate is stirred daily, continues 10 days, is completely dried to the fermentation material in pallet, to fermentation Object is crushed, is sieved to obtain trichoderma harzianum powder or Parasitism broom bacterium bacterium powder, takes trichoderma harzianum powder or Parasitism broom respectively Bacterium bacterium powder and sterile water mixed preparing contain the Trichoderma harzianum bacteria suspension or Parasitism broom bacterium bacteria suspension of certain viable count, by certain Trichoderma harzianum bacteria suspension is mixed with Parasitism broom bacterium bacteria suspension and obtains compounding biocontrol agent by volume ratio.

Further, wheat bran, rice chaff described in step 1, wheat mass ratio be 1.67:1:7.23；The solid fermentation training The initial water content for supporting base is 75%；The temperature of the steam pressure sterilizing is 121 DEG C, pressure 20psig, and sterilization time is 30min repeats sterilizing 3 times；The inoculum concentration of Trichoderma harzianum fermentation seed liquid described in step 2 or Parasitism broom bacterium fermentation seed liquid For 3mL/40g.

Further, fermentation material described in step 3 crush gained bacterium powder be 200 mesh bacterium powder, the Trichoderma harzianum bacteria suspension or Viable count contained by Parasitism broom bacterium bacteria suspension is 10⁶Cfu/mL, the Trichoderma harzianum bacteria suspension and Parasitism broom bacterium bacteria suspension Volume ratio is 0.5~1.5:1.

A kind of application of the compounding biocontrol agent provided by the invention in terms of Strategies of Agricultural Bio-control.

Further, including to ash arrhizus bacteria, Pathogen, rhizoctonia solani, anthrax bacteria, sickle-like bacteria and wilt Prevention and treatment.

Further, including the application in terms of improving agro-ecology disease resistance.

A kind of application of the compounding biocontrol agent provided by the invention in terms of crop seeds and seedling growth-promoting.

A kind of application of the compounding biocontrol agent provided by the invention in terms of fruit and vegetable anti-corrosive fresh-keeping.

Beneficial effects of the present invention:

The present invention mutually compounds Trichoderma harzianum with two kinds of biocontrol fungis of Parasitism broom bacterium, obtains that a kind of antimicrobial spectrum is wide, antagonism Effect is strong, the lasting period is long, production cost is low, the compounding biocontrol agent of the simple compound biocontrol fungicide of zymotechnique, can be extensive Applied to Strategies of Agricultural Bio-control field, ash arrhizus bacteria, Pathogen, rhizoctonia solani, anthrax bacteria, sickle-like bacteria can be prevented and treated and withered The fungus-caused crop disease such as germ.

Compounding biocontrol agent provided by the invention is not only able to effectively inhibit the growth of crop disease fungal pathogens, moreover it is possible to promote Make the more salicylic acids of induction biosynthesis, to enhance the disease resistance of plant, can be obtained more comprehensively from different prevention and treatment mechanism Efficient control efficiency, prevention and treatment range are wider.

Compounding biocontrol agent provided by the invention has growth-promoting functions to crop seeds and seedling, and seed base-root can be improved Increment improves Seedling Height and stem thickness.Meanwhile compounding biocontrol microorganisms machine also has the function of anti-corrosive fresh-keeping, is applied to fruit Vegetable reduces rotting rate and weightlessness.

Detailed description of the invention

Fig. 1 is the growing state picture that 3 different disposal method of experimental example acts on lower tomato seedling, in figure from left to right successively For Parasitism broom bacterium biocontrol agent processing group, Trichoderma harzianum biocontrol agent processing group, compounding biocontrol agent processing group, apply a times strong place Reason group and clear water processing group；

Fig. 2 is that experimental example 8 first applies pair that biocontrol microorganisms inoculate PRO contained by the tomato leaf that graw mold of tomato is handled Than figure；

Fig. 3 is pair that the first inoculating tomato gray mold of experimental example 8 applies PRO contained by the tomato leaf that biocontrol microorganisms are handled again Than figure；

Fig. 4 is that experimental example 8 first applies pair that biocontrol microorganisms inoculate POD contained by the tomato leaf that graw mold of tomato is handled Than figure；

Fig. 5 is pair that the first inoculating tomato gray mold of experimental example 8 applies POD contained by the tomato leaf that biocontrol microorganisms are handled again Than figure；

Fig. 6 is that experimental example 8 first applies pair that biocontrol microorganisms inoculate MDA contained by the tomato leaf that graw mold of tomato is handled Than figure；

Fig. 7 is pair that the first inoculating tomato gray mold of experimental example 8 applies MDA contained by the tomato leaf that biocontrol microorganisms are handled again Than figure；

Fig. 8 is that experimental example 8 first applies pair that biocontrol microorganisms inoculate APX contained by the tomato leaf that graw mold of tomato is handled Than figure；

Fig. 9 is pair that the first inoculating tomato gray mold of experimental example 8 applies APX contained by the tomato leaf that biocontrol microorganisms are handled again Than figure；

Figure 10 first applies biocontrol microorganisms for experimental example 8 and inoculates SOD's contained by the tomato leaf that graw mold of tomato is handled Comparison diagram；

Figure 11 is that the first inoculating tomato gray mold of experimental example 8 applies SOD's contained by the tomato leaf that biocontrol microorganisms are handled again Comparison diagram；

Figure 12 is the comparison diagram that 9 different disposal method of experimental example acts on lower tomato seedling salicylic acid content.

Specific embodiment

Below with reference to embodiment, the following further describes the technical solution of the present invention, and however, it is not limited to this, all right Technical solution of the present invention is modified or replaced equivalently, and without departing from the spirit and scope of the technical solution of the present invention, should all be contained Lid is within the protection scope of the present invention.