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CN-122012246-A - Paenispira rosea MLY32, microbial inoculum, preparation method and application thereof

CN122012246ACN 122012246 ACN122012246 ACN 122012246ACN-122012246-A

Abstract

The invention discloses a strain of Paenispira rosea MLY32, a microbial inoculum, a preparation method and application thereof. The MLY32 strain is obtained by multi-generation co-culture domestication breeding of a wild type P.rosea strain CR032 separated from rhizosphere soil of a healthy tobacco strain in Guizhou Xingyi and a wild type P.nicotianae pathogenic strain PpN03, can be attached to a P.nicotianae hypha through parasitism and shows unique explosive growth, and after the strain is parasitized, the proliferation speed of the wild type P.nicotianae strain PpN03 is greatly slowed down, so that the MLY32 strain has strong bacteriostasis effect on the P.nicotianae, and the strain is obtained by co-culture of a non-pathogenic P.nicotianae MLY61 to obtain a bacterial agent P.nicotianae 3261, and still has strong parasitism and no pathogenic risk on the wild type P.nicotianae strain 3703. The invention has wide application potential in the field of agricultural biological control bactericides.

Inventors

  • YANG LEI
  • YI KE
  • XIANG DONG
  • WANG CONG
  • CHEN LIPING
  • CHENG ZHIJUN
  • XING YUNFEI
  • ZHOU CHANGJIN

Assignees

  • 湖南中烟工业有限责任公司

Dates

Publication Date
20260512
Application Date
20241108

Claims (10)

  1. 1. A strain of Paenispira rosea (Clonostachys rosea) MLY32 has a preservation number of CGMCC No.40347.
  2. 2. Use of the trichoderma reesei MLY32 according to claim 1 for inhibiting fungi, in particular phytophthora.
  3. 3. The use according to claim 2 for inhibiting phytophthora parasitica.
  4. 4. A phytophthora-inhibiting microbial agent is characterized by being prepared by culturing the Pachyspira rosea MLY32 according to claim 1.
  5. 5. The microbial inoculum according to claim 4, wherein the microbial inoculum is composed of the Saprolegnia rosea MLY32 and the non-pathogenic phytophthora parasitica (Phytophthora parasitica var. Nicotianae) MLY61, and the preservation number of the MLY61 is CGMCC No.40348.
  6. 6. The method for preparing the phytophthora-inhibiting microbial agent according to claim 4 or 5, wherein the leptospirillum roseum MLY32, the phytophthora parasitica MLY61 and auxiliary materials are mixed and cultured.
  7. 7. The method according to claim 6, wherein the auxiliary materials comprise at least one of bran, corn starch, corn flour, soybean flour, tapioca flour, and xanthan gum.
  8. 8. The method of manufacturing according to claim 6, comprising the steps of: Firstly, bran 50-150 g, wood dust 100-800 g, distilled water 0.1-1.0L, uniformly stirring, placing into a high-temperature sterilization pot at 120-135 ℃, sterilizing for 5-30 min, sterilizing and cooling; Step two, respectively cutting out small bacterial blocks with the length and width of 3.0-5.0 mm from a PDYH solid culture medium with a Paenibacillus roseus strain MLY32 and a Phytophthora nicotianae MLY61, respectively adding 5-20 small bacterial blocks into the solid fermentation materials in the step one of every 800 g, putting the solid fermentation materials in a constant-temperature incubator at 25-35 ℃, and fermenting and culturing for 15-60 days; The PDYH solid culture medium is prepared by dissolving potato powder 6.0 g, glucose 20.0 g, agar 20.0 g and yeast extract 6.0 g in 1000 mL distilled water and humic acid 50.0 g in 250 mL distilled water, sterilizing at high temperature, and mixing; thirdly, after fermentation is finished, taking 100-300 g of MLY32-61 solid fermentation materials, and mixing with 50-100 g of corn flour; fourthly, mixing 300-700 g tapioca flour with 150-400 mL boiling water at 100 ℃, and gelatinizing the tapioca flour to form dough; fifthly, mixing the mixed material in the third step with the cooled tapioca flour dough in the fourth step, and kneading and pressing the dough to wrap the mixed material; And sixthly, putting the dough which is wrapped with the mixed material and obtained in the fifth step into a pill, and extruding the pill into 0.5-10 cm granules to obtain the microbial inoculum.
  9. 9. The method of using a phytophthora inhibiting microbial agent according to claim 4 or 5, for inhibiting fungi, in particular phytophthora.
  10. 10. The use according to claim 9 for inhibiting phytophthora parasitica.

Description

Paenispira rosea MLY32, microbial inoculum, preparation method and application thereof Technical Field The invention relates to the field of agricultural biological control bactericides, in particular to a strain of Paenispira rosea MLY32, a bactericides, a preparation method and application thereof. Background Phytophthora nicotianae (Phytophthora nicotianae) is a pathogenic bacterium which seriously threatens crop health, has very wide hosts, can infect 255 plant genera of 90 plant families and can cause crop diseases such as crown rot, root rot, fruit rot, retardation, leaf rot, stem infection and the like according to statistics. It can survive in soil for a long period (4-6 years) even without host plants, and can reproduce through sexual and asexual reproduction with a reproduction period of 72 hours, and is a fungal pathogen that is difficult to treat. Paenispira rosea (Clonostachys rosea) is an endophytic filamentous fungus that is distributed throughout the world. In nature, it mainly acts as a saprophyte in soil, an epiphyte on plants, and exists as a branching parasitic fungus, exhibiting a wide range of nutritional plasticity. Because the plant growth-promoting agent has the capability of parasitizing and promoting plant growth, can generate various volatile organic compounds, has certain toxicity to other fungi, bacteria and insects, and has good application prospect in the fields of biological control, biodegradation and the like. In recent years, the solid fermentation technology of fungi is gradually applied to agricultural production, and as an environment-friendly pesticide preparation, the fungus can not only prevent and treat diseases, but also promote crop growth, increase yield and efficiency, improve soil health, facilitate sustainable development of agriculture and provide important guarantee for realizing green agriculture and ecological agriculture. The spore yield and disease resistance of the Saprolegnia rosea MLY32 can be obviously improved by culturing the Saprolegnia rosea MLY32 through a solid fermentation technology. In the solid fermentation process, proper solid matrixes (such as wood chips, bran and the like) and nutrition additives are selected, so that a good growth environment can be provided for the Saprolegnia rosea MLY32, and mass propagation and generation of metabolic active products are promoted. These active products, including antibiotics, enzymes and secondary metabolites, are effective in inhibiting phytophthora parasitica growth and infestation, protecting crops from disease attack. In addition, compared with a liquid fermentation method, the solid fermentation method is more economical and environment-friendly, and is suitable for large-scale production and application. The biological control effect of the Paenispira rosea MLY32 can be further improved by optimizing fermentation conditions such as temperature, humidity, ventilation and the like. However, parasitic fungi nutrition metabolism is often specific and there is a high risk of parasitic capacity degeneration after multiple generations of non-parasitic culture and expansion culture. This is also one of the main reasons for the instability of the activity and application of some parasitic agents. Co-cultivation with host pathogenic bacteria (Phytophthora nicotianae) is a good method for maintaining the parasitism of the Pachyrhizus crassipes, however, the finished product carries pathogenic Phytophthora nicotianae and presents a certain risk of pathogenicity during application. Therefore, if the non-pathogenic phytophthora parasitica and the trichosporon roseum can be used for co-culture, the parasitic activity can be effectively maintained, and the pathogenic risk can be avoided. The biological control agent prepared by utilizing the solid fermentation co-culture technology of the Paenibacillus roseus MLY32 and the non-pathogenic phytophthora parasitica MLY61 is used for controlling the phytophthora parasitica, so that the efficient parasitic activity of the MLY32 is maintained, the pathogenic risk of the phytophthora parasitica is avoided, and the technical product has wide application prospect and important practical significance. Disclosure of Invention The invention aims to provide a Paenispira rosea, a microbial inoculum, a preparation method and application thereof. As a branching parasitic fungus, the trichoderma reesei not only can generate secondary metabolites and degrading enzymes to directly participate in the control of pathogenic bacteria, but also can indirectly improve the survival ability of plants by inducing the systemic resistance of the plants, thereby being a biocontrol strain with great attention. If the biological control agent is prepared by utilizing the method, the method is in good agreement with the current sustainable development concept of green. The microbial inoculum is prepared by a low-cost high-yield solid fermentation method, and the solid fermentation material c