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Trichoderma koningiopsis ReTk1 bacterial strain and bacterial agent preparation method and application

A technology of Trichoderma pseudocorningii and bacterial strains, applied in the fields of botany equipment and methods, biochemical equipment and methods, applications, etc., can solve problems that have not been seen before, and achieve the effects of easy material collection, labor saving, and simple preparation process

Inactive Publication Date: 2015-01-14
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

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  • Trichoderma koningiopsis ReTk1 bacterial strain and bacterial agent preparation method and application
  • Trichoderma koningiopsis ReTk1 bacterial strain and bacterial agent preparation method and application
  • Trichoderma koningiopsis ReTk1 bacterial strain and bacterial agent preparation method and application

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Embodiment 1

[0048] A preparation method of Trichoderma pseudokoningeni ReTk1 bacterial strain, the steps are:

[0049] 1. Isolation location and isolation method of the strain:

[0050] Healthy rapeseed root tissues from diseased rapeseed fields in Zhijiang area, Hubei Province were collected, cleaned, soaked in 70% (volume ratio) ethanol for 1 min, soaked in 5% NaOCl for 5 min, and then washed 3 times with sterile water. Cut the roots into 0.5cm small pieces and put them on the PDA medium, culture them at 25°C for 14 days, dilute the grown fungal spores on the PDA plate, and perform single spore purification; obtain a fungal strain and number it as For ReTk1, its colony, spore peduncle and spore morphology are shown in figure 1 , further combined with ITS DNA sequence for identification.

[0051] 2. Strains of 16S rDNA Identification:

[0052] The above strain ReTk1 was inoculated on a PDA plate covered with cellophane, cultured at 28°C for 1 day, and the hyphae were scraped of...

Embodiment 2

[0061] The isolated bacterial strain of the present invention ReTk1 Endophytic Verification in Rapeseed Roots:

[0062] A verification of the endophyticity of Trichoderma pseudokoningeni ReTk1 strain: the RFP expression vector was constructed with pCAMBIA1301 as the backbone, the conidia were collected after culturing the Trichoderma pseudokoningeni ReTk1 strain on PDA for 5-7 days, and the transformation method mediated by Agrobacterium Transform Trichoderma pseudokoningeni ReTk1 strain with RFP vector, select transformants on PDA containing 50ug / mL hygromycin, and conduct a second round of screening on PDA containing 100ug / mL hygromycin, and finally select the one that can emit red fluorescence Strains with no change in biological performance ( figure 2 A). Inoculate 1×10 strains capable of emitting red fluorescence in 200 mL of PDB medium 6 The spores were cultured at 28°C and 200rpm for 15h, centrifuged at 4000rpm for 5min to collect the spores, and washed 3 times w...

Embodiment 3

[0064] A preparation method of biocontrol bacterium Trichoderma pseudokoningeni ReTk1 inoculum, the steps are:

[0065] A. The ReTk1 strain was cultured in PDA based on activation at 28°C for 3 days, and then inoculated on the PDA slant;

[0066] B. Cultivate again at 28°C for 5 days in step A, add sterilized deionized water, wash the spore suspension, add sterilized deionized water, and adjust the concentration of spore liquid to 5×10 5 Spores / mL.

[0067] C. Add 1mL of spore liquid to 250mL of PDB medium, culture at 180rpm and 25°C for 6 days, and the concentration can reach 1×10 9 Spores / mL, the spores were collected to prepare a spore suspension preparation (microbial agent).

[0068] The components and proportioning of the PDA and PDB medium in the above steps are as follows:

[0069] PDA medium: 200 g of potatoes, 20 g of glucose, 10 g of agar, add distilled water to 1000 mL, adjust the pH to 7.0, and sterilize at 121°C for 30 minutes;

[0070] PDB medium: 200 g of p...

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Abstract

The invention discloses a trichoderma koningiopsis ReTk1 bacterial strain and a bacterial agent preparation method and an application thereof. The preparation method comprises the following steps: A, activating and culturing a ReTv2 bacterial strain in a PDA culture medium at 28DEG C for three days; B, then inoculating the ReTv2 bacterial strain on a PDA bevel, culturing at 28 DEG C for five days, and adding sterile deionized water; and C, adjusting the concentration of a spore solution to 5*10<5> spores / mL, adding 1mL of spore solution into 250mL of a PDB culture medium, culturing for six days under the condition of 180rpm and 25 DEG C, and collecting spores to prepare a spore suspension preparation. The invention discloses the application of the trichoderma koningiopsis ReTk1 bacterial agent to promoting oilseed rape growth and preparing medicines for treating or preventing plasmodiophora brassicae of oilseed rape as well as the application of the trichoderma koningiopsis ReTk1 bacterial strain on germination inhibition of resting spores of plasmodiophora brassicae of oilseed rape in a fermentation filtrate. The bacterial strain has a remarkable growth-promoting effect to oilseed rape and meanwhile has a remarkable preventing and treating effect on plasmodiophora brassicae of oilseed rape. The control effect of the biological agent on plasmodiophora brassicae of oilseed rape under an indoor cultivation condition reaches 55.2%, and the bacterial strain can be directly applied during seeding. The bacterial strain disclosed by the invention is simple in application method and labor force-saving and is safe to human and livestock serving as a biopesticide.

Description

technical field [0001] The invention belongs to the technical field of biological control of plant diseases, and more specifically relates to a bacterial strain Trichoderma pseudocorningi ReTk1 used for the biological control of rapeseed clubroot. The preparation method of the fungus ReTk1 strain also relates to the application of a Trichoderma pseudokoningeni ReTk1 strain in the preparation of medicines for treating or preventing clubroot of cruciferous plants such as rapeseed. Background technique [0002] Rapeseed is one of the important oil crops in my country, and it is threatened by various pathogenic bacteria in the production process of rapeseed. Clubroot of rapeseed is caused by Plasmobacterium brassicae ( Plasmodiophora brassicae) An important disease caused by this disease, which mainly damages the host root and causes parenchyma cells in the root to proliferate and form tumors. The disease also harms other cruciferous plants. At present, there is no effective c...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12N3/00A01N63/04A01P21/00A01P3/00C12R1/885
CPCA01N63/30C12N3/00C12N1/145C12R2001/885
Inventor 程家森穆罕默德·哈森王庆庆姜道宏付艳苹谢甲涛
Owner HUAZHONG AGRI UNIV
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