Fungus cross detoxification method and application thereof
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A technology of fungi and strains, applied in the fungal cross detoxification method and its application field, can solve the problems of long detoxification cycle and weak strain activity.
Pending Publication Date: 2022-03-01
HUAZHONG AGRI UNIV
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The fungal cross detoxification method provided by the present invention can achieve the effect of removing multiple viruses at the same time and / or can complete the detoxification and rejuvenation work within two weeks, thus solving the problem that the existing fungal detoxification technology generally can only remove a single virus , long detoxification period or the technical problem that the activity of the strain is not strong after detoxification
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Embodiment 1
[0057] Embodiment 1: Cross-detoxification result of Lentinus edodes Z28 bacterial strain
Embodiment 2
[0058] Embodiment 2: cross detoxification result of Pleurotus ostreatus T2 bacterial strain
[0059] The cross detoxification of test bacterial strain in the above embodiment all carries out according to the following steps:
[0060] (1) Detoxification of test strains
[0061] S1 Configure the medium containing ribavirin: take the MYG medium without adding chemical reagents as the blank control, configure the MYG medium containing ribavirin, and the configuration steps are as follows:
[0062] ①Weigh 0.02g ribavirin, dissolve in 1ml ddH 2 O, prepared as a mother liquor.
[0063] ② The ribavirin solution obtained in ① was filtered through a bacterial filter and dissolved in 200 ml of MYG solid medium to obtain a MYG medium containing 100 μg / mL ribavirin.
[0064] S2 Prepare 20mg / mL lysozyme solution: weigh 0.02g lysozyme, dissolve in 1ml of 0.6M mannitol solution, and use a bacterial filter to filter and sterilize.
[0065] (1) Cross-detoxification of the test strain in S3:...
Embodiment 3
[0073] Example 3 Verifies the detoxification effect in the detoxification regeneration strain Z28
[0074] Part of the detoxified strains in the detoxified and regenerated Z28 strains obtained in Example 1 were selected for qRT-PCR. The results of the changes in the expression of the Z28 strain LePV1 before and after detoxification are shown in image 3 A, the results of the changes in the expression level of the Z28 strain LeV-HKB before and after detoxification are shown in image 3 b.
[0075] Depend on image 3 The test results showed that in the Z28 virus-free strain, LePV1 was not detected in VC3, VC4, and VC5, and LeV-HKB was not detected in VC1, VC2, and VC5. Screening requirements for virus-free regenerating strains.
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Abstract
The invention discloses a fungus cross detoxification method, which comprises the following steps of: obtaining a part of fungus tissues which can be cultured into a complete individual under the condition that a to-be-detoxified fungus strain in a development state acts for a long enough time under the action of a chemical reagent with an effective concentration and the virus carried by the strain is in a proliferation inhibition state, and detecting and screening a non-toxic strain after regeneration culture; the fungal tissue is a fungal part containing at least one whole genome copy number; the fungi in the development state are in the development state before the sporocarp of the fungi; the chemical reagent comprises one or a combination of more of 5-dihydrouracil DHT, virazole, actinomycete ketone, rifampicin and moroxydine hydrochloride, the fungal tissue comprises one of protoplast, hypha tip and hypha fragment, the fungus cross detoxification method provided by the invention can remove various viruses in the same strain, the detoxification period is short, the rejuvenation effect is good, and the method is suitable for industrial production. The method is also suitable for removing unknown viruses, and can be applied to strain degeneration caused by virus infection and / or preparation of strains with strong rejuvenation.
Description
technical field [0001] The invention belongs to the technical field of virus detoxification methods, and more specifically relates to a fungal cross-detoxification method and its application. Background technique [0002] At present, mycoviruses have been found in dozens of edible fungi, and most of the mycoviruses carried in edible fungi are combined infections. 23. [0003] Existing studies on mycoviruses are mainly focused on the detection of viruses and the discovery of new viruses, while there are few studies on detoxification of mycoviruses. Fungal virions mostly exist in spherical shape, and a few of them are club-shaped, linear and rod-shaped. It can be (+) ssRNA or (-) ssRNA. Viruses are different in shape, size and genome, and the same virus will also be different in different hosts or in the same host. The diversity of mycoviruses brings many difficulties to fungal detoxification, which is an aspect of mycovirus detoxification research that is less reported. I...
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