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Application of a wine strain of Hansenula spp. in the control of postharvest fruit diseases

A technology of wine and yeast, applied in application, preservation of fruits and vegetables, and methods based on microorganisms, etc., can solve the problems of lack of antibacterial spectrum strains, biocontrol effects are only verified on a few fruits, and achieve significant social and ecological benefits , avoid harm to people, and have a wide antibacterial spectrum effect

Active Publication Date: 2021-03-05
山东凯普菲特生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, although there are nearly a hundred species of antagonistic yeasts reported at home and abroad, the biocontrol effects of most antagonistic yeasts have only been verified on a few fruits.
However, since the biocontrol effects of different strains of the same yeast are very different (Filonow et al., 1996), most of the antagonistic yeasts lack strains with broad antibacterial spectrum and stable effect.

Method used

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  • Application of a wine strain of Hansenula spp. in the control of postharvest fruit diseases
  • Application of a wine strain of Hansenula spp. in the control of postharvest fruit diseases
  • Application of a wine strain of Hansenula spp. in the control of postharvest fruit diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1: Biological characteristics of Hanseniaspora vineae BY19 1. Morphological characteristics

[0020] (1) YPDA medium (1% yeast extract powder, 2% peptone, 2% glucose, 1.8% agar, sterilized at 121°C for 20 minutes) was cultured at 26°C for 48h, and the colonies were round and white with smooth and round edges. The cell shape is ellipsoidal.

[0021] (2) After culturing in YPDA liquid medium for 24 hours, no mold was formed, the bacterial solution was turbid, and there was precipitation. Microscopically, the yeast cells were oval and budded.

[0022] 2. Molecular biological identification

[0023] Use the universal forward primer NL-1 (5'-GCATATCAATAAGCGGAGGAAAAG-3') and the reverse primer NL-4 (5'-GGTCCGTGTTTCAAGACGG-3') to PCR amplify the yeast 26S rDNA D1 / D2 region nucleic acid sequence, and PCR The sequencing results of the product were entered into the website www.NCBI.nlm.nih.gov for BLAST, the homologous sequences were downloaded from the GenBank database...

Embodiment 2

[0024] Example 2 Inhibitory effect of Hansella sporogenes BY19 on apple penicillium and botrytis cinerea

[0025] 1. Experimental protocol

[0026] Take Han. spp. BY19 out of the refrigerator at -80°C, activate it with YPDA medium (10g of yeast extract powder, 20g of peptone, 20g of glucose, 18g of agar, 1000ml of deionized water, natural pH, sterilized at 121°C for 30min), Pick a single colony into the YPD liquid medium, culture it at 26°C and 200r / min for 24h, centrifuge at 4000rpm for 5min, discard the supernatant, wash the collected bacteria repeatedly with sterile water for 3 times, and count on a hemocytometer to prepare Prepared to a concentration of 1 x 10 8 Cells / mL of Hansula sporogenes BY19 suspension in wine.

[0027] Activate Penicillium expansum or Botrytis cinerea on a PDA medium plate, culture at 26°C for 7-14 days, scrape appropriate amount of spores, and prepare a concentration of 5×10 with sterile water. 4 cells / mL of Penicillium or Botrytis cinerea spore...

Embodiment 3

[0035] Example 3 Inhibitory effect of Hansspora yeast BY19 on pear fruit blue mold and gray mold

[0036] 1. Experimental protocol

[0037] Take Han. spp. BY19 out of the refrigerator at -80°C, activate it with YPDA medium (10g of yeast extract powder, 20g of peptone, 20g of glucose, 18g of agar, 1000ml of deionized water, natural pH, sterilized at 121°C for 30min), Pick a single colony into the YPD liquid medium, culture it at 26°C and 200r / min for 24h, centrifuge at 4000rpm for 5min, discard the supernatant, wash the collected bacteria repeatedly with sterile water for 3 times, and count on a hemocytometer to prepare Prepared to a concentration of 1 x 10 8 Cells / mL of Hansula sporogenes BY19 suspension in wine.

[0038] Activate Penicillium expansum or Botrytis cinerea on a PDA medium plate, culture at 26°C for 7-14 days, scrape appropriate amount of spores, and prepare a concentration of 5×10 with sterile water. 4 cells / mL of Penicillium or Botrytis cinerea spore suspens...

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Abstract

The invention discloses Hanseniaspora vineae BY19, a wine spore yeast Hanseniaspora vineae BY19 with wide antibacterial spectrum and stable effect, which is used for the prevention and treatment of post-harvest diseases of fruits and vegetables and a use method thereof. The preservation strain number of the strain in the General Microbiology Center of the China Committee for Culture Collection of Microorganisms is CGMCC No.14903. The method of using the wine Hansenula spp.: activate the strain, ferment and cultivate it with YPD, centrifuge, and mix the bacteria with sterile water to make 1×10 8 cells / mL bacterial suspension; put apples, pears, grapes, strawberries, citrus or cherry tomatoes and other fruits and vegetables into the bacterial suspension, soak for 30 seconds, take out, and air-dry; put them in a fresh-keeping box and store at room temperature. The wine Hansenula spp. strain simultaneously controls apple Penicillium and Botrytis cinerea, pear Penicillium and Botrytis cinerea, grapevine Botrytis, Aspergillus, black spot, Fusarium and anthracnose, strawberry Botrytis cinerea, citrus penicillium, and tomato cinerea and aspergillosis can reduce losses caused by postharvest diseases and have good application prospects.

Description

technical field [0001] The invention relates to the field of biological control of fruit postharvest diseases, in particular to a strain of Hanseniaspora vineae used for biological control of fruit postharvest diseases. The main postharvest diseases of the female fruit have significant control effects. Background technique [0002] The quality deterioration of fresh fruit products is affected by many factors, but the rot and deterioration caused by diseases, especially fungal diseases, is the most serious factor in the postharvest loss of fruits. Although postharvest diseases of fruits can be controlled through many methods such as agricultural control, physical control, chemical control and biological control, the main measure to control postharvest diseases of fruits is chemical control (Eckert & Ogawa, 1985, 1988). However, long-term use of chemical pesticides not only leads to the development of drug resistance of pathogenic bacteria and reduces the bactericidal effect ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/16A23B7/155C12R1/645
CPCA23B7/155C12N1/16C12N1/145C12R2001/645
Inventor 王友升姚婷陈滢黄津津
Owner 山东凯普菲特生物科技有限公司