Antagonistic yeast Pichia deserticola and preparation and use method thereof

A Pichia pastoris and strain technology, applied in the field of Pichia pastoris, can solve the problems of lack of bacteriostatic spectrum strains, biocontrol effects are only verified on a few fruits, etc., achieving significant social and ecological benefits, low cost of use, good quality The effect of the application foreground

Active Publication Date: 2018-05-04
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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  • Antagonistic yeast Pichia deserticola and preparation and use method thereof
  • Antagonistic yeast Pichia deserticola and preparation and use method thereof
  • Antagonistic yeast Pichia deserticola and preparation and use method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1: Biological properties of Pichia deserticola strain BY25

[0018] 1. Morphological features

[0019] (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.

[0020] (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.

[0021] 2. Molecular biological identification

[0022] 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 datab...

Embodiment 2

[0024] The inhibitory effect of implementation example 2 Pichia BY25 to apple blue mold, botrytis cinerea and black spot

[0025] 1. Experimental protocol

[0026] Pichia BY25 was taken out from the -80°C refrigerator, activated 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), and picked a single Colony into YPD liquid medium, culture 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, count on a hemocytometer to prepare the concentration 1×10 8 cells / mL of Pichia BY25 suspension.

[0027] Activate Penicillium expansum, Botrytis cinerea or Alternaria tenuissima on PDA medium plate, culture at 26°C for 7-14 days, scrape appropriate amount of spores, wash with sterile water Prepared to a concentration of 5 x 10 4 cells / mL of Penicillium, Botrytis, or Alternar...

Embodiment 3

[0037] The inhibitory effect of implementation example 3 Pichia BY25 on pear fruit blue mold and black spot

[0038] 1. Experimental protocol

[0039] Pichia BY25 was taken out from the -80°C refrigerator, activated 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), and picked a single Colony into YPD liquid medium, culture 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, count on a hemocytometer to prepare the concentration 1×10 8 cells / mL of Pichia BY25 suspension.

[0040] 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 suspension. Healthy and undamaged pe...

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Abstract

The invention discloses a strain BY25 of Pichia deserticola and a use method and applications thereof. The strain has a wide antibacterial spectrum, is used to control diseases of picked vegetables and fruits, and has a stable effect. The strain is preserved in China General Microbiological Culture Collection Center, CGMCC, and the preservation number is CGMCC No. 14906. The use method of Pichia deserticola comprises following steps: activating strains, using YPD to carry out fermentation culture, carrying out centrifugation, preparing bacterial suspension (1*10<8> cells / mL) from bacteria andsterile water; soaking fruits and vegetables (the apple, pear, grape, strawberry, orange, cherry tomato, etc.) in the bacterial suspension for 30 seconds, drying fruits and vegetables in the air, placing the fruits and vegetables in a fresh-keeping box, and storing the fruits and vegetables at a room temperature. The Pichia deserticola strain can control blue mold, gray mold, and black spot of theapple, blue mode and gray mold of the pear fruit, gray mold, aspergillosis, black spot, pink rot, and phoma disease of the grape, grey mold and aspergillosis of the straw berry, grey mold of orange,and grey mold and aspergillosis of the cherry tomato, reduces the loss caused by diseases after picking, and has a good application prospect.

Description

technical field [0001] The invention relates to the field of biological control of postharvest diseases of fruits and vegetables, in particular to a strain of Pichia deserticola used for biological control of postharvest diseases of fruits and vegetables. The main postharvest diseases have significant control effect. Background technique [0002] Although the quality deterioration of fresh fruits and vegetables is affected by many factors, disease is the main reason. Among them, rot and deterioration caused by fungal diseases is the most serious factor in postharvest fruit loss. Although postharvest diseases of fruits and vegetables can be prevented and controlled in many ways such as agricultural control, physical control, chemical control and biological control, the main measure at present 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 but also red...

Claims

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Application Information

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IPC IPC(8): C12N1/16A23B7/155A01N63/04A61P3/00C12R1/84
CPCA23B7/155A23V2002/00A01N63/30C12N1/16C12N1/165C12R2001/84A23V2200/10
Inventor 王友升黄津津姚婷马琳任向峰李丽萍
Owner 山东凯普菲特生物科技有限公司
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