Antagonistic yeast Pichia galeiformis as well as preparation and application method thereof

A technology of Pichia pastoris and strains, which is applied in the field of Pichia pastoris, can solve the problems of lack of antibacterial spectrum strains, and the biocontrol effect is only verified on a few fruits, so as to achieve significant social and ecological benefits and avoid harm to people , easy to cultivate effect

Active Publication Date: 2018-04-20
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

Method used

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  • Antagonistic yeast Pichia galeiformis as well as preparation and application method thereof
  • Antagonistic yeast Pichia galeiformis as well as preparation and application method thereof
  • Antagonistic yeast Pichia galeiformis as well as preparation and application method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1: Biological properties of Pichia galeiformis strain BY27

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

Embodiment 2

[0024] The inhibitory effect of implementation example 2 Pichia BY27 on apple blue mold, botrytis cinerea and black spot

[0025] 1. Experimental protocol

[0026] Pichia BY27 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 BY27 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 BY27 on pear fruit blue mold and black spot

[0038] 1. Experimental protocol

[0039] Pichia BY27 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 BY27 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 Pichia galeiformis bacterial strain BY27 used for control of postharvest diseases of fruits and vegetables and with broad antibacterial spectrum and stable effect and an application method thereof. The bacterial strain is preserved in CGMCC with the preservation number of CGMCC No.14907. The application method of the pichia pastoris comprises the following steps: activating the bacterial strain, carrying out fermental cultivation with YPD, centrifuging, and preparing bacterium suspension with the concentration of 1*10<8> cells/mL from thalli with sterile water; putting fruits and vegetables such as apples, pears, grapes, strawberries, citrus and cherry tomatoes into the bacterium suspension, soaking for 30 seconds, then taking out, and carrying out air drying; andputting into a preservation box, and storing at room temperature. The Pichia galeiformis bacterial strain disclosed by the invention can control penicilliosis, botrytis and black spot of the apples,penicilliosis and botrytis of pear fruits, botrytis, aspergillosis, black spot, fusariosis and cephalothecium roseum of the grapes, botrytis and aspergillosis of the strawberries, penicilliosis of thecitrus as well as botrytis and aspergillosis of the cherry tomatoes at the same time, and loss caused by the postharvest diseases is reduced, so that the Pichia galeiformis bacterial strain disclosedby the invention has a good application prospect.

Description

technical field [0001] The invention relates to the field of biological control of post-harvest diseases of fruits and vegetables, in particular to a strain of Pichia galeiformis used for the biological control of post-harvest diseases of fruits and vegetables. The main postharvest diseases have significant control effects. 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 a...

Claims

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Application Information

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IPC IPC(8): C12N1/16A23B7/155C12R1/84
CPCA23B7/155C12N1/16C12N1/165C12R2001/84
Inventor 王友升黄津津王颖任向峰姚婷李丽萍
Owner 山东凯普菲特生物科技有限公司
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