Pichia cactophila capable of effectively controlling fruit postharvest diseases as well as preparation and using method of pichia cactophila

A technology for postharvest fruit diseases and Pichia pastoris, applied in botanical equipment and methods, microbial-based methods, biochemical equipment and methods, etc., can solve the problem of lack of antibacterial spectrum strains, and the biocontrol effect is only on a small number of fruits Obtain verification and other issues, achieve significant social and ecological benefits, avoid harm to people, and have good application prospects

Active Publication Date: 2018-06-05
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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  • Pichia cactophila capable of effectively controlling fruit postharvest diseases as well as preparation and using method of pichia cactophila
  • Pichia cactophila capable of effectively controlling fruit postharvest diseases as well as preparation and using method of pichia cactophila
  • Pichia cactophila capable of effectively controlling fruit postharvest diseases as well as preparation and using method of pichia cactophila

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Embodiment 1: Biological characteristics of Pichia plesiosae BY35

[0017] 1. Morphological characteristics

[0018] (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.

[0019] (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. 2. Molecular biological identification

[0020] 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 da...

Embodiment 2

[0022] Implementation Example 2 The Inhibition Effect of Pichia cactus Pichia BY35 on Apple Penicillium and Botrytis Botrytis

[0023] 1. Experimental protocol

[0024] Pichia cactus BY35 was taken out from the -80°C refrigerator, activated with YPDA medium (yeast extract powder 10g, peptone 20g, glucose 20g, agar 18g, deionized water 1000ml, natural pH, sterilized at 121°C for 30min) to activate, pick Take 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 the preparation into a concentration of 1×10 8 cells / mL of Pichia cactus BY35 bacterial suspension.

[0025] 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 Peni...

Embodiment 3

[0032] Implementation example 3 Pichia cactus Pichia BY35 inhibitory effect on pear fruit blue mold and gray mold

[0033] 1. Experimental protocol

[0034] Pichia cactus BY35 was taken out from the -80°C refrigerator, activated with YPDA medium (yeast extract powder 10g, peptone 20g, glucose 20g, agar 18g, deionized water 1000ml, natural pH, sterilized at 121°C for 30min) to activate, pick Take 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 the preparation into a concentration of 1×10 8 cells / mL of Pichia cactus BY35 bacterial suspension.

[0035] 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 B...

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Abstract

The invention discloses a pichia cactophila strain BY35 for effectively controlling fruit postharvest diseases with advantages of wide antibacterial spectrum and stable effect and a using method of pichia cactophila. The strain has a strain collection number of CGMCC No. 14909 in China General Microbiological Culture Collection Center. The using method of the pichia cactophila strain comprises thefollowing steps: activating the strain, performing fermentation culture by YPD, centrifuging, and preparing the culture into bacterium suspension of 1*10<8>cells per mL with sterile water; adding apples, pears, grapes or citrus and the like into the bacterium suspension, soaking for 30 seconds, taking out the fruits, and airing; and putting into a preservation box, and storing at a normal temperature. The pichia cactophila is capable of simultaneously controlling apple penicillium disease and botrytis cinerea, penicilliosis and botrytis cinerea of pear fruits and fusarium wilt disease and anthracnose of grapes as well as blue mold of citrus fruits. Losses caused by the postharvest diseases are decreased, and the strain has excellent application prospects.

Description

technical field [0001] The invention relates to the field of biological control of postharvest diseases of fruits, in particular to a strain of Pichia cactophila used for biological control of postharvest diseases of fruits. Has a significant control effect. 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, the long-term use of chemical pesticides not only leads to the development of drug resistance of pathogenic bacteria and reduces the bactericidal effect (Prusky et al., 1985; as et al., 1991; ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/16A23B7/155A01G13/00C12R1/84
CPCA01G13/00A23B7/155A23V2002/00C12N1/16C12N1/165C12R2001/84A23V2250/76
Inventor 王友升任向峰马国为黄津津李丽萍王颖
Owner 山东凯普菲特生物科技有限公司
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