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Pichia manshurica for controlling postharvest diseases of oranges and grapes

A technology of Pichia pastoris and Pichia pastoris, applied in the direction of microorganism-based methods, biochemical equipment and methods, food preservation, etc., can solve the problems of lack of bio-control strains, fruit rot, large losses, etc., and achieve significant economic benefits and social benefits, wide antibacterial spectrum, and the effect of avoiding harm to people

Active Publication Date: 2016-08-17
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

(2) Citrus postharvest rot mainly includes Penicillium italicum caused by Penicillium italicum and green mold caused by Penicillium digitatum, among which the rot caused by Penicillium digitatum is particularly serious, This kind of pathogenic bacteria is widely distributed, and spreads a large number of conidia produced in the air in the form of saprophytic organic matter, and then spreads to the fruit, causing the fruit to rot and causing great losses
The main reason is the lack of highly effective biocontrol strains

Method used

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  • Pichia manshurica for controlling postharvest diseases of oranges and grapes
  • Pichia manshurica for controlling postharvest diseases of oranges and grapes
  • Pichia manshurica for controlling postharvest diseases of oranges and grapes

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1: the microbiological characteristics of Pichia manshurica (Pichia manshurica) Y4 strain are as follows:

[0027] 1. Morphological characteristics

[0028] (1) NYDA culture medium (0.8% beef extract, 0.5% yeast extract, 1% glucose, 2% agar, 121°C damp heat sterilization for 20min) cultured at 28°C for 48h, the colonies were round, smooth and moist, Easy to provoke. The cells are oblong in shape and scattered individually.

[0029] (2) After culturing in NYDB liquid medium for 24 hours, no mold was formed, and the bacterial solution was turbid with precipitation. Microscopically, the yeast cells were oval and budded.

[0030] 2. Molecular biological identification

[0031] Sequence analysis of the 5.8S rDNA-ITS region of the screened strain Pichia manshurica was performed by searching on GenBank and identified as Pichia manshurica. According to the retrieved homologous strains, the Mege5.1 program of DNAStar software was used to construct the biological e...

Embodiment 2

[0033] Embodiment 2: the control effect of Pichia chastori to citrus green mold

[0034] 1) Test plan

[0035] The tested fruits have been carefully selected, and the fruit should be selected as uniform as possible, without scratches or mechanical damage on the surface and damage by diseases and insect pests. Use a sterile puncher to punch 3 holes at the equator of the fruit, and the surface wound of the citrus is 5mm (diameter) × 3mm (depth). Add 30 μL of the following treatment solution equally to each wound: (1) 1×10 8 cells / mL of Pichia chaltiformis suspension; (2) Sterile distilled water. After 2 hours, 30 μL of 1×10 5 spores / mL of Penicillium digitatum (P.digitatum) spore suspension. After drying naturally, put the fruit into a plastic basket and seal it with a plastic wrap, cultivate it in a constant temperature incubator (humidity 95%), and place it at room temperature (20°C). Each treatment was replicated 3 times, and 12 fruits were replicated each time. The who...

Embodiment 3

[0042] Embodiment 3: the control effect of Pichia crestella to Botrytis cinerea and Aspergillus niger

[0043] 1) Test plan

[0044] The activated yeast was inoculated in a conical flask filled with 50 mL of NYDB (5 g of yeast extract, 8 g of beef extract, 10 g of glucose, and 1000 mL of distilled water), and cultured on a shaking table at 28°C and 180 r / min for 20 h. Take the bacterial solution and centrifuge it at 8000r / min for 10min, discard the supernatant, wash it repeatedly with sterile water for 3 times, and count it on a hemocytometer to obtain a concentration of 1×10 8 cells / mL of bacterial suspension, and the control group was sterile water. A hole with a diameter of 3m (width)×3mm (depth) was punched at the waist of the fruit, and then 15 μL of the yeast suspension of the above concentration was injected into each hole, and 15 μL of sterile water was injected into the control fruit. After 2 hours, 10 μL of 1×10 5 spores / mL of Botrytis cinerea (B.cinerea) spore su...

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Abstract

The invention discloses a Pichia manshurica for controlling postharvest diseases of oranges and grapes, and belongs to the field of biological control of postharvest diseases of fruits. A strain is identified as Pichia manshurica, and is named as Pichia manshurica Y4, and the preservation number of the Pichia manshurica Y4 is CGMCC 2.5415. When the Pichia manshurica is used, the Pichia manshurica is activated, cultured and centrifuged to obtain thalli, and the thalli are diluted with sterile water to prepare a thallus suspension with the concentration of 1.0*10<8> / mL. The suspension is sprinkled to the exocuticle of the fruits through using a watering pot, the fruits are air-dried, the dried fruits are put in a plastic basket, the plastic basket is sealed with a preservative film, and the sealed fruits are stored under room temperature conditions. The incidence of orange Penicillium digitatum, grape Aspergillus niger and gray molds is substantially reduced after treatment with the Pichia manshurica. The Pichia manshurica can substitute chemical bactericides to prevent and control postharvest diseases of oranges and grapes, avoids harms of the chemical bactericides to people, and has substantial economic and social benefits.

Description

technical field [0001] The invention relates to a strain of Pichia manshurica Y4, which can control postharvest green mold of citrus and postharvest grape aspergillus niger and gray mold, and belongs to the field of biological control of fruit postharvest diseases. Background technique [0002] The serious loss of fruits due to post-harvest rot has long been a global problem. According to conservative estimates, 10-20% of fresh fruits are lost to post-harvest rot in developed countries, and in developing countries that lack storage and transportation refrigeration equipment, their rot The loss rate is as high as 40-50%, and the fruit rot in my country is generally 20-30%. [0003] my country is the largest fruit production and sales country in the world, with a wide variety of fruits, and its output accounts for 80% of the world's fruit output. Because the fruit production area is far away from the city, and the ripening period is relatively concentrated, a large amount of p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/16A23B7/155C12R1/84
CPCA23B7/155C12N1/165C12R2001/84
Inventor 张红印李万海张晓云任晓锋朱淑云
Owner JIANGSU UNIV
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