A kind of fermentation Pichia pastoris and its application

A technology of Pichia pastoris and Gibberella moniliforme, which is applied in fermenting Pichia pastoris and its application fields, can solve problems such as difficult pollination, affecting hybrid pollination, and reducing fruit quality, so as to increase pollen germination rate, promote pollination efficiency, Reduce the effect of falling flowers and fruits

Active Publication Date: 2021-11-23
ZHEJIANG FORESTRY ACAD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has been more than 100 years since the introduction of pecans in my country. The main problem faced by the existing pecan production areas is that the output is unstable, fluctuating, and the overall output is low.
There are many reasons for this phenomenon, such as pollination difficulties caused by improper variety allocation, poor pollination effect caused by low pollen vigor and then low seed setting rate, and fruit filling rate affected by diseases and insect pests. Although this method has played a role in controlling diseases, it affects cross pollination, reduces fruit quality, and increases food safety hazards

Method used

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  • A kind of fermentation Pichia pastoris and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0011] Example 1: Isolation and identification of fermenting Pichia pastoris 15B1 strain

[0012] Take fresh wild grapes, disinfect the surface with 70% alcohol, then disinfect with 50% antiformin (sodium hypochlorite solution) for half an hour, and finally soak in sterile water and rinse for 4-5 times. Cut the above-mentioned treated material on the ultra-clean bench, take the inner epidermis and inoculate it on the PDA medium (solid potato medium) plate, and when bacteria grow out, they are respectively streaked and purified, and the DNA is extracted for molecular identification , thus obtaining and confirming the strain. The specific steps for strain confirmation are as follows:

[0013] 1) Sample collection: Scrape a small amount of bacteria to be tested from the plate into a 200 μL PCR tube, add 100 μL of sterile water, centrifuge at high speed for a short time, and remove the supernatant;

[0014] 2) Sample lysis: Add glass beads with a diameter of 0.1mm-0.5mm to 50μL ...

Embodiment 2

[0020] Example 2: Morphological and biological characteristics of fermenting Pichia pastoris 15B1 strain

[0021] Use the plate dilution culture method to detect the optimal growth conditions of 15B1. Take 50 μL of the spare bacterial solution grown on the PDB medium, dilute it to 100 μL with water, and then take 10 μL of it and dilute it to 100 μL, and so on to form 4 gradient dilutions. , take 5 μL of each gradient dilution and spread it on the PDA medium plate and the LB medium plate, and compare their growth conditions. It was found that fermenting Pichia pastoris 15B1 was difficult to grow on LB medium, but grew better on PDA medium, and its optimum temperature range was 32-34°C (such as figure 1 shown), in addition, it can also grow slowly at 37 °C.

Embodiment 3

[0022] Embodiment 3: the bacteriostasis of fermenting Pichia pastoris 15B1

[0023] Pick 15B1 single clone and inoculate it in PDB medium, culture it at 32°C and 150rpm for 48h, then centrifuge at 4000rpm for 10min, filter the supernatant through a sterile filter membrane with a pore size of 0.22μm, take 0.2mL of the filtrate and evenly spread it on a PDA plate On the culture medium, an equal volume of sterile water was used as a control, and 0.5 cm of glial cell anthracnose bacteria ( Colletotrichum gloeosporioides ), Botrytis ( Botryosphaeria dothidea ) and Gibberella moniliforme ( Gibberella moniliformis) Bacterial block, the growth diameter of mycelium was measured after cultivating at 28°C for 3 days. Each treatment was repeated 3 dishes. The fungal colony diameter is measured by the cross method, and the mycelial growth inhibition rate is calculated, wherein the inhibition rate (%)=[(control fungal colony diameter-treatment fungal colony diameter) / (control fungal c...

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Abstract

A fermented Pichia pastoris and its application belong to the field of biotechnology. On the one hand, the present invention provides a new fermenting Pichia pastoris strain, and on the other hand, it provides the application of the strain, especially the effect of the strain on maintaining and improving the vigor of plant pollen. The fermented Pichia pastoris and its fermented liquid of the present invention have a better promoting effect on the germination activity of fruit tree pollen, and have a certain disease resistance effect on plants, and can be used in agricultural production to improve pollination efficiency, increase seed setting rate, and reduce flower drop Fruit drop, thereby increasing the yield and improving the economic benefits of fruit trees. The fermented Pichia pastoris and its fermented product of the present invention come from nature and belong to nature, are environmentally friendly and compatible, have no pollution and residue, and can be used as a new type of pollution-free microbial agent for the harmless production of fruit trees.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a fermented Pichia pastoris and its application. Background technique [0002] In the process of planting fruit trees, various reasons often lead to severe flower and fruit drop of fruit trees. This phenomenon is not only unfavorable for the healthy growth of fruit trees, but also reduces fruit yield, which seriously affects the economic benefits of fruit tree planting, especially the precious economy. species. Taking pecans as an example, the economic benefits of cultivating pecans are very obvious, and they are the most economically valuable tree species in the genus Carya. Thin-shelled hickory is pollinated by wind and is a typical outcrossing plant. Existing production practices believe that the main factors affecting the yield of pecans are high-efficiency pollination and fruit-setting rate. The introduction of pecans in my country has a history of more than 100 y...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/16A01N63/30A01P21/00A01P3/00C12R1/84
CPCA01N63/30
Inventor 彭华正金群英朱汤军张飞英叶华琳
Owner ZHEJIANG FORESTRY ACAD
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