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Biocontrol yeast active lyophilized powder and preparation method thereof

A bio-control yeast and freeze-dried powder technology, applied in biochemical equipment and methods, microorganism-based methods, microorganisms and other directions, can solve the problems of not many and low control efficiency, and achieves improvement of vitality and inhibition of anthracnose rot. Diseases and the effect of improving the survival rate

Pending Publication Date: 2018-12-21
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are not many biocontrol agents that can be used to effectively control the anthracnose and rot of nectarines in the process of picking, transporting, and storing, and the control efficiency is not high.

Method used

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  • Biocontrol yeast active lyophilized powder and preparation method thereof
  • Biocontrol yeast active lyophilized powder and preparation method thereof
  • Biocontrol yeast active lyophilized powder and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1: Isolation and identification of bacterial strain

[0027] Pichia pastoris Y35-1 ( Pichia guilliermondii ) was screened by our laboratory and was deposited in China Center for Type Culture Collection (Address: Wuhan, China, Wuhan University; deposit number CCTCCNO: M2016719) on December 5, 2016.

[0028] The main steps of strain screening and identification process are as follows:

[0029] Isolation of antagonistic yeast: collect mature and healthy loquat fruit from the agricultural ecological orchard of Gaoqiao Town, Zhenjiang City, isolate and screen the strains on the surface of the intact fruit, select yeast single colonies with different culture characteristics and separate and purify them on the NYDA plate, pick each single Colonies were cryopreserved.

[0030] Preparation of antagonistic yeast suspension: resuspend the cultured yeast to form a suspension, calculate the concentration of the yeast suspension with a hemocytometer, adjust the suspensi...

Embodiment 2

[0040] Embodiment 2: Preparation of biocontrol yeast active freeze-dried powder

[0041] Enrichment culture of yeast: first activate the yeast strain, thaw the deep-frozen yeast in a water bath, insert it into NYDB liquid medium, culture it in a shaker flask at 28°C and 200r / min for 24-48h, and repeat the culture once. Then, take two loops of the activated biocontrol yeast and inoculate them in NYDB seed medium (50ml / 250ml), and culture them at 28°C and 200r / min for 24h; take 100μl of the seed solution and insert it into the NYTB fermentation medium (50ml / 250ml), cultured at 28℃, 200r / min for 48h; NYDB medium includes 8g of beef extract, 5g of yeast extract, 10g of glucose, 1000ml of distilled water, natural pH; NYTB medium includes beef extract 8g, yeast extract 5g, trehalose 10g, distilled water 1000ml, natural pH;

[0042] The conventional NYDB seed medium described therein includes beef extract 8g, yeast extract 5g, glucose 10g, distilled water 1000ml, natural pH; NYTB f...

Embodiment 3

[0050] Embodiment 3: the influence of each added ingredient in the freeze-dried powder on effect

[0051] In this embodiment, the effect of different addition amounts of each component has been verified, and the specific method is as follows:

[0052] Screening of lyoprotectants: the concentration screening of each protectant is shown in Table 1, and the protection results for Pichia mongolica are shown in Table 2. Mix the yeast suspensions added with each freeze-drying protective agent evenly, take 1ml samples for gradient dilution and coating, and culture the plates in a constant temperature incubator at 28°C for 48 hours to determine the number of viable bacteria. When the concentration of trehalose was 5wt%, the survival rate of Pichia mongoliana Y35-1 cells reached the highest value, which was 25.6%; when the concentration was 2.5wt% and 10wt%, the survival rates were 15.5% and 12.0%, respectively, so choosing 5wt% trehalose is the optimal protective agent.

[0053] Tab...

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Abstract

The invention relates to biocontrol yeast active lyophilized powder and a preparation method thereof, which belongs to the technical field of biological prevention. The lyophilized powder in the invention comprises pichia guilliermondii Y35-1, trehalose and glutathione. The preparation method comprises the following steps: firstly inoculating a NYDB seed culture medium with activated pichia guilliermondii, culturing the pichia guilliermondii, then inoculating a NYTB fermentation culture medium with a seed solution, and fermenting the seed solution; centrifuging a yeast fermentation solution, washing by using sterile water, washing the obtained precipitates by using sterile water, and preparing a yeast suspension; and adding trehalose and glutathione as a lyophilized protection agent, uniformly mixing, loading into a flat plate, vacuum freeze drying, and obtaining the lyophilized powder. The biocontrol yeast active lyophilized powder provided by the invention is good in biocontrol effect, the yeast after the lyophilized powder is rehydrated is suspended, and the nectarine fruit disease rate and the fruit disease speckle diameter can be apparently lower than that of a control group and are significantly different from the control group, so that the lyophilized powder preparation can effectively inhibit the anthracnose rot disease of the nectarine fruit after being rehydrated.

Description

technical field [0001] The invention relates to a biocontrol yeast active freeze-dried powder and a preparation method thereof, belonging to the technical field of biocontrol. Background technique [0002] Nectarine fruit has thin skin and is ripe and picked in early summer with high temperature and humidity, so it is very vulnerable to mechanical damage and microbial attack during picking, transportation and storage, and rots and deteriorates, causing huge economic losses. Anthracnose is one of the main diseases that are prone to occur in the post-harvest transportation and storage of various fruits. Colletotrichum gloeosporioides ) and Anthracnose oxysporum ( Colletotrichum acutatum ) two kinds. For a long time, the control of fruit diseases has mainly relied on chemical fungicides, but with the strengthening of people's awareness of food safety and environmental protection, the use of chemical fungicides has been more restricted. Therefore, it is necessary to explore n...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/16C12N1/04A23L3/3571C12R1/84
CPCA23L3/3571C12N1/04C12N1/165C12R2001/84
Inventor 孙卫红周秋阳杨慧慧孙玲王玲温新宇
Owner JIANGSU UNIV
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