CN105145637A - Production process for plant compound immune product - Google Patents

Production process for plant compound immune product Download PDF

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Publication number
CN105145637A
CN105145637A CN201510439088.1A CN201510439088A CN105145637A CN 105145637 A CN105145637 A CN 105145637A CN 201510439088 A CN201510439088 A CN 201510439088A CN 105145637 A CN105145637 A CN 105145637A
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plant
complex immunity
virus
minutes
viral
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CN201510439088.1A
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李素影
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Individual
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  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)

Abstract

The invention relates to a production process for a plant compound immune product. The production process comprises: firstly taking a plant virus source; artificially inoculating the plant virus source to a special production plant; then collecting diseased plant tissues which are crushed for fermented treatment; then carrying out separation; and preparing the plant compound immune product through secondary attenuation or disinfection. The production process provided by the invention can be used for factory-like large-scaled production and has the characteristics of being low in cost, strong in product function and fast in production.

Description

One Plants complex immunity product producing process
Technical field
The invention belongs to the production technology of Plants complex immunity goods, specifically a kind of have the production technology comprehensively improving the complex immunity goods that plant immune function inhibitio plant disease occurs.
Background technology
Plant is the same with people and animal, and itself, by stress reaction, has immunologic function to disease.For adapting to current needs of preserving the ecological environment; and then protection human body is healthy; the object that some countries and regions start to take chemistry to lure anti-method cures the disease to the immunologic function strengthening plant to reach plant disease-proof, not only but effect is not obvious, and still there is the problem of chemical contamination.Also some countries are had to carry out plant vaccine research, and start to carry out tentative application, but these plant vaccines are all generally independent vaccines, only have certain control and therapeutic action to a kind of disease, and what take is that the preparation method of conventional animal immune formulation carries out producing, the limited amount once produced, cost is higher, not only fundamentally can not solve Pesticides Residues, and because of the restriction of preparation method, the demand in market cannot be met in a large number.
Summary of the invention
Object of the present invention, is to provide rapidly and efficiently a kind of and the plant complex immunity product producing process of energy batch production large-scale production.
The present invention takes following technical scheme, a Plants complex immunity product producing process, and step is as follows:
The first step, collects viral source, gathers the germ on this kind of morbidity plant or virus in advance, carry out separating-purifying in field;
Second step, sets up the special malicious garden of phytopathy, inoculates above-mentioned germ or virus in special plant virus garden, allows a large amount of bacteria infection of this Plants and virus;
3rd step, enters the high-incidence season, pathological tissues of gathering in the morbidity of viral garden plant, pulverizes with 100-200 order common grinder, send into fermentation vat, stir, carry out fermentation attenuation.During the fermentation, fermentation temperature will reach more than 36 DEG C, and the time is 5-7 days.
4th step, after fermentation completes, the water that feeding 150-250 order seperators add equivalent is separated; For increasing operation rate, the water that the residue after separation continues to add isodose continues to stir, and carries out second time and is separated; The mixing material containing germ or virus that twice is separated is concentrated in 1/3rd rear loading commodity bottles of original content by Hollow fiber systems, namely becomes plant complex immunity goods.
Further, the viral vaccine after bottling can carry out secondary ultraviolet and to go out poison, makes plant complex immunity goods.
Further, the method for purification in the first step is:
The viral source gathered is organized in bruisher and smashs to pieces, adds in buffer solution (1: 2W/V), then adds 0.01mol DDTC and 0.5% mercaptoethanol, homogenate 3-5 minute; Add cooling chloroform (1: 1W/V) again, speed change grinds 1 minute; Initial centrifugation, 4 DEG C leave standstill 15 minutes, stay upper strata aqueous phase, obtain just extract;
Add 4% polyethylene glycol and 0.25mol sodium chloride in first extract, stir 1 hour; Centrifugal 10000g, lower 30 minutes of 4 DEG C of temperature, stay precipitation; By 1ml/100ml buffer solution buffer solution settling flux precipitation, stir 2 hours; Centrifugal 3000g, 4 DEG C leave standstill 10 minutes, stay supernatant; Centrifugal 10000g again, 4 DEG C leave standstill 30 minutes, stay precipitation; Again with concentration be 1ml/100ml buffer solution suspend, make extract again;
Last centrifugal 3000g, 4 DEG C leave standstill 10 minutes, stay supernatant, be the viral suspension after purification, 4 DEG C of preservations.
The present invention is applicable to the production of the complex immunity goods of all plant varieties such as fruit tree, vegetables, cereal crops, forest, flowers.
The present invention is particularly useful for the complex immunity production of articles of white rot, anthrachose of grape, downy mildew or anthracnose in grape.
The present invention; production technology is simple; production cost is lower; not only can produce the immune compound formulation of various plants; and can industrialization, large-scale production; output is large; the demand in market can be met fast; because these complex immunity goods contain a large amount of attenuations or degree of going out germ virus and a large amount of plant serum; not only can strengthen the immunologic function of plant; and to morbidity plant, there is certain therapeutic action, to the quality safety of agricultural product of improving the ecological environment, ensure and protection human health, there is very important effect.
Embodiment
Embodiment 1
To make grape complex immunity goods
The first step, gathers viral source, and gather the morbidity germ on grape, white rot, anthrachose of grape, downy mildew or anthracnose, carry out suitable separating-purifying with the method for purifying virus described in technique scheme, obtains a kind of plyability viral source.The viral source gathered is organized in bruisher and smashs to pieces, adds in buffer solution (1: 2W/V), then adds 0.01mol DDTC and 0.5% mercaptoethanol, homogenate 3-5 minute; Add cooling chloroform (1: 1W/V) again, speed change grinds 1 minute; Initial centrifugation, 4 DEG C leave standstill 15 minutes, stay upper strata aqueous phase, obtain just extract; Add 4% polyethylene glycol and 0.25mol sodium chloride in first extract, stir 1 hour; Centrifugal 10000g, lower 30 minutes of 4 DEG C of temperature, stay precipitation; By 1ml/100ml buffer solution buffer solution settling flux precipitation, stir 2 hours; Centrifugal 3000g, 4 DEG C leave standstill 10 minutes, stay supernatant; Centrifugal 10000g again, 4 DEG C leave standstill 30 minutes, stay precipitation; Again with concentration be 1ml/100ml buffer solution suspend, make extract again; Last centrifugal 3000g, 4 DEG C leave standstill 10 minutes, stay supernatant, be the viral suspension after purification, 4 DEG C of preservations.
Second step, carrying out viral source expands numerous, set up the plant virus garden specializing in breeding germ, virus inoculation use, in the season of applicable germ, viral growth, the viral suspension collected, the kindred plant (grape in if tree) of orientation is inoculated by aphis propagation, friction, grafting or the artificial methods such as Wound spray that manufacture, and heated by greenhouse facility and pour water and increase the way of humidity, create the growing environment of suitable morbidity, allow a large amount of bacteria infection of this directed plant, virus.
3rd step, the incidence tissue that gathers carries out fermentation attenuation, and after morbidity peaks, the pathological tissues of this directed plant of gathering, pulverizes with 100-200 order common grinder, sends into special fermenting tank, stirs, carry out fermentation attenuation.During the fermentation, fermentation temperature will reach more than 36 DEG C, and the time is 5-7 days.
4th step, separating-purifying makes commodity, and after having fermented, the water that feeding 150-250 order seperators add equivalent is separated.For increasing operation rate, the water that the residue after separation continues to add isodose continues to stir, and carries out second time and is separated.Load in commodity bottle after the mixing material containing germ, virus that twice is separated is concentrated to 1/3rd of original content by Hollow fiber systems, namely become plant complex immunity goods.
In implementation column, the germ also twice can separated or viral mixing material load in commodity bottle after concentrated, and send the ultraviolet room ultraviolet lamp of 30-40 watts to and irradiate, irradiation time is generally 30-40 minute.Namely germ after Ultraviolet radiation, viral mixing material are loaded in commodity bottle by (concentration ratio is the same) after concentrated becomes immune product.
In implementation column, the germ twice can also separated, viral mixing material are sent in pressure cooker and carry out high temperature virus killing.Virus killing temperature is 100 DEG C, and the time is 30 minutes.Namely germ after high temperature virus killing, viral mixing material are loaded in commodity bottle by (concentration ratio is the same) after concentrated becomes immune product.
In implementation column, this production technology, except above-mentioned lifted grape, is applicable to the production of the complex immunity goods of all plant varieties such as fruit tree, vegetables, cereal crops, forest, flowers, and its production technology, production procedure, production method are identical with above-mentioned.
In actual production is used, dress seed better with the malicious immune product that goes out, general application concentration is 1/50th of commodity immune product, i.e. the ratio of 1:50; Woody plant stoste, takes to carry out the mode of tree root or the inoculation of the tender tip; Herbaceous plant takes to spray before virus is suitable for the emergence period, and application concentration is immune product 600-1/700th, i.e. 1:600-700.

Claims (5)

1. a Plants complex immunity product producing process, it is characterized in that, step is as follows:
The first step, collects viral source, gathers the germ on this kind of morbidity plant or virus in advance, carry out separating-purifying in field;
Second step, sets up the special malicious garden of phytopathy, inoculates above-mentioned germ or virus in special plant virus garden, allows a large amount of bacteria infection of this Plants and virus;
3rd step, enters the high-incidence season, pathological tissues of gathering in the morbidity of viral garden plant, pulverizes with 100-200 order common grinder, send into fermentation vat, stir, carry out fermentation attenuation; During the fermentation, fermentation temperature will reach more than 36 DEG C, and the time is 5-7 days;
4th step, after fermentation completes, the water that feeding 150-250 order seperators add equivalent is separated; For increasing operation rate, the water that the residue after separation continues to add isodose continues to stir, and carries out second time and is separated; The mixing material containing germ or virus that twice is separated is concentrated in 1/3rd rear loading commodity bottles of original content by Hollow fiber systems, namely becomes plant complex immunity goods.
2. plant complex immunity product producing process as claimed in claim 1, is characterized in that, the viral vaccine after bottling can carry out secondary ultraviolet and to go out poison, makes plant complex immunity goods.
3. plant complex immunity product producing process as claimed in claim 1, it is characterized in that, the method for purification in the first step is:
The viral source gathered is organized in bruisher and smashs to pieces, adds in buffer solution (1: 2W/V), then adds 0.01mol DDTC and 0.5% mercaptoethanol, homogenate 3-5 minute; Add cooling chloroform (1: 1W/V) again, speed change grinds 1 minute; Initial centrifugation, 4 DEG C leave standstill 15 minutes, stay upper strata aqueous phase, obtain just extract;
Add 4% polyethylene glycol and 0.25mol sodium chloride in first extract, stir 1 hour; Centrifugal 10000g, lower 30 minutes of 4 DEG C of temperature, stay precipitation; By 1ml/100ml buffer solution buffer solution settling flux precipitation, stir 2 hours; Centrifugal 3000g, 4 DEG C leave standstill 10 minutes, stay supernatant; Centrifugal 10000g again, 4 DEG C leave standstill 30 minutes, stay precipitation; Again with concentration be 1ml/100ml buffer solution suspend, make extract again;
Last centrifugal 3000g, 4 DEG C leave standstill 10 minutes, stay supernatant, be the viral suspension after purification, 4 DEG C of preservations.
4. the plant complex immunity product producing process as described in claim 1-3, is characterized in that, is applicable to the production of the complex immunity goods of all plant varieties such as fruit tree, vegetables, cereal crops, forest, flowers.
5. the plant complex immunity product producing process as described in claim 1-3, is characterized in that, is applicable to the complex immunity production of articles of the white rot in grape, anthrachose of grape, downy mildew or anthracnose.
CN201510439088.1A 2015-07-24 2015-07-24 Production process for plant compound immune product Pending CN105145637A (en)

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1085387A (en) * 1992-08-24 1994-04-20 迈可根公司 The application of bacillus thuringiensis separator on the control pests in the family aphididae
CN101496527A (en) * 2009-03-09 2009-08-05 浙江省农业科学院 Method for preventing and treating virus disease of petunia with mild virus
CN102121039A (en) * 2010-11-01 2011-07-13 北京丰原康泰生物科技有限公司 Preparation method of plant immune protein
CN102308738A (en) * 2011-07-11 2012-01-11 云南省烟草公司红河州公司 Method of replacing tobacco aphids and with cabbage aphids to reproduce tobacco aphidius gifuensis and prevent and control tobacco pests
US20140287919A1 (en) * 2013-03-14 2014-09-25 Algal Scientific Corporation Modulation of plant immune system function

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1085387A (en) * 1992-08-24 1994-04-20 迈可根公司 The application of bacillus thuringiensis separator on the control pests in the family aphididae
CN101496527A (en) * 2009-03-09 2009-08-05 浙江省农业科学院 Method for preventing and treating virus disease of petunia with mild virus
CN102121039A (en) * 2010-11-01 2011-07-13 北京丰原康泰生物科技有限公司 Preparation method of plant immune protein
CN102308738A (en) * 2011-07-11 2012-01-11 云南省烟草公司红河州公司 Method of replacing tobacco aphids and with cabbage aphids to reproduce tobacco aphidius gifuensis and prevent and control tobacco pests
US20140287919A1 (en) * 2013-03-14 2014-09-25 Algal Scientific Corporation Modulation of plant immune system function

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
刘成亨: "木毒蛾复合病毒制剂研究", 《华东昆虫学报》 *
张亦冰: "抗植物病毒病疫苗――ZYMV-2002", 《世界农药》 *
胡伟贞等: "番茄环斑病毒的提纯", 《植物检疫》 *

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Application publication date: 20151216