CN109776648A - It is a kind of for preventing and treating the Charantin extraction process and device of TYLCV infection insect - Google Patents
It is a kind of for preventing and treating the Charantin extraction process and device of TYLCV infection insect Download PDFInfo
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- CN109776648A CN109776648A CN201910228337.0A CN201910228337A CN109776648A CN 109776648 A CN109776648 A CN 109776648A CN 201910228337 A CN201910228337 A CN 201910228337A CN 109776648 A CN109776648 A CN 109776648A
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Abstract
The invention discloses a kind of for preventing and treating the Charantin extraction process and device of TYLCV infection insect, extraction process is the following steps are included: step 1: removing after balsam pear is cleaned end to end, separate bitter melon seed and balsam pear meat, by bitter melon seed crushed after being dried, bitter melon seed powder and ethanol solution are homogenized, balsam pear meat is then added and continues to be homogenized；Step 2: preparing buffer, and cellulase reaction ultrasonic treatment is added, adjusts supernatant pH to neutrality；Step 3: Thief zone balsam pear solution will be obtained by filtration after balsam pear slurries steam blasting；Step 4: by Thief zone balsam pear solution heating water bath, being added the fermentation of modified cellulose enzyme solutions, will take supernatant liquid filtering after fermentation liquid sonic oscillation, by filtrate by obtaining balsam pear extract after freeze drier drying.In short, extracting method of the invention has recovery rate high, extraction rate is fast, the high advantage of product purity.
Description
Technical field
The present invention relates to technical field of biological extraction, are specifically related to a kind of for preventing and treating the balsam pear of TYLCV infection insect
Plain extraction process and device.
Background technique
Tomato be the most commonly used vegetable variety of the current cultivated area in China and our most important vegetable species it
One.The annual cultivated area in the whole nation reaches 800,000 hm2 since 2015 according to statistics, accounts for about the 4.5% of national sowing of vegetable area.Kind
Eggplant tomato yellow leaf curl China virus (TYLCV) is the tomato destructive disease occurred recent years, and the virus Yangtze river basin in 2006 originates
Raw, north area sprawling occurs rapidly later, present Shandong, Hebei, Jiangsu, Henan, Beijing, Xinjiang, southern Liaoning Province
Have been found that tomato yellow leaf curl virus disease, which is to endanger disease the most serious to tomato at present.
The tomato plant for infecting tomato yellow leaf curl China viral disease is downgraded, slow growth or stagnation, top vane often slightly chlorisis jaundice,
Become smaller, blade thickness, leaf roll qualitative change is hard on edge, and leaf back vein often shows purple.Growth and development early stage infected plant is seriously short
Contracting, only upper leaf and sprouting performance symptom, number of results are reduced the growth and development later period infected plant that can not normally yield positive results, fruit
Become smaller, maturity period fruit color is uneven (red impermeable), loses commodity control measure substantially.
Studies have found that infection insect Bemisia tabaci can be walked quickly and kept away by being sprayed on tomato plant after the dilution of balsam pear extract
Feeding, thus achieve the purpose that prevent and treat tomato yellow leaf curl virus disease, but the extraction process of balsam pear extract is more at present
Simply, extraction efficiency is low, at high cost, present invention finds a kind of new Charantin extraction process and device, the balsam pear extracted
Extract is high compared with the recovery rate that the prior art is extracted, and extraction rate is fast, and product purity is high.
Summary of the invention
In view of the above problems, the Charantin extraction that the present invention provides a kind of for preventing and treating TYLCV infection insect
Technique and device.
The technical scheme is that it is a kind of for preventing and treating the Charantin extraction process of TYLCV infection insect, mainly include
Following steps:
Step 1: balsam pear is cleaned with ultrapure water, is removed end to end, separation bitter melon seed and balsam pear meat, and bitter melon seed is sent to dry
With the dry 2-3h of 58-60 DEG C of temperature in dry device, the bitter melon seed after drying is sent and is crushed into Kind of Mini-powder Pulverizer, hardship is obtained
Then squash seed meal the bitter melon seed powder is added into homogenization apparatus, being added to homogenization apparatus with bitter melon seed powder mass ratio is 1:5
97% concentration ethanol solution, open homogenization apparatus be homogenized 20-30min, then by the balsam pear meat be segmented be added homogenate dress
In setting, continues to be homogenized 20-30min, obtain balsam pear slurries；
Step 2: take glycerine, acetic acid, zinc chloride, 2 hydroxy ethylamine, ultrapure water according to molal weight ratio 5:1:2:1:10
Ratio be uniformly mixed and obtain buffer, the cellulase of quality such as be added in Xiang Suoshu buffer, stir evenly reaction 1-2d,
It is 24-37 DEG C that reaction temperature is kept during reaction, and end of reaction carries out in ultrasonic unit with 4500- cellulase solution
5000MHz is ultrasonically treated 1-2min, takes supernatant, adjusts the supernatant pH to neutrality, obtain modified cellulose enzyme solutions；
Step 3: the balsam pear slurries are added into steam blasting machine, then are added into steam blaster and are starched with balsam pear
The deionized water of the quality such as liquid presoaks 40-50min, and being then turned on steam blasting machine makes pressure rise in steam blasting machine extremely
Thief zone balsam pear solution is obtained by filtration in balsam pear solution after cooling by 5MPa, explosion of then deflating；
Step 4: the Thief zone balsam pear solution is sent into installation for fermenting, 65-75 DEG C of heating water bath 15-20min, so
The modified cellulose enzyme solutions are added in installation for fermenting afterwards, the quality of modified cellulose enzyme solutions and Thief zone balsam pear solution
Than controlling 38-42 DEG C of fermentation temperature, fermentation time 1-2d obtains fermentation liquid after fermentation, fermentation liquid is sent for 1:2
30-50s is vibrated with 6000-6200Hz into sonic oscillation machine, takes supernatant by filtering in vacuum filter, obtains containing hardship
The filtrate of melon element, by the filtrate by obtaining balsam pear extract after freeze drier drying.
Further, the drying mode of the drying device is that hot-air convection is dried, the dry dependence pair of hot-air convection
Heat is spread to material heat supply, and the steam in material turns to vapor, and vapor is then taken away by air-flow, and rate of drying is fast and mode
Mildly, Charantin is not destroyed.
Further, bitter melon seed is crushed to 60-80 mesh in the step 1, increases bitter melon seed and divides in steam blasting machine
Efficiency from Charantin.
Further, supernatant pH is adjusted with the sodium hydroxide solution of 1mol/L in the step 2.
Further, which is characterized in that the rate of pressure rise of steam blasting machine is 0.5MPa/min in the step 3, quickly
Boosting explosion can be such that the supramolecular structure of balsam pear destroys, convenient for extracting Charantin.
It is a kind of for preventing and treating the Charantin extraction element of TYLCV infection insect, including drying device, Kind of Mini-powder Pulverizer,
Homogenization apparatus, ultrasonic unit, steam blasting machine, installation for fermenting, sonic oscillation machine, vacuum filter and freeze drier, institute
It states drying device, Kind of Mini-powder Pulverizer, homogenization apparatus, steam blasting machine to be sequentially connected, the installation for fermenting surpasses with described respectively
Acoustic wave device, steam blasting machine, the connection of sonic oscillation machine, the vacuum filter are connect with sonic oscillation machine, and the freezing is dry
Dry machine is connect with vacuum filter, and it is high and at low cost to extract convenience, extraction efficiency using the device.
Further, the balsam pear extract is sprayed directly on tomato plant after diluting 50 times, can walk quickly and keep away biography poison
Insect Bemisia tabaci feeding tomato plant.
The beneficial effects of the present invention are: a kind of Charantin for preventing and treating TYLCV infection insect provided by the invention extracts
Technique and device first separate bitter melon seed with balsam pear meat, will extract, avoid together with balsam pear meat after bitter melon seed drying and crushing
The waste behavior for abandoning bitter melon seed in traditional extraction technique, and the extraction efficiency of Charantin in bitter melon seed is improved, first
Ethanol solution is added after dry, pulverize bitter melon seed by the way that hot-air convection is dry and balsam pear meat is homogenized, carries out steam after homogenate
Explosion obtains Thief zone balsam pear solution, and modified cellulase solution is then carried out fermentation with Thief zone balsam pear solution and is mentioned
It takes, obtains balsam pear extract.In short, extracting method of the invention has recovery rate high, extraction rate is fast, and product purity is high
Advantage.
Detailed description of the invention
Fig. 1 is extraction element structural schematic diagram of the invention.
Wherein, 1- drying device, 2- Kind of Mini-powder Pulverizer, 3- homogenization apparatus, 4- ultrasonic unit, 5- steam blasting machine,
6- installation for fermenting, 7- sonic oscillation machine, 8- vacuum filter, 9- freeze drier
Specific embodiment
For convenient for the understanding to technical solution of the present invention, with reference to the accompanying drawing 1 and specific embodiment the present invention is done into one
The explanation of step, embodiment do not constitute the restriction to invention protection scope.
Embodiment 1
It is a kind of for preventing and treating the Charantin extraction process of TYLCV infection insect, mainly comprise the steps that
Step 1: balsam pear is cleaned with ultrapure water, is removed end to end, separation bitter melon seed and balsam pear meat, and bitter melon seed is sent to dry
With the dry 2h of 58 DEG C of temperature convection in dry device 1, the bitter melon seed after drying is sent and is crushed to 60 into Kind of Mini-powder Pulverizer 2
Mesh obtains bitter melon seed powder, and then bitter melon seed powder is added into homogenization apparatus 3, is added and bitter melon seed silty amount to homogenization apparatus 3
It than the ethanol solution of 97% concentration for 1:5, opens homogenization apparatus 3 and is homogenized 20min, then balsam pear meat is segmented, homogenate dress is added
It sets in 3, continues to be homogenized 20min, obtain balsam pear slurries；
Step 2: take glycerine, acetic acid, zinc chloride, 2 hydroxy ethylamine, ultrapure water according to molal weight ratio 5:1:2:1:10
Ratio be uniformly mixed obtain buffer, into buffer be added etc. quality cellulase, stir evenly reaction 1d, react the phase
Between keep the reaction temperature to be 24 DEG C, end of reaction be ultrasonically treated in ultrasonic unit 4 with 4500MHz to cellulase solution
1min takes supernatant, adjusts supernatant pH to neutrality with the sodium hydroxide solution of 1mol/L, obtains modified cellulose enzyme solutions；
Step 3: balsam pear slurries are added into steam blasting machine 5, then are added and balsam pear slurries into steam blaster 5
Etc. quality deionized water, presoak 40min, being then turned on steam blasting machine 5 makes the pressure in steam blasting machine 5 with 0.5MPa/
The rate of pressure rise of min is increased to 5MPa, explosion of then deflating, and Thief zone balsam pear solution is obtained by filtration in balsam pear solution after cooling；
Step 4: Thief zone balsam pear solution is sent into installation for fermenting 6,65 DEG C of heating water bath 15min, then will be modified fine
It ties up plain enzyme solutions to be added in installation for fermenting 6, the mass ratio of modified cellulose enzyme solutions and Thief zone balsam pear solution is 1:2, control
38 DEG C of fermentation temperature, fermentation time 1d obtains fermentation liquid after fermentation, by fermentation liquid send into sonic oscillation machine 7 with
6000Hz vibrates 30s, takes supernatant by filtering in vacuum filter 8, obtains the filtrate containing Charantin, filtrate is passed through cold
Balsam pear extract is obtained after lyophilizer 9 is dry, melon extract is sprayed directly on tomato plant after diluting 50 times.
As shown in Figure 1, a kind of for preventing and treating the Charantin extraction element of TYLCV infection insect, including it is drying device 1, micro-
Powder grinding device 2, homogenization apparatus 3, ultrasonic unit 4, steam blasting machine 5, installation for fermenting 6, sonic oscillation machine 7, vacuum filter
Machine 8 and freeze drier 9, drying device 1, Kind of Mini-powder Pulverizer 2, homogenization apparatus 3, steam blasting machine 5 are sequentially connected, fermentation dress
It sets 6 to connect with ultrasonic unit 4, steam blasting machine 5, sonic oscillation machine 7 respectively, vacuum filter 8 and sonic oscillation machine 7 connect
It connects, freeze drier 9 is connect with vacuum filter 8.
Embodiment 2
It is a kind of for preventing and treating the Charantin extraction process of TYLCV infection insect, mainly comprise the steps that
Step 1: balsam pear is cleaned with ultrapure water, is removed end to end, separation bitter melon seed and balsam pear meat, and bitter melon seed is sent to dry
With the dry 2.5h of 59 DEG C of temperature convection in dry device 1, the bitter melon seed after drying is sent and is crushed to 70 into Kind of Mini-powder Pulverizer 2
Mesh obtains bitter melon seed powder, and then bitter melon seed powder is added into homogenization apparatus 3, is added and bitter melon seed silty amount to homogenization apparatus 3
It than the ethanol solution of 97% concentration for 1:5, opens homogenization apparatus 3 and is homogenized 25min, then balsam pear meat is segmented, homogenate dress is added
It sets in 3, continues to be homogenized 25min, obtain balsam pear slurries；
Step 2: take glycerine, acetic acid, zinc chloride, 2 hydroxy ethylamine, ultrapure water according to molal weight ratio 5:1:2:1:10
Ratio be uniformly mixed obtain buffer, into buffer be added etc. quality cellulase, stir evenly reaction 1.5d, reaction
Period keep reaction temperature be 30 DEG C, end of reaction to cellulase solution carry out ultrasonic unit 4 in 4700MHz ultrasound at
1.5min is managed, supernatant is taken, supernatant pH is adjusted to neutrality with the sodium hydroxide solution of 1mol/L, it is molten to obtain modified cellulose enzyme
Liquid；
Step 3: balsam pear slurries are added into steam blasting machine 5, then are added and balsam pear slurries into steam blaster 5
Etc. quality deionized water, presoak 45min, being then turned on steam blasting machine 5 makes the pressure in steam blasting machine 5 with 0.5MPa/
The rate of pressure rise of min is increased to 5MPa, explosion of then deflating, and Thief zone balsam pear solution is obtained by filtration in balsam pear solution after cooling；
Step 4: Thief zone balsam pear solution is sent into installation for fermenting 6,70 DEG C of heating water bath 18min, then will be modified fine
It ties up plain enzyme solutions to be added in installation for fermenting 6, the mass ratio of modified cellulose enzyme solutions and Thief zone balsam pear solution is 1:2, control
40 DEG C of fermentation temperature, fermentation time 1.5d obtains fermentation liquid after fermentation, and fermentation liquid is sent into sonic oscillation machine 7
40s is vibrated with 6100Hz, takes supernatant by filtering in vacuum filter 8, obtains the filtrate containing Charantin, filtrate is passed through
Balsam pear extract is obtained after freeze drier 9 is dry, melon extract is sprayed directly on tomato plant after diluting 50 times.
Embodiment 3
It is a kind of for preventing and treating the Charantin extraction process of TYLCV infection insect, mainly comprise the steps that
Step 1: balsam pear is cleaned with ultrapure water, is removed end to end, separation bitter melon seed and balsam pear meat, and bitter melon seed is sent to dry
With the dry 3h of 60 DEG C of temperature convection in dry device 1, the bitter melon seed after drying is sent and is crushed to 80 into Kind of Mini-powder Pulverizer 2
Mesh obtains bitter melon seed powder, and then bitter melon seed powder is added into homogenization apparatus 3, is added and bitter melon seed silty amount to homogenization apparatus 3
It than the ethanol solution of 97% concentration for 1:5, opens homogenization apparatus 3 and is homogenized 30min, then balsam pear meat is segmented, homogenate dress is added
It sets in 3, continues to be homogenized 30min, obtain balsam pear slurries；
Step 2: take glycerine, acetic acid, zinc chloride, 2 hydroxy ethylamine, ultrapure water according to molal weight ratio 5:1:2:1:10
Ratio be uniformly mixed obtain buffer, into buffer be added etc. quality cellulase, stir evenly reaction 2d, react the phase
Between keep the reaction temperature to be 37 DEG C, end of reaction be ultrasonically treated in ultrasonic unit 4 with 5000MHz to cellulase solution
2min takes supernatant, adjusts supernatant pH to neutrality with the sodium hydroxide solution of 1mol/L, obtains modified cellulose enzyme solutions；
Step 3: balsam pear slurries are added into steam blasting machine 5, then are added and balsam pear slurries into steam blaster 5
Etc. quality deionized water, presoak 50min, being then turned on steam blasting machine 5 makes the pressure in steam blasting machine 5 with 0.5MPa/
The rate of pressure rise of min is increased to 5MPa, explosion of then deflating, and Thief zone balsam pear solution is obtained by filtration in balsam pear solution after cooling；
Step 4: Thief zone balsam pear solution is sent into installation for fermenting 6,75 DEG C of heating water bath 20min, then will be modified fine
It ties up plain enzyme solutions to be added in installation for fermenting 6, the mass ratio of modified cellulose enzyme solutions and Thief zone balsam pear solution is 1:2, control
42 DEG C of fermentation temperature, fermentation time 2d obtains fermentation liquid after fermentation, by fermentation liquid send into sonic oscillation machine 7 with
6200Hz vibrates 50s, takes supernatant by filtering in vacuum filter 8, obtains the filtrate containing Charantin, filtrate is passed through cold
Balsam pear extract is obtained after lyophilizer 9 is dry, melon extract is sprayed directly on tomato plant after diluting 50 times.
Comparative example 1
Charantin technique is extracted using traditional soda and extracts Charantin.
Experimental example 1
Balsam pear extract extraction efficiency research of the present invention.
Experiment condition: weighing the identical balsam pear of four parts of weight, the extracting method provided respectively with embodiment 1-3, comparative example 1
Charantin is extracted, measures the Charantin extraction efficiency of embodiment 1-3, comparative example 1, Charantin extraction efficiency=final extraction respectively
Charantin quality/balsam pear quality.
Experimental result: as shown in table 1.
1 balsam pear extract extraction efficiency table of table
| Embodiment 1 | Embodiment 2 | Embodiment 3 | Comparative example 1 | |
| Extraction efficiency (%) | 0.019 | 0.020 | 0.023 | 0.011 |
A kind of conclusion: Charantin extraction process extraction Charantin for preventing and treating TYLCV infection insect provided by the invention
The efficiency of extract is apparently higher than the prior art, and the method efficiency highest that embodiment 3 provides.
Experimental example 2
Balsam pear extract DNA purity research of the present invention.
Experiment condition: are extracted by obtained Charantin and is measured with ultraviolet spectrophotometry for embodiment 1-3, comparative example 1 respectively
The purity of its respective Charantin effective component.
Experimental result: as shown in table 2.
2 Charantin extract purity of table detects table
| Embodiment 1 | Embodiment 2 | Embodiment 3 | Comparative example 1 | |
| Purity (%) | 91.1 | 94.3 | 96.5 | 73.8 |
A kind of conclusion: balsam pear that the Charantin extraction process for preventing and treating TYLCV infection insect is extracted provided by the invention
Extract purity is higher than the prior art, and the technique DNA purity highest that embodiment 3 provides.
Finally, it should be noted that the above embodiments are merely illustrative of the technical solutions of the present invention, rather than its limitations；Although
Present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that: it still may be used
To modify to technical solution documented by previous embodiment or equivalent replacement of some of the technical features；And
These are modified or replaceed, the spirit and model of technical solution of the embodiment of the present invention that it does not separate the essence of the corresponding technical solution
It encloses.
Claims (8)
1. a kind of for preventing and treating the Charantin extraction process of TYLCV infection insect, which is characterized in that mainly comprise the steps that
Step 1: balsam pear is cleaned with ultrapure water, is removed end to end, separation bitter melon seed and balsam pear meat, and bitter melon seed is sent to dry and is filled
It sets in (1) with the dry 2-3h of 58-60 DEG C of temperature, the bitter melon seed after drying is sent and is crushed into Kind of Mini-powder Pulverizer (2), is obtained
Then bitter melon seed powder the bitter melon seed powder is added into homogenization apparatus (3), homogenization apparatus (3) are added and bitter melon seed silty
Amount opens homogenization apparatus (3) and is homogenized 20-30min than the ethanol solution for 97% concentration for being 1:5, then by the balsam pear meat point
Section is added in homogenization apparatus (3), continues to be homogenized 20-30min, obtains balsam pear slurries；
Step 2: take glycerine, acetic acid, zinc chloride, 2 hydroxy ethylamine, ultrapure water according to molal weight ratio 5:1:2:1:10 ratio
Example, which is uniformly mixed, obtains buffer, and the cellulase of the quality such as addition in Xiang Suoshu buffer stirs evenly reaction 1-2d, reaction
It is 24-37 DEG C that period, which keeps reaction temperature, and end of reaction carries out in ultrasonic unit (4) with 4500- cellulase solution
5000MHz is ultrasonically treated 1-2min, takes supernatant, adjusts the supernatant pH to neutrality, obtain modified cellulose enzyme solutions；
Step 3: the balsam pear slurries are added into steam blasting machine (5), then addition and balsam pear into steam blaster (5)
The deionized water of the quality such as slurries presoaks 40-50min, and being then turned on steam blasting machine (5) makes pressure in steam blasting machine (5)
Power is increased to 5MPa, explosion of then deflating, and Thief zone balsam pear solution is obtained by filtration in balsam pear solution after cooling；
Step 4: the Thief zone balsam pear solution is sent into installation for fermenting (6), 65-75 DEG C of heating water bath 15-20min, then
The modified cellulose enzyme solutions are added in installation for fermenting (6), the matter of modified cellulose enzyme solutions and Thief zone balsam pear solution
Amount controls 38-42 DEG C of fermentation temperature, fermentation time 1-2d, fermentation liquid is obtained after fermentation, by fermentation liquid than being 1:2
It send into sonic oscillation machine (7) and 30-50s is vibrated with 6000-6200Hz, take supernatant by filtering in vacuum filter (8), obtain
To the filtrate containing Charantin, by the filtrate by obtaining balsam pear extract after freeze drier (9) drying.
2. according to claim 1 a kind of for preventing and treating the Charantin extraction process of TYLCV infection insect, feature exists
In the drying mode of the drying device (1) is dry for hot-air convection.
3. according to claim 1 a kind of for preventing and treating the Charantin extraction process of TYLCV infection insect, feature exists
In bitter melon seed is crushed to 60-80 mesh in the step 1.
4. according to claim 1 a kind of for preventing and treating the Charantin extraction process of TYLCV infection insect, feature exists
In with the sodium hydroxide solution adjusting supernatant pH of 1mol/L in the step 2.
5. according to claim 1 a kind of for preventing and treating the Charantin extraction process of TYLCV infection insect, feature exists
In the rate of pressure rise of steam blasting machine (5) is 0.5MPa/min in the step 3.
6. a kind of for preventing and treating the Charantin extraction element of TYLCV infection insect, which is characterized in that including drying device (1), micro-
Powder grinding device (2), homogenization apparatus (3), ultrasonic unit (4), steam blasting machine (5), installation for fermenting (6), sonic oscillation machine
(7), vacuum filter (8) and freeze drier (9), the drying device (1), Kind of Mini-powder Pulverizer (2), homogenization apparatus (3),
Steam blasting machine (5) is sequentially connected, the installation for fermenting (6) respectively with the ultrasonic unit (4), steam blasting machine (5), super
Sound oscillation machine (7) connection, the vacuum filter (8) connect with sonic oscillation machine (7), the freeze drier (9) and vacuum
Filter (8) connection.
7. according to claim 1 a kind of for preventing and treating the Charantin extraction process of TYLCV infection insect, feature exists
In the balsam pear extract is sprayed directly on tomato plant after diluting 50 times.
8. according to claim 6 a kind of for preventing and treating the Charantin extraction process of TYLCV infection insect, feature exists
In the drying mode of the drying device (1) is dry for hot-air convection.
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