CN104255384A - Method for cultivating botrytised grapes - Google Patents
Method for cultivating botrytised grapes Download PDFInfo
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- CN104255384A CN104255384A CN201410523702.8A CN201410523702A CN104255384A CN 104255384 A CN104255384 A CN 104255384A CN 201410523702 A CN201410523702 A CN 201410523702A CN 104255384 A CN104255384 A CN 104255384A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G17/00—Cultivation of hops, vines, fruit trees, or like trees
- A01G17/02—Cultivation of hops or vines
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G7/00—Botany in general
- A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
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Abstract
The invention provides a method for cultivating botrytised grapes. The method comprises the following steps of (1) preparation of grey botrytis cinerea bacteria, (2) cultivation of raw materials of the grapes, (3) artificial inoculation of grey botrytis cinerea, and (4) harvesting of the botrytised grapes. The grapes are artificially inoculated by the grey botrytis cinerea, the botrytis cinerea can normally infect fruits of the grapes under the condition of low air humidity, and a botrytised effect is achieved; and requirements of the botrytised grapes in a natural planting environment are reduced, a cultivation region in which the botrytised grapes can be cultivated is expanded, so that the high-quality botrytised grapes can be produced in lots of places, and requirements of high-grade grape wine on high-quality grape raw materials are met.
Description
Technical field
The present invention relates to viticulture technical field, be specially the method for the expensive rotten grape of a kind of artificial culture.
Background technology
Botrytised wine is one of the most high-grade in the world wine, and it is the high-grade sweet white wine that raw material is produced by the grape after there is your corruption effect.Your corruption effect be due to grape infected close to the maturing stage Botrytis cinerea (
botytis cinerea) caused by, Botrytis cinerea infect and grow the moisture evaporation that can make grape material, cause the compositions such as carbohydrate in grape fruit, organic acid, polyalcohol to change, final form the very high and fragrant strong expensive rotten grape of sugar content.After Botrytis cinerea infects grape fruit, generally can cause grape grey mould thus cause the underproduction, but then can cause your corruption effect under the particular surroundings of sombrely dry sweltering heat in moistening, afternoon sooner or later.Analyzing its main cause is, clammy condition is sooner or later conducive to the growth of Botrytis cinerea and infects the generation of grape, and the hot dry weather after noon just can make the moisture in grape fruit evaporate from infection, and dehydration improves the sugariness of grape.Because the condition your corruption effect occurs is very harsh, only have a few viny region, Europe to obtain expensive rotten grape by abiogenous approach at present, its output can not meet the demand of high-grade sweet wine to the expensive rotten grape of high-quality far away.
Central and west regions, Guangxi belong to subtropics monsoon climate, between annual October-December, rainfall is less, dry, illumination is sufficient, temperature is substantially at 10-25 DEG C, very advantageously by rain shelter cultivation, term delaying the planting technology high-quality grapes such as adjustment, wherein also comprise the white grape variety of making wine of high-quality.This provides the most basic raw material basis for obtaining expensive rotten grape.
But because weather continues dry, air humidity is stablized on the low side, and Botrytis cinerea does not possess the condition infecting ripe grape, cannot rely on natural conditions and obtain natural expensive rotten grape as expensive rotten viny region, Europe.
Summary of the invention
The object of this invention is to provide a kind of by artificial infection Botrytis cinerea, and can normally infect grape fruit under low air humidity conditions, your corruption effect occurs, the expensive rotten grape of final acquisition high-quality, meets the continuous demand of high-grade wine to the expensive rotten grape of high-quality.
Technical scheme of the present invention: a kind of breeding method of expensive rotten grape, comprises the steps:
1) preparation of Botrytis cinerea bacterial classification: Botrytis cinerea bacterial strain is inoculated, liquid state fermentation to concentration is 10
6-10
8cFU/mL, filters, washing, dry, and is ground into particle diameter and is less than 200 object the pathogen of Botrytis cinerea powder;
2) cultivation of grape: with the white grape variety of making wine of maturation for plantation object, control grape fruit in sunny, temperature is fully ripe the month of 10-25 DEG C;
3) artificial infection Botrytis cinerea:
The preparation of A, bacterium powder suspension: the pathogen of Botrytis cinerea powder is suspended in clear water by the amount of mass volume ratio 0.1%-5%, adds the polyglutamic acid of 0.1%-2% by mass volume ratio, the fructo-oligosaccharide (FOS) of 0.5%-5%, fully mix for subsequent use;
B, selection weather are fine, mean temperature of air is evenly spray in ripe grape fruit ear surface by the pathogen of Botrytis cinerea powder suspension by the amount of 10mL-100mL/ fringe fruit the opportunity of 10 DEG C-25 DEG C, treat that grape fruit surface has obvious white thread bacterial plaque to generate, illustrate and inoculate successfully;
4) to gather expensive rotten grape: inoculate successfully, period sampling measuring fruit juice sugar content, when sugar content reaches 300g/L-340g/L, expensive rotten grape finished product of can gathering.
Preferably, described Botrytis cinerea bacterial classification liquid-state fermentation technology is as follows:
1) from original seed inclined-plane scraping Botrytis cinerea spore or mycelium, preparing biomass is 10
6-10
8the original seed suspension of CFU/mL, and the amount of 4%-10% is inoculated in seed culture medium by volume;
2) under 10 DEG C of-20 DEG C of conditions, oscillation and fermentation or stirring fermentation reach 5%-8% to thalline weight and culture volume ratio and stop fermenting.
Preferably, the preparation technology of described seed culture medium is as follows:
1) get potato and fructus hordei germinatus that mass ratio is 1:1, dehydrated potato powder is broken to the fritter that diameter is not more than 1cm, and fructus hordei germinatus is crushed to particle diameter 20 order;
2) thin up, the quality of described water is 2.5 times of potato and fructus hordei germinatus gross mass;
3) 60 DEG C of saccharification 8h;
4) through 40 mesh filter screens or screen filter cleaner;
5) supplementing water, makes the volume of medium consistent with the volume before saccharification;
6) 110-121 DEG C of moist heat sterilization 20min.
Preferably, described bacterial strain is selected from Botrytis cinerea ACCC36257 or ACCC36258 of Chinese agriculture Microbiological Culture Collection administrative center preservation, or is drawn by wild Botrytis cinerea bacterial strain screening.
Preferably, described white grape variety of making wine comprises match beauty treatment, rope taste dense, Chardonnay, Riesling etc.
Preferably, by rain shelter cultivation and/term regulation technology control grape fruit maturation time.
Preferably, the spraying frequency of described the pathogen of Botrytis cinerea powder suspension is: often within 1-3 days, spray 1 time, sprays 3-6 time continuously.
Breeding method of the present invention has following significant beneficial effect:
1, the present invention is by artificial infection Botrytis cinerea, improves the probability that Botrytis cinerea infects grape, improves the productive rate of expensive rotten grape;
2, moisture-keeping function is played on the polyglutamic acid in the bacterium powder suspension sprayed and the surface of fructo-oligosaccharide (FOS) to grape, suppress varied bacteria growing, promote that Botrytis cinerea surely grows and infects grape fruit, Botrytis cinerea is surely grown under the meteorological condition of sunny drying, infects ripe grape fruit, and your corruption effect occurs;
3, breeding method of the present invention reduces the requirement of expensive rotten grape to plantation environment, expands and can cultivate the cultural area obtaining expensive rotten grape, and making more can the expensive rotten grape of production high-quality, meets the demand of high-grade wine to high-quality grape material.
Embodiment
Below in conjunction with specific embodiment, set forth the present invention further.Should be understood that these embodiments are only not used in for illustration of the present invention to limit the scope of the invention.In addition should be understood that those skilled in the art can do various amendment or change to the present invention, and these equivalent form of values fall within the application's appended claims limited range equally after the content of having read the present invention's instruction.
Embodiment 1
1) Botrytis cinerea bacterial classification preparation
A, seed culture medium: potato 200g is crushed to diameter and is not more than 1cm fritter, about fructus hordei germinatus 200g is crushed to particle diameter 20 order, add in 1L running water, 60 DEG C of saccharification 8h, 40 mesh filter screens or screen filter cleaner, filtrate complements to 1L volume with running water, 121 DEG C of moist heat sterilization 20min;
B, bacterial strain: select by the Botrytis cinerea ACCC36257 of Chinese agriculture Microbiological Culture Collection administrative center preservation;
C, fermentation: inoculation fermentation is from original seed inclined-plane scraping Botrytis cinerea spore or mycelium, and preparing biomass is 10
8the original seed suspension of CFU/mL, the amount of 10% is inoculated in seed culture medium by volume, under 20 DEG C of conditions in shaking flask oscillation and fermentation, or the 10d that ferments in fermentation tank mechanical agitating fermentation tank, reaches 8% termination to thalline weight and culture volume ratio and ferments;
Prepared by D, bacterial classification pulvis: will ferment gained thalline collecting by filtration, after washing 3 times, be less than 30% in 20 DEG C of vacuum dryings to water content with sterile water, be ground into particle diameter and be less than 200 object the pathogen of Botrytis cinerea powder.
2) cultivation of grape
With the white grape variety of making wine of maturation match beauty treatment for raw material, in central and west regions, Guangxi, by rain shelter cultivation and term regulation technology, control grape fruit fully ripe in late October.
3) artificial infection Botrytis cinerea
The pathogen of Botrytis cinerea powder is suspended in clear water by the amount of mass volume ratio 5%, the polyglutamic acid of 2% is added by mass volume ratio, the fructo-oligosaccharide (FOS) of 5%, after fully mixing, selection weather is fine, mean temperature of air is evenly spray in grape fruit ear surface by the pathogen of Botrytis cinerea powder suspension by the amount of 100mL/fringe fruit the opportunity of 25 DEG C, within every 1 day, spray 1 time, continuous spraying 6 times, during spraying and observe the generation situation of fruit surface Botrytis cinerea bacterial plaque after spraying, treat that grape fruit surface has obvious white thread bacterial plaque to generate, illustrate and inoculate successfully.
4) to gather expensive rotten grape
Inoculate successfully, period sampling measuring fruit juice sugar content, when sugar content reaches 340g/L, expensive rotten grape finished product of can gathering.
Embodiment 2
1) Botrytis cinerea bacterial classification preparation
A, seed culture medium: potato 200g is crushed to diameter and is not more than 1cm fritter, about fructus hordei germinatus 200g is crushed to particle diameter 20 order, add in 1L running water, 60 DEG C of saccharification 8h, 40 mesh filter screens or screen filter cleaner, filtrate complements to 1L volume with running water, 121 DEG C of moist heat sterilization 20min.Seed culture medium can expand amount of preparation in proportion.
B, bacterial strain: select by the Botrytis cinerea ACCC36258 of Chinese agriculture Microbiological Culture Collection administrative center preservation as fermentation strain;
C, fermentation: inoculation fermentation is from original seed inclined-plane scraping Botrytis cinerea spore or mycelium, and preparing biomass is 10
6the original seed suspension of CFU/mL, the amount of 4% is inoculated in seed culture medium by volume, under 10 DEG C of conditions in shaking flask oscillation and fermentation, or in fermentation tank mechanical agitating fermentation tank ferment 5, reach 5% termination to thalline weight and culture volume ratio and ferment;
Prepared by D, bacterial classification pulvis: will ferment gained thalline collecting by filtration, after washing 3 times, be less than 30% in 20 DEG C of vacuum dryings to water content with sterile water.Be ground into particle diameter and be less than 200 object the pathogen of Botrytis cinerea powder.
2) cultivation of grape
With the white grape variety of making wine Chardonnay of maturation for raw material.In central and west regions, Guangxi, by rain shelter cultivation and term regulation technology, control grape fruit fully ripe in early November.
3) artificial infection Botrytis cinerea
The pathogen of Botrytis cinerea powder is suspended in clear water by the amount of mass volume ratio 1%, the polyglutamic acid of 1% is added by mass volume ratio, the fructo-oligosaccharide (FOS) of 2%, after fully mixing, selection weather is fine, and mean temperature of air is evenly sprayed in grape fruit ear surface by the amount of 10mL/ fringe fruit by the pathogen of Botrytis cinerea powder suspension the opportunity of 20 DEG C.Within every 2 days, spray 1 time, spray 4 times continuously, during spraying and observe the generation situation of fruit surface Botrytis cinerea bacterial plaque after spraying.Treat that grape fruit surface has obvious white thread bacterial plaque to generate, illustrate and inoculate successfully.
4) to gather expensive rotten grape
Inoculate successfully, period sampling measuring fruit juice sugar content, when sugar content reaches 300g/L, expensive rotten grape finished product of can gathering.
Embodiment 3
1) Botrytis cinerea bacterial classification preparation
A, seed culture medium: potato 300g is crushed to diameter and is not more than 1cm fritter, about fructus hordei germinatus 300g is crushed to particle diameter 20 order, add in 1.5L running water, 60 DEG C of saccharification 8h, 40 mesh filter screens or screen filter cleaner, filtrate complements to 1.5L volume with running water, 120 DEG C of moist heat sterilization 20min.Seed culture medium can expand amount of preparation in proportion;
B, bacterial strain: select by the Botrytis cinerea ACCC36258 of Chinese agriculture Microbiological Culture Collection administrative center preservation as fermentation strain;
C, fermentation: inoculation fermentation is from original seed inclined-plane scraping Botrytis cinerea spore or mycelium, and preparing biomass is 10
7the original seed suspension of CFU/mL, the amount of 8% is inoculated in seed culture medium by volume, under 15 DEG C of conditions in shaking flask oscillation and fermentation, or the 5d that ferments in fermentation tank mechanical agitating fermentation tank, reaches 6% termination to thalline weight and culture volume ratio and ferments;
Prepared by D, bacterial classification pulvis: will ferment gained thalline collecting by filtration, after washing 3 times, be less than 30% in 20 DEG C of vacuum dryings to water content with sterile water.Be ground into particle diameter and be less than 200 object the pathogen of Botrytis cinerea powder.
2) cultivation of grape
Dense for raw material with the white grape variety of making wine rope taste of maturation.In central and west regions, Guangxi, by rain shelter cultivation and term regulation technology, control grape fruit fully ripe mid-November.
3) artificial infection Botrytis cinerea
The pathogen of Botrytis cinerea powder is suspended in clear water by the amount of mass volume ratio 2.5%, the polyglutamic acid of 1.5% is added by mass volume ratio, the fructo-oligosaccharide (FOS) of 2.5%, after fully mixing, selection weather is fine, and mean temperature of air is evenly sprayed in grape fruit ear surface by the amount of 80mL/ fringe fruit by the pathogen of Botrytis cinerea powder suspension the opportunity of 18 DEG C.Within every 3 days, spray 1 time, spray 3 times continuously, during spraying and observe the generation situation of fruit surface Botrytis cinerea bacterial plaque after spraying.Treat that grape fruit surface has obvious white thread bacterial plaque to generate, illustrate and inoculate successfully.
4) to gather expensive rotten grape
Inoculate successfully, period sampling measuring fruit juice sugar content, when sugar content reaches 330g/L, expensive rotten grape finished product of can gathering.
Embodiment 4
Embodiment 4 is similar to the breeding method of embodiment 1, its difference is: the grape variety cultivated is Riesling, the time controlling grape maturity is late November, and the addition of bacterium powder is 0.5%(w/v), the addition of polyglutamic acid is 0.5%(w/v), the addition of fructo-oligosaccharide (FOS) is 1%(w/v), temperature when spraying bacterium powder is 15 DEG C, and spraying frequency is 2 days/time, sprays 5 times continuously, when sugar content reaches 320g/L, expensive rotten grape finished product of can gathering.
Embodiment 5
Embodiment 5 is similar to the breeding method of embodiment 1, its difference is: the grape variety cultivated is Chardonnay, the time controlling grape maturity is early December, and the addition of bacterium powder is 0.1%(w/v), the addition of polyglutamic acid is 0.1%(w/v), the addition of fructo-oligosaccharide (FOS) is 0.5%(w/v), temperature when spraying bacterium powder is 10 DEG C, and spraying frequency is 1 day/time, sprays 3 times continuously, when sugar content reaches 310g/L, expensive rotten grape finished product of can gathering.
The relevant experimental data of above-described embodiment is as shown in the table:
Claims (6)
1. a breeding method for expensive rotten grape, is characterized in that, comprise the steps:
1) preparation of Botrytis cinerea bacterial classification: Botrytis cinerea bacterial strain is inoculated, liquid state fermentation to concentration is 10
6-10
8cFU/mL, filters, washing, dry, and is ground into particle diameter and is less than 200 object the pathogen of Botrytis cinerea powder;
2) cultivation of grape: with the white grape variety of making wine of maturation for plantation object, control grape fruit in sunny, temperature is fully ripe the month of 10-25 DEG C;
3) artificial infection Botrytis cinerea:
The preparation of A, bacterium powder suspension: the pathogen of Botrytis cinerea powder is suspended in clear water by the amount of mass volume ratio 0.1%-5%, adds the polyglutamic acid of 0.1%-2% by mass volume ratio, the fructo-oligosaccharide (FOS) of 0.5%-5%, fully mix for subsequent use;
B, selection weather are fine, mean temperature of air is evenly spray in ripe grape fruit ear surface by the pathogen of Botrytis cinerea powder suspension by the amount of 10mL-100mL/ fringe fruit the opportunity of 10 DEG C-25 DEG C, treat that grape fruit surface has obvious white thread bacterial plaque to generate, illustrate and inoculate successfully;
4) to gather expensive rotten grape: inoculate successfully, period sampling measuring fruit juice sugar content, when sugar content reaches 300g/L-340g/L, expensive rotten grape finished product of can gathering.
2. the breeding method of expensive rotten grape as claimed in claim 1, is characterized in that, described Botrytis cinerea bacterial classification liquid-state fermentation technology is as follows:
1) from original seed inclined-plane scraping Botrytis cinerea spore or mycelium, preparing biomass is 10
6-10
8the original seed suspension of CFU/mL, and the amount of 4%-10% is inoculated in seed culture medium by volume;
2) under 10 DEG C of-20 DEG C of conditions, oscillation and fermentation or stirring fermentation reach 5%-8% to thalline weight and culture volume ratio and stop fermenting.
3. the breeding method of expensive rotten grape as claimed in claim 2, it is characterized in that, the preparation technology of described seed culture medium is as follows:
1) get potato and fructus hordei germinatus that mass ratio is 1:1 and pulverize, dehydrated potato powder is broken to the fritter that diameter is not more than 1cm, and fructus hordei germinatus is crushed to particle diameter 20 order;
2) thin up, the quality of described water is 2.5 times of potato and fructus hordei germinatus gross mass;
3) 60 DEG C of saccharification 8h;
4) through 40 mesh filter screens or screen filter cleaner;
5) supplementing water, makes the volume of medium consistent with the volume before saccharification;
6) 110-121 DEG C of moist heat sterilization 20min.
4. the method for the expensive rotten grape of preparation as claimed in claim 1 artificial, is characterized in that, by rain shelter cultivation and/or term regulation technology control grape fruit maturation time.
5. the breeding method of expensive rotten grape as claimed in claim 1, is characterized in that, the spraying frequency of described the pathogen of Botrytis cinerea powder suspension is: often within 1-3 days, spray 1 time, sprays 3-6 time continuously.
6. the breeding method of expensive rotten grape as claimed in claim 1, it is characterized in that, described the pathogen of Botrytis cinerea is selected good strains in the field for seed from Botrytis cinerea ACCC36257 or ACCC36258 of Chinese agriculture Microbiological Culture Collection administrative center preservation, or is drawn by wild Botrytis cinerea bacterial strain screening.
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| CN201410523702.8A CN104255384B (en) | 2014-10-08 | 2014-10-08 | A kind of breeding method of expensive rotten grape |
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|---|---|---|---|
| CN201410523702.8A CN104255384B (en) | 2014-10-08 | 2014-10-08 | A kind of breeding method of expensive rotten grape |
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| CN104255384B CN104255384B (en) | 2016-03-30 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109055234A (en) * | 2018-08-08 | 2018-12-21 | 西北农林科技大学 | The preparation method and its application method of a kind of expensive rotten bacterium dry powder of grape |
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| JP2003070360A (en) * | 2001-08-31 | 2003-03-11 | Ryoji Watabe | Cultivation system for fruits and vegetables |
| CN101288375A (en) * | 2008-05-22 | 2008-10-22 | 辽宁太阳谷庄园葡萄酒业有限公司 | Ice grape planting process |
| CN101379939A (en) * | 2008-01-31 | 2009-03-11 | 纪俭 | New fruit culture technique of fresh grape and ice wine material |
| CN102260611A (en) * | 2010-05-31 | 2011-11-30 | 单小兵 | Method for brewing fruit noble rot and noble rot wine |
-
2014
- 2014-10-08 CN CN201410523702.8A patent/CN104255384B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2003070360A (en) * | 2001-08-31 | 2003-03-11 | Ryoji Watabe | Cultivation system for fruits and vegetables |
| CN101379939A (en) * | 2008-01-31 | 2009-03-11 | 纪俭 | New fruit culture technique of fresh grape and ice wine material |
| CN101288375A (en) * | 2008-05-22 | 2008-10-22 | 辽宁太阳谷庄园葡萄酒业有限公司 | Ice grape planting process |
| CN102260611A (en) * | 2010-05-31 | 2011-11-30 | 单小兵 | Method for brewing fruit noble rot and noble rot wine |
Non-Patent Citations (3)
| Title |
|---|
| 崔志峰等: "灰葡萄孢产孢和孢子萌发条件", 《浙江农业学报》, vol. 20, no. 6, 25 November 2008 (2008-11-25), pages 441 - 445 * |
| 赵红霞等: "灰葡萄孢菌致病机理研究进展", 《生物技术》, vol. 24, no. 1, 15 February 2014 (2014-02-15), pages 100 - 103 * |
| 邢守营: "贵腐葡萄及贵腐葡萄酒的酿造工艺", 《中国林副特产》, no. 8, 15 August 2013 (2013-08-15), pages 45 - 46 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109055234A (en) * | 2018-08-08 | 2018-12-21 | 西北农林科技大学 | The preparation method and its application method of a kind of expensive rotten bacterium dry powder of grape |
| CN109055234B (en) * | 2018-08-08 | 2022-02-25 | 西北农林科技大学 | Preparation method and use method of grape noble rot fungus dry powder |
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