CN112578041A - Grape rootstock evaluation method based on characteristic aroma substance GC-IMS fingerprint - Google Patents
Grape rootstock evaluation method based on characteristic aroma substance GC-IMS fingerprint Download PDFInfo
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- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/72—Mass spectrometers
- G01N30/7206—Mass spectrometers interfaced to gas chromatograph
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/86—Signal analysis
- G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
- G01N30/8686—Fingerprinting, e.g. without prior knowledge of the sample components
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Abstract
The invention provides a grape rootstock evaluation method based on a characteristic aroma substance GC-IMS fingerprint, which comprises the following steps: s1, standard sample treatment; s2, processing a sample to be detected; s3, analyzing a gas phase-ion mobility spectrum; s4, data processing and analyzing; s5, constructing a selection and evaluation method; and S6, judging a sample to be detected. The method analyzes the grape aroma substances of different stocks by combining a simple and rapid gas chromatography-ion mobility spectrometry technology with chemical analysis, and rapidly evaluates and judges the quality of fruits grafted on the different stocks based on the fingerprint spectrum of the characteristic aroma substances. The method is simple to operate, short in sample detection time, free of complex sample pretreatment and high in application value.
Description
Technical Field
The invention relates to the technical field of grape planting, in particular to a grape rootstock evaluation method based on a characteristic aroma substance GC-IMS fingerprint.
Background
Grape grafting cultivation began in the 70's of the 19 th century, and GastonBazille proposed grafting of European grapes onto American grapes to combat the aphid root cause of the Ozus tumefaciens. With the development of grape rootstock research and the popularization of grafting cultivation technology, besides the utilization of the resistance of the rootstock, the improvement of the growth result and the fruit quality of the grapes by the grafting technology is also a continuous research focus. The mutual influence of the rootstock and the scion is a relatively complex process, the root system of the rootstock can influence the absorption of nutrient elements of the scion, the growth vigor, the fruiting habit and the fruit quality of the scion are affected differently, the scion variety has certain influence on the root configuration of the same rootstock, particularly the root distribution and the root quantity, but the interaction mechanism of the rootstock and the scion is still unclear at present. The research at home and abroad finds that the rootstock has different influences on the aspects of phenological period, yield, ear quality, grain quality, fruit sugar acid content, soluble solid, fruit hardness, coloring, fruit resveratrol and other secondary metabolite content, maturity, storage period and the like of the grafted variety.
Before grafting, rootstock screening is needed for a specific grape variety. Screening rootstock resistance according to environmental factors of planting regions, such as cold resistance, salt and alkali resistance and the like; according to the characteristics of the variety to be grafted, carrying out stock and spike affinity evaluation; finally, the suitability of the grafted rootstock is comprehensively evaluated through the growth and development, fruiting characteristics and fruit quality after grafting. The result characteristic evaluation is an index for terminal evaluation, and is particularly important. In the existing research and technical application, the rootstock has relatively single evaluation standard for grape growth and development and fruiting characteristics, is usually evaluated and judged based on basic physicochemical indexes such as fruit grain size and sugar acid composition, and has certain randomness. The fruit character and the fruit quality of grapes grafted on different stocks by scholars in China are evaluated and analyzed, physical and chemical indexes such as fruit grain size and fruit shape index, fruit ear size and ear weight, soluble solid, acidity, fruit hardness, total phenol, total anthocyanin content, sugar composition and organic acid composition are measured, the evaluation indexes are various, the advantages of the grafted stocks are often judged according to the level of one or more basic physical and chemical indexes, the results are too level, and the conclusion cannot be well applied to production.
The GC-IMS analysis technology is widely applied to the field of food flavor research, and has the advantages of high sensitivity, good stability, high response speed, simple data processing and the like. In the current research, the aroma-based fingerprint spectrum is applied to the technical fields of origin discrimination, rapid detection, adulteration inspection, rapid classification, processing technology evaluation and the like. At present, no application example in the aspect of fruit tree stock grafting effect evaluation is seen. Therefore, the characteristic aroma substance composition can be used as an important index for evaluating the suitability of the stock of the variety. The method for efficiently evaluating the growth result and the fruit quality effect of the grafted grape has important application reference value and industrial significance.
The grape is 1 red bud change found in a high grafting test of 'Ruitongxiangyu' (female parent is 'Jingxiu' and male parent is 'Xiangfei') of forestry fruit tree research of agriculture and forestry academy of sciences of Beijing city, is approved by a crop variety approval committee of Beijing city in 12 months in 2014, and has the characteristics of precocity, red peel, crisp and sweet fruit pulp, rich rose fragrance and the like. Ruidehongyu is a typical rose-flavored grape variety, and the aroma characteristics are important factors for evaluating the fruit quality of the variety. The method can be used for evaluating the suitability of the variety stock of the RuiDou red grape based on the characteristic aroma substance composition, and has a very high practical application prospect.
Disclosure of Invention
The invention aims to solve the problems and provides a grape rootstock evaluation method based on a characteristic aroma substance GC-IMS fingerprint, which is simple to operate, short in sample detection time, free of complex sample pretreatment and high in application value.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows:
a grape rootstock evaluation method based on a characteristic aroma substance GC-IMS fingerprint comprises the following steps,
s1, standard sample treatment:
s11, standard sample selection: the method comprises the following steps that samples are from the same cultivation area, grapes are grafted on different stocks which are planted according to the same cultivation management measures, and the number N of the samples is 3-20 different stocks; selecting 1-3 samples of the variety from the root seedlings;
s12, sampling: mixing and sampling selected samples, cutting the samples by a blade, taking out grape seeds, crushing the grape seeds into powder to obtain a standard sample, filling the standard sample into an injection bottle, sealing the injection bottle by a sealing film, and freezing the sample at-80 ℃ for later use;
s2, treating a sample to be detected:
processing the rootstock to be detected according to the method of the step S1 to obtain a sample to be detected;
s3, gas phase-ion mobility spectrometry analysis: detecting volatile substances of the standard sample by a gas chromatography-ion mobility spectrometry instrument to obtain a fingerprint spectrogram;
s4, data processing and analysis:
extracting a characteristic value area on the fingerprint obtained in the step 2 by adopting a comparison method, wherein the comparison method is to select spectrogram characteristic areas with deep color change or large peak area difference of volatile matter mass spectrogram points of different rootstocks and self-rooted seedling grape fruit samples according to the color difference change and the peak area of characteristic matters in a fingerprint information spectrogram;
s5, constructing a selection and evaluation method:
according to the difference of the substance peak intensity signals and the retention time and migration time corresponding to the substances, 2-5 types of characteristic aroma substances which can obviously distinguish different rootstocks and self-rooted grapes are selected from the characteristic areas and used as the standard for evaluating the rootstocks of the grape variety;
s6, judging a sample to be detected:
and (4) detecting the sample to be detected by adopting the same instrument conditions in the step S3 to obtain a fingerprint, and judging according to the evaluation standard in the step S5.
Preferably, in step S1, the specific sampling step includes: processing and sampling each sample of the step S11 respectively, wherein 5 strains in each processing are 1 repetition, and 2-6 repetitions can be set; in each repeated experiment, 1-3 clusters of grapes are randomly sampled from each tree, 1 fruit grain is respectively sampled from the upper, middle and lower parts of a single grape, and the mixed sample is a repeated sample.
Preferably, in step S1, the grape seeds are removed, the grape seeds are added with liquid nitrogen and crushed into powder, and 1-10g of the sample is sampled in a frozen state and is filled into a headspace sampling bottle.
Preferably, in step S2, the sample is analyzed by gas-ion mobility spectrometry with a column type FS-SE-54-CB-1 at a column temperature of 40 deg.C and a carrier/drift gas N2IMS temperature is 45 ℃; and (3) headspace sample introduction unit conditions: the sample injection volume is 500ul, the incubation time is 15min, the incubation temperature is 40 ℃, the sample injection needle temperature is 85 ℃, and the incubation rotation speed is 500 rpm; the drift gas flow rate is 150mL/min, and the gas phase carrier gas flow rate is 2-20 mL/min.
Preferably, in step S4, when the grape variety is rdish red grape, the specific evaluation criteria are as follows:
(1) screening out a first class of substances, wherein the gas phase retention time of Pentanal and a polymer thereof is 3.760min, the ion drift time is 1.1935ms, the fragrance of green grass and bananas is reflected, the first class of substances is used for distinguishing rootstock grafting Ruiyu red jade grapes and self-rooted seedling grapes in main component analysis, when the peak area is higher than 350au, the first class of substances is self-rooted seedling Ruiyu red jade grapes, and otherwise, the first class of substances is rootstock grafting Ruiyu red jade grapes;
(2) a second class of substances was selected, including 3 substances including Linalool,2-Propanol,3-methylbutanal, with gas phase retention times: 16.575min, 2.191min and 3.302min, wherein the ion drift time is respectively as follows: 1.217ms, 1.169ms and 1.405ms, the critical peak area is judged to be 1000au, the method has the characteristics of flower fragrance and fruit fragrance of roses, lily of the valley, apples and the like, and the method is used for judging the typicality and the richness of the fragrance quality of the Ruitoyu grape fruit grafted on the rootstock in the analysis of main components;
the second type of substances are obviously contained, have typical fragrance of the variety, and do not have typical fragrance; the second type of substance has high peak response intensity and reflects high aromatic fullness, otherwise, the second type of substance has low aromatic fullness; whether the rootstock is suitable for selection can be directly judged according to the second substances;
(3) and screening out a third class of substances, which comprises 11 substances, wherein the substance number, the gas phase retention time and the ion drift time are as follows: the number 1 peak has gas phase retention time of 4.937min, ion drift time of 1.097ms, and critical peak area of 100 au;
the number 3 peak has gas phase retention time of 3.643min, ion drift time of 1.263ms, and critical peak area of 100 au;
the number 10 peak is gas phase retention time 3.523min, ion drift time 1.059ms, and critical peak area 500au is judged;
the No. 11 peak has the gas phase retention time of 3.510min, the ion drift time of 1.229ms, and the critical peak area of 100 au;
3-Pentanone, gas phase retention time 3.838min, ion drift time 1.108ms, critical peak area 300 au;
n-Nonanal, gas phase retention time 16.640min, ion drift time 1.477ms, and critical peak area 500 au;
furfurol, gas phase retention time 6.071min, ion drift time 1.083ms, and critical peak area 150 au;
2,3-Pentanedione, gas phase retention time is 3.812min, ion drift time is 1.224ms, and critical peak area is judged to be 100 au;
gamma-Butyrolactone, gas phase retention time 8.671min, ion drift time 1.080ms, critical peak area 100 au;
the 11 substances are used for judging the fullness or complexity of the aroma quality of the fruits of the RuiDou Hongyu grapes grafted on the rootstocks in the principal component analysis, 6-11 substances in the 11 substances indicate that the sample has high aroma fullness and strong complexity, and indicate that the rootstocks grafted on the rootstocks have superiority; on the contrary, less than 6 types of the samples indicate that the sample has a single aroma structure and low fullness, which indicates that the rootstocks used for grafting have no superiority.
Wherein, the inclusion of the substance means that the peak area for detection exceeds the critical peak area corresponding to each substance in the following table.
Due to the adoption of the technical scheme, the invention has the following beneficial effects:
1. the grape rootstock evaluation method based on the characteristic aroma substance GC-IMS fingerprint provided by the invention is used for analyzing the aroma substances of grapes of different rootstocks by combining a simple and rapid gas phase-ion mobility spectrometry technology with chemical analysis, and the quality of fruits grafted on the grapes of different rootstocks is rapidly evaluated and judged based on the characteristic aroma substance GC-IMS fingerprint. The method is simple to operate, short in sample detection time, free of complex sample pretreatment and high in application value.
2. According to the grape rootstock evaluation method based on the characteristic aroma substance GC-IMS fingerprint, aroma components of fresh grapes mainly comprise chemical components such as alcohol, aldehyde, ester and terpenes, and the volatile substances are secondary grape metabolites. Typical aroma characteristics of a particular species are made up of characteristic volatile components. The accumulated level of the types and the contents of the characteristic aroma substances can fully reflect the maturity and the fruit quality of the variety to a certain extent. Therefore, the characteristic of distinguishing the RuiDou ruby grafting stock based on the characteristic aroma substance fingerprint spectrum has scientific basis.
3. The grape rootstock evaluation method based on the characteristic aroma substance GC-IMS fingerprint greatly shortens and simplifies the fruit tree rootstock selection process, can be used for reference in the research and production of other fruit trees, and has very high practical application prospect.
Drawings
FIG. 1 is a comparison of fingerprints of volatile organic compounds in grape samples grafted on different rootstocks;
FIG. 2 shows the variation of the difference of volatile organic compounds in grape samples grafted on different rootstocks.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Example 1
A grape rootstock evaluation method based on a characteristic aroma substance GC-IMS fingerprint comprises the following steps,
s1, standard sample treatment:
s11, standard sample selection: the method comprises the following steps that samples are from the same cultivation area, grapes are grafted on different stocks which are planted according to the same cultivation management measures, and the number N of the samples is 3-20 different stocks; selecting 1-3 samples of the variety from the root seedlings;
s12, sampling: mixing and sampling the selected samples, wherein 5 strains in each treatment are 1 repetition, and 2-6 repetitions can be set; in each repeated experiment, 1-3 clusters of grapes are randomly sampled from each tree, 1 fruit grain is respectively sampled from the upper, middle and lower parts of a single grape, and the mixed sample is a repeated sample. Cutting with a blade, taking out grape seeds, crushing the grape pulp into powder to obtain a standard sample, filling the standard sample into a sample injection bottle, sealing with a sealing film, and freezing at-80 ℃ for sample storage;
s2, treating a sample to be detected:
processing the rootstock to be detected according to the method of the step S1 to obtain a sample to be detected;
s3, gas phase-ion mobility spectrometry analysis: detecting volatile substances of the standard sample by a gas chromatography-ion mobility spectrometry instrument to obtain a fingerprint spectrogram;
s4, data processing and analysis:
extracting a characteristic value area on the fingerprint obtained in the step 2 by adopting a comparison method, wherein the comparison method is to select spectrogram characteristic areas with deep color change or large peak area difference of volatile matter mass spectrogram points of different rootstocks and self-rooted seedling grape fruit samples according to the color difference change and the peak area of characteristic matters in a fingerprint information spectrogram; performing orthogonal transformation and dimension reduction on the material peak data of all the characteristic regions to realize rapid clustering on the NxM grape fruit samples;
s5, constructing a selection and evaluation method:
according to the difference of the substance peak intensity signals and the retention time and migration time corresponding to the substances, 2-5 types of characteristic aroma substances which can obviously distinguish different rootstocks and self-rooted grapes are selected from the characteristic areas and used as the standard for evaluating the rootstocks of the grape variety;
s6, judging a sample to be detected:
and (4) detecting the sample to be detected by adopting the same instrument conditions in the step S3 to obtain a fingerprint, and judging according to the evaluation standard in the step S5.
Example 2
A grape stock evaluation method based on characteristic aroma substance GC-IMS fingerprint spectrum, wherein the grape variety is Rui Du Hongyu grape, comprises the following steps,
s1, standard sample treatment:
s11, standard sample selection: the sample is from the same cultivation area, different stocks planted according to the same cultivation management measure are grafted with Ruilu red jade grapes, and stocks of varieties 11R, 5BB, 3309M, S04 and HY are selected; selecting 2 samples of the variety self-rooted seedlings;
s12, sampling: mixing and sampling the selected samples, wherein 5 strains in each treatment are 1 repetition, and 2-6 repetitions can be set; in each repeated experiment, 1-3 clusters of grapes are randomly sampled from each tree, 1 fruit grain is respectively sampled from the upper, middle and lower parts of a single grape, and the mixed sample is a repeated sample.
Cutting with a blade, taking out grape seeds, crushing the grape pulp into powder to obtain a standard sample, filling the standard sample into a sample injection bottle, sealing with a sealing film, and freezing at-80 ℃ for sample storage;
s2, treating a sample to be detected:
processing the rootstock to be detected according to the method of the step S1 to obtain a sample to be detected;
s3, gas phase-ion mobility spectrometry analysis: detecting volatile substances of the standard sample by a gas chromatography-ion mobility spectrometry instrument to obtain a fingerprint spectrogram;
analyzing the sample by gas-ion mobility spectrometry, wherein the chromatographic column type is FS-SE-54-CB-1, the column temperature is 40 ℃, and the carrier gas/drift gas is N2See IMS temperature 45 ℃; and (3) headspace sample introduction unit conditions: the sample injection volume is 500ul, the incubation time is 15min, the incubation temperature is 40 ℃, the sample injection needle temperature is 85 ℃, and the incubation rotation speed is 500 rpm; float for anglingThe gas flow rate is 150mL/min, and the gas flow rate of the gas-phase carrier gas is 2-20 mL/min.
TABLE 1 gas chromatography conditions
| Time | E1 (drifting airflow rate) | E2 (gas-phase carrier gas flow rate) | R |
| 00:00,000 | 150mL/min | 2ml/min | rec |
| 02:00,000 | 150mL/min | 2ml/min | - |
| 10:00,000 | 150mL/min | 20ml/min | - |
| 20:00,000 | 150mL/min | 100mL/min | - |
| 25:00,000 | 150mL/min | 150mL/min | stop |
| 25:00,000 | 150mL/min | 150mL/min | - |
S4, data processing and analysis:
extracting a characteristic value area on the fingerprint information spectrogram obtained in the step 2 by adopting a comparison method, wherein the comparison method is based on the color difference change and the peak area of the characteristic substance in the fingerprint information spectrogram; selecting spectrogram characteristic areas with deep color change or large peak area difference of volatile matter mass spectrogram material points of different rootstocks and self-rooted seedling grape fruit samples; and performing orthogonal transformation and dimension reduction on the material peak data of all the characteristic regions to realize rapid clustering on the NxM grape fruit samples.
S5, constructing a selection and evaluation method:
according to the difference of the substance peak intensity signals and the retention time and migration time corresponding to the substances, a plurality of characteristic fragrant substances which can obviously distinguish different rootstocks and self-rooted grapes are selected from the characteristic regions to evaluate and distinguish the grape variety rootstocks.
In fig. 1, each row represents all signal peaks selected in one grape sample; each column represents a signal peak of the same volatile organic compound in grape samples grafted on different rootstocks; from the figure, the complete volatile information for each sample and the difference in volatile organic compounds between samples can be seen
As can be seen from fig. 1, the whole fingerprint spectrum is 110R, 5BB, S04, 3309m and HY grape sample from top to bottom, wherein: the substances in the red frame are respectively substances with higher content in each grape sample relative to other samples;
it can be clearly seen that:
the substances with higher content in the 110R sample and lower content in other samples are substances such as 2, 3-pentaneedione (2, 3-acetylacetone), gamma-Butyrolactone (gamma-Butyrolactone), n-Nonanal (Nonanal), furfururol (furfural), 3-Pentanone (3-Pentanone), and the like.
Among the higher levels of material in the S04 sample and the lower levels of material in the other samples were 1-Hexanol (1-Hexanol), methyl propanal (isobutyraldehyde), Ethyl Acetate (ethylacetate), and 3-methylbutanal (3-methylbutanal).
The substances with higher content in the 5BB sample and lower content in other samples are acetone and the like.
"13" to "14" are substances having a high content in a 3309m sample and a low content in other samples.
The materials with higher content in the HY sample and lower content in other samples were Pentanal (Pentanal).
The specific evaluation criteria are as follows:
(1) screening out a first class of substances, wherein the gas phase retention time of Pentanal and a polymer thereof is 3.760min, the ion drift time is 1.1935ms, the fragrance of green grass and bananas is reflected, the first class of substances is used for distinguishing rootstock grafting Ruiyu red jade grapes and self-rooted seedling grapes in main component analysis, when the peak area is higher than 350au, the first class of substances is self-rooted seedling Ruiyu red jade grapes, and otherwise, the first class of substances is rootstock grafting Ruiyu red jade grapes;
(2) a second class of substances was selected, including 3 substances including Linalool,2-Propanol,3-methylbutanal, with gas phase retention times: 16.575min, 2.191min and 3.302min, wherein the ion drift time is respectively as follows: 1.217ms, 1.169ms and 1.405ms, the critical peak area is judged to be 1000au, the method has the characteristics of flower fragrance and fruit fragrance of roses, lily of the valley, apples and the like, and the method is used for judging the typicality and the richness of the fragrance quality of the Ruitoyu grape fruit grafted on the rootstock in the analysis of main components;
the second type of substances are obviously contained, have typical fragrance of the variety, and do not have typical fragrance; the second type of substance has high peak response intensity and reflects high aromatic fullness, otherwise, the second type of substance has low aromatic fullness; whether the rootstock is suitable for selection can be directly judged according to the second substances;
(3) the third class of substances, including 11 substances, whose qualitative parameters (substance number, gas phase retention time and ion drift time) are shown in table 2 below, were screened, and the inclusion of the substance means that the detected peak area exceeded the critical peak area corresponding to each substance in the table below.
TABLE 2 qualitative parameter List
| Material number or name | Gas phase retention time/min | Ion drift time/ms | Discrimination Critical Peak area/ |
| Peak number | |||
| 1 | 4.937 | 1.097 | 100 |
| Peak No. 3 | 4.599 | 1.360 | 50 |
| Peak No. 7 | 3.643 | 1.263 | 100 |
| |
3.523 | 1.059 | 500 |
| |
3.510 | 1.229 | 100 |
| |
2.795 | 1.152 | 500 |
| 3-Pentanone | 3.838 | 1.108 | 300 |
| n-Nonanal | 16.640 | 1.477 | 500 |
| Furfurol | 6.071 | 1.083 | 150 |
| 2,3-Pentanedione | 3.812 | 1.224 | 100 |
| gamma-Butyrolactone | 8.671 | 1.080 | 100 |
The 11 substances are used for judging the fullness or complexity of the aroma quality of the fruits of the RuiDou Hongyu grapes grafted on the rootstocks in the principal component analysis, 6-11 substances in the 11 substances indicate that the sample has high aroma fullness and strong complexity, and indicate that the rootstocks grafted on the rootstocks have superiority; on the contrary, less than 6 types of the samples indicate that the sample has a single aroma structure and low fullness, which indicates that the rootstocks used for grafting have no superiority.
According to the three selected substances, the Rui-Du red jade grape sample is rapidly analyzed through GC-IMS, so that the grape of the self-rooted seedling and the grape of the grafted seedling can be distinguished based on the characteristic fragrant substances of the first kind, the suitability of the rootstock is evaluated based on the second kind of substances, and the superiority of the third kind of substances to the rootstock is further distinguished and screened, so that the purpose of rapidly screening the rootstock is achieved.
S6, judging a sample to be detected:
and (4) detecting the sample to be detected by adopting the same instrument conditions in the step S3 to obtain a fingerprint, and judging according to the evaluation standard in the step S5.
The above description is intended to describe in detail the preferred embodiments of the present invention, but the embodiments are not intended to limit the scope of the claims of the present invention, and all equivalent changes and modifications made within the technical spirit of the present invention should fall within the scope of the claims of the present invention.
Claims (5)
1. A grape rootstock evaluation method based on a characteristic aroma substance GC-IMS fingerprint is characterized by comprising the following steps,
s1, standard sample treatment:
s11, standard sample selection: the method comprises the following steps that samples are from the same cultivation area, grapes are grafted on different stocks which are planted according to the same cultivation management measures, and the number N of the samples is 3-20 different stocks; selecting 1-3 samples of the variety from the root seedlings;
s12, sampling: mixing and sampling selected samples, cutting the samples by a blade, removing grape seeds, crushing pulp into powder to obtain a standard sample, filling the standard sample into a sample injection bottle, sealing the sample injection bottle by a sealing film, and freezing the sample at-80 ℃ for later use;
s2, treating a sample to be detected:
processing the rootstock to be detected according to the method of the step S1 to obtain a sample to be detected;
s3, gas phase-ion mobility spectrometry analysis: detecting volatile substances of the standard sample by a gas chromatography-ion mobility spectrometry instrument to obtain a fingerprint spectrogram;
s4, data processing and analysis:
extracting a characteristic value area on the fingerprint obtained in the step 2 by adopting a comparison method, wherein the comparison method is to select spectrogram characteristic areas with deep color change or large peak area difference of volatile matter mass spectrogram points of different rootstocks and self-rooted seedling grape fruit samples according to the color difference change and the peak area of characteristic matters in a fingerprint information spectrogram;
s5, constructing a selection and evaluation method:
according to the difference of the substance peak intensity signals and the retention time and migration time corresponding to the substances, 2-5 types of characteristic aroma substances which can obviously distinguish different rootstocks and self-rooted grapes are selected from the characteristic areas and used as the standard for evaluating the rootstocks of the grape variety;
s6, judging a sample to be detected:
and (4) detecting the sample to be detected by adopting the same instrument conditions in the step S3 to obtain a fingerprint, and judging according to the evaluation standard in the step S5.
2. The method for evaluating the grape rootstock based on the GC-IMS fingerprint of the characteristic aroma substance according to claim 1, wherein in the step S1, the specific sampling steps are as follows: processing and sampling each sample of the step S11 respectively, wherein 5 strains in each processing are 1 repetition, and 2-6 repetitions can be set; in each repeated experiment, 1-3 clusters of grapes are randomly sampled from each tree, 1 fruit grain is respectively sampled from the upper, middle and lower parts of a single grape, and the mixed sample is a repeated sample.
3. The method for evaluating grape rootstock based on the GC-IMS fingerprint of the characteristic aroma substance according to claim 1, wherein in the step S1, grape seeds are taken out, liquid nitrogen is added to the grape seeds and the grape seeds are crushed into powder, and 1 to 10g of a sample is sampled in a frozen state and then the sample is filled into a headspace sampling bottle.
4. The method for evaluating grape rootstock based on the characteristic aroma substance GC-IMS fingerprint according to claim 1, wherein in step S2, a gas phase-ion mobility spectrometry is used for analyzing the sample, the type of a chromatographic column is FS-SE-54-CB-1, the column temperature is 40 ℃, and the carrier gas/drift gas is N2IMS temperature is 45 ℃; and (3) headspace sample introduction unit conditions: the sample injection volume is 500ul, the incubation time is 15min, the incubation temperature is 40 ℃, the sample injection needle temperature is 85 ℃, and the incubation rotation speed is 500 rpm; the drift gas flow rate is 150mL/min, and the gas phase carrier gas flow rate is 2-20 mL/min.
5. The method for evaluating grape rootstock based on the GC-IMS fingerprint of the characteristic aroma substance according to claim 1, wherein in the step S4, when the grape variety is Vitis vinifera, the specific evaluation criteria are as follows:
(1) screening out a first class of substances, wherein the gas phase retention time of Pentanal and a polymer thereof is 3.760min, the ion drift time is 1.1935ms, the fragrance of green grass and bananas is reflected, the first class of substances is used for distinguishing rootstock grafting Ruiyu red jade grapes and self-rooted seedling grapes in main component analysis, when the peak area is higher than 350au, the first class of substances is self-rooted seedling Ruiyu red jade grapes, and otherwise, the first class of substances is rootstock grafting Ruiyu red jade grapes;
(2) a second class of substances was selected, including 3 substances including Linalool,2-Propanol,3-methylbutanal, with gas phase retention times: 16.575min, 2.191min and 3.302min, wherein the ion drift time is respectively as follows: 1.217ms, 1.169ms and 1.405ms, the critical peak area is judged to be 1000au, the method has the characteristics of flower fragrance and fruit fragrance of roses, lily of the valley, apples and the like, and the method is used for judging the typicality and the richness of the fragrance quality of the Ruitoyu grape fruit grafted on the rootstock in the analysis of main components;
the second type of substances are obviously contained, have typical fragrance of the variety, and do not have typical fragrance; the second type of substance has high peak response intensity and reflects high aromatic fullness, otherwise, the second type of substance has low aromatic fullness; whether the rootstock is suitable for selection can be directly judged according to the second substances;
(3) and screening out a third class of substances, which comprises 11 substances, wherein the substance number, the gas phase retention time and the ion drift time are as follows: the number 1 peak has gas phase retention time of 4.937min, ion drift time of 1.097ms, and critical peak area of 100 au;
the number 3 peak has gas phase retention time of 3.643min, ion drift time of 1.263ms, and critical peak area of 100 au;
the number 10 peak is gas phase retention time 3.523min, ion drift time 1.059ms, and critical peak area 500au is judged;
the No. 11 peak has the gas phase retention time of 3.510min, the ion drift time of 1.229ms, and the critical peak area of 100 au;
peak 16, gas phase retention time 2.795min, ion drift time 1.152ms, and critical peak area 500 au;
3-Pentanone, gas phase retention time 3.838min, ion drift time 1.108ms, critical peak area 300 au;
n-Nonanal, gas phase retention time 16.640min, ion drift time 1.477ms, and critical peak area 500 au;
furfurol, gas phase retention time 6.071min, ion drift time 1.083ms, and critical peak area 150 au;
2,3-Pentanedione, gas phase retention time is 3.812min, ion drift time is 1.224ms, and critical peak area is judged to be 100 au;
gamma-Butyrolactone, gas phase retention time 8.671min, ion drift time 1.080ms, critical peak area 100 au;
the 11 substances are used for judging the fullness or complexity of the aroma quality of the fruits of the RuiDou Hongyu grapes grafted on the rootstocks in the principal component analysis, 6-11 substances in the 11 substances indicate that the sample has high aroma fullness and strong complexity, and indicate that the rootstocks grafted on the rootstocks have superiority; on the contrary, less than 6 types of the samples indicate that the sample has a single aroma structure and low fullness, which indicates that the rootstocks used for grafting have no superiority.
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