CN109187673A - A kind of plant Relationship iden- tification method based on pollen - Google Patents
A kind of plant Relationship iden- tification method based on pollen Download PDFInfo
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- CN109187673A CN109187673A CN201811242374.9A CN201811242374A CN109187673A CN 109187673 A CN109187673 A CN 109187673A CN 201811242374 A CN201811242374 A CN 201811242374A CN 109187673 A CN109187673 A CN 109187673A
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- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
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Abstract
The plant Relationship iden- tification method based on pollen that the invention discloses a kind of.This includes the following steps: that the plant pollen by n kind sample is added separately to extract in m kind different solvents, obtains nm kind pollen extract liquor；Wherein, n >=2, m >=2；The nm kind pollen extract liquor is mixed with graphene dispersing solution respectively and is embedded, nm kind embedding system is obtained；Nm kind embedding system is respectively coated in screen printing electrode surface, nm kind modified electrode is obtained；The nm kind modified electrode is respectively placed in progress Pulse Voltammetry scanning in phosphate buffer, constructs to obtain electrochemistry map by the multi-loop electric chemical signal that nm kind modified electrode respectively obtains；The similarity of the electrochemistry map of different samples in homogeneous solvent is compared, electrochemistry map registration is higher, and plant affiliation is closer.The advantages that present invention has qualification process simple and easy, and identification speed is fast, at low cost.
Description
Technical field
The invention belongs to plant identification technical fields, more particularly to a kind of plant Relationship iden- tification side based on pollen
Method.
Background technique
Plant Taxonomy is the origin of the Different groups for mainly studying entire plant kingdom, affiliation, and evolution
One basic subject of the rule of development.Namely numerous and complicated plant kingdom is classified and differentiates to plant always, and presses system
It lines up, in order to which people recognize and utilize plant.
According to incompletely statistics, about 500,000 kinds or more of the plant to survive on earth (250000 kinds of left sides of seed plant
It is right).And the affiliation about plant, it mainly conducts a research so far from plant morphology and DNA molecular marker etc..And
Start with from morphology, the error of naked eyes judgement is too big, and efficiency and accuracy be not high and imperfect to form when identifying affiliation
The plant of even only pollen samples is helpless；And DNA molecular marker method can DNA sequencing to two kinds of plants and comparison, but
This obvious process of method is cumbersome and cost is excessively high.
Seed is phanerogamous breeding system, mostly remains the inhereditary material of plant to the greatest extent, but much natural hybrization or
The plant of artificial hybridization breeding is triploid plant, and by taking Lycoris as an example, they normally bloom but seeds abortion.Therefore with plant flowers
It is accurate for the more science of the affiliation between plant identification that powder is started with.
For example, CN 107488712A discloses the polychrome of a kind of quickly analysis and detection Tribe Triticeae chromosome configuration
Mc-GISH is applied to point of chromosome pairing behavior during pollen mother cells by in-situ hybridization method, this method
In analysis, primary hybridization can effectively identify chromosome pairing situation of Tribe Triticeae different genes group during meiosis, energy
The affiliation being enough between analysis Tribe Triticeae different genes group.However this method qualification process is cumbersome, cost is excessively high, unfavorable
In Large scale identification.
To sum up, the present invention in view of the above-mentioned defects in the prior art and the market demand, develops a kind of plant based on pollen
Relationship iden- tification method, this method realize the fast of plant affiliation with electrochemical means, by simple and easy method
Speed identification.
Summary of the invention
The plant Relationship iden- tification method based on pollen that the purpose of the present invention is to provide a kind of.The present invention has identification
The advantages that process is simple and easy, and identification speed is fast, at low cost.
To achieve the above object, the present invention takes following technical scheme:
A kind of plant Relationship iden- tification method based on pollen, includes the following steps:
(1) plant pollen of n kind sample is added separately to extract in m kind different solvents, obtains nm kind pollen extraction
Take liquid；Wherein, n >=2, m >=2.Pollen grain does not have particular/special requirement, and the higher the better for integrated degree；
(2) the nm kind pollen extract liquor is mixed with graphene dispersing solution respectively and is embedded, obtain nm kind embedding body
System.In the present invention, the specific surface area of the graphene dispersing solution is far longer than plant tissue, can be realized to plant tissue
Embedding, forms uniform dispersion liquid；
(3) nm kind embedding system is respectively coated in screen printing electrode surface, obtains nm kind modified electrode.It is described
The embodiment of coating does not have particular/special requirement, and dry modified electrode is obtained after finishing to solvent natural evaporation；
(4) the nm kind modified electrode is respectively placed in progress Pulse Voltammetry scanning in phosphate buffer, by nm kind
The multi-loop electric chemical signal that modified electrode respectively obtains constructs to obtain electrochemistry map；
(5) similarity of the electrochemistry map of different samples in homogeneous solvent is compared, electrochemistry map registration is higher, plants
Object affiliation is closer.
Preferably, the ratio between the plant pollen grain number amount of sample and the volume of solvent are 1:(30~200 in the step (1))
μL。
Preferably, solvent includes water, dehydrated alcohol, methanol, ethylene glycol, n,N-Dimethylformamide in the step (1)
(DMF) or one of n-hexane.
Preferably, the concentration of step (2) graphene dispersing solution is 0.1~1mg/mL.
Preferably, the volume ratio of pollen extract liquor and graphene dispersing solution is (50~120) in the step (2): (5~
12)。
Preferably, each screen electrode surface coats the embedding system of 1~8 μ L in the step (3).
Preferably, step (4) phosphate buffer is dissolved in water by potassium dihydrogen phosphate and sodium dihydrogen phosphate and is formulated,
The concentration of middle potassium dihydrogen phosphate and sodium dihydrogen phosphate independently is 0.03~0.3mol/L.
Preferably, the sweep interval of pulse voltammetric scan is 1.2V~-0.2V in the step (4)；Scanning speed is 80
~150mV/s；Sampling interval is 1~5mV.
The present invention provides one kind based on electrochemical analysis, is the plant relationship for implementing object with different plant pollens
Relationship analysis method.Different extracts are obtained with different solvents extraction plant pollen, by extract modification in electrode surface, to repairing
The electrode adornd carries out Pulse Voltammetry scanning, and obtained electrochemical signals are converted into electrochemistry map, by comparing not same
The similarity of product electrochemistry map in homogeneous solvent is far and near come the affiliation for judging the two.Identification method provided by the invention
It is simple and easy, the affiliation of the close plant of energy Rapid identification form.
The invention has the advantages that:
1. entire analytic process proposed by the present invention is not necessarily to expensive reagent and instrument, congener relationship pass is greatly reduced
It is the cost identified.
2. using plant pollen as test sample, largely avoid because plant tissue caused by growing environment at
Divide difference.
3. plant parentage analysis method proposed by the present invention potential can be fabricated to mini kit product, has and examine on the spot
The potentiality of survey.
Detailed description of the invention
Fig. 1 is the standard electrochemical of Lycoris, straw short-tube lycoris, Lycoris aurea, Guangxi short-tube lycoris and rose short-tube lycoris in ethylene glycol
Learn map.
Fig. 2 is Lycoris, straw short-tube lycoris, Lycoris aurea, the standard electrochemical figure of Guangxi short-tube lycoris and rose short-tube lycoris in water
Spectrum.
Fig. 3 is the standard electric of Bulbus Lycoridis longitubae, yellow Bulbus Lycoridis longitubae, fragrant short-tube lycoris, red blue stone garlic and Anhui short-tube lycoris in ethylene glycol
Chemical profile.
Fig. 4 is Bulbus Lycoridis longitubae, yellow Bulbus Lycoridis longitubae, fragrant short-tube lycoris, the standard electrochemical of red blue stone garlic and Anhui short-tube lycoris in water
Map.
Specific embodiment
Following specific embodiments are the further explanations to method provided by the invention and technical solution, but are not construed as
Limitation of the present invention.
Embodiment 1: identify the affiliation of lycoris plants known to 10 various names.
(1) preparation of buffer:
By potassium dihydrogen phosphate and disodium hydrogen phosphate preparation phosphate buffer soluble in water；Potassium dihydrogen phosphate and sodium dihydrogen phosphate
Concentration be 0.1mol/L；
(2) prepared by plant pollen extract liquor:
In Lycoris, straw short-tube lycoris, Lycoris aurea, Guangxi short-tube lycoris, rose short-tube lycoris, Bulbus Lycoridis longitubae, yellow Bulbus Lycoridis longitubae, fragrant stone
In ten garlic, red blue stone garlic and Anhui short-tube lycoris pollen samples, two pollen grains is respectively taken to be added separately to 200 μ L ethylene glycol, 200 μ L
In water, sufficiently after oscillation, 20 parts of above-mentioned 10 kinds of Lycoris pollen pollen extract liquor in water and methanol respectively is obtained.
(3) graphene embeds:
The 100 μ L of pollen extract liquor for taking above-mentioned Lycoris, is added in 10 μ L graphene dispersing solutions each independently, shakes rapidly
It is even, it is embedded into pollen extract liquor sufficiently in graphene dispersing solution, obtains the embedding system of 20 kinds of Lycoris pollen.
(4) electrode modification:
Each 2 μ L of embedding system for taking above-mentioned 20 kinds of Lycoris pollen, is added dropwise each independently in screen printing electrode surface,
20 modified electrodes that different embedding systems have been added dropwise are obtained after it is spontaneously dried.
(5) electrochemical analysis:
20 sheet modified electrodes are independently respectively placed in phosphate buffer and are formed into a loop, Pulse Voltammetry scanning is carried out.Its
In, the sweep interval of the Pulse Voltammetry scanning is 1.2V~-0.2V；Scanning speed is 100mV/s；Sampling interval is 1mV.
Every kind of Lycoris blade obtains 2 groups of different electrochemical signals (with ethylene glycol extraction, with each one group of water extraction),
The electrochemistry map for constructing sample respectively, obtains electrochemistry map, as a result as shown in attached drawing 1~4.
Wherein, Fig. 1,2 and Fig. 3,4 comparisons are it is found that the electrification that short-tube lycoris pollen of the same race obtains after the processing of different extract liquors
It is different to learn map.In addition, by short-tube lycoris electrograph spectrum comparison not of the same race in group picture it is found that not of the same race after extract liquor of the same race is processed
The getable electrochemistry map of short-tube lycoris pollen institute is similar without identical, and affiliation is closer, and electrochemistry map registration is got over
Greatly.Such as there are two current peaks between 0.0-0.2V for rose short-tube lycoris and straw short-tube lycoris as can see from Figure 1, and in 0.9V
Between have a current peak, although peak height is different, peak shape is similar.Meanwhile it can be seen that rose short-tube lycoris and straw stone in Fig. 2
Garlic has a current peak at the place 0.25V or so.In conjunction with the result of two figures, it is believed that the electrochemistry of rose short-tube lycoris and straw short-tube lycoris
Active material is similar, therefore affiliation is closer each other.For another example Bulbus Lycoridis longitubae and yellow Bulbus Lycoridis longitubae are between 0-0.4V in Fig. 3
There is apparent current peak, also there is apparent current peak between 0.8-1.0.Meanwhile in Fig. 4 it can be seen that Bulbus Lycoridis longitubae and
Yellow Bulbus Lycoridis longitubae has similar current peak between 0.1-0.8V.In conjunction with the result of two figures, it is believed that Bulbus Lycoridis longitubae and Huang
The electroactive substance of Bulbus Lycoridis longitubae is similar, therefore affiliation is closer each other.
As seen from the above embodiment, the present invention provides one kind based on electrochemical analysis, is with different plant pollens
Implement the plant Genetic relationship method of object.Different extracts are obtained with different solvents extraction plant pollen, by extract
It modifies in electrode surface, Pulse Voltammetry scanning is carried out to the electrode of modified, obtained electrochemical signals are converted into electrochemistry
Map judges that the affiliation of the two is far and near by comparing the similarity of two sample electrochemistry maps in homogeneous solvent.
Identification method provided by the invention is simple and easy, the affiliation of the close plant of energy Rapid identification form.
The method of the present invention that the above embodiments are only used to help understand and its core concept.It should be pointed out that for
For those skilled in the art, without departing from the principle of the present invention, if can also be carried out to the present invention
Dry improvement and modification, these improvement and modification are also fallen into the claims in the present invention protection scope.
Claims (8)
1. a kind of plant Relationship iden- tification method based on pollen, which comprises the steps of:
(1) plant pollen of n kind sample is added separately to extract in m kind different solvents, obtains nm kind pollen extract liquor；
Wherein, n >=2, m >=2；
(2) the nm kind pollen extract liquor is mixed with graphene dispersing solution respectively and is embedded, obtain nm kind embedding system；
(3) nm kind embedding system is respectively coated in screen printing electrode surface, obtains nm kind modified electrode；
(4) the nm kind modified electrode is respectively placed in progress Pulse Voltammetry scanning in phosphate buffer, is modified by nm kind
The multi-loop electric chemical signal that electrode respectively obtains constructs to obtain electrochemistry map；
(5) similarity of the electrochemistry map of different samples in homogeneous solvent is compared, electrochemistry map registration is higher, plant parent
Edge relationship is closer.
2. a kind of plant Relationship iden- tification method based on pollen according to claim 1, which is characterized in that the step
Suddenly the ratio between the plant pollen grain number amount of sample and the volume of solvent are 1:(30~200 in (1)) μ L.
3. a kind of plant Relationship iden- tification method based on pollen according to claim 1, which is characterized in that the step
Suddenly solvent includes one of water, dehydrated alcohol, methanol, ethylene glycol, N,N-dimethylformamide (DMF) or n-hexane in (1).
4. a kind of plant Relationship iden- tification method based on pollen according to claim 1, which is characterized in that the step
Suddenly the concentration of (2) graphene dispersing solution is 0.1~1mg/mL.
5. a kind of plant Relationship iden- tification method based on pollen according to claim 1, which is characterized in that the step
Suddenly the volume ratio of pollen extract liquor and graphene dispersing solution is (50~120): (5~12) in (2).
6. a kind of plant Relationship iden- tification method based on pollen according to claim 1, which is characterized in that the step
Suddenly each screen electrode surface coats the embedding system of 1~8 μ L in (3).
7. a kind of plant Relationship iden- tification method based on pollen according to claim 1, which is characterized in that the step
Suddenly (4) phosphate buffer is dissolved in water by potassium dihydrogen phosphate and sodium dihydrogen phosphate and is formulated, wherein potassium dihydrogen phosphate and di(2-ethylhexyl)phosphate
The concentration of hydrogen sodium independently is 0.03~0.3mol/L.
8. a kind of plant Relationship iden- tification method based on pollen according to claim 1, which is characterized in that the step
Suddenly the sweep interval of pulse voltammetric scan is 1.2V~-0.2V in (4)；Scanning speed is 80~150mV/s；Sampling interval is 1
~5mV.
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| CN201811242374.9A CN109187673B (en) | 2018-10-24 | 2018-10-24 | Plant genetic relationship identification method based on pollen |
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| CN201811242374.9A CN109187673B (en) | 2018-10-24 | 2018-10-24 | Plant genetic relationship identification method based on pollen |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111128310A (en) * | 2019-12-24 | 2020-05-08 | 国家林业和草原局桉树研究开发中心 | Method for identifying eucalyptus tree species and genetic relationship between species |
| CN111398398A (en) * | 2020-04-01 | 2020-07-10 | 杭州电子科技大学 | Method for constructing camellia nitidissima phylogenetic tree |
| CN111413392A (en) * | 2020-03-25 | 2020-07-14 | 杭州电子科技大学 | Method for collecting electrochemical spectrum of lycoris seeds |
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| CN105936936A (en) * | 2016-06-22 | 2016-09-14 | 陕西师范大学 | Method for detecting Psa in kiwi fruit pollens |
| CN107488712A (en) * | 2017-08-07 | 2017-12-19 | 山东农业大学 | A kind of quick analysis and the polychrome in-situ hybridization method of detection Tribe Triticeae chromosome configuration |
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| CN105936936A (en) * | 2016-06-22 | 2016-09-14 | 陕西师范大学 | Method for detecting Psa in kiwi fruit pollens |
| CN107488712A (en) * | 2017-08-07 | 2017-12-19 | 山东农业大学 | A kind of quick analysis and the polychrome in-situ hybridization method of detection Tribe Triticeae chromosome configuration |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111128310A (en) * | 2019-12-24 | 2020-05-08 | 国家林业和草原局桉树研究开发中心 | Method for identifying eucalyptus tree species and genetic relationship between species |
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| CN111413392A (en) * | 2020-03-25 | 2020-07-14 | 杭州电子科技大学 | Method for collecting electrochemical spectrum of lycoris seeds |
| CN111413392B (en) * | 2020-03-25 | 2022-05-13 | 杭州电子科技大学 | Method for collecting electrochemical spectrum of lycoris seeds |
| CN111398398A (en) * | 2020-04-01 | 2020-07-10 | 杭州电子科技大学 | Method for constructing camellia nitidissima phylogenetic tree |
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Effective date of registration: 20220217 Address after: 230000 Anhui Hefei high tech Zone Innovation Industrial Park two phase J2 District C block 18 floor. Patentee after: HEFEI LONGTUTEM INFORMATION TECHNOLOGY Co.,Ltd. Address before: 310018 no.1158, No.2 street, Baiyang street, Hangzhou Economic and Technological Development Zone, Zhejiang Province Patentee before: HANGZHOU DIANZI University |