CN105028492A - Application of rhodiola rosea extract to botrytis cinerea and alternaria oleracea inhibition - Google Patents
Application of rhodiola rosea extract to botrytis cinerea and alternaria oleracea inhibition Download PDFInfo
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Abstract
The invention discloses an application of a rhodiola rosea extract to botrytis cinerea and alternaria oleracea inhibition. A preparation method of the rhodiola rosea extract comprises the steps of extracting the root of rhodiola rosea by using 95 percent ethanol water, collecting an extracting solution, and removing ethanol in the extracting solution, so as to obtain a 95 percent ethanol extract of the rhodiola rosea, namely the rhodiola rosea extract. Experiments prove that a rhodiola crenulata 95 percent ethanol extract, a small flower rhodiola rosea 95 percent ethanol extract and a rhodiola elongata 95 percent ethanol extract with the concentration of 1mg/ml have obvious inhibition functions on botrytis cinerea and alternaria oleracea, indicating that rhodiola crenulata, rhodiola elongata and/or small flower rhodiola rosea can be used for preparing botrytis cinerea and alternaria oleracea inhibitors.
Description
The divisional application that the application is application number is 201410069489.8, the applying date is on 02 27th, 2014, invention and created name is " gadol extract and preparation method thereof and application ".
Technical field
The present invention relates to gadol extract and suppress the application in botrytis cinerea and Alternaria brassicae.
Background technology
Plant pathogenic fungi is distributed widely in all over the world, cause crops, forest, vegetables, economic crops, ornamental crops disease, often cause that the early stage chlorosis of plant, leaf are withered, fallen leaves, withered and rot, have a strong impact on growth and development of plants, Yield and qualities, bring heavy losses to agricultural production and people's lives.Current fungal diseases of plants control is still based on the Agro-chemicals control of organic synthesis, but the use of a large amount of chemical pesticide, bring very large negative environmental impact, and easily make plant produce poisoning and pesticide resistance, have a strong impact on the application of anti-plant pathogenic fungi preparation in agricultural production.In recent years; along with the improvement of the safeguard measures such as greenhouse by solar heat, plastic tunnel, plastic mulching; the fungal disease of fruits and vegetables is serious, has become the main cause that restriction China fruits and vegetables is efficient, keep the safety in production, has often made national economy and people's lives sustain losses severely.Along with Agricultural product without pollution scale progressively expand and chemical pesticide use the problem such as pesticide resistance, residue of pesticide produced day by day seriously, find and use nontoxic, efficient novel plant source pesticide is more and more subject to people's attention.
Crassulaceae (Crassulaceae) rhodiola (Rhodiola) plant, the whole world about has 90 kinds, is mainly distributed in Northern Hemisphere severe cold areas.Be distributed in Himalaya Region, western part of Asia to northern, through Korea, Japan to North America, China is the distribution center of Rhodida plant, and about more than 70 plant more.Rhodida plant is perennial tool root-like stock draft, and root-like stock has scale; The different strain of flower unisexuality or polygamy, line up corymb, and usual 4 numbers, male flower has the gynoecium of degeneration, and female flower is without the stamen of degenerating, and petal is very little or disappear completely.Pharmacological research shows, rhodiola root has antitumor, anti-oxidant, the function such as antifatigue, radioresistance, is the important medicinal material of Aeronautics and Astronautics, military medicine, sports medical science and health care.Wherein the more clearly major physiological active substance being also everybody comparatively admits of research is rhodioside (salidroside) at present.
Summary of the invention
Technical problem to be solved by this invention is to provide gadol extract and preparation method thereof and application.
The invention provides three kinds of gadol extracts.
The first gadol extract provided by the present invention is rhodiola root 95% ethanol extract, its preparation method comprises: extract root of Rhodiola sachalinensis with 95% ethanol water, collect extract and the ethanol removed in described extract, obtain rhodiola root 95% ethanol extract; Described rhodiola root 95% ethanol extract is the first gadol extract; Described rhodiola root is rhodiola, sachalin rhodiola rhizome and/or little premium red-spotted stonecrop.
The second gadol extract provided by the present invention is rhodiola root acetic acid ethyl ester extract, its preparation method comprises: 1) extract root of Rhodiola sachalinensis with 95% ethanol water, collect extract and the ethanol removed in described extract, obtain rhodiola root 95% ethanol extract;
2) aqueous dispersion of described rhodiola root 95% ethanol extract is become suspension, with suspension described in petroleum ether extraction, collect aqueous phase, be extracted with ethyl acetate described aqueous phase, collect ethyl acetate phase, remove described ethyl acetate mutually in ethyl acetate namely obtain rhodiola root acetic acid ethyl ester extract; Described rhodiola root acetic acid ethyl ester extract is the second gadol extract;
Described rhodiola root is rhodiola, sachalin rhodiola rhizome and/or little premium red-spotted stonecrop.
The third gadol extract provided by the present invention is rhodiola root eluent ethyl acetate thing, and its preparation method comprises:
1) extract root of Rhodiola sachalinensis with 95% ethanol water, collect extract and the ethanol removed in described extract, obtain rhodiola root 95% ethanol extract;
2) aqueous dispersion of described rhodiola root 95% ethanol extract is become suspension, with suspension described in petroleum ether extraction, collect aqueous phase, be extracted with ethyl acetate described aqueous phase, collect ethyl acetate phase, remove described ethyl acetate mutually in ethyl acetate namely obtain rhodiola root acetic acid ethyl ester extract;
3) by step 2) rhodiola root acetic acid ethyl ester extract carry out silica gel column chromatography and obtain rhodiola root eluent ethyl acetate thing; The wash-out of described silica gel column chromatography first carries out wash-out with ethyl acetate-light petrol, then carries out wash-out with ethyl acetate, the eluent that collection eluent ethyl acetate obtains, obtain rhodiola root eluent ethyl acetate thing; Described ethyl acetate-light petrol is the mixture that ethyl acetate and benzinum are mixed to get according to the volume ratio of 1:1; Described rhodiola root eluent ethyl acetate thing is the third gadol extract;
Described rhodiola root is rhodiola.
Experiment proves, rhodiola 95% ethanol extract, when concentration is 1mg/ml, reaches 0.523 to the bacteriostasis rate of graw mold of tomato germ, reaches 0.490 to the bacteriostasis rate of Alternaria brassicae; Little premium red-spotted stonecrop 95% ethanol extract, when concentration is 1mg/ml, reaches 0.404 to the bacteriostasis rate of graw mold of tomato germ, reaches 0.449 to the bacteriostasis rate of Alternaria brassicae; Sachalin rhodiola rhizome 95% ethanol extract, when concentration is 1mg/ml, reaches 0.510 to the bacteriostasis rate of graw mold of tomato germ, reaches 0.337 to the bacteriostasis rate of Alternaria brassicae.Rhodiola acetic acid ethyl ester extract reaches 0.580 when concentration is 0.1mg/ml to the bacteriostasis rate of graw mold of tomato germ, and rhodiola eluent ethyl acetate thing reaches 0.690 when concentration is 0.1mg/ml to the bacteriostasis rate of graw mold of tomato germ; Rhodiola acetic acid ethyl ester extract reaches 0.406 when concentration is 0.1mg/ml to the bacteriostasis rate of Alternaria brassicae, and rhodiola eluent ethyl acetate thing reaches 0.376 when concentration is 0.1mg/ml to the bacteriostasis rate of Alternaria brassicae.Illustrate that rhodiola, sachalin rhodiola rhizome and/or little premium red-spotted stonecrop can be used for preparing graw mold of tomato germ and/or Cabbage Leaf Spot bacteria inhibitor.
The first gadol extract above-mentioned, the second gadol extract and the third gadol extract are preparing the application in graw mold of tomato germ and/or Cabbage Leaf Spot bacteria inhibitor, and rhodiola, sachalin rhodiola rhizome and/or little premium red-spotted stonecrop also belong to protection scope of the present invention preparing the application in graw mold of tomato germ and/or Cabbage Leaf Spot bacteria inhibitor.
The plant pathogenic fungi inhibitor being active component with above-mentioned gadol extract also belongs to protection scope of the present invention; Described plant pathogenic fungi is two kinds or a kind of in botrytis cinerea and Alternaria brassicae.
Above, described 95% ethanol water to be ethanol contend percentage composition be 95% the aqueous solution.
The present invention, with the potential novel harmonious agricultural chemicals of Chinese herbal medicine rhodiola root a kind of environmental friendliness that has been development of raw materials, efficient, low toxicity, has profound significance to the health of ecological long term growth, the mankind and the comprehensive regulation of disease.Compared with traditional agricultural chemicals, the present invention has following feature and advantage: 1, gadol extract of the present invention, has obvious inhibitory action to graw mold of tomato germ and Alternaria brassicae, can be used as the exploitation of a kind of botanical pesticide.2, gadol extract of the present invention and polar fraction not overlapping (the tough brightness of Tai in health-care components rhodioside place in rhodiola root, the Study on Preparation of high-purity salidroside, R&D of modern TCM and practice, 2004,18 (4), 58-59.) do not affect the use of rhodiola root effective component, can become and well exploit natural resources.
Accompanying drawing explanation
Fig. 1 is that 95% ethanol extract of rhodiola, little premium red-spotted stonecrop and sachalin rhodiola rhizome is to the inhibitory action of Alternaria brassicae.
In figure, No. 1, left side picture is that control medium is dull and stereotyped, the band medicine culture medium flat plate of No. 2 pictures in left side to be the content of rhodiola 95% ethanol extract be 1mg/mL, the band medicine culture medium flat plate of No. 3 pictures to be the content of little premium red-spotted stonecrop 95% ethanol extract be 1mg/mL, the band medicine culture medium flat plate of No. 4 pictures to be the content of sachalin rhodiola rhizome 95% ethanol extract be 1mg/mL.
Fig. 2 is that 95% ethanol extract of rhodiola, little premium red-spotted stonecrop and sachalin rhodiola rhizome is to the inhibitory action of botrytis cinerea.
In figure, No. 1, left side picture is that control medium is dull and stereotyped, the band medicine culture medium flat plate of No. 2 pictures in left side to be the content of rhodiola 95% ethanol extract be 1mg/mL, the band medicine culture medium flat plate of No. 3 pictures to be the content of little premium red-spotted stonecrop 95% ethanol extract be 1mg/mL, the band medicine culture medium flat plate of No. 4 pictures to be the content of sachalin rhodiola rhizome 95% ethanol extract be 1mg/mL.
Embodiment
Below in conjunction with embodiment, the present invention is further described in detail, the embodiment provided only in order to illustrate the present invention, instead of in order to limit the scope of the invention.Experimental technique in following embodiment, if no special instructions, is conventional method.Material used in following embodiment, reagent etc., if no special instructions, all can obtain from commercial channels.
Botrytis cinerea (Botrytiscinerea) in following embodiment (Zhu Jianlan. the biological characteristic research of botrytis cinerea, Gansu Agriculture University's journal, 1995,30 (1), 73-78.), the public from field acquisition, also can obtain from Beijing Agricultural College, this biomaterial related experiment of the present invention of only attaching most importance to again is used, not can be used as other purposes and uses.Botrytis cinerea is called for short in the application.
Monilinia fructicola (Moniliniafructicola) (Hu Haiwen, Yang Haiqing, Wang Peng in following embodiment, Liu Suhua, Zhao Xiaoyan, Liu Zhengping. the separation screening of Monilinia fructicola antagonistic bacterium and qualification, Chinese agronomy circular, 2009,25 (12): 195-200), the public from field acquisition, also can obtain from Beijing Agricultural College, this biomaterial related experiment of the present invention of only attaching most importance to again is used, not can be used as other purposes and uses.Monilinia fructicola is called for short in the application.
Prune maize ear rot bacterium (Polvporushirsutus) (Ren Namin in following embodiment, Ma Huanpu, Liu Zhimin. Antagonistic Actinomycetes F1 zymotic fluid antimicrobial spectrum and stability analysis, Beijing Agricultural College's journal, 2012,27 (1), 32-35.), the public from field acquisition, also can obtain from Beijing Agricultural College, this biomaterial related experiment of the present invention of only attaching most importance to again is used, not can be used as other purposes and uses.Prune maize ear rot bacterium is called for short in the application.
Alternaria brassicae (Alternariabrassicae) (Ren Namin in following embodiment, Ma Huanpu, Liu Zhimin. Antagonistic Actinomycetes F1 zymotic fluid antimicrobial spectrum and stability analysis, Beijing Agricultural College's journal, 2012,27 (1), 32-35.), the public from field acquisition, also can obtain from Beijing Agricultural College, this biomaterial related experiment of the present invention of only attaching most importance to again is used, not can be used as other purposes and uses.Alternaria brassicae is called for short in the application.
Potato dry rot bacterium (Pythiumcoeruleum) in following embodiment (Bu Chunya etc. suppress plant pathogenic fungi active substance to be studied in Chinese Stellera Root. plant protection. northern gardening .2013 (19): 120-123); the public can from field acquisition; also can obtain from Beijing Agricultural College; this biomaterial related experiment of the present invention of only attaching most importance to again is used, not can be used as other purposes and uses.Potato dry rot bacterium is called for short in the application.
Amur honeysuckle in following embodiment false tail spore leaf spot fungi (Pseudocercosporalonicericola) (Ren Namin, Ma Huanpu, Liu Zhimin. Antagonistic Actinomycetes F1 zymotic fluid antimicrobial spectrum and stability analysis, Beijing Agricultural College's journal, 2012,27 (1), 32-35.), the public from field acquisition, also can obtain from Beijing Agricultural College, this biomaterial related experiment of the present invention of only attaching most importance to again is used, not can be used as other purposes and uses.Amur honeysuckle false tail spore leaf spot fungi is called for short in the application.
Cotton red spot disease bacterium (Fusariummonihforme) (Ren Namin in following embodiment, Ma Huanpu, Liu Zhimin. Antagonistic Actinomycetes F1 zymotic fluid antimicrobial spectrum and stability analysis, Beijing Agricultural College's journal, 2012,27 (1), 32-35.), the public from field acquisition, also can obtain from Beijing Agricultural College, this biomaterial related experiment of the present invention of only attaching most importance to again is used, not can be used as other purposes and uses.Cotton red spot disease bacterium is called for short in the application.
The dry root of the rhodiola (Rhodiolacrenulata) in following embodiment, Tibet, the place of production, buys from Hui nationality's Chinese Medicinal Materials Markets.
The dry root of the sachalin rhodiola rhizome (Rhodiolasachalinensis) in following embodiment, Changchun, the place of production, buys from Hui nationality's Chinese Medicinal Materials Markets.
The dry root of the little premium red-spotted stonecrop in following embodiment, Tibet, the place of production, buys from Hui nationality's Chinese Medicinal Materials Markets.
95% ethanol water in following embodiment to be ethanol contend percentage composition be 95% the aqueous solution.In following embodiment, the preparation of gadol extract is all carried out at 20-25 DEG C.
The preparation of embodiment 1, gadol extract and the experiment of suppression plant pathogenic fungi thereof
1, the preparation of gadol extract
This step has prepared following 14 kinds of gadol extracts: rhodiola 95% ethanol extract, little premium red-spotted stonecrop 95% ethanol extract, sachalin rhodiola rhizome 95% ethanol extract, rhodiola petroleum ether extract, rhodiola acetic acid ethyl ester extract, the rhodiola hydrotrope, rhodiola ethyl acetate-light petrol eluate, rhodiola eluent ethyl acetate thing, little premium red-spotted stonecrop petroleum ether extract, little premium red-spotted stonecrop acetic acid ethyl ester extract, the little premium red-spotted stonecrop hydrotrope, sachalin rhodiola rhizome petroleum ether extract, sachalin rhodiola rhizome acetic acid ethyl ester extract, the sachalin rhodiola rhizome hydrotrope.Concrete preparation method is as follows:
The preparation of 1.1 Rhodiola crenulata extracts
1) after being ground in liquid nitrogen by the dry root of rhodiola, add 95% ethanol water and extract, collect extract, the ethanol that reduced pressure concentration removes in described extract obtains medicinal extract, and this medicinal extract is rhodiola 95% ethanol extract;
2) by step 1) the aqueous dispersion of rhodiola 95% ethanol extract become suspension, with this suspension of petroleum ether extraction, collect benzinum phase and aqueous phase respectively, reduced pressure concentration remove this benzinum mutually in benzinum namely obtain rhodiola petroleum ether extract;
3) be extracted with ethyl acetate step 2) aqueous phase, collect ethyl acetate phase and aqueous phase respectively, reduced pressure concentration remove this ethyl acetate mutually in ethyl acetate namely obtain rhodiola acetic acid ethyl ester extract, namely the water that reduced pressure concentration removes in this aqueous phase obtain the rhodiola hydrotrope;
4) by step 3) rhodiola acetic acid ethyl ester extract carry out silica gel column chromatography and obtain rhodiola eluent ethyl acetate thing; The wash-out of described silica gel column chromatography first carries out wash-out with ethyl acetate-light petrol and then carries out wash-out with ethyl acetate, the eluent that collection eluent ethyl acetate obtains, and obtains rhodiola eluent ethyl acetate thing; Described ethyl acetate-light petrol is the mixture that ethyl acetate and benzinum are mixed to get according to the volume ratio of 1:1.Concrete grammar is as follows:
The chromatographic column adopt the chromatograph (SP1-B2B0) of biotage company of Sweden, adopting the catalog number (Cat.No.) of biotage company of Sweden to be the SNAPCartridgeKP-Sil100g of FSK0-1107-0100 carries out silica gel column chromatography.Wherein, chromatograph is with UV-detector, catalog number (Cat.No.) is the column length of the chromatographic column of the SNAPCartridgeKP-Sil100g of FSK0-1107-0100 is 16cm, internal diameter is 4cm, chromatography media is silica gel (200-300 order), silica gel quality in this chromatographic column is 100g, and the bed volume of this chromatographic column is 145ml.
By step 3) rhodiola acetic acid ethyl ester extract and silica gel (200-300 order) in mass ratio 1:1 mix sample, after sample complete on this sample, be eluted to 254nm with above-mentioned ethyl acetate-light petrol and no longer occur chromatographic peak, the ethyl acetate-light petrol sharing 20 column volumes washes post, flow velocity is 20ml/min, the liquid that the ethyl acetate-light petrol collecting these 20 column volumes elutes, this liquid is called rhodiola ethyl acetate-light petrol eluent, reduced pressure concentration removes the ethyl acetate-light petrol in this rhodiola ethyl acetate-light petrol eluent, obtain rhodiola ethyl acetate-light petrol eluate, then no longer there is chromatographic peak with eluent ethyl acetate to 254nm again, share the eluent ethyl acetate of 7 column volumes, flow velocity is 20ml/min, the liquid that the eluent ethyl acetate collecting these 7 column volumes goes out, this liquid is called rhodiola eluent ethyl acetate liquid, reduced pressure concentration removes the ethyl acetate in this rhodiola eluent ethyl acetate liquid, obtains rhodiola eluent ethyl acetate thing.
The preparation of 1.2 little Hua gadol extracts
1) after being ground in liquid nitrogen by the dry root of little premium red-spotted stonecrop, add 95% ethanol water and extract, collect extract, the ethanol that reduced pressure concentration removes in described extract obtains medicinal extract, and this medicinal extract is little premium red-spotted stonecrop 95% ethanol extract;
2) by step 1) the aqueous dispersion of little premium red-spotted stonecrop 95% ethanol extract become suspension, with this suspension of petroleum ether extraction, collect benzinum phase and aqueous phase respectively, reduced pressure concentration remove this benzinum mutually in benzinum namely obtain little premium red-spotted stonecrop petroleum ether extract;
3) be extracted with ethyl acetate step 2) aqueous phase, collect ethyl acetate phase and aqueous phase respectively, reduced pressure concentration remove this ethyl acetate mutually in ethyl acetate namely obtain little premium red-spotted stonecrop acetic acid ethyl ester extract, namely the reduced pressure concentration water removed in this aqueous phase obtain the little premium red-spotted stonecrop hydrotrope.
The preparation of 1.3 Actions of Extracts of Rhodiola Sachalinensis
1) after being ground in liquid nitrogen by the dry root of sachalin rhodiola rhizome, add 95% ethanol water and extract, collect extract, the ethanol that reduced pressure concentration removes in described extract obtains medicinal extract, and this medicinal extract is sachalin rhodiola rhizome 95% ethanol extract;
2) by step 1) the aqueous dispersion of sachalin rhodiola rhizome 95% ethanol extract become suspension after, with this suspension of petroleum ether extraction, collect benzinum phase and aqueous phase respectively, reduced pressure concentration remove this benzinum mutually in benzinum namely obtain sachalin rhodiola rhizome petroleum ether extract;
3) be extracted with ethyl acetate step 2) aqueous phase, collect ethyl acetate phase and aqueous phase respectively, reduced pressure concentration remove this ethyl acetate mutually in ethyl acetate namely obtain sachalin rhodiola rhizome acetic acid ethyl ester extract, namely the water that reduced pressure concentration removes in this aqueous phase obtain the sachalin rhodiola rhizome hydrotrope.
2, gadol extract suppresses the activity of plant pathogenic fungi
Respectively with a kind of bacterial strain in botrytis cinerea, Monilinia fructicola, prune maize ear rot bacterium, Alternaria brassicae, potato dry rot bacterium, the false tail spore leaf spot fungi of Amur honeysuckle and these 7 kinds of fungal bacterial strains of cotton red spot disease bacterium for strains tested, the activity according to the suppression plant pathogenic fungi of 14 kinds of gadol extracts of following determination of experimental method step 1:
Rhodiola 95% ethanol extract is removed by step 1, each gadol extract in 11 kinds of gadol extracts of little premium red-spotted stonecrop 95% ethanol extract and sachalin rhodiola rhizome 95% alcohol extract beyond the region of objective existence is individually dissolved in dimethyl sulfoxide (DMSO) (DMSO), the concentration of each gadol extract is made to be 10mg/mL, obtain the rhodiola petroleum ether extract dimethyl sulphoxide solution that concentration is following 11 kinds of gadol extract dimethyl sulphoxide solution: 10mg/mL of 10mg/mL, 10mg/mL rhodiola acetic acid ethyl ester extract dimethyl sulphoxide solution, 10mg/mL rhodiola hydrotrope dimethyl sulphoxide solution, 10mg/mL rhodiola ethyl acetate-light petrol eluate dimethyl sulphoxide solution, 10mg/mL rhodiola eluent ethyl acetate thing dimethyl sulphoxide solution, 10mg/mL little premium red-spotted stonecrop petroleum ether extract dimethyl sulphoxide solution, 10mg/mL little premium red-spotted stonecrop acetic acid ethyl ester extract dimethyl sulphoxide solution, 10mg/mL little premium red-spotted stonecrop hydrotrope dimethyl sulphoxide solution, 10mg/mL sachalin rhodiola rhizome petroleum ether extract dimethyl sulphoxide solution, 10mg/mL sachalin rhodiola rhizome acetic acid ethyl ester extract dimethyl sulphoxide solution, 10mg/mL sachalin rhodiola rhizome hydrotrope dimethyl sulphoxide solution.
Rhodiola 95% ethanol extract in step 1, little premium red-spotted stonecrop 95% ethanol extract and sachalin rhodiola rhizome 95% ethanol extract are individually dissolved in dimethyl sulfoxide (DMSO) (DMSO), make the concentration of each gadol extract be 100mg/mL, obtain 100mg/mL rhodiola 95% ethanol extract dimethyl sulphoxide solution, 100mg/mL little premium red-spotted stonecrop 95% ethanol extract dimethyl sulphoxide solution and 100mg/mL sachalin rhodiola rhizome 95% ethanol extract dimethyl sulphoxide solution.
Drawing concentration is that a kind of gadol extract dimethyl sulphoxide solution 1mL of 10mg/mL is added in the triangular flask that the PDA medium (45 DEG C) that 100mL melts is housed and mixes, obtain 101ml medicine medium, this 101ml medium on average pours (diameter 9cm) in 5 sterilizing culture dishes into, and the content making this kind of gadol extract is the band medicine culture medium flat plate of 0.1mg/mL.
Drawing concentration is that a kind of gadol extract dimethyl sulphoxide solution 1mL of 100mg/mL is added in the triangular flask that the PDA medium (45 DEG C) that 100mL melts is housed and mixes, obtain 101ml medicine medium, this 101ml medium on average pours (diameter 9cm) in 5 sterilizing culture dishes into, and the content making this kind of gadol extract is the band medicine culture medium flat plate of 1mg/mL.
Absorption 1mL dimethyl sulfoxide (DMSO) is added in the triangular flask that the PDA medium (45 DEG C) that 100mL melts is housed and mixes, obtain 101ml control medium, this 101ml control medium on average pours (diameter 9cm) in 5 sterilizing culture dishes into, makes control medium flat board.
The assay method of each strains tested is all as follows: strains tested buys at colony edge mycelial growth vigorous place card punch the bacterium cake that cut-off footpath is 0.8cm cultivate 5 days in 25 DEG C in PDA medium after, is inoculated in each culture dish central authorities (having of mycelia to face down) respectively.Each band medicine culture medium flat plate and each control medium flat board respectively put a bacterium cake, often 3 repetitions (each repetition 1 flat board) are established in process, cultivate in 25 DEG C of incubators after 7 days, adopt right-angled intersection method to measure colony diameter, calculate bacteriostasis rate as follows:
Bacteriostasis rate=(control medium flat-plate bacterial colony diameter-0.8)-(band medicine culture medium flat plate colony diameter-0.8)/(control medium flat-plate bacterial colony diameter-0.8)
Wherein, PDA medium is prepared as follows: potato 200g (potato of peeling is cut into small pieces, and boils and crosses elimination filtrate in 30 minutes), glucose 20g, agar 18-20g, is settled to 1000ml with distilled water, 121 DEG C of steam sterilization 20min.
Result is as shown in table 1-4, show rhodiola 95% ethanol extract, little premium red-spotted stonecrop 95% ethanol extract and sachalin rhodiola rhizome 95% ethanol extract are when concentration is 1mg/ml, to graw mold of tomato germ and Alternaria brassicae, there is obvious inhibitory action, but to potato dry rot bacterium, prune maize ear rot bacterium, the false tail spore leaf spot fungi of Amur honeysuckle and cotton red spot disease bacterium all do not show inhibitory action: rhodiola 95% ethanol extract is when concentration is 1mg/ml, 0.523 is reached to the bacteriostasis rate of graw mold of tomato germ, 0.490 is reached to the bacteriostasis rate of Alternaria brassicae, little premium red-spotted stonecrop 95% ethanol extract, when concentration is 1mg/ml, reaches 0.404 to the bacteriostasis rate of graw mold of tomato germ, reaches 0.449 to the bacteriostasis rate of Alternaria brassicae, sachalin rhodiola rhizome 95% ethanol extract, when concentration is 1mg/ml, reaches 0.510 to the bacteriostasis rate of graw mold of tomato germ, reaches 0.337 to the bacteriostasis rate of Alternaria brassicae.Rhodiola acetic acid ethyl ester extract reaches 0.580 when concentration is 0.1mg/ml to the bacteriostasis rate of graw mold of tomato germ, and rhodiola eluent ethyl acetate thing reaches 0.690 when concentration is 0.1mg/ml to the bacteriostasis rate of graw mold of tomato germ; Rhodiola acetic acid ethyl ester extract reaches 0.406 when concentration is 0.1mg/ml to the bacteriostasis rate of Alternaria brassicae, and rhodiola eluent ethyl acetate thing reaches 0.376 when concentration is 0.1mg/ml to the bacteriostasis rate of Alternaria brassicae.
Table 1, rhodiola root 95% ethanol extract (concentration=1mg/ml) bacteriostasis rate to plant pathogenic fungi
Table 2, gadol extract are to the bacteriostasis rate of botrytis cinerea
| Sample | Concentration (mg/ml) | Bacteriostasis rate |
| Rhodiola 95% ethanol extract | 1.0 | 0.523±0.019 |
| Rhodiola petroleum ether extract | 0.1 | 0.094±0.009 |
| Rhodiola acetic acid ethyl ester extract | 0.1 | 0.580±0.007 |
| The rhodiola hydrotrope | 0.1 | 0.008±0.003 |
| Rhodiola ethyl acetate-light petrol eluate | 0.1 | 0.127±0.003 |
| Rhodiola eluent ethyl acetate thing | 0.1 | 0.690±0.001 |
| Little premium red-spotted stonecrop 95% ethanol extract | 1.0 | 0.404±0.009 |
| Little premium red-spotted stonecrop petroleum ether extract | 0.1 | 0.143±0.015 |
| Little premium red-spotted stonecrop acetic acid ethyl ester extract | 0.1 | 0.167±0.038 |
| The little premium red-spotted stonecrop hydrotrope | 0.1 | 0.007±0.003 |
| Sachalin rhodiola rhizome 95% ethanol extract | 1.0 | 0.510±0.009 |
| Sachalin rhodiola rhizome acetic acid ethyl ester extract | 0.1 | 0.225±0.008 |
| The sachalin rhodiola rhizome hydrotrope | 0.1 | 0.005±0.038 |
Table 3, gadol extract are to the bacteriostasis rate of Alternaria brassicae
| Sample | Concentration (mg/ml) | Bacteriostasis rate |
| Rhodiola 95% ethanol extract | 1.0 | 0.490±0.029 |
| Rhodiola petroleum ether extract | 0.1 | 0.015±0.007 |
| Rhodiola acetic acid ethyl ester extract | 0.1 | 0.406±0.007 |
| The rhodiola hydrotrope | 0.1 | 0.015±0.007 |
| Rhodiola ethyl acetate-light petrol eluate | 0.1 | 0.064±0.012 |
| Rhodiola eluent ethyl acetate thing | 0.1 | 0.376±0.014 |
| Little premium red-spotted stonecrop 95% ethanol extract | 1.0 | 0.449±0.029 |
| Little premium red-spotted stonecrop petroleum ether extract | 0.1 | 0.065±0.007 |
| Little premium red-spotted stonecrop acetic acid ethyl ester extract | 0.1 | 0.115±0.007 |
| The little premium red-spotted stonecrop hydrotrope | 0.1 | 0.007±0.007 |
| Sachalin rhodiola rhizome 95% ethanol extract | 1.0 | 0.337±0.245 |
| Sachalin rhodiola rhizome petroleum ether extract | 0.1 | 0.137±0.005 |
| Sachalin rhodiola rhizome acetic acid ethyl ester extract | 0.1 | 0.337±0.245 |
| The sachalin rhodiola rhizome hydrotrope | 0.1 | 0.033±0.005 |
Table 4, gadol extract are to the bacteriostasis rate of Monilinia fructicola
| Sample | Concentration (mg/ml) | Bacteriostasis rate |
| Rhodiola 95% ethanol extract | 1.0 | 0.125±0.007 |
| Rhodiola petroleum ether extract | 0.1 | 0.019±0.006 |
| Rhodiola acetic acid ethyl ester extract | 0.1 | 0.176±0.031 |
| The rhodiola hydrotrope | 0.1 | 0.019±0.007 |
| Rhodiola ethyl acetate-light petrol eluate | 0.1 | 0.214±0.031 |
| Rhodiola eluent ethyl acetate thing | 0.1 | 0.113±0.031 |
| Little premium red-spotted stonecrop 95% ethanol extract | 1.0 | 0.138±0.007 |
| Little premium red-spotted stonecrop petroleum ether extract | 0.1 | 0.029±0.007 |
| Little premium red-spotted stonecrop acetic acid ethyl ester extract | 0.1 | 0.241±0.015 |
| The little premium red-spotted stonecrop hydrotrope | 0.1 | 0.022±0.015 |
| Sachalin rhodiola rhizome 95% ethanol extract | 1.0 | 0.046±0.007 |
| Sachalin rhodiola rhizome petroleum ether extract | 0.1 | 0.032±0.015 |
| Sachalin rhodiola rhizome acetic acid ethyl ester extract | 0.1 | 0.067±0.015 |
| The sachalin rhodiola rhizome hydrotrope | 0.1 | 0.079±0.005 |
Claims (10)
1. gadol extract is suppressing the application in plant pathogenic fungi, the preparation method of described gadol extract comprises: extract root of Rhodiola sachalinensis with 95% ethanol water, collect extract and the ethanol removed in described extract, obtain rhodiola root 95% ethanol extract, described rhodiola root 95% ethanol extract is gadol extract;
Described plant pathogenic fungi is botrytis cinerea and Alternaria brassicae or described plant pathogenic fungi is botrytis cinerea; Described rhodiola root is rhodiola, sachalin rhodiola rhizome and/or little premium red-spotted stonecrop.
2. gadol extract is suppressing the application in plant pathogenic fungi, and the preparation method of described gadol extract comprises:
1) extract root of Rhodiola sachalinensis with 95% ethanol water, collect extract and the ethanol removed in described extract, obtain rhodiola root 95% ethanol extract;
2) aqueous dispersion of described rhodiola root 95% ethanol extract is become suspension, with suspension described in petroleum ether extraction, collect aqueous phase, be extracted with ethyl acetate described aqueous phase, collect ethyl acetate phase, remove described ethyl acetate mutually in ethyl acetate namely obtain rhodiola root acetic acid ethyl ester extract; Described rhodiola root acetic acid ethyl ester extract is gadol extract;
Described plant pathogenic fungi is two kinds or a kind of in botrytis cinerea and Alternaria brassicae; Described rhodiola root is rhodiola, sachalin rhodiola rhizome and/or little premium red-spotted stonecrop.
3. gadol extract is suppressing the application in plant pathogenic fungi, and the preparation method of described gadol extract comprises:
1) extract root of Rhodiola sachalinensis with 95% ethanol water, collect extract and the ethanol removed in described extract, obtain rhodiola root 95% ethanol extract;
2) aqueous dispersion of described rhodiola root 95% ethanol extract is become suspension, with suspension described in petroleum ether extraction, collect aqueous phase, be extracted with ethyl acetate described aqueous phase, collect ethyl acetate phase, remove described ethyl acetate mutually in ethyl acetate namely obtain rhodiola root acetic acid ethyl ester extract;
3) by step 2) rhodiola root acetic acid ethyl ester extract carry out silica gel column chromatography and obtain rhodiola root eluent ethyl acetate thing; The wash-out of described silica gel column chromatography first carries out wash-out with ethyl acetate-light petrol then to carry out wash-out with ethyl acetate, the eluent that collection eluent ethyl acetate obtains, and namely the ethyl acetate in removing eluent obtain rhodiola root eluent ethyl acetate thing; Described ethyl acetate-light petrol is the mixture that ethyl acetate and benzinum are mixed to get according to the volume ratio of 1:1; Described rhodiola root eluent ethyl acetate thing is gadol extract;
Described plant pathogenic fungi is two kinds or a kind of in botrytis cinerea and Alternaria brassicae; Described rhodiola root is rhodiola.
4. the preparation method of gadol extract, comprise: extract root of Rhodiola sachalinensis with 95% ethanol water, collect extract and the ethanol removed in described extract, obtain rhodiola root 95% ethanol extract, described rhodiola root 95% ethanol extract is gadol extract;
Described rhodiola root is rhodiola, sachalin rhodiola rhizome and/or little premium red-spotted stonecrop.
5. the preparation method of gadol extract, comprising:
1) extract root of Rhodiola sachalinensis with 95% ethanol water, collect extract and the ethanol removed in described extract, obtain rhodiola root 95% ethanol extract;
2) aqueous dispersion of described rhodiola root 95% ethanol extract is become suspension, with suspension described in petroleum ether extraction, collect aqueous phase, be extracted with ethyl acetate described aqueous phase, collect ethyl acetate phase, remove described ethyl acetate mutually in ethyl acetate namely obtain rhodiola root acetic acid ethyl ester extract; Described rhodiola root acetic acid ethyl ester extract is gadol extract;
Described rhodiola root is rhodiola, sachalin rhodiola rhizome and/or little premium red-spotted stonecrop.
6. the preparation method of gadol extract, comprising:
1) extract root of Rhodiola sachalinensis with 95% ethanol water, collect extract and the ethanol removed in described extract, obtain rhodiola root 95% ethanol extract;
2) aqueous dispersion of described rhodiola root 95% ethanol extract is become suspension, with suspension described in petroleum ether extraction, collect aqueous phase, be extracted with ethyl acetate described aqueous phase, collect ethyl acetate phase, remove described ethyl acetate mutually in ethyl acetate namely obtain rhodiola root acetic acid ethyl ester extract;
3) by step 2) rhodiola root acetic acid ethyl ester extract carry out silica gel column chromatography and obtain rhodiola root eluent ethyl acetate thing; The wash-out of described silica gel column chromatography first carries out wash-out with ethyl acetate-light petrol then to carry out wash-out with ethyl acetate, the eluent that collection eluent ethyl acetate obtains, and obtains rhodiola root eluent ethyl acetate thing; Described ethyl acetate-light petrol is the mixture that ethyl acetate and benzinum are mixed to get according to the volume ratio of 1:1; Described rhodiola root eluent ethyl acetate thing is gadol extract;
Described rhodiola root is rhodiola.
7. the gadol extract prepared by described preparation method arbitrary in claim 4-6.
8. the application of rhodiola root in preparation plant pathogenic fungi inhibitor, described plant pathogenic fungi is two kinds or a kind of in botrytis cinerea and Alternaria brassicae; Described rhodiola root is rhodiola, sachalin rhodiola rhizome and/or little premium red-spotted stonecrop.
9. application according to claim 8, is characterized in that: the active component of described plant pathogenic fungi inhibitor is gadol extract according to claim 7.
10. plant pathogenic fungi inhibitor, its active component is gadol extract according to claim 7;
Described plant pathogenic fungi is two kinds or a kind of in botrytis cinerea and Alternaria brassicae; Described rhodiola root is rhodiola, sachalin rhodiola rhizome and/or little premium red-spotted stonecrop.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201510490827.XA CN105028492B (en) | 2014-02-27 | 2014-02-27 | Application of the gadol extract in botrytis cinerea and Alternaria brassicae is suppressed |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201510490827.XA CN105028492B (en) | 2014-02-27 | 2014-02-27 | Application of the gadol extract in botrytis cinerea and Alternaria brassicae is suppressed |
| CN201410069489.8A CN103814954B (en) | 2014-02-27 | 2014-02-27 | Gadol extract and preparation method thereof and application |
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| CN106857808A (en) * | 2017-04-09 | 2017-06-20 | 湖南易科生物工程有限公司 | A kind of compound method of leaf vegetables antistaling agent |
| CN107048147A (en) * | 2017-04-09 | 2017-08-18 | 湖南易科生物工程有限公司 | A kind of compound method of chopped hot pepper antistaling agent |
| CN107087674A (en) * | 2017-04-24 | 2017-08-25 | 湖南易科生物工程有限公司 | A kind of compound method of fruit antistaling agent |
| CN107258417A (en) * | 2016-04-06 | 2017-10-20 | 万素梅 | The occurrence regularity of jujube alternaria |
| CN107410463A (en) * | 2017-04-09 | 2017-12-01 | 湖南易科生物工程有限公司 | A kind of compound method of fruit vegetables antistaling agent |
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| CN107048147A (en) * | 2017-04-09 | 2017-08-18 | 湖南易科生物工程有限公司 | A kind of compound method of chopped hot pepper antistaling agent |
| CN107410463A (en) * | 2017-04-09 | 2017-12-01 | 湖南易科生物工程有限公司 | A kind of compound method of fruit vegetables antistaling agent |
| CN107087674A (en) * | 2017-04-24 | 2017-08-25 | 湖南易科生物工程有限公司 | A kind of compound method of fruit antistaling agent |
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| CN103814954A (en) | 2014-05-28 |
| CN105028492B (en) | 2018-04-13 |
| CN103814954B (en) | 2015-08-19 |
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