CN102180857A

CN102180857A - Xanthene derivative, and preparation method and application thereof

Info

Publication number: CN102180857A
Application number: CN2011100633829A
Authority: CN
Inventors: 李春远; 丁唯嘉; 林永成; 佘志刚
Original assignee: South China Agricultural University
Current assignee: South China Agricultural University
Priority date: 2011-03-16
Filing date: 2011-03-16
Publication date: 2011-09-14

Abstract

The invention relates to a xanthene derivative 8-hydroxy-3-methyl-9-oxo-9H-xanthene-1-carboxylicacidmethylether extracted and separated from coculture fermentation liquid of two strains of marine plant endophytic fungi, a preparation method thereof, and use of the xanthene derivative as a pesticide for preventing and controlling plant pathogenic fungi. The preparation method comprises the following steps of: cultivating the two strains of marine plant endophytic fungi in a culture medium respectively; mixing and fermenting the two strains of marine plant endophytic fungi; filtering the obtained fermentation liquid; removing thalli; concentrating the fermentation liquid at a low temperature and under reduced pressure; extracting the concentrated fermentation liquid by using an organic solvent; concentrating the extract liquid; performing chromatographic separation; concentrating the obtained eluent; and recrystallizing by using the organic solvent to obtain the compound. The xanthene derivative derived from metabolin through mixed fermentation of the two strains of marine plant endophytic fungi has antifungal activity, can effectively inhibit growth of plant pathogenic fungi hyphae, can be applied to developing pesticide antibiotics and has a wide application prospect, wherein the raw materials can be produced on a large scale.

Description

A kind of xanthene derivative and its production and application

Technical field

The present invention relates to a kind of from the common cultivation and fermentation liquid of two strain marine plant endogenetic fungus extraction separation xanthene derivative 8-hydroxy-3-methyl-9-oxo-9H-xanthene-1-carboxylic acid methyl ether and preparation method thereof and as the application of the agricultural bactericide of control phytopathogen.

Background technology

Since nineteen twenty-nine was prepared penicillin from fungi since, the meta-bolites of fungi became the abundant source of medicine, and most microbiotic of using all derive from fungi and bacterium.Unique metabolic way because the singularity of ocean environment, thalassiomycetes develop, the meta-bolites that did not run in the fungi of land is provided.Plant endogenesis epiphyte in evolutionary process with host's coevolution, its secondary metabolites is the result that the adaptation of endogenetic fungus and plant common defence and disease develops coevolution, has good disease resistance activity more.And the marine plant endogenetic fungus has thalassiomycetes and plant endogenesis epiphyte physilogical characteristics concurrently, and the prospect of therefrom seeking new microbial pesticide is very wide.Mixed fermentation is mainly used in and improves enzyme and antibiotic output, has not yet to see the agricultural chemicals that utilizes mixed fermentation to obtain valuable control phytopathogen from the marine plant endogenetic fungus.

Summary of the invention

The object of the present invention is to provide the application of a kind of xanthene derivative 8-hydroxy-3-methyl-9-oxo-9H-xanthene-1-carboxylic acid methyl ether that derives from the common cultivation and fermentation liquid of marine plant endogenetic fungus and preparation method thereof and conduct control phytopathogen agricultural chemicals.It can satisfy the demand of prior art.

A kind of xanthene derivative is characterized in that molecular formula is C ₁₆H ₁₂O ₅, structural formula is:

The preparation method of above-mentioned xanthene derivative is characterized in that in substratum two strain marine plant endogenetic fungus being carried out spawn culture respectively earlier, then two strain bacterium is carried out mixed fermentation, with the gained filtering fermentation liquor, remove thalline, the concentrate under reduced pressure at low temperature fermented liquid is used organic solvent extraction; Extraction liquid concentrates laggard circumstances in which people get things ready for a trip spectrum to be separated, and the gained elutriant is concentrated, and obtains yellow crystals with the organic solvent recrystallization, is this xanthene derivative.

The application of above-mentioned xanthene derivative on control phytopathogen agricultural bactericide.

Compared with prior art, the present invention has following beneficial effect:

Xanthene derivative of the present invention has very strong inhibition oranges and tangerines mould, soybean phytophthora, banana anthrax, the mould effect of lichee frost epidemic disease, can be used as the microbial pesticide sterilant, and raw material can scale operation, has a extensive future.

Embodiment

Take following each step during the preparation The compounds of this invention

(1) in substratum, respectively autumn eggplant endogenetic fungus K38 and Eucheuma muricatum (Gmel.) Web endogenetic fungus E33 are carried out spawn culture earlier, used substratum is for containing glucose 0.8-3.0%(weight percentage, down together), yeast extract paste 0.1-1.5%, peptone 0.2-1.5%, agar 1.0-1.5%, sodium-chlor 0.3-1.5%, all the other are water, are made into the test tube slant during use, and above-mentioned bacterial classification was cultivated 3-10 days down at 28 ℃.

(2) again autumn eggplant endogenetic fungus K38 and Eucheuma muricatum (Gmel.) Web endogenetic fungus E33 are carried out mixing fermentation culture, used substratum is for containing glucose 1.5-5.0%, yeast extract paste 0.5-2.5%, peptone 0.5-2.5%, sodium-chlor 1-3.5%, all the other are water, and two bacterial classifications that slant culture is good are separated by respectively and are chosen into fermention medium in 0-7 days, leave standstill under room temperature 25-30 ℃ and cultivate 25-40 days.

(3) above-mentioned cultured filtering fermentation liquor is obtained nutrient solution, be no more than under 50 ℃ of conditions in temperature, the nutrient solution heating is condensed into the 1/10-1/5 of original volume, with ethyl acetate extraction repeatedly (ethyl acetate can use chloroform, sherwood oil, propyl carbinol to replace), concentrated extract, carrying out chromatographic separation in silicagel column, is the eluent gradient elution with the petroleum ether-ethyl acetate.

(4) in the gradient elution process, collect 2% ~ 40% petrol ether/ethyl acetate elutriant, concentrate, then in silicagel column with 10% petrol ether/ethyl acetate wash-out, collect elutriant, concentrate and obtain solid, use chloroform (available acetone, ethyl acetate replace) to carry out recrystallization purifying again, obtain yellow needle crystal, be 8-hydroxy-3-methyl-9-oxo-9H-xanthene-1-carboxylic acid methyl ether.

The spectral data of gained compound:

8-hydroxy-3-methyl-9-oxo-9H-xanthene-1-carboxylic?acid?methyl?ether:?FABMS:285,?283;?Elemental?anal.?C?67.59%,?H?4.26%,?N?0.?calcd?for?C ₁₆H ₁₂O ₅,?C?67.60%,?H?4.25%,?N?0.?IR?bands(neat):?3438,?2923,?2853,?1705,?1634,?1610,?1272,?1238,?822cm ^-1;? ¹HNMR?(600?MHz?CDCl ₃):?7.15(d,?1.2Hz,?1H), 7.35(d,?1.2Hz,?1H),?6.91(dd,?1.2,?8.4Hz,?1H),?7.59(dd,?7.8,?8.4Hz,?1H),?6.80(dd,?1.2,?7.8Hz,?1H),?2.52(s,?3H),?4.03(s,?3H),?12.31(br?s,?1H)?;? ¹³C?NMR?(150?MHz?CDCl ₃):?133.3(C),?124.2(CH),?147.0(C),?119.2(CH),?106.9(CH),?136.9(CH),?110.8(CH),?161.7(C),?180.8(C),?115.3(C),?156.2(C),?155.8(C),?108.9(C),?22.0(CH ₃),?169.7(C),?53.2(CH ₃)。

Compound of the present invention is to oranges and tangerines mould, soybean phytophthora, banana anthrax, the mould bacteriostatic activity test of lichee frost epidemic disease

Adopt the mycelial growth rate method to measure The compounds of this invention to the oranges and tangerines mould, soybean phytophthora, the banana anthrax, the mould bacteriostatic activity of lichee frost epidemic disease, take by weighing compound 15mg, with DMSO it is dissolved constant volume to 10 mL, drawing 1 mL with liquid-transfering gun joins and is equipped with in the triangular flask that 15 mL have melted PDA substratum (about 60 ℃ of temperature), fully shake up, pour into again in the culture dish of diameter 90 mm and make the pastille culture medium flat plate that concentration is 100 μ g/mL, 6mm is placed on the flat board through three cultured phytopathogen cakes of continuous passage, with the PDA substratum that contains equivalent DMSO in contrast, cultivate under 28 ℃ of conditions, measure for the colony diameter that tries bacterial classification with the right-angled intersection method behind the cultivation 72h, calculate the mycelial growth bacteriostasis rate as follows: relative bacteriostasis rate=(contrast colony diameter-processing colony diameter) * 100%/contrast colony diameter.Each handles 3 repetitions.Experimental result sees Table 1, and data are represented with the mean value ± standard error of three measurements in the table.

Table 1 The compounds of this invention is to the mycelial growth inhibition rate (100 μ g/mL) of phytopathogen

Experimental result shows that The compounds of this invention has the effect of very strong inhibition mycelial growth to the pathogenic bacteria of oranges and tangerines mould, soybean phytophthora, banana anthrax, the mould harm of lichee frost epidemic disease cash crop, can be made into agricultural chemicals, and liquid spray or pulvis all can.

Claims

1. xanthene derivative, the molecular formula that it is characterized in that described derivative is C ₁₆H ₁₂O ₅, structural formula suc as formula ( ) shown in:

Formula (

).

2. the preparation method of the described xanthene derivative of claim 1 is characterized in that comprising the steps:

(1) in substratum, respectively autumn eggplant endogenetic fungus K38 and Eucheuma muricatum (Gmel.) Web endogenetic fungus E33 are carried out spawn culture earlier, used substratum is for containing glucose 0.8 ~ 3.0wt%, yeast extract paste 0.1 ~ 1.5wt%, peptone 0.2 ~ 1.5wt%, agar 1.0 ~ 1.5wt%, sodium-chlor 0.3 ~ 1.5wt%, all the other are water, be made into the test tube slant during use, above-mentioned bacterial classification was cultivated 3-10 days down at 28 ℃;

(2) again autumn eggplant endogenetic fungus K38 and Eucheuma muricatum (Gmel.) Web endogenetic fungus E33 are carried out mixing fermentation culture, used substratum is for containing glucose 1.5 ~ 5.0wt%, yeast extract paste 0.5 ~ 2.5wt%, peptone 0.5 ~ 2.5wt%, sodium-chlor 1 ~ 3.5wt%, all the other are water, and two bacterial classifications that slant culture is good are separated by respectively and are chosen into fermention medium in 0 ~ 7 day, leave standstill under 25 ~ 30 ℃ of room temperatures and cultivate 25 ~ 40 days;

(3) above-mentioned cultured filtering fermentation liquor is obtained nutrient solution, be no more than under 50 ℃ of conditions in temperature, the nutrient solution heating is condensed into 1/10 ~ 1/5 of original volume, use organic solvent extraction, concentrated extract, carrying out chromatographic separation in silicagel column, is the eluent gradient elution with the petroleum ether-ethyl acetate;

(4) in the gradient elution process, collect 2% ~ 40% petrol ether/ethyl acetate elutriant, concentrate, then in silicagel column with 10% petrol ether/ethyl acetate wash-out, collect elutriant, concentrate and obtain solid, carry out recrystallization purifying with chloroform again, obtain yellow needle crystal, be 8-hydroxy-3-methyl-9-oxo-9H-xanthene-1-carboxylic acid methyl ether.

3. the application of xanthene derivative according to claim 1 in control phytopathogen agricultural chemicals.