CN100389111C - Large ring lactone compound containing meta-phenol ring and its application - Google Patents

Abstract

The present invention relates to a large ring lactone compound containing a meta-phenol ring and an application thereof, which belongs to the technical field of the biological pesticide. The active compound is obtained by a producing strain in the method of solid fermentation and extraction separation, the production strain is caryospora carllicarpa ZD2 which is preserved at the common microorganism center of the CCCCM on 28th July, 2003, and the preservation number is CGMCC No. 0983; the active compound comprises three compounds, can efficiently poison pine wood nematode, and can be used for preparing biological nematicide.

Description

The Macrocyclic lactone compounds that phenol ring incorporates between one class

Technical field:

The present invention relates to the Macrocyclic lactone compounds that phenol ring incorporates between a class, belong to biological pesticide technical field.

Background technology:

The pine nematode of pine tree is by pine wood nematode (Bursaphelenchus xylophilus (Steiner﹠amp; Buhrer) Nickle) colonize in a kind of especially big destructive disease of getting food nutrition in Pinus seeds (Pinus sp.) body and causing the rapid death of trees, belong to international important Quarantine Objects, classify as first of the forest disease and pest, be called as " smokeless hill fire ".This disease now has been distributed widely in Japan, the U.S., Korea S, Canada, Mexico, Greece, Portugal and China etc. since finding, be in the news in 1934 from 1905.Japan is used for this sick expenses for prevention and control and accounts for 93.6% of forest pest control expense every year, accounts for 20% of forestry gross investment, but still reach 100 ten thousand ms because of this disease loss timber every year ³About.Since China finds this disease in nineteen eighty-two, now expanded to ground such as Jiangsu, Zhejiang, Anhui, Guangdong, Shandong, Hubei, Shanghai, Hong Kong and Taiwan, area has taken place surpassed 7.3 * 10hm ², dead pine tree 2 * 10 ⁷Strain causes the massive losses on economy of forestry, the forest ecology and the havoc of natural landscape, and the pine forest of the vast normal region of China is constituted a serious threat.Since the eighties, the method for preventing and treating of pine wood nematode has been mainly contained:

1. use the communication media worm that can kill pine wood nematode--the sterilant Sumithion (MEP) of Monochamus alternatus (Monochamusalternatus), Tiguvon (MPP), carbaryl (NAC) etc.This characteristics of preventing and treating method are that the passive method of preventing and treating that vector is protected pine tree is killed in a kind of dependence, and it is the big generation of disease controlling effectively, and have the pharmaceutical chemicals of certain degree to pollute to ground vegetation.

2. (mesulfenphos tartalicacid) prevents and treats pine wood nematode directly to inject nematocides in the tree body.This method only is applicable to the protection rare tree species, and effect is undesirable, and big to the person poultry toxicity.

The method of control pine nematode is mainly controlled Monochamus alternatus based on spraying insecticide at present, but because the Monochamus alternatus imago phase is long, and resistance is strong, prevention effect is very undesirable.So far still do not work out the special-purpose agricultural chemicals of the high-efficiency low-toxicity that kills pine wood nematode of energy practical application, also not seeing has the double medicament of pine wood nematode and Monochamus alternatus or the research report of method controlled.

Summary of the invention:

The object of the present invention is to provide the Macrocyclic lactone compounds that phenol ring incorporates between a class.

Production bacterial strain of the present invention was Caryospora carllicarpa ZD2, and this bacterial strain has been kept at China Committee for Culture Collection of Microorganisms common micro-organisms center on July 28th, 2003; Preserving number is CGMCCNo.0983.

The structural framework that a class of the present invention has a pine wood nematode poisoning activity be between the Macrocyclic lactone compounds that incorporates of phenol ring be that extraction separation obtains 3 kinds of new compounds from the fermenting culture of producing bacterial strain, the structural formula of these 3 kinds of compounds is (the following compound 1-3 that abbreviates as respectively) as follows respectively:

Compound 1 compound 2 compounds 3

The physico-chemical constant of active compound 1-3 of the present invention, hydrogen spectrum data and carbon spectrum data see Table 1 respectively, table 2.

The physico-chemical constant of table 1. compound 1-3

	1	2	3
				Molecular formula	C 23H 30O 8	C 20H 26O 8	C 20H 24O 8
Molecular weight	434	394	392
				[α] D (methanol)	[α] D 271+62.1°(c0.31)	[α] D 215+30.0°(c0.76)	[α] D 271+58.1°(c0.54)
UV (CH 3OH) λ max(logε)	320.2(4.01)，267.0(4.22)， 229.8(4.70)，204.4(4.71)	319.0(3.83)，267.2(4.01)， 225.4(4.54)	320.3(3.97)，267.0(4.07)， 225.3(4.60)
				IR(KBr)	3441，2959，2931，2875，1728， 1648，1598，1472，1448，1381， 1360，1316，1246，1224，1169， 1126，1057，1041，1018，969， 834，743，606	3424，2929，2856，1726， 1648，1597，1474，1447， 1383，1358，1308，1248， 1226，1202，1169，1136，1063， 1021，965，887，610	3431，2939，2865，1728， 1648，1598，1472，1448， 1381，1360，1306，1245， 1225，1200，1166，1059， 1021，969，854，609

The NMR (acetone) of table 2. compound 1-3

Active compound of the present invention is that 36 hours, compound 1-3 are respectively 100,80,80ug/mL to the semilethal rate of pine wood nematode in the treatment time in the nematicide active testing of soaked with liquid method.Therefore, compound of the present invention can be used as the application of preparation agricultural chemicals nematode killing agent.

Embodiment:

Embodiment 1: the separation preparation of compound 1-3

A. the seed culture of fungi Caryospora carllicarpa ZD2:

The PDA substratum of improvement: potato 200g, glucose 20g, yeast extract 1.0g, glycerine 6.0g, agar 17g, each 1 of medical vitamin B complexes and C, water 1000mL makes flat board, and the picking bacterial strain inserts dull and stereotyped, cultivates 15 days for 25 ℃;

B. with the mycelium freeze-drying of above-mentioned cultured C.carllicarpa ZD2, use randomly organic solvent (for example methyl alcohol, ethanol, propyl alcohol again, ethyl acetate, tetrahydrofuran (THF), acetonitrile) cold soaking extracts 4 times, spissated crude extract is absorbed in the solid absorbing material, and (natural materials of Fen Suiing for example is as kaolin, clay, talcum, quartz etc.; Or the synthetic materials of pulverizing, as microgranular silica gel, aluminum oxide etc.) on, in silicagel column, carrying out chromatographic separation, sample on the dry method is the eluent gradient elution with petrol ether/ethyl acetate=0-100%.

C. collect the petrol ether/ethyl acetate elutriant of 80%-90%, concentrating under reduced pressure.

D. methyl alcohol or the acetone gel post of above-mentioned concentrated solution through Sephedex-LH20 separated, can obtain compound 1-3.

Embodiment 2:

Adopt conventional liquid immersed method to detect the active test of nematicide of The compounds of this invention.

1, test with medicament:

To test with sample and be dissolved in a small amount of organic reagent DMSO earlier, add the sample test solution that certain density Tween solution prepares concentration as shown in table 3 respectively again.In this solution, the final concentration of the final concentration of DMSO＜3%, tween＜5 ‰.The aqueous solution that in addition DMSO of same concentrations is dissolved in the tween 20 that contains 0.5% (V/V) in contrast.

2. cultivation of pine wood nematode and preparation

The cultural method of pine wood nematode B.xylophilus: Botrytis cinerea (Botrytis cinerea) is inserted the PDA flat board, under 25 ℃, be cultured to and cover with flat board, insert a substratum that has B.xylophilus, be cultured to mycelia and disappear under 25 ℃, nematode is covered with flat board.

The substratum that will be loaded with nematode during use is chosen, and is put in the nematode separator, and after with sterilized water nematode being washed out, centrifugal concentrating is prepared into nematode suspension (about 500 every milliliter).

3. test method

It is the culture dish of 5cm that The compounds of this invention 200ppm, 100ppm, 80ppm, 50ppm concentration are placed diameter for reagent liquid 2ml, adds the suspension of living nematode 20ul (about 300) nematode again, behind the mixing culture dish is positioned over gently in 25 ℃ the incubator.Respectively at the mortality ratio of checking the calculating nematode in 12,24 and 48 hours.And under binocular microscope, observe observation line polypide wall changing conditions.

The method of identifying nematode death is: add 1-5 and drip 5%NaCl solution in handling flat board, observe after 2 minutes, dead worm is stiff, and the worm that lives is then curled or twisting.

Mortality ratio %=dead wire borer population/bus borer population * 100

With the test soln that does not add sample is contrast, and whole experiment triplicate is got three mean value calculation and gone out average mortality.

4. test-results

Table 3. sample is to the cytotoxicity test result of pine wood nematode

The result shows: compound 1-3 of the present invention has toxic action preferably to pine wood nematode, can be used for preparing the agricultural chemicals nematode killing agent and uses.

Claims (1)

1. the Macrocyclic lactone compounds that phenol ring incorporates between a class by producing bacterial strain, adopts the method for conventional solid fermentation and extraction separation to obtain, and it is characterized in that:

A. producing bacterial strain is Caryospora carllicarpa ZD2, the bacterial strain preservation date: on July 28th, 2003, be kept at China Committee for Culture Collection of Microorganisms common micro-organisms center; Preserving number is CGMCC No.0983;

B. such active compound is 3 kinds of compounds, wherein:

The structural formula of compound 1 is:

The structural formula of compound 2 is:

The structural formula of compound 3 is: