阅读说明：本技术 大戟因子l3在制备防治植物卵菌病害农药制剂中的应用 (Application of euphorbia factor L3 in preparation of pesticide preparation for preventing and treating plant oomycetes diseases ) 是由 陈雨 王碧 冯煦 徐曙 李丕睿 刘飞 赵兴增 田梅 李林蔚 单宇 印敏 管福琴 于 2020-04-16 设计创作，主要内容包括：本发明涉及植物源农药技术领域,公开了大戟因子L3(Euphorbia Factor L3)在制备防治植物卵菌病害农药制剂中的新用途。本发明中的大戟因子L3是从药用植物续随子(Euphorbia lathylris)中提取分离得到的二萜醇酯类化合物,对多种植物病原卵菌具有抑菌活性,可与辅料结合制备成防治卵菌病害的农药制剂。(The invention relates to the technical field of plant-derived pesticides, and discloses a new application of Euphorbia Factor L3(Euphorbia Factor L3) in preparation of a pesticide preparation for preventing and treating plant oomycetes diseases. The Euphorbia factor L3 in the invention is a diterpene alcohol ester compound extracted and separated from medicinal plant Euphorbia lathyris (Euphorbia lathyris), has bacteriostatic activity on various plant pathogenic oomycetes, and can be combined with auxiliary materials to prepare a pesticide preparation for preventing and treating oomycete diseases.)

1. An application of euphorbia factor L3 in preparing the agricultural chemical for preventing and treating the oomycete diseases is disclosed.

2. As shown in claim 1, the compound can be used for preventing and treating plant diseases caused by oomycete pathogenic bacteria, such as late blight, damping-off, pythium, epidemic disease, root rot, phytophthora blight, downy mildew and the like.

3. As shown in claim 1, the compound can be processed into conventional pesticide formulations such as wettable powder, suspending agent, powder, aqueous agent, missible oil, aqueous emulsion, large granule, oil agent and the like.

Technical Field

The invention relates to the technical field of plant-derived pesticides, in particular to application of a diterpene alcohol ester compound Euphorbia Factor L3(Euphorbia Factor L3) extracted and separated from seeds of Euphorbia lathyris (Euphorbia lathyris) of medicinal plants in preventing and treating plant oomycete diseases.

Background

The plant pathogenic oomycetes are a kind of important pathogenic bacteria threatening the global agricultural production. At present, chemical prevention and control are main means for effectively treating plant oomycete diseases, but the prevention and control effect of the chemical bactericide is not ideal, and meanwhile, a series of negative effects are brought to the ecological environment, the living quality of people and livestock, the survival of non-target organisms and the like, and the problem of drug resistance is increasingly highlighted along with long-term large-area application. Therefore, there is an urgent need to develop new preventive agents to alleviate the problem of resistance to drugs and reduce the loss of agricultural production. The botanical pesticide has the characteristics of rich source, specific action mode, environmental friendliness, low toxicity, relative safety to non-target organisms and the like, is an ideal object for developing novel control agents, and has great application value and popularization prospect.

Euphorbia lathyris (Euphorbia lathyris) is a plant of Euphorbia of Euphorbiaceae, mainly distributed in North China and Jiangzhe China, and mainly produced in Shandong, Hebei and Henan provinces. The seeds of Euphorbiae Lathyridis semen, also known as Qianjin seed, collected in the first part of the Chinese pharmacopoeia, have the effects of purging and expelling water, and breaking blood and relieving swelling; it can be used for treating tinea and wart. Euphorbia lathyris seed mainly contains euphorbia lathyris alkyl type and ingenol type diterpene ester components, and is a main raw material for industrially preparing medicine raw material ingenol. While euphorbia factor L3 belongs to euphorbia lathyris alkane type diterpene, is one of the main diterpene components of euphorbia lathyris seeds, and the content of euphorbia lathyris factor L3 accounts for 40% of the total diterpene components of seeds (phytochem. anal.2001, 12, 255-262). The euphorbia factor L3 has been reported to have the effect of resisting tumor multidrug resistance, but the research is still in the experimental stage at present. In summary, Euphorbia lathyris is widely distributed and has sufficient resources, the seeds are common traditional Chinese medicinal materials and raw materials for preparing medicine raw materials ingenol, the Euphorbia factor L3 is a main diterpene alcohol ester compound in Euphorbia lathyris seeds, the content is high, the preparation process is simple and easy, but the medicinal products are not seen on the market. The invention discloses a new application of euphorbia factor L3 in preparing a pesticide preparation for preventing and treating plant oomycetes diseases, which provides a pesticide molecule for preventing and treating plant oomycetes diseases on one hand and is beneficial to the comprehensive utilization of euphorbia lathyris plant resources on the other hand.

Disclosure of Invention

The invention aims to provide pesticide application of euphorbia factor L3, and is characterized in that euphorbia factor L3 is applied to preparation of pesticide preparations for preventing and treating oomycete diseases.

The technical scheme adopted by the invention is as follows: euphorbia factor L3 is a euphorbia lathyris alkyl diterpenoid ester compound, and the molecular formula is as follows: c₃₁H₃₈O₇The chemical structural formula is as follows:

further, said euphorbia factor L3 can be extracted and isolated from euphorbia lathyris seeds. The preparation process mainly comprises the following steps: crushing euphorbia lathyris seeds, performing reflux extraction with 95% ethanol, extracting with an organic solvent, performing silica gel column chromatography, recrystallizing and other conventional natural product chemical methods to obtain a pure product.

The compound can be processed into conventional pesticide formulations, such as wettable powder, suspending agent, powder, aqueous agent, missible oil, aqueous emulsion, large granule and oil agent.

The pesticide formulation processed by the compound can be used for preventing and treating plant diseases caused by oomycetes pathogenic bacteria, including late blight, damping-off, cotton rot, epidemic disease, root rot, cotton blight and downy mildew.

Detailed Description

The present invention will be described in further detail with reference to specific embodiments, but the present invention is not limited to only the illustrated embodiments.

Example 1: dissolving 15.0g (purity > 98%) of euphorbia factor L3 of the invention in 55.0g of acetone solvent, slowly heating to 60 ℃, and continuously stirring until uniform; then 20.0g of surfactant Tween 80 is added and continuously stirred until the mixture is uniform; and finally, adding 10.0g of a penetrating agent JEC, and continuously stirring until the mixture is uniform to obtain a final product.

Example 2: dissolving 10.0g (purity > 98%) of euphorbia factor L3 of the invention in 80.0g of dimethyl sulfoxide solvent, slowly heating to 40 ℃, and continuously stirring until uniform; then 8.0g of anionic surfactant calcium dodecyl benzene sulfonate is added and continuously stirred until the mixture is uniform; and finally, adding 2.0g of penetrant azone, and continuously stirring until the mixture is uniform to obtain a final product.

Example 3: indoor virulence determination of an agricultural oomycete containing euphorbia factor L3 formulated as in example 1:

the plant pathogenic oomycetes to be tested comprise Phytophthora sojae (Phytophthora sojae), Phytophthora infestans (Phytophthora infestans), Phytophthora capsici (Phytophthora capsici), Phytophthora nicotianae (Phytophthora nicotiana), Pythium aphanidermatum (Pythium aphanidermatum) and Pythium ultimum (Pythium ultimum). All tested pathogens were field-collected isolates. Indoor toxicity of 6 plant pathogenic oomycetes is determined by a hyphal growth rate method. Diluting the mother liquid of the bactericide to be tested with sterile water, adding V8 culture medium, mixing, preparing V8 drug-containing culture medium plates with 5 concentration gradients by using a multiple dilution method, and adding sterile water as a control. And (3) punching a bacterial cake on the edge of an activated pathogenic bacteria colony by using a sterilization puncher with the diameter of 5mm, reversely inoculating a culture medium containing a bactericide, placing the culture medium in a constant-temperature incubator at 25 ℃ for culturing for 48h, measuring the diameter of the bacterial colony by using a cross method, calculating the average diameter of the bacterial colony, and calculating the inhibition rate of the bactericide to be tested on the growth of the bacterial colony. Each group was treated in 3 replicates.

And (3) test results: the agricultural fungicide containing euphorbia factor L3 prepared according to example 1 has 1000-fold, 5000-fold, 10000-fold, 20000-fold and 50000-fold dilution concentration, and the results of the inhibitory activity assay on various plant pathogenic oomycetes for test are shown in Table 1. As can be seen from the results in the table, the agricultural fungicide containing the euphorbia factor L3 produced by the technology of the invention has better drug effect on the pathogenic oomycetes of the test plants.

TABLE 1 inhibition of plant pathogenic oomycetes by an agricultural fungicide containing euphorbia factor L3 formulated in example 1

Example 4:

the in vivo efficacy of the agricultural fungicide containing euphorbia factor L3 formulated as in example 2 on several plant pathogenic oomycetes was determined: the test plant pathogenic oomycetes include phytophthora sojae, phytophthora capsici and pythium aphanidermatum; the crops tested were soybean, pepper and cucumber seedlings. The stock solution of the bactericide to be tested was diluted 1000 times, 2000 times and 5000 times with sterile water, and all the agents were sprayed using a knapsack sprayer using clear water as a blank control. The artificial spraying inoculation is carried out by using the pathogenic bacteria spore suspension liquid respectively one day before the medicine application, and the second spraying is carried out 7 days after the medicine application. After inoculation, the cells were cultivated in a greenhouse. When the blank control morbidity reaches more than 50%, checking the morbidity of each treatment, carrying out graded statistics on the pathogenic leaves, and calculating the disease index and the prevention and treatment effect of each treatment. Each group was treated in 3 replicates. And (3) test results: the agricultural fungicide containing the euphorbia factor L3 prepared according to the example 2 has no adverse effect on crops when the dilution concentration is 1000 times, 2000 times and 5000 times, has good control effect on diseases caused by phytophthora sojae, phytophthora capsici and pythium aphanidermatum, and has control effect on three diseases of more than 75% under the dilution dosage of 1000 times and control effect on more than 50% under the dilution dosage of 5000 times.