CN113519541B

CN113519541B - Application of euphorbia factor L3 in preparation of pesticide preparation for preventing and treating plant oomycetes diseases

Info

Publication number: CN113519541B
Application number: CN202010297444.1A
Authority: CN
Inventors: 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; 王奇志
Original assignee: Institute of Botany of CAS
Current assignee: Institute of Botany of CAS
Priority date: 2020-04-16
Filing date: 2020-04-16
Publication date: 2022-09-13
Anticipated expiration: 2040-04-16
Also published as: CN113519541A

Abstract

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.

Description

Application of euphorbia factor L3 in preparation of pesticide preparation for preventing and treating plant oomycetes diseases

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 control is a main means for effectively treating plant oomycetes diseases, but the control effect of chemical bactericides is not ideal, 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 prominent 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. The euphorbia factor L3 belongs to euphorbia lathyris alkyl diterpenoid and is one of the main diterpenoid components of euphorbia lathyris seeds, and the content of the euphorbia lathyris factor L3 accounts for 40 percent of the total diterpenoid components of the 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 the pesticide application of euphorbia factor L3, and is characterized in that euphorbia factor L3 is applied to the preparation of pesticide preparations for preventing and treating oomycete diseases.

The technical scheme adopted by the invention is as follows: euphorbia factor L3 is euphorbia lathyris diterpene ester compound, and its molecular formula is: c ₃₁ H ₃₈ O ₇ The 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 is more than 98%) of the euphorbia factor L3 related to 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; finally, 2.0g of penetrant azone is added, and the mixture is continuously stirred until the mixture is uniform, thus obtaining the 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-harvested isolates. Indoor toxicity of 6 plant pathogenic oomycetes is determined by a hyphal growth rate method. Diluting the mother solution of the test bactericide with sterile water, adding V8 culture medium, mixing, preparing 5 concentration gradient V8 medicated culture medium plates by multiple dilution method, and adding sterile water as 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 inhibitory activity measurement results for various plant pathogenic oomycetes 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:

in vivo efficacy of the agricultural fungicide containing euphorbia factor L3 formulated as in example 2 against several plant pathogenic oomycetes was determined: the test plant pathogenic oomycetes include phytophthora sojae, phytophthora capsici and pythium aphanidermatum; the test crops were soybean, pepper and cucumber seedlings. The mother 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 clean 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.

Claims

1. The application of the euphorbia factor L3 in preventing and treating plant diseases is characterized in that the plant diseases are caused by phytophthora sojae, phytophthora infestans, phytophthora capsici, phytophthora nicotianae, pythium aphanidermatum and pythium ultimum.

2. The use according to claim 1, characterized in that euphorbia factor L3 is processed into suspension, powder, aqueous emulsion and oil.