阅读说明：本技术 鼠李糖脂的应用及其制备方法 (Application of rhamnolipid and preparation method thereof ) 是由 徐兆林 廖美德 石梦滢 赵鹏飞 王诗琦 马自杰 张婉婷 于 2019-06-11 设计创作，主要内容包括：本发明公开了一种鼠李糖脂的制备方法,其中,使用胶孢炭疽菌次生代谢产物制备鼠李糖脂。拓展了鼠李糖脂的制备途径,为该化合物功能的进一步研究奠定了基础。本发明中还公开了鼠李糖脂、其农用盐或酯作为除草活性成分的应用。双鼠李糖脂对多种杂草均具有显著的除草活性,不管是单独作为除草活性成分还是与其他常用化学除草剂混用均能高效抑制其种子萌发和生长,可用于杂草的防除,可将其开发出高效、无污染的绿色除草剂组合物,或者为新型除草剂组合物的创制提供先导化合物,具有显著的社会效益、经济效益和生态效益。(the invention discloses a preparation method of rhamnolipid, wherein the rhamnolipid is prepared from colletotrichum gloeosporioides secondary metabolites. Expands the preparation way of rhamnolipid and lays a foundation for further research of the functions of the compound. The invention also discloses application of the rhamnolipid and the agriculturally acceptable salt or ester thereof as a weeding active ingredient. The dirhamnolipid has obvious herbicidal activity on various weeds, can effectively inhibit the germination and growth of seeds of the weeds whether being used as a herbicidal active ingredient alone or being mixed with other common chemical herbicides, can be used for preventing and removing the weeds, can develop an efficient and pollution-free green herbicide composition, or provides a lead compound for creating a novel herbicide composition, and has obvious social benefit, economic benefit and ecological benefit.)

1. A preparation method of rhamnolipid is characterized in that: rhamnolipids are prepared using colletotrichum gloeosporioides secondary metabolites.

2. the method of claim 1, wherein: the rhamnolipid is a dual rhamnolipid Rha-Rha-C10-C10.

3. Use of rhamnolipids, their agriculturally acceptable salts or esters, as herbicidal active ingredients.

4. Use according to claim 3, characterized in that: the rhamnolipid is a dual rhamnolipid Rha-Rha-C10-C10.

5. a herbicidal composition characterized by: the only weeding active component is rhamnolipid and agricultural salt or ester thereof.

6. A herbicidal composition according to claim 5, characterized in that: the rhamnolipid is a dual rhamnolipid Rha-Rha-C10-C10.

7. A herbicidal composition according to claim 5 or 6, characterized in that: the herbicide composition also contains an agricultural additive or a carrier.

8. a method of controlling undesirable vegetation, comprising: applying to the vegetation the herbicide composition of any one of claims 5-7.

9. The method of claim 8, wherein: the vegetation comprises any one or more of herba seu radix Cirsii Japonici, semen Celosiae, herba Bidentis Bipinnatae, herba Capsellae, Echinochloa crusgalli, Amaranthus retroflexus, flos Osmundae, herba Desmodii Multifloi, radix seu folium Cayratiae Oligocarpae, Bromus margaritae, and herba Euphorbiae Humifusae.

Technical Field

The invention relates to the field of biological pesticide research, and particularly relates to application of rhamnolipid and a preparation method thereof.

Background

With the progress of social civilization and the improvement of public health consciousness, the negative effects brought by the wide, long-term and large-scale use of chemical pesticides are increasingly shown and are concerned by the world. On one hand, the application of a large amount of chemical pesticide brings environmental pollution, and especially the application of long-residual pesticide causes residual toxicity and phytotoxicity, which leads to the reduction of yield of the next crop and even poisoning of people and livestock. On the other hand, excessive dependence and long-term use of a large amount of single pesticides result in generation and development of drug resistance of the diseases, pests and weeds, so that the pesticide effect is reduced, the dosage is increased, the cost is increased, the pollution is also increased, and the continuous use of the existing pesticide varieties and the safety of agricultural production are directly threatened. Therefore, the development of novel pesticides with broad spectrum, high efficiency and low toxicity, especially the development of biogenic pesticides, is very urgent.

the rhamnolipid has more agricultural application researches, can be used as an additive for production of related products in the agricultural field, and has the effects of promoting nutrient absorption of crops, improving soil environment, inhibiting fungi and mold, preventing and treating partial plant diseases and insect pests, and increasing yield and income. CN105454258A discloses an insecticidal composition containing rhamnolipid and matrine, which is prepared by reasonably compounding rhamnolipid and matrine to obtain the insecticidal composition with obvious synergy, thereby enhancing the quick action and reducing the cost of pesticides. CN105964676A discloses a method for reducing the accumulation of heavy metals in rice plants by using a biodegradable chelating agent. Diluting the degradable chelating agent rhamnolipid with water and uniformly stirring to obtain a diluent; the diluent is applied to the paddy field twice, thereby achieving the purpose of reducing the heavy metal in the paddy seeds. The application of the rhamnolipid in weeding is not reported temporarily.

disclosure of Invention

The invention aims to provide a preparation method of rhamnolipid.

The invention aims to provide an application of rhamnolipid, in particular to an application of rhamnolipid in preparation of a herbicide composition.

The technical scheme adopted by the invention is as follows:

A method for preparing rhamnolipid, wherein the rhamnolipid is prepared from secondary metabolite of colletotrichum gloeosporioides.

further, the method specifically comprises the following steps:

1) Extracting a secondary metabolite of colletotrichum gloeosporioides with ethyl acetate, and then carrying out rotary evaporation and concentration under reduced pressure to obtain an extract;

2) Performing silica gel column chromatography on the extract obtained in the step 1) by using a dichloromethane/methanol system, combining by TLC, and performing activity determination;

3) Further separating the active product obtained in 2) by reverse phase silica gel column chromatography to obtain the crude component of the compound;

4) purifying the crude component of the compound obtained in the step 3) by adopting a crystallization method to obtain the compound.

further, the rhamnolipid is a dirhamnolipid Rha-Rha-C10-C10.

use of rhamnolipids, their agriculturally acceptable salts or esters, as herbicidal active ingredients.

Further, the rhamnolipid is a dirhamnolipid Rha-Rha-C10-C10.

A herbicide composition has a herbicidal active ingredient which is only rhamnolipid, an agriculturally acceptable salt or ester thereof.

further, the rhamnolipid is a dirhamnolipid Rha-Rha-C10-C10.

Furthermore, the herbicide composition also contains an agricultural adjuvant or a carrier.

A method for controlling undesirable vegetation, applying to the vegetation the above herbicide composition.

Further, the vegetation comprises any one or more of ageratum conyzoides, feather cockscomb, sticktight, shepherd's purse, barnyard grass, amaranth retroflexus, mikania micrantha, descurainia sophia, ardisia crenata, sparrow wheat and alopecurus.

The invention has the beneficial effects that:

The invention also provides a preparation method of the novel biosurfactant, namely the dirhamnolipid, expands the preparation ways of the rhamnolipid and lays a foundation for further research on the functions of the compound.

the invention provides application of rhamnolipid in preparation of a herbicide composition. The dirhamnolipid has obvious herbicidal activity on various weeds, can effectively inhibit the germination and growth of seeds of the weeds whether being used as a herbicidal active ingredient alone or being mixed with other common chemical herbicides, can be used for preventing and removing the weeds, can develop an efficient and pollution-free green herbicide composition, or provides a lead compound for creating a novel herbicide composition, and has obvious social benefit, economic benefit and ecological benefit.

Drawings

FIG. 1 is colletotrichum gloeosporioides;

FIG. 2 shows fermentation broth of colletotrichum gloeosporioides;

FIG. 3 shows a crude extract of ethyl acetate from fermentation broth of colletotrichum gloeosporioides;

FIG. 4 shows the determination of silica gel column chromatography fraction 82 as the active fraction;

FIG. 5 shows the determination of the active fraction as R127 in reversed-phase column chromatography;

FIG. 6 is a visual representation of the behavior of the compound Rha-Rha-C10-C10;

FIG. 7 is an HPLC check map of compound Rha-Rha-C10-C10;

FIG. 8 is a ¹ H-NMR spectrum of a compound Rha-Rha-C10-C10;

FIG. 9 is a ¹³ C-NMR spectrum of a compound Rha-Rha-C10-C10;

FIG. 10 is a mass spectrum of compound Rha-Rha-C10-C10;

FIG. 11 is an IR spectrum of a compound Rha-Rha-C10-C10;

FIG. 12 shows the herbicidal activity of compounds Rha-Rha-C10-C10 against weeds;

FIG. 13 is a scanning electron micrograph of the mature region of the Bidens pilosa root tip damaged by the compound Rha-Rha-C10-C10.

Detailed Description

the present invention will be described in further detail with reference to examples. It will also be understood that the following examples are included merely for purposes of further illustrating the invention and are not to be construed as limiting the scope of the invention, as the invention extends to insubstantial modifications and adaptations of the invention following in the light of the principles set forth herein. The specific process parameters and the like of the following examples are also only one example of suitable ranges, and the skilled person can make a selection within the suitable ranges through the description herein, and are not limited to the specific data of the following examples.