CN102524255A - Application of linoleic acid in aspect of preventing and controlling Meloidogyne incognita - Google Patents
Application of linoleic acid in aspect of preventing and controlling Meloidogyne incognita Download PDFInfo
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- CN102524255A CN102524255A CN2011104449105A CN201110444910A CN102524255A CN 102524255 A CN102524255 A CN 102524255A CN 2011104449105 A CN2011104449105 A CN 2011104449105A CN 201110444910 A CN201110444910 A CN 201110444910A CN 102524255 A CN102524255 A CN 102524255A
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Abstract
The invention provides an application of linoleic acid in the aspect of preventing and controlling Meloidogyne incognita. The application comprises the steps: linoleic acid is dissolved in methanol to linoleic acid concentration being 0.50-4.00 mM; and when the linoleic acid concentration range is 1.00-4.00 mM, linoleic acid has a dispelling effect to Meloidogyne incognita. According to the invention, used linoleic acid is rich in sources, low in toxicity, and safe to human, livestock and environment; and the preventing and controlling effect is significantly improved.
Description
Technical field
The invention belongs to agricultural technology field.Relate to the application of a kind of fatty acid cpds-linoleic acid aspect the control Meloidogyne incognita.
Background technology
Root-knot nematode is one of worldwide important causal organism, and production has brought heavy losses to world agriculture for it, and the agricultural economy loss that therefore the annual whole world cause is up to 157,000,000,000 (Abad; P, Gouzy, J; Aury, J.M., et al.Genome sequence of the metazoan plant-parasitic nematode Meloidogyne incognita.Nat.Biotechnol.2008; 26,909-915).Kind surplus the root-knot nematode of having reported in the world at present has 90; And Meloidogyne incognita (Meloidogyne incognita), javanese root knot nematode (M.javanica); Peanut root-knot nematode (M.arenaria) and northern root-knot nematode (M.hapla) account for whole root-knot nematode 95% (force is raised; Zheng Jingwu, Shang Hanwu. root-knot nematode classification and evaluation approach and progress. Zhejiang agricultural journal .2005,17:106-110).Meloidogyne incognita causes a series of problems of facility production and makes its a large amount of economic crops underproduction.And in northern China, be main with Meloidogyne incognita, the host plant of root-knot nematode is in extensive range; Surpass plant in 3000, comprise remunerative crops such as Solanaceae in monocotyledon, dicotyledon, herbaceous plant, the woody plant, particularly vegetables, Curcurbitaceae; Especially tomato and cucumber after being injured, generally can be caused the underproduction 30%～40%; Even total crop failure (Peng Deliang. the generation of vegetables nematodiasis and control. China's Vegetable .1998,4:57-58).Current, the control of root-knot nematode is main with the chemicals control mainly, still, because its harm to environment, person poultry safety is cancelled by legislative branch gradually.So safe and effective, the medicine of low toxicity and the method for preventing and treating are extremely urgent.
Root-knot nematode mainly comprises six breeding times; And in two length of times of nematode; Utilize the flourishing cerebral organ that innervates; Olfactory nerves through head is first, and (AWA, the semiochemicals that AWC) comes the perception root system to give out carry out the taxis migration to host plant; Infect plant; Thereby accomplish its life cycle (Cortese M R.The expression of the homologue of the Caenorhabditis elegans lin-45 raf is regulated in the motile stages of the plant parasitic nematode Meloidogyne artiellia.Molecular and Biochemical Parasitology, 2006,149:38～47).Therefore, in two length of times of root-knot nematode, can play the purpose of nematoda control through disturbing the taxis migration of root-knot nematode.And our previous work finds, E-value≤1 o'clock; Beautiful nematode and Meloidogyne incognita homologous gene are near 6000, and wherein the mpk-1 gene is one of major function gene in the Ras approach, and be relevant with the taxis migration of nematode; And mpk-1, higher (Huang Li, Wang Yu of autoploidy in the Meloidogyne incognita of beautiful nematode; Dong Linlin, etc. Meloidogyne incognita and homogenic evaluation of beautiful nematode and analysis thereof. China Agricultural University's journal, 2010; 15:45-50), so the taxis migration of Meloidogyne incognita can supported and reflect to mpk-1 expression of gene situation.
" plant protection " 2011 the 37th volumes the 2nd phase 99-102 page or leaf discloses the article of " the organic acid compound is to the biologically active of Meloidogyne incognita " by name.Adopt indoor isolated measuring method in this article, tested 15 kinds of organic acids Meloidogyne incognita (Meloidogyne incognita) 2 instar larvae lethality rates and ovum hatching inhibiting rate.The result shows that formic acid and propionic acid are the highest to the biologically active of 2 instar larvaes and ovum, obviously be superior to other and supply the examination organic acid, but activity is lower than contrast medicament AVM.To the discovery of comparing of biologically active result and organic acid chemical constitution, small molecular weight compounds has than high bioactivity Meloidogyne incognita.Yet, though micromolecular compound has higher biologically active to Meloidogyne incognita, micromolecular compound and instability thereof, volatile, the growth of crop is caused great harm, and quicken the acidifying of soil environment.And macromolecular fatty acid, for example, linoleic acid is relatively stable, has the various plants body, and comparatively safe to people and animals, environment, shows more potentiality at Meloidogyne incognita aspect preventing and treating.
Linoleic acid is an essential fatty acid; Peanut oil, walnut oil contain a large amount of linoleic acid, and very favourable to the control of hypertension, angina pectoris, atherosclerotic, senile obesity etc., therefore; Linoleic acid is as the material of control nematode, to environment, person poultry safety close friend.
Summary of the invention
For overcoming above-mentioned defective, the object of the present invention is to provide the application of a kind of linoleic acid aspect the control Meloidogyne incognita.
Said being applied as is dissolved in linoleic acid in the methyl alcohol, and the linoleic acid concentration range is 0.5mM to 4.00mM.
Said linoleic acid concentration is when 1.00mM to 4.00mM, as repellant.
Linoleic acid is an essential fatty acid, and is very favourable to the control of hypertension, angina pectoris, atherosclerotic, senile obesity etc., and therefore, linoleic acid is as the material of control nematode, to environment, person poultry safety close friend.In addition, the linoleic acid aboundresources extensively is distributed in the grease of plant, and institute is essential for mammal.And linoleic synthetic method is convenient, and simple, material is abundant.Linoleic acid is as the repellant of Meloidogyne incognita.
The present invention, under laboratory conditions, the checking linoleic acid moves and prevents and treats nematode through the taxis of regulation and control Meloidogyne incognita, and strong data support and new material is provided for the Meloidogyne incognita control efficiency.
Description of drawings
Fig. 1 shows the influence of linoleic acid to the Meloidogyne incognita taxis.
Fig. 2 shows the influence of linoleic acid to the relative expression quantity of mpk-1 gene.
Embodiment
Below in conjunction with specific embodiment, the present invention is described in further detail.Should be understood that these embodiment just in order to demonstrate the invention, but not limit scope of the present invention by any way.
The linoleic acid that uses among the embodiment (CAS:60-33-3) is bought from sigma company, and the methyl alcohol of employing is chromatographically pure methyl alcohol.
The following example adopts the second instar larvae of the linoleic acid solution-treated Meloidogyne incognita of variable concentrations, utilizes the taxis index, from physiological data the effect of linoleic acid as the Meloidogyne incognita repellant is described.
Embodiment 1
Chromatographically pure methyl alcohol is made into the storage liquid that concentration is 4.00mM, and uses the dilution of chromatographically pure methyl alcohol to be the solution of linoleic acid concentration, and be stored in-20 ℃ of refrigerators as 0.50mM.
Breadboard constructed experiment; According to Ward (Ward Samuel.Chemotaxis by the nematode Caenorhabditis elegans:Identification of attractants and analysis of the response by use of mutants.Proc.Nat.Acad.Sci.USA.1973,70,817-821) with (Bargmann C.I. such as Bargmann; Hartweig E.; Horvitz H.R.Odorant selective genes and neurons mediate olfaction in C.elegans.Cell, 1993,74; The method that 515-527) is provided; Adopting diameter is the culture dish of 6cm, in diagonal positions, is being the circle of 1cm apart from edge 1cm label diameter respectively.The agar that adds 10ml 0.8% in each culture dish.In the culture dish of the two circles of mark, add the chromatographically pure methyl alcohol of 10ml in the side circle, opposite side adds the linoleic acid solution of the 0.5mM for preparing of 10ml.Culture dish behind the adding compound; Put into 25 ℃ of incubator lucifuges hatching 2h, in the middle part of each culture dish, with the position of the round equidistance of both sides; The second instar larvae that adds 50 Meloidogyne incognitas that just hatched; 25 ℃ of incubator lucifuge hatching 3h are according to Bargmann and Horvitz (Bargmann C.I., Horvitz H.R.Chemosensory neurons with overlapping functions direct chemotaxis to multiple chemicals in C.elegans.Neuron; 1991; 7, method statistic taxis index (Chemotaxis Index) 729-742), its computational methods are: the quantity and the ratio that adds nematode population in the methanol solvate circle of adding nematode in the linoleic acid solution circle; Taxis index>=0.2 o'clock is to attract; 0≤taxis index＜0.2 o'clock is faint attraction, and taxis index≤-0.02 o'clock is to repel.And, the lethality of statistics second instar larvae, the computational methods of lethality are: move to and add in the variable concentrations linoleic acid solution circle ratio of line insect number in the nematode death toll and circle.
Embodiment 2
Except the concentration of linoleic acid solution of preparation is the 1mM, all the other are with embodiment 1.
Embodiment 3
Except the concentration of linoleic acid solution of preparation is the 2mM, all the other are with embodiment 1.
Embodiment 4
Except the concentration of the linoleic acid solution that uses as the 4.00mM, all the other are with embodiment 1.
Embodiment 5
According to the method for Ward and Bargmann etc., collect the second instar larvae of 2.000 different disposal simultaneously, be used for the analysis of Mi-mpk-1 gene expression.The primer that gene information and quantitative fluorescent PCR adopt, as shown in table 1.
Table 1.Actin, mpk-1 gene and primer information
RNease Kit (Tiangen) is adopted in the extraction of the total RNA of nematode, and total RNA adopts RNase-free DNase (Takara) to carry out purifying, utilizes M-MLV (Promega) to reverse, and obtains article one cDNA chain.With the cDNA chain is template (2ug), carries out Real-time PCR, and fluorescent dye adopts 2 * SYBR Green mix (Toyobo), and each 1ul of 10uM upstream primer and downstream primer supplies 25ul with high purity water.The PCR reaction condition is a two step method:
The first step: 50 ℃ of 2min, 95 ℃ of 1min
Second step: 95 ℃ of 15s, 52 ℃ of 15s, 72 ℃ of 45s, 40 circulations, and at 72 ℃ of collection fluorescence.
Result by Figure of description 1 shows that linoleic acid concentration is the second instar larvae of 1.00mM to 4.00mM repellent Meloidogyne incognita, and the scope of anti-repelling properties index is-0.12～-0.05, and repellent effect strengthens along with the increase of linoleic acid concentration; When linoleic acid concentration is 0.05mM; The taxis index is-0.01; Motion has faint influence to the taxis of Meloidogyne incognita, and negative control (only using methyl alcohol) result shows, the taxis index has only and is-0.008; Between-0.02～0, get rid of the influence of solvent methanol to Meloidogyne incognita.
2 results show by Figure of description, and linoleic acid concentration is 0.05mM to 4.00mM, reduce the relative expression quantity of mpk-1 gene; The reduction scope is: 17%-49%; And the reduction amplitude increases along with linoleic acid concentration, and molecular data is consistent with the taxis index, and the result shows; In our test, linoleic acid can the repellent Meloidogyne incognita.
Physiological data and molecular data show: the linoleic acid in 1.00mM to the 4.00mM scope is as repellant, and the repellent Meloidogyne incognita makes it away from host plant, and the repellent effect strengthens along with the increase of linoleic acid concentration.
Claims (3)
1. the application of linoleic acid aspect the control Meloidogyne incognita.
2. the application of linoleic acid according to claim 1 aspect the control Meloidogyne incognita is characterized in that linoleic acid is dissolved in the methyl alcohol, and the linoleic acid concentration range is 0.5mM to 4.00mM.
3. the application of linoleic acid according to claim 1 aspect the control Meloidogyne incognita is characterized in that linoleic acid concentration is when 1.00mM to 4.00mM, as repellent agent.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105377034A (en) * | 2013-07-18 | 2016-03-02 | Fmc有限公司 | Nematode repellent composition |
| CN108142415A (en) * | 2017-12-26 | 2018-06-12 | 河北农业大学 | A kind of method for preparing entomopathogenic nematode capsule |
| CN110436990A (en) * | 2018-09-03 | 2019-11-12 | 中国农业大学 | A kind of linoleic acid micro emulsion agent and its preparation method |
Citations (1)
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| CN101167452A (en) * | 2006-10-23 | 2008-04-30 | 黄永 | Double effective fatty acid preparation for improving plant growing and protecting plant |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101167452A (en) * | 2006-10-23 | 2008-04-30 | 黄永 | Double effective fatty acid preparation for improving plant growing and protecting plant |
Non-Patent Citations (2)
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| 刘丹丹 等: "有机酸类化合物对南方根结线虫的生物活性", 《植物保护》, vol. 37, no. 2, 8 April 2011 (2011-04-08), pages 99 - 102 * |
| 黄利 等: "南方根结线虫与秀丽线虫同源基因的鉴定及其应用分析", 《中国农业大学学报》, vol. 15, no. 4, 31 December 2010 (2010-12-31), pages 45 - 50 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105377034A (en) * | 2013-07-18 | 2016-03-02 | Fmc有限公司 | Nematode repellent composition |
| JP2016527233A (en) * | 2013-07-18 | 2016-09-08 | エフ エム シー コーポレーションＦｍｃ Ｃｏｒｐｏｒａｔｉｏｎ | Nematode control composition |
| CN108142415A (en) * | 2017-12-26 | 2018-06-12 | 河北农业大学 | A kind of method for preparing entomopathogenic nematode capsule |
| CN108142415B (en) * | 2017-12-26 | 2020-09-04 | 河北农业大学 | Method for preparing entomopathogenic nematode capsule |
| CN110436990A (en) * | 2018-09-03 | 2019-11-12 | 中国农业大学 | A kind of linoleic acid micro emulsion agent and its preparation method |
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