一种植物源精制木醋液对根结线虫防治制剂的制备方法

本发明公开了一种植物源精制木醋液对根结线虫防治制剂的制备方法，其中精制木醋液采用无菌水稀释至1:25-150。

精制木醋液中含有对根结线虫卵具抑孵活性及杀线虫活性的 ... CN105211100A-一种植物源精制木醋液对根结线虫防治制剂的制备方法 -GooglePatents 一种植物源精制木醋液对根结线虫防治制剂的制备方法 DownloadPDF Info Publicationnumber CN105211100A CN105211100A CN201510593160.6A CN201510593160A CN105211100A CN105211100A CN105211100A CN105211100A CN201510593160A CN201510593160A CN201510593160A CN105211100A CN105211100A CN105211100A Authority CN China Priorartkeywords acetumpyrolignosum pyrolignosumrectificatum rectificatumliquid root preparation Priorartdate 2015-09-18 Legalstatus(Thelegalstatusisanassumptionandisnotalegalconclusion.Googlehasnotperformedalegalanalysisandmakesnorepresentationastotheaccuracyofthestatuslisted.) Pending Applicationnumber CN201510593160.6A Otherlanguages English(en) Inventor 何英杰 袁拥福 王扬 韦少江 蒙春高 符锦 尚钰 薛淑昆 李继文 尧弟龙 李迅东 徐云 彭秀玲 CurrentAssignee(Thelistedassigneesmaybeinaccurate.Googlehasnotperformedalegalanalysisandmakesnorepresentationorwarrantyastotheaccuracyofthelist.) KUNMINGXIANDIBIOTECHNOLOGYCoLtd OriginalAssignee KUNMINGXIANDIBIOTECHNOLOGYCoLtd Prioritydate(Theprioritydateisanassumptionandisnotalegalconclusion.Googlehasnotperformedalegalanalysisandmakesnorepresentationastotheaccuracyofthedatelisted.) 2015-09-18 Filingdate 2015-09-18 Publicationdate 2016-01-06 2015-09-18 ApplicationfiledbyKUNMINGXIANDIBIOTECHNOLOGYCoLtd filed Critical KUNMINGXIANDIBIOTECHNOLOGYCoLtd 2015-09-18 PrioritytoCN201510593160.6A priority Critical patent/CN105211100A/zh 2016-01-06 PublicationofCN105211100A publication Critical patent/CN105211100A/zh Status Pending legal-status Critical Current Links Espacenet GlobalDossier Discuss 239000007788 liquid Substances 0.000 title claims abstract description 44 241000243785 Meloidogynejavanica Species 0.000 title claims abstract description 30 238000002360 preparationmethod Methods 0.000 title claims abstract description 23 XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemicalcompound 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OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1 150000002989 phenols Chemicalclass 0.000 description 1 230000035479 physiologicaleffects,processesandfunctions Effects 0.000 description 1 238000006116 polymerizationreaction Methods 0.000 description 1 239000002244 precipitate Substances 0.000 description 1 230000001737 promoting Effects 0.000 description 1 238000000746 purification Methods 0.000 description 1 238000000197 pyrolysis Methods 0.000 description 1 239000002994 rawmaterial Substances 0.000 description 1 238000002310 reflectometry Methods 0.000 description 1 210000002345 respiratorysystem Anatomy 0.000 description 1 238000000926 separationmethod Methods 0.000 description 1 230000035943 smell Effects 0.000 description 1 239000012085 testsolution Substances 0.000 description 1 230000035899 viability Effects 0.000 description 1 Abstract 本发明公开了一种植物源精制木醋液对根结线虫防治制剂的制备方法，其中精制木醋液采用无菌水稀释至1:25-150。

精制木醋液中含有对根结线虫卵具抑孵活性及杀线虫活性的成分，因而精制木醋液具有长效的抑孵活性及杀线活性，在防治根结线虫病害领域具有研究及应用价值和开发前景，精制木醋液对土壤没有污染，其中的有益元素在植物完成生长阶段后仍在起作用，它还吸附了土壤中的重金属元素，对改良土壤起了积极的作用。

Description 一种植物源精制木醋液对根结线虫防治制剂的制备方法 技术领域 [0001] 本发明涉及植物病害防治技术领域，具体是一种植物源精制木醋液对根结线虫防治制剂的制备方法。

背景技术 [0002] 根结线虫是危害农作物的重要病原生物之一。

农业上主要采用化学防治来控制植物线虫病害.可是由于大多数杀线虫剂有较高的毒性，对人畜不安全，有的还有普遍杀生性，长期使用对环境有很大影响。

根结线虫是一个重要的植物病原线虫，在世界各地均有分布，根结线虫寄生于各种高等植物根部，当其在植物的根内取食与繁殖时，能诱导产生球状体或根结，扰乱被侵染植物的生理，降低产量及产品质量。

[0003] 为了控制线虫病害，一些生物制剂及有机物试剂及其安全使用方法方面的研究已经取得了一定的进展，但由于土壤环境十分复杂并且土壤抑菌作用的普遍存在，于很大程度上限制了微生物制剂作用的发挥。

因此目前仍需要开发一些有效、对环境无害的产品或化合物来用于防治植物线虫病害。

自20世纪60年代开始从植物中提取天然杀线虫活性物质以来，开发植物源杀线虫剂的工作引起研究人员极大兴趣。

[0004] 粗木醋液作为木材炭化的副产物，在废弃物的综合利用以及环境保护和可持续发展农业中的应用前景吸引了研究学者们的关注，木醋液是由木材干馏而得到的含有多种有机化合物的酸性淡褐色透明液，木醋液中除了主要成分乙酸外，还含有含量较少、种类繁多的酮类化合物、酚类化合物、醛类和醇类等化合物，木醋液在国际上被用于农业、畜牧业，目前为止国内对木醋液的研究较少，对木醋液促进植物生长方面有一些初步的研究，但就木醋液对根结线虫卵的抑孵作用的方面鲜有报道。

发明内容 [0005] 本发明的目的在于公开了一种植物源精制木醋液对根结线虫防治制剂的制备方法。

[0006] 为实现上述目的，本发明提供如下技术方案: 一种植物源精制木醋液对根结线虫防治制剂的制备方法，其中精制木醋液采用无菌水稀释至1:25-150。

[0007] 作为本发明进一步的方案:所述精制木醋液采用无菌水稀释至1:75-120。

[0008] 作为本发明再进一步的方案:所述精制木醋液采用无菌水稀释至1:100。

[0009] 与现有技术相比，本发明的有益效果是: 精制木醋液中含有对根结线虫卵具抑孵活性及杀线虫活性的成分，因而精制木醋液具有长效的抑孵活性及杀线活性，在防治根结线虫病害领域具有研究及应用价值和开发前景，精制木醋液对土壤没有污染，其中的有益元素在植物完成生长阶段后仍在起作用，它还吸附了土壤中的重金属元素，对改良土壤起了积极的作用。

具体实施方式 [0010] 下面结合具体实施方式对本专利的技术方案作进一步详细地说明。

[0011] —种植物源精制木醋液对根结线虫防治制剂的制备方法，其中精制木醋液采用无菌水稀释至1:75-100。

[0012] 采用不同浓度的精制木醋液对根结线虫卵进行抑制孵化试验，把精制木醋液用无菌水稀释至1:25、1:50、1:75、1:100、1:150、及1:200并与无菌水进行对照，分别在12h、24h、48h、72h、96h、144h进行观察，试验结果表明，不同稀释度木醋液表现的抑孵活性不同，低于100倍的稀释度时，根结线虫卵未孵化率较高，稀释150倍以上抑孵活性开始降低；木醋液抑制效果理想的稀释度在75倍与100倍之间，实际应用效果最好的稀释度是100倍。

在低于100倍稀释度时，24h后刚刚孵化的二龄幼虫存活率几乎为零，在同一稀释倍数下，抑制活性随着时间的延长逐渐增加；实验如下: 1.材料与方法: 1.1材料: 1.1.1供试试剂 粗木醋液从昆明线敌生物科技有限公司采取，炭化用木材以柞木为主的阔叶树木材，采取后的木醋液在贮存罐中静置6个月以后用作本实验的原料。

PH值为4.0左右。

[0013] 1.1.2供试线虫:根结线虫由农业生物多样性工程中心植物线虫研究室培养，寄主为易感品种番前。

[0014] 1.2方法: 1.2.1供试线虫卵的制备:取卵囊较多的番茄根10g剪碎，置于搅拌器中加入200ml的l%NaOCL溶液，离散用于测定抑孵活性的卵。

开启搅拌器45s，立即用无菌水洗入15L的大水桶中进行稀释，将悬浮液快速经过孔径分别为75μπι和26μπι的筛网，将26μπι筛网倒置于另一个15L大水桶进行冲洗，将悬浮液再经过孔径26μm筛网过滤，在孔径26μm筛网上即可收集到游离卵，将游离卵悬于20ml无菌水中，取出5ml悬浮液配置成虫体密度为100粒/ml的悬浮液体系，全部存于4°C的冰箱中备用。

[0015] 1.2.2精制木醋液的制备:静置法是制备精制木醋液方法中最简便有效的方法，将粗木醋液倒入2L的容器，静置六个月左右。

在静置的过程中，木醋液中的不稳定成分发生氧化、聚合等作用而沉淀。

取上部的澄清液即为精制木醋液。

由于长期放置，需要选用耐酸性的材料。

如果想短时间内获得则需要应用一系列的诸如蒸馏、有机络合萃取等分离提纯方法。

[0016] 1.2.3精制木醋液的稀释:以150ml为配制溶液的体系，设置6个稀释梯度，即分别以25倍、50倍、75倍、100倍、150倍、200倍进行稀释配制，配置好的供试溶液置于棕色广口瓶中，避光保存备用，最好是现配现用，配制好的溶液保存时间不易超过15d。

[0017] 1.2.4活性测定:采用浸渍法，选用直径为6cm的质透明表面皿，每个表面皿中用移液枪加入100粒/ml的菌根结线虫卵悬液1ml，再往表面皿中加入3ml稀释好的配制溶液，将表面皿置于培养皿中，放于26°C恒温培养箱中培养，以无菌水为对照，本实验包括对照有7个处理，每个处理设5次重复，共35个样品，分别在12h、24h、48h、72h、96h、144h将表面皿置于体视显微镜下观察卵孵化的情况并计数，同时将孵化出的二龄幼虫挑出，在清水中复活一定时间，分别计算精制木醋液不同稀释度下根结线虫二龄幼虫的存活率。

计算孵化率时，每个处理卵数为100粒，计算公式如下: 孵化率=100处理中孵化出的虫量X100%未孵化率=100-孵化率校正未孵化率=1-对照未孵化率处理未孵化率-对照未孵化率X100%存活率=处理中的孵化出的二龄幼虫孵化出的二龄幼虫活虫X100%。

[0018] 1.2.5数据统计分析:采用SAS8.0进行单因素方差分析。

[0019] 2.结果与分析: 2.1精制木醋液对根结线虫卵的抑孵作用:在精制木醋液100倍稀释度以下的处理仅有极少数的卵孵化出二龄幼虫，一部分幼虫还未破卵壳而出时已经死亡，呈环抱状，在显微镜下可以看出卵内部有些变化，未破壳的线虫幼虫体腔内部结构部分消解，同时产生液泡。

在用75倍与50倍精制木醋液稀释液处理时，极少有幼虫破卵壳，即使刚刚孵化出，虫体也是极细，且很快死亡，虫体体腔内部结构大量消解，体壁与肠分离。

[0020] 精制木醋液在144h后对根结线虫卵孵化的抑制作用按其稀释倍数顺序减小。

在低于100倍稀释度时根结线虫卵孵化率较低，从稀释25倍到100倍之间，它们对根结线虫卵的抑孵作用差异不显著(P〈0.05)，处于一个水平，实际应用效果最好的稀释度是在100倍时；高于150倍稀释度时，根结线虫卵的抑制作用在快速降低，在稀释200倍时校正未孵化率低至46.71%。

低于100倍时则增加应用成本同时对根结线虫卵的抑制作用并无实质提高。

本实验所做的处理与对照相比，都有明显的抑制孵化的效果，并且有显著的差异，试验证明精制木醋液具有抑制根结线虫卵孵化的性能。

[0021] 所述精制木醋液改变了根线虫生存的土壤环境，接触半径PH会变化，精制木醋液不断地释放乙酸味，此释放靠借助水分实现，植物根部吸入氧气的同时吸入了芳香氨醑，刺激呼吸道造成反射性地兴奋作用使线虫不断地大量地吸入愈创木酚，外加安息香酸的作用而死亡，精制木醋液的作用机理是抑制土壤中病原物的繁殖和对植物根部的侵袭，及外围根线虫难以侵入施药部位，精制木醋液的有机成分中乙醛类、苯酚类、酮类对根部四周进行很好地杀菌、消毒，创造了良好的土壤环境，使根线虫在这片土壤难于生存。

[0022] 精制木醋液中含有对根结线虫卵具抑孵活性及杀线虫活性的成分，因而精制木醋液具有长效的抑孵活性及杀线活性，在防治根结线虫病害领域具有研究及应用价值和开发前景，精制木醋液对土壤没有污染，其中的有益元素在植物完成生长阶段后仍在起作用，它还吸附了土壤中的重金属元素，对改良土壤起了积极的作用。

[0023] 上面对本专利的较佳实施方式作了详细说明，但是本专利并不限于上述实施方式，在本领域的普通技术人员所具备的知识范围内，还可以在不脱离本专利宗旨的前提下作出各种变化。

Claims(3) 1.一种植物源精制木醋液对根结线虫防治制剂的制备方法，其特征在于，其中精制木醋液采用无菌水稀释至1:25-150。

2.根据权利要求1所述的植物源精制木醋液对根结线虫防治制剂的制备方法，其特征在于，所述精制木醋液采用无菌水稀释至1:75-120。

3.根据权利要求1所述的植物源精制木醋液对根结线虫防治制剂的制备方法，其特征在于，所述精制木醋液采用无菌水稀释至1:100。

CN201510593160.6A 2015-09-18 2015-09-18 一种植物源精制木醋液对根结线虫防治制剂的制备方法 Pending CN105211100A (zh) PriorityApplications(1) ApplicationNumber PriorityDate FilingDate Title CN201510593160.6A CN105211100A (zh) 2015-09-18 2015-09-18 一种植物源精制木醋液对根结线虫防治制剂的制备方法 ApplicationsClaimingPriority(1) ApplicationNumber PriorityDate FilingDate Title CN201510593160.6A CN105211100A (zh) 2015-09-18 2015-09-18 一种植物源精制木醋液对根结线虫防治制剂的制备方法 Publications(1) PublicationNumber PublicationDate CN105211100A true CN105211100A (zh) 2016-01-06 Family ID=54980717 FamilyApplications(1) ApplicationNumber Title PriorityDate FilingDate CN201510593160.6A Pending CN105211100A (zh) 2015-09-18 2015-09-18 一种植物源精制木醋液对根结线虫防治制剂的制备方法 CountryStatus(1) Country Link CN (1) CN105211100A (zh) CitedBy(2) *Citedbyexaminer,†Citedbythirdparty Publicationnumber Prioritydate Publicationdate Assignee Title CN106490046A (zh) * 2016-10-31 2017-03-15 郑州思辩科技有限公司 一种防治根结线虫病的农药及其制备方法 CN108432819A (zh) * 2018-02-13 2018-08-24 平南县亿翔环保能源有限公司 一种木醋液根结线虫杀虫剂 Citations(4) *Citedbyexaminer,†Citedbythirdparty Publicationnumber Prioritydate Publicationdate Assignee Title CN101606536A (zh) * 2008-06-20 2009-12-23 中国科学院沈阳应用生态研究所 木醋液土壤消毒剂及其制备方法 CN101967380A (zh) * 2010-10-22 2011-02-09 岳永德 一种竹醋土壤调理剂的制备工艺 CN102674974A (zh) * 2012-05-29 2012-09-19 河北玖农生物肥料有限公司 一种含木醋液的生物肥及其制备方法 JP2013184890A (ja) * 2012-03-06 2013-09-19 PanefuriKogyoKk ネコブセンチュウ防除剤 2015 2015-09-18 CN CN201510593160.6A patent/CN105211100A/zh active Pending PatentCitations(4) *Citedbyexaminer,†Citedbythirdparty Publicationnumber Prioritydate Publicationdate Assignee Title CN101606536A (zh) * 2008-06-20 2009-12-23 中国科学院沈阳应用生态研究所 木醋液土壤消毒剂及其制备方法 CN101967380A (zh) * 2010-10-22 2011-02-09 岳永德 一种竹醋土壤调理剂的制备工艺 JP2013184890A (ja) * 2012-03-06 2013-09-19 PanefuriKogyoKk ネコブセンチュウ防除剤 CN102674974A (zh) * 2012-05-29 2012-09-19 河北玖农生物肥料有限公司 一种含木醋液的生物肥及其制备方法 Non-PatentCitations(2) *Citedbyexaminer,†Citedbythirdparty Title 李维蛟等:"木醋液的杀线活性及对根结线虫病的防治效果研究",《中国农业科学》 * 李维蛟等:"精制木醋液对根结线虫卵的抑孵作用",《道客巴巴》 * CitedBy(2) *Citedbyexaminer,†Citedbythirdparty Publicationnumber Prioritydate Publicationdate Assignee Title CN106490046A (zh) * 2016-10-31 2017-03-15 郑州思辩科技有限公司 一种防治根结线虫病的农药及其制备方法 CN108432819A (zh) * 2018-02-13 2018-08-24 平南县亿翔环保能源有限公司 一种木醋液根结线虫杀虫剂 SimilarDocuments Publication PublicationDate Title CN103518750B (zh) 2015-01-28 一种高效复配生物杀菌剂及其应用 CN103392745A (zh) 2013-11-20 一种利用夹竹桃提取物合成的环保型杀虫剂及其制备方法 CN104521635A (zh) 2015-04-22 一种生态环保型有害生物的防治方法 US20190037849A1 (en) 2019-02-07 Compositioncontaininganorganosilaneandaphotocatalyst,andmethodsoftreatingfloweringplantsinfectedwithabacterialdiseaseusingthecomposition CN106508970A (zh) 2017-03-22 一种复配杀菌组合物及其应用 CN105746585B (zh) 2018-11-30 一种蚜虫捕食性天敌昆虫的引诱剂 CN102499266A (zh) 2012-06-20 水溶性木焦油复合消毒剂 CN105211100A (zh) 2016-01-06 一种植物源精制木醋液对根结线虫防治制剂的制备方法 SrinivasaRaoetal. 2011 OccurrenceofblackrotinJatrophacurcasL.plantationsinIndiacausedbyBotryosphaeriadothidea Yeonetal. 2019 Controlofroot-knotnematodesbyamixtureofmaleicacidandcoppersulfate CN103493832B (zh) 2014-10-22 一种高效复配生物杀菌剂及其应用 CN104412968A (zh) 2015-03-18 香草醛在制备南方根结线虫杀虫抑制剂中的应用 CN101502275A (zh) 2009-08-12 一种植物源农药 WO2010150034A2 (en) 2010-12-29 Plantconditioningcompositionofnaturaloriginandmethodofapplicationthereof JP5372066B2 (ja) 2013-12-18 線虫防除剤 CN103548849B (zh) 2015-01-14 一种协同增效复配生物杀菌剂及其应用 CN102037964B (zh) 2014-01-22 杜鹃素在制备杀菌剂中的应用 CN104754945B (zh) 2017-11-24 用于抑制和/或控制生物系统生长的含有寄生性、致病性或杂草生物系统的核酸的组合物 JP2012201596A (ja) 2012-10-22 センチュウ抵抗性誘導剤及びセンチュウ防除方法 ABDAL-TAMIMIetal. 2020 TheefficacytestofnanochitosanandphylexinresistanceearlyblightdiseaseintomatocausedbyAlternariaSolanifungus CN103053480A (zh) 2013-04-24 一种柑橘全爪螨若螨的rna干扰方法 D'Souzaetal. 2021 EarlyinfectionbyPhytophthoraagathidicidaup‐regulatesphotosyntheticactivityinAgathisaustralisseedlings Powelletal. 2013 Assessingtheriskofphylloxerasurvivalduringwhitegrapeprocessing DK2493317T3 (en) 2015-04-13 Treatmentofplantsandsåmaterialeagainsta/byanattackofbacterialpathogens Loffredoetal. 2012 Benefitsfromcompostuseinthepreparationofgrowingsubstratesforplantsincontainer:inhibitionofSclerotiniasclerotiorumandprotectionofPhilodendron LegalEvents Date Code Title Description 2016-01-06 C06 Publication 2016-01-06 PB01 Publication 2016-03-02 C10 Entryintosubstantiveexamination 2016-03-02 SE01 Entryintoforceofrequestforsubstantiveexamination 2019-04-19 RJ01 Rejectionofinventionpatentapplicationafterpublication 2019-04-19 RJ01 Rejectionofinventionpatentapplicationafterpublication Applicationpublicationdate: 20160106