CN112423592A - 防治植物病原性真菌的生物学方法 - Google Patents
防治植物病原性真菌的生物学方法 Download PDFInfo
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- CN112423592A CN112423592A CN201980047295.9A CN201980047295A CN112423592A CN 112423592 A CN112423592 A CN 112423592A CN 201980047295 A CN201980047295 A CN 201980047295A CN 112423592 A CN112423592 A CN 112423592A
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Abstract
本发明涉及使用生物防治剂防治植物病原性真菌的方法。更具体地,本发明涉及防治具有长潜伏期的植物病原性真菌的方法。特别地,本发明的方法特别适合于防治葡萄白粉病的病原体——真菌葡萄白粉菌。
Description
本发明涉及使用生物防治剂防治植物病原性真菌的方法。更具体地,本发明涉及防治具有长潜伏期的植物病原性真菌的方法。特别地,本发明的方法特别适合于防治葡萄白粉病的病原体——真菌葡萄白粉菌(Erysiphe necator)。
背景技术
已经开发了几种合成的化学溶液,并已将其商业化以防治真菌病害。然而,消费者对在其生产期间较少使用合成的化学产品处理的食品的需求与日俱增。为满足这种需求,正在开发生物防治剂。
大多数具有杀真菌活性的生物防治剂通常被推荐给种植者以在真菌病害实际上发生之前——即在种植者观察到该病害的可见症状(visual symptoms)之前——施用于作物植物上。
然而,许多植物病原性真菌具有长的潜伏期,即病害的可见症状在真菌已经实际上感染植物数周之后才出现。对于这类真菌,在显现症状之前的施用可能实际上与最佳施用时间不对应,因为尽管症状尚未可见,但病害可能已经在植物中很好地存在。
由葡萄白粉菌(也称为葡萄白粉菌(Uncinula necator))引起的葡萄树白粉病是世界范围内最普遍的葡萄树(葡萄(Vitis vinifera L.))病害之一。葡萄白粉菌属于子囊菌纲(ascomycete),并且是专性的活体营养真菌,即其生长和繁殖完全依赖于其存活的葡萄树宿主(葡萄和叶子)。葡萄白粉菌是一种具有长潜伏期的植物病原性真菌,潜伏期可持续数周。
WO 00/58442中记载了生物菌株短小芽胞杆菌(Bacillus pumilus)QST2808(也称为NRRL号B-30087)对某些植物病害具有活性。更具体地,当在其中病原体和处理伴随施用的实验控制条件下进行测试时,短小芽胞杆菌菌株QST2808已经显示具有一定的防治葡萄白粉病葡萄白粉菌的能力。这些数据显示,生物菌株短小芽胞杆菌QST2808具有一定的防治葡萄白粉病葡萄白粉菌的能力,但其没有提供关于这种能力可能根据葡萄树植物的侵染阶段更高效或更低效的任何信息。因此,从WO 00/58442可以假定,无论侵染水平如何,短小芽胞杆菌菌株QST2808防治葡萄白粉菌的能力都是相似的,并且在侵染的任何时间的处理都是有效的。
本发明的发明人已经确定,如果在潜伏期开始时施用生物防治剂,则所述生物防治剂确实更好地防治具有长潜伏期的真菌。更具体地,发明人已经确定,如果在真菌潜伏期早期施用,则生物菌株短小芽胞杆菌QST2808在田间条件下更好地防治葡萄白粉病葡萄白粉菌。对于一些其他生物防治剂也已经注意到了同样的情况。已经表明这些生物防治剂在侵染之前——即病害的首发症状在葡萄树植物上变得可见之前——更好地起作用。然而,对于植物种植者来说,在见到病害侵染迹象之前施用产品是一个挑战,因为他们除了观察他们的田地或邻近田地中植物上病害的首发症状以外,几乎没有工具。当种植者必须面对具有长潜伏期的真菌的侵染时,如果真菌已经处于潜伏期后期的侵染阶段,即当所述真菌已经很好地在作物中存在时,则在可见症状之前施用生物防治剂对于这种产品的最佳功效来说可能实际上已经太迟。此外,处理对种植者来说是昂贵的,并且他们优选仅在他们确信有必要这样做时施用一次。实际上,在没有任何可见迹象的情况下进行处理要么承担因为田间没有侵染而白费处理的风险,要么承担在田间处理过早而看到处理在侵染到来之前被雨水冲洗的风险。
当涉及由具有长潜伏期的真菌(例如葡萄树中的真菌葡萄白粉菌)引起的病害时,这些风险变得甚至更加真实。如大多数具有长潜伏期的真菌一样,真菌葡萄白粉菌确实具有这样的特殊性,即其在葡萄树上的症状通常仅在实际侵染植物数周之后才变得对人眼可见。因此,葡萄种植者难以在他们可以在植物上观察到可见症状之前评估他们的葡萄树是否实际上被侵染,但当这种症状变得可见时,侵染已经在被侵染的植物中很好地存在了。已知当在田地中一些葡萄树植株上的首发症状变得可见这样的时间施用时,生物防治剂如生物菌株短小芽胞杆菌QST2808仍然具有一些功效。通常,种植者从邻近田地收集信息,与天气数据一起,用于评估其感染的风险。当在邻近田地中已经实际上观察到可见症状,并且天气条件对应于有利于真菌发育的那些条件时,他们确实认为他们自己的田地有被侵染的风险,并且他们认为他们的作物处于预侵染的状态。然而,对于真菌如葡萄白粉菌,这种阶段通常对应于真菌实际侵染作物的情况,所述真菌已经处于其潜伏期的晚期,并且接近于对作物造成损害,以至于可见症状将很快变得可以观察到。
为尝试提高种植者在病害的可见症状出现之前知晓他们的葡萄园是否被具有长潜伏期的真菌侵染的能力,从而可能更早地进行一些处理,已经开发出了某些精确的检测工具如qPCR,其能够检测到植物上极少量的真菌。用于真菌葡萄白粉菌的这样的工具例如记载于WO2017/009251中。
然而,尽管已知生物菌株短小芽胞杆菌QST2808在被葡萄白粉菌侵染之前——理解为意指在田间观察到首发可见症状之前,与侵染在田间大多数植物上明显可见时相比——更加有效,但没有证据表明在侵染过程中更早地施用这种产品进行处理将会一样有效。
发明人使用精确的检测工具已经确定,当其在葡萄白粉菌的侵染过程中非常早地施用时,生物防治剂如短小芽胞杆菌菌株QST2808甚至更加有效,因此,使用这种精确的检测工具用于在早期侵染阶段定位处理的方法能够优化产品如短小芽胞杆菌菌株QST2808的功效。该发现被认为可类似地适用于使用生物防治剂处理由具有长潜伏期的真菌引起的其他真菌病害。
发明内容
本发明提供使用生物防治剂防治具有长潜伏期的植物真菌病原体的方法,其特征在于,当植物真菌病原体的感染处于潜伏期开始时,将有效量的生物防治剂施用于所述植物上。
类似地,当考虑由数百种或数千种植物组成的作物田地时,本发明提供一种使用生物防治剂防治具有长潜伏期的植物真菌病原体的方法,其特征在于,当作物田地上植物真菌病原体的感染平均来说处于潜伏期开始时,将有效量的生物防治剂施用于所述作物田地上。
在侵染性病害中,“潜伏期”是从宿主被病原体感染的时刻直至首发症状变得可观察到的时刻的持续时间。因此,在植物真菌病原体的情况下,潜伏期对应于宿主植物(对于给定的宿主-病原体关系)与病原体接触的时刻与病害的首发症状在某些植物部位上变得可观察到的时刻之间的时间段。对于植物病原体,该潜伏期通常被认为对应于一个时间段,在此期间,病原体在某些植物部位(例如叶、根、果实……)的表面上繁殖,使用特殊的侵染器官(吸器)穿透此类植物部位,在植物细胞之间生长并以它们为食,至由这样的侵染引起的生物退化产生特定于该病害的某些症状的程度,变得可通过人眼在被感染的植物部位上观察到。因此,可以理解的是,可观察到的症状不是指定为潜伏期的时间段的一部分,并且其观察时刻实际上是潜伏期的终点。宿主植物与病原体接触的那一刻通常被称为感染阶段,并且可以由与植物部位物理接触的单个病原体传染体组成。对于真菌病原体,这种单个的病原体感染体通常是病原体如孢子的生殖体和繁殖体。根据所考虑的宿主-病原体关系,症状可呈现各种形式,影响植物生长的症状(某些器官的发育不足或过度发育,正常外观的改变,例如叶斑、叶脉带、叶变形、菌丝体或孢子包衣……),或某些植物器官的坏死(某些植物部位的腐烂和腐朽)。在Sambamurty(2006),Textbook of plant pathology,IKInternational Pvt Ltd,第416页中给出了某些植物-病原体关系的症状的实例。
具有长潜伏期的植物真菌病原体是对某些植物致病的真菌,其潜伏期持续数天、优选数周。更优选地,具有长潜伏期的植物真菌病原体是具有至少一周、至少两周、至少三周、最优选至少一个月的潜伏期的真菌。潜伏期高度取决于植物病原体及其感染的宿主植物,但也可能受环境因素如温度、日光或湿度的影响。因此,潜伏期理解为给定的植物病原体在给定植物上的平均潜伏期。平均长潜伏期是约2周至约16周的潜伏期,更确切地约2周至约12周、约2周至约8周、约2周至约4周、约3周至约4周的潜伏期。
具有长潜伏期的植物真菌病原体的非限制性实例为:葡萄白粉病(葡萄白粉菌)、灰霉病(灰霉菌(Botrytis cinerea)壳针孢属(Septoria)叶疱病(禾生球腔菌(Mycosphaerella graminicola))、颖枯壳针孢斑(Septoria nodorum blotch)(颖枯壳针孢(Phaeosphaeria nodorum))。
根据本发明的方法,当植物真菌病原体的感染处于潜伏期开始时,将生物防治剂施用于植物或作物田地上。可以理解的是,对于给定的真菌病原体,应在潜伏期开始时考虑感染,所述潜伏期是从真菌病原体的一些感染体(例如其孢子)与宿主植物接触持续到直至真菌开始在植物表面上生长的一段时间。由于这些事件对人眼来说是不可见的,因此当植物真菌病原体的侵染处于潜伏期开始时,需要某些检测手段来识别这样的时间。在P.Narayanasamy,2011,Microbial Plant Pathogens-Detection and DiseaseDiagnosis:Fungal Pathogens,第1卷,编辑Springer Science+Business Media:5-199中对合适的检测手段进行了说明。
优选的检测方法是qPCR,如例如在国际PCT专利申请WO95/29260或WO2017/009251中所记载。
无论使用何种检测方法,确定植物真菌病原体感染处于潜伏期开始时都可能取决于多种因素,并且因此可以通过不同的方法评估。
优选地,在作物田地水平上确定真菌病原体对植物的感染水平。在这种情况下,通过在整个田地上随机取样并评估一定数量的植株来统计确定感染水平。所取样品的数量取决于田地中植株的总数。统计方法使得能够确定代表整个田地的样本大小。随机取样可通过收集至少10株植物、至少20株植物、至少50株植物或至少100株植物的植物材料来进行。收集的植物部位取决于田地中种植的作物的性质,以及意图评估的病原体。某些真菌病原体实际上仅影响植物的某些部位。在测量葡萄树田地中葡萄白粉病葡萄白粉菌的可能的早期感染的情况下,优选收集叶作为材料,并且优选在植物开花前收集它们。根据该取样方法,如果在使用检测方法测量真菌病原体之后,检测结果表明受感染样本少于50%,优选少于40%、30%、20%、10%,更优选少于5%,并且甚至更优选少于1%,则认为感染处于潜伏期开始时。
在某些情况下,可能需要确定单个植株的感染水平。对于这样的测定,优选允许定量测量真菌病原体的感染水平的检测工具。在这种情况下,可以在植物的不同部位取样,这也取决于植物的性质和意图测试的真菌病原体的性质。在进行定量测量时,认为在潜伏期开始时的感染水平非常依赖于所使用的检测方法。然而,可以认为与潜伏期开始时的感染水平相对应的措施是低检测水平。因此,考虑到待测试的植物和真菌病原体以及待使用的检测方法,技术人员会知道将何种测定视为在潜伏期开始时的感染水平。
因此,本发明还提供使用生物防治剂防治具有长潜伏期的植物真菌病原体侵染植物的方法,其包括以下步骤:
(a)在可观察到可见症状之前使用检测植物真菌病原体的手段确定植物真菌病原体对植物的感染阶段;
(b)当步骤(a)的测定显示植物真菌病原体的感染处于潜伏期开始时,将有效量的生物防治剂施用于植物上。
除使用检测方法以外,还可考虑已知与真菌病原体的感染风险有关的其他信息来确定在潜伏期开始时的感染水平,或确定使用检测方法之一评估感染水平的理想时刻。实际上,已知真菌病原体的感染受某些环境因素例如湿度、空气温度或日光的影响。此外,关于给定真菌病原体在邻近田地中的感染状态的信息可能与确定给定田地中的感染风险相关。所有这些另外的信息均可用于确定在给定田地中的感染风险,从而指导在这样的给定田地中使用检测方法之一来评估感染水平的最佳时机。因此,应认为当给定真菌病原体的其他信息集有助于确定在给定田地中即将发生的感染风险时,则使用检测方法之一来评估田地中的实际感染水平是合适的。对于给定的真菌病原体和作物,相关的所有其他信息均可通过技术人员可用的适当手段来测量,技术人员然后还能够解释这样的附加信息来确定在给定田地中的感染风险。这些附加信息还可从各种来源收集,并被输入到包括一个或多个旨在基于这些附加信息计算在给定田地中的感染风险的计算机程序的数字工具中。
因此,本发明还提供一种使用生物防治剂防治具有长潜伏期的植物真菌病原体侵染作物田地的方法,其包括以下步骤:
(a)通过测量与植物真菌病原体的发育相关的某些参数来确定作物田地是否处于被植物真菌病原体感染的风险中,然后使用这些测量所得信息来计算作物田地中这种发展的可能性;
(b)如果步骤(a)的测定发现作物田地处于风险中,则在可观察到可见症状之前使用检测植物真菌病原体的手段确定植物真菌病原体对田地的感染阶段;
(c)当步骤(a)的测定显示植物真菌病原体的感染处于潜伏期开始时,将有效量的生物防治剂施用于田地上。
根据一个特定的实施方案,本发明提供一种使用生物防治剂防治真菌病原体葡萄白粉病的方法,其特征在于,当植物真菌病原体的感染处于潜伏期开始时,将有效量的生物防治剂施用于葡萄树上。葡萄白粉病是真菌葡萄白粉菌(也称为葡萄白粉菌(Uncinulanecator))。该真菌影响物种葡萄的各种葡萄树品种或栽培种。
如本文中所使用的,术语“防治”或“防治”本质上是指降低给定真菌病原体在给定植物、优选作物植物上生长或发育的能力,从而减少由该病害引起的症状在有关植物上的发展。根据一个特定的实施方案,所述真菌病原体是真菌葡萄白粉菌,并且所述作物植物是物种葡萄的任意品种或栽培种。
在农业中,“生物防治剂”通常定义为用于防治影响植物、更特别地作物植物的生长的害虫、病害或杂草的试剂,并且其由活生物体,或从活生物体获得的产物或组合物,或其混合物组成。根据本发明,生物防治剂是微生物、更特别地是细菌和/或由其获得的产物或组合物和/或其混合物。从微生物获得的产物或组合物可以是从这种微生物的细胞中提取的产物。更优选地,从微生物获得的产物或组合物是由这种微生物合成并分泌到培养基中的产物或组合物,其中这种微生物在所述培养基中发酵以存活和生长,例如肉汤培养基。后一种类型的产物也被称为发酵产物。
可用于本发明方法中的生物防治剂的实例为:
细菌,例如(B1.1)枯草芽孢杆菌(Bacillus subtilis)、特别是菌株QST713/AQ713(可作为SERENADE OPTI或SERENADE ASO从Bayer CropScience LP,US获得,NRRL登记号B21661并记载US专利号6060051中);(B1.2)短小芽孢杆菌、特别是菌株QST2808(可作为从Bayer CropScience LP,US获得,登记号NRRL B-30087并记载于US专利号6245551中);(B1.3)短小芽孢杆菌、特别是菌株GB34(可作为Yield从Bayer AG,DE获得);(B1.4)短小芽孢杆菌、特别是菌株BU F-33(NRRL登记号50185);(B1.5)解淀粉芽孢杆菌(Bacillus amyloliquefaciens)、特别是菌株D747(可作为Double NickelTM从Certis,US获得,登记号FERM BP-8234,并公开于US专利号7094592中);(B1.6)枯草芽孢杆菌Y1336(可作为WP从Bion-Tech,中国台湾获得,在中国台湾注册为生物杀真菌剂,注册号4764、5454、5096和5277);(B1.7)解淀粉芽孢杆菌菌株MBI 600(可作为SUBTILEX从BASF SE获得);(B1.8)枯草芽孢杆菌菌株GB03(可作为从Bayer AG,DE获得);(B1.9)解淀粉枯草芽孢杆菌变种(Bacillus subtilis var.amyloliquefaciens)菌株FZB24(可作为杀真菌剂或ECO(EPA注册号70127-5)从Novozymes Biologicals Inc.,Salem,Virginia或Syngenta Crop Protection,LLC,Greensboro,North Carolina获得);(B1.10)蕈状芽胞杆菌(Bacillus mycoides),分离株J(可作为BmJ TGAI或WG从Certis USA获得);(B1.11)地衣芽孢杆菌(Bacilluslicheniformis)、特别是菌株SB3086(可作为EcoGuard TM Biofungicide和Green Releaf从Novozymes获得);(B1.12)类芽孢杆菌属(Paenibacillus sp.)菌株,登记号NRRL B-50972或登记号NRRL B-67129,并记载于国际专利公开号WO 2016/154297中。
在一些实施方案中,所述生物防治剂为产生泛革素(fengycin)型化合物或制磷脂菌素(plipastatin)型化合物、伊枯草素(iturin)型化合物和/或表面活性素(surfactin)型化合物的枯草芽孢杆菌或解淀粉芽孢杆菌菌株。关于背景,参见以下综述文章:Ongena,M.等人,“Bacillus Lipopeptides:Versatile Weapons for Plant DiseaseBiocontrol,”Trends in Microbiology,第16卷,第3期,2008年3月,第115-125页。能够产生脂肽的芽孢杆菌菌株包括枯草芽孢杆菌QST713(可作为SERENADE OPTI或SERENADE ASO从Bayer CropScience LP,US获得,NRRL登记号B21661并记载于US专利号6060051中)、解淀粉芽孢杆菌菌株D747(可作为Double NickelTM从Certis,US获得,登记号FERM BP-8234并公开于US专利号7094592中);枯草芽孢杆菌MBI600(可作为从BeckerUnderwood,US EPA获得,登记号71840-8);枯草芽孢杆菌Y1336(可作为WP从Bion-Tech,中国台湾获得,在中国台湾注册为生物杀真菌剂,注册号4764、5454、5096和5277);解淀粉芽孢杆菌、特别是菌株FZB42(可作为从ABiTEP,DE获得);和解淀粉枯草芽孢杆菌变种FZB24(可作为杀真菌剂或ECO(EPA注册号70127-5)从Novozymes Biologicals Inc.,Salem,Virginia或SyngentaCrop Protection,LLC,Greensboro,North Carolina获得);和
真菌,例如:(B2.1)盾壳霉(Coniothyrium minitans)、特别是菌株CON/M/91-8(登记号DSM-9660;例如购自Bayer的);(B2.2)核果梅奇酵母(Metschnikowiafructicola)、特别是菌株NRRL Y-30752(例如);(B2.3)小球壳孢(Microsphaeropsis ochracea)(例如购自Prophyta的);(B2.5)木霉属(Trichoderma spp.),包括深绿木霉(Trichoderma atroviride),记载于国际申请号PCT/IT2008/000196中的菌株SC1;(B2.6)哈茨木霉(Trichoderma harzianum rifai)菌株KRL-AG2(也称作菌株T-22,/ATCC 208479,例如购自BioWorks,US的PLANTSHIELD T-22G、和TurfShield);(B2.14)粉红粘帚霉(Gliocladium roseum),购自W.F.Stoneman Company LLC的菌株321U;(B2.35)黄色蠕形霉(Talaromyces flavus),菌株V117b;(B2.36)棘孢木霉(Trichoderma asperellum),购自Isagro的菌株ICC 012;(B2.37)棘孢木霉(Trichoderma asperellum),菌株SKT-1(例如购自Kumiai Chemical Industry的);(B2.38)深绿木霉(Trichoderma atroviride),菌株CNCM I-1237(例如购自Agrauxine,FR的WP);(B2.39)深绿木霉,菌株号V08/002387;(B2.40)深绿木霉,菌株NMI号V08/002388;(B2.41)深绿木霉,菌株NMI号V08/002389;(B2.42)深绿木霉,菌株NMI号V08/002390;(B2.43)深绿木霉,菌株LC52(例如Agrimm Technologies Limited的Tenet);(B2.44)深绿木霉,菌株ATCC 20476(IMI 206040);(B2.45)深绿木霉,菌株T11(IMI352941/CECT20498);(B2.46)钩状木霉(Trichoderma harmatum);(B2.47)哈茨木霉(Trichoderma harzianum);(B2.48)哈茨木霉T39(例如购自Makhteshim,US的);(B2.49)哈茨木霉、特别是菌株KD(例如购自Biological ControlProducts,SA(被Becker Underwood收购)的Trichoplus);(B2.50)哈茨木霉,菌株ITEM 908(例如购自Koppert的Trianum-P);(B2.51)哈茨木霉,菌株TH35(例如Mycontrol的Root-Pro);(B2.52)绿木霉(Trichoderma virens)(也称作绿粘帚霉(Gliocladium virens))、特别是菌株GL-21(例如Certis,US的SoilGard 12G);(B2.53)绿色木霉(Trichodermaviride),菌株TV1(例如Koppert的Trianum-P);(B2.54)白粉寄生孢(Ampelomycesquisqualis)、特别是菌株AQ 10(例如IntrachemBio Italia的AQ);(B2.56)出芽短梗霉(Aureobasidium pullulans)、特别是菌株DSM14940的芽生孢子;(B2.57)出芽短梗霉(Aureobasidium pullulans)、特别是菌株DSM14941的芽生孢子;(B2.58)出芽短梗霉、特别是菌株DSM14940和DSM 14941的芽生孢子的混合物(例如bio-ferm,CH的);(B2.64)芽枝状枝孢(Cladosporium cladosporioides),菌株H39(Stichting DienstLandbouwkundig Onderzoek);(B2.69)链孢粘帚菌(Gliocladium catenulatum)(同义词:Clonostachys rosea f.catenulate)菌株J1446(例如AgBio Inc.的以及例如Kemira Agro Oy的);(B2.70)蜡蚧轮枝菌(Lecanicillium lecanii)(先前称为蜡蚧轮枝孢(Verticillium lecanii)),菌株KV01的分生孢子(例如Koppert/Arysta的);(B2.71)蠕形青霉(Penicillium vermiculatum);(B2.72)异常毕赤酵母(Pichia anomala),菌株WRL-076(NRRL Y-30842);(B2.75)深绿木霉,菌株SKT-1(FERM P-16510);(B2.76)深绿木霉,菌株SKT-2(FERM P-16511);(B2.77)深绿木霉,菌株SKT-3(FERMP-17021);(B2.78)盖姆斯木霉(Trichoderma gamsii)(原绿色木霉(T.viride)),菌株ICC080(IMI CC 392151CABI,例如AGROBIOSOL DE MEXICO,S.A.DE C.V.的BioDerma);(B2.79)哈茨木霉,菌株DB 103(例如Dagutat Biolab的T-Gro 7456);(B2.80)多孢木霉(Trichoderma polysporum),菌株IMI 206039(例如BINAB Bio-Innovation AB,Sweden的Binab TF WP);(B2.81)子座木霉(Trichoderma stromaticum)(例如Ceplac,Brazil的Tricovab);(B2.83)奥德曼细基格孢(Ulocladium oudemansii)、特别是菌株HRU3(例如Botry-Zen Ltd,NZ的);(B2.84)黑白轮枝孢(Verticillium albo-atrum)(原大丽轮枝菌(V.dahliae)),菌株WCS850(CBS 276.92;例如Tree Care Innovations的Dutch Trig);(B2.86)厚垣轮枝孢菌(Verticillium chlamydosporium);(B2.87)棘孢木霉菌株ICC 012和盖姆斯木霉菌株ICC 080的混合物(称作例如BIO-TAMTM的产品,购自BayerCropScience LP,US)。
根据一个优选的实施方案,生物防治剂包括细菌菌株短小芽胞杆菌QST2808、其突变体、其发酵产物或其混合物。细菌菌株短小芽胞杆菌QST2808记载于公开为WO 00/58442的国际PCT专利申请中。在WO 00/58442中,该菌株被称为NRRL号B-30087,其与短小芽胞杆菌QST2808同义。依据布达佩斯条约关于国际承认用于专利程序目的的微生物保藏的条款(the provisions of the Budapest Treaty on the International Recognition ofthe Deposit of Microorganisms for the Purpose of Patent Procedure),于1999年1月14日将短小芽胞杆菌QST2808以NRRL保藏,登记号B-30087。这样的生物防治剂可以商标名市售获得。
根据另一个实施方案,生物防治剂是包含从活生物体获得的化合物的组合物。这种组合物的一个实例是包含可从真菌细胞壁或甲壳纲动物外骨骼获得的壳聚糖(chito-oligo-saccharide)(也称为COS)的组合物。另一个实例是包含可从各种植物细胞壁获得的包含果胶衍生的寡聚半乳糖醛酸苷(oligo-galacturonide)(也称为OGA)的组合物。已知COS和OGA是植物天然防御机制的激发子。优选的组合物是包含壳聚糖和寡聚半乳糖醛酸苷的组合物,如欧洲专利申请EP2115066中记载的所述组合物。这样的生物防治剂可以商标名市售获得。
根据另一个实施方案,生物防治剂包含酵母酿酒酵母(Saccharomycescerevisiae)、更具体地酿酒酵母菌株LAS117的提取物、其突变体、其发酵产物或其混合物。这样的生物防治剂记载于公开为WO2007/074303的国际PCT专利申请中。这样的生物防治剂可以商标名市售获得。
根据另一个实施方案,生物防治剂包括真菌菌株白粉寄生孢(Ampelomycesquisqualis)AQ10、其突变体、其发酵产物或其混合物。真菌菌株白粉寄生孢AQ10记载于欧洲专利申请EP0353662中。这样的生物防治剂可以商标名AQ市售获得。
根据另一个实施方案,生物防治剂包括细菌菌株解淀粉芽孢杆菌植物亚种(Bacillus amyloliquefaciens subsp.plantarum)菌株D747、其突变体、其发酵产物或其混合物。这样的生物防治剂可以商标名市售获得。
根据另一个实施方案,生物防治剂包括茶树植物(互叶白千层(Melaleucaalternifolia))的提取物。这样的生物防治剂记载于公开为WO2011/140309的国际PCT专利申请中。这样的生物防治剂可以商标名市售获得。
根据另一个实施方案,生物防治剂包括植物大虎杖(Reynoutria sachalinensis)的提取物。这样的生物防治剂记载于公开为WO2011/014596的国际PCT专利申请中。这样的生物防治剂可以商标名市售获得。
关于生物防治剂,术语“突变体”是指衍生自制备生物防治剂的有关活生物体的遗传变体。在一个实施方案中,突变体具有生物防治剂的有关活生物体的一个或多个或所有识别特征(其生物防治功能)。在特定的情况下,突变体或其发酵产物具有至少与有关亲本活生物体一样好的生物防治特性。这样的突变体可以是具有基因组序列的遗传变体,其与有关的活生物体具有大于约85%、大于约90%、大于约95%、大于约98%或大于约99%的序列同一性。突变体可通过用化学品或辐射处理相关的活生物体细胞或通过从这样的细胞群中选择自发突变体(如噬菌体抗性或抗生素抗性突变体)或本领域技术人员熟知的其他方式来获得。
本发明的产品或组合物可根据本领域中众所周知的方法(包括例如关于生物防治剂短小芽胞杆菌菌株QST2808,使用培养基和记载于公开为WO 00/58442的国际PCT专利申请中的其他方法)通过培养制备所述生物防治剂的有关活生物体获得。
当生物防治剂由微生物、更具体地细菌制成时,常规的大规模微生物培养方法包括深层发酵、固态发酵或液体表面培养。在发酵快结束时,随着营养物被耗尽,细胞开始从生长期向孢子形成期过渡,使得发酵的最终产物主要为孢子、代谢物和残留的发酵培养基。孢子形成是大多数微生物、特别是细菌的自然生命周期的一部分,并且通常由细胞响应于营养限制而引发。配置发酵以获得细菌的高水平的菌落形成单位(“cfu”)和促进孢子形成。培养基中通过发酵产生的细菌细胞、孢子和代谢物可直接使用或通过常规工业方法如离心、切向流过滤、深度过滤和蒸发浓缩。发酵液和发酵液浓缩物在本文中均被称为“发酵产物”。本公开的组合物包括发酵产物。在一些实施方案中,例如通过渗滤法洗涤浓缩的发酵液以除去残留的发酵液和代谢物。
发酵液或发酵液浓缩物可在添加或不添加载体的情况下使用常规干燥过程或方法进行干燥,包括但不限于喷雾干燥、冷冻干燥、盘式干燥、流化床干燥、转鼓式干燥或蒸发。
所得干燥产物可以被进一步加工,如通过研磨或造粒,例如以达到特定的粒径(例如约1至约5 000、约1至约2 500、约1至约500、约1至约250、约1至约100、约1至约50、约1至约25、约1至约10μm的平均粒径,或本领域中期望且已知的任何其他粒径或其范围)或物理形式。也可在干燥后加入下文中记载的载体。
本发明芽孢杆菌属的新的变体和菌株的发酵液的无细胞制剂可通过本领域中已知的任何手段获得,如发酵液的萃取、离心和/或过滤。本领域技术人员将会理解,所谓的无细胞制剂可能并非没有细胞,而是很大程度上没有细胞或基本上没有细胞,这取决于为除去细胞而使用的技术(例如离心速度)。可将所得无细胞制剂干燥和/或与有助于其施用至植物或植物生长介质的组分一起配制。上文中记载的用于发酵液的浓缩方法和干燥技术也适用于无细胞制剂。
在短小芽胞杆菌菌株QST2808的情况下,可根据公开为WO 00/58442的国际PCT专利申请中列出的方法获得代谢物。如本文中使用的术语“代谢物”可指半纯的和纯的或基本上纯的代谢物,或指尚未从有关活生物体中分离的代谢物。
上文记载的用于发酵液制剂的浓缩方法和干燥技术也适用于代谢物。
本发明的组合物可包括制剂惰性物质,其添加到包含细胞、无细胞制剂或代谢物的组合物中以改善功效、稳定性和可用性和/或促进加工、包装和最终用途应用。这样的制剂惰性物质和成分可包括载体、稳定剂、营养物或物理性质改性剂,其可单独添加或组合添加。在一些实施方案中,载体可包括液体材料如水、油和其他有机或无机溶剂,以及固体材料,如生物来源的或通过化学合成的矿物、聚合物或聚合物配合物。在一些实施方案中,载体是促进组合物粘附到植物部位如种子或根的粘结剂或粘合剂。参见例如Taylor,A.G.等人,"Concepts and Technologies of Selected Seed Treatments",Annu.Rev.Phytopathol.28:321-339(1990)。稳定剂可包括抗结块剂、抗氧化剂、干燥剂、保护剂或防腐剂。营养物可包括碳、氮和磷源,如糖、多糖、油、蛋白质、氨基酸、脂肪酸和磷酸盐。物理性质改性剂可包括填充剂、润湿剂、增稠剂、pH改性剂、流变改性剂、分散剂、佐剂、表面活性剂、防冻剂或着色剂。在一些实施方案中,包含细胞、无细胞制剂或通过发酵产生的代谢物的组合物可以在有或没有水作为稀释剂的情况下直接使用而无需任何其他制剂。在一些实施方案中,在浓缩发酵液之后以及在干燥期间和/或干燥之后添加制剂惰性物质。
本公开的组合物可包括载体,其为添加到发酵产物或无细胞制剂中以提高回收率、功效或物理特性和/或帮助包装和施用的惰性制剂成分。这样的载体可单独添加或组合添加。
本公开的组合物可与其他化学和非化学添加剂、佐剂和/或处理剂混合,其中这样的处理剂包括但不限于化学和非化学杀真菌剂、杀昆虫剂、杀螨剂、杀线虫剂、肥料、营养物、矿物质、生长素、生长刺激剂等。
可与本发明的生物防治剂混合的杀真菌剂是化学或生物杀真菌剂。
在一些实施方案中,所述化学或生物杀真菌剂是市售可得的配制的产品,并且与本公开的组合物桶混。在其他实施方案中,在配制之前将化学或生物杀真菌剂与生物防治剂混合,使得所述组合物形成一种配制的产品。
“有效量”是足以实现有益的或期望的结果的量。有效量可在一个或多个应用中施用。就处理和保护而言,“有效量”是足以防治病害,更具体地足以改善、稳定、逆转、减缓或延迟病害的进展的量。
包含生物防治剂的组合物的应用可以是作为叶面喷雾剂、作为植物的繁殖部位(例如种子或块茎)的处理剂和/或作为土壤处理剂施用。
根据一个具体的实施方案,将本文中记载的组合物施用至葡萄树植物或植物部位,优选地上植物部位如芽、花、叶、葡萄、树干(或臂)或茎(藤)。优选将本发明的组合物喷洒在整个葡萄树植物上,以便与植物的所有地上部位接触。
作为处理方案的一部分,可将包含生物防治剂的组合物施用于田地中的作物植物上。通常,作物植物田地不仅接受一种作物保护产品的处理,而是接受数种相同或不同的作物保护产品的处理,计划将所述作物保护产品在作物发育期间依次施用于作物田地上,使作物保护最佳。这样的最佳处理计划是处理方案。更具体地,处理方案可包括使用不同的作物保护产品,其可以是化学产品或生物防治剂或两者。为减少化学产品的使用,处理方案优选包括化学产品和生物防治剂。当生物防治剂被整合到处理方案中时,必须将它们最佳地置于这样的处理方案中,即在作物发育阶段或意图防治的给定害虫的发育阶段(例如植物真菌病原体的感染阶段)允许其最佳的防治功效的处理方案。
根据一个具体的实施方案,所述生物防治剂在根据本发明的方法中用作处理方案的一部分,所述处理方案可包括使用化学作物保护产品和/或其他生物防治剂进行的额外处理。当将生物防治剂包括在旨在防治某些植物真菌病原体的处理方案中时,其优选位于此类植物真菌病原体的潜伏期的开始处。
在其中某些植物真菌病原体尚未被成功防治并因此已能够在作物田地中发育的情况下,则处理方案可以预见使用已知能够防治完全存在(fully-installed)的植物真菌病原体的作物保护产品。当进行这样的处理时,通常可从有关的植物真菌病原体中治愈所述作物。然而,第二次或再次被植物真菌病原体侵染的风险可能仍然存在。因此,如果在潜伏期开始时检测到植物真菌病原体的第二次或再次感染,则根据本发明的方法也可在这样的时间施用生物防治剂。
可包括在处理方案中的作物保护产品可以是化学产品或生物防治剂或两者。
可用于防治植物病原体的化学产品是化学杀真菌剂。此类化学杀真菌剂的实例可以是:
1)麦角固醇生物合成抑制剂,例如(1.001)环丙唑醇(cyproconazole)、(1.002)苯醚甲环唑(difenoconazole)、(1.003)氟环唑(epoxiconazole)、(1.004)环酰菌胺(fenhexamid)、(1.005)苯锈啶(fenpropidin)、(1.006)丁苯吗啉(fenpropimorph)、(1.007)胺苯吡菌酮(fenpyrazamine)、(1.008)氟喹唑(fluquinconazole)、(1.009)粉唑醇(flutriafol)、(1.010)抑霉唑(imazalil)、(1.011)抑霉唑硫酸盐(imazalil sulfate)、(1.012)种菌唑(ipconazole)、(1.013)叶菌唑(metconazole)、(1.014)腈菌唑(myclobutanil)、(1.015)多效唑(paclobutrazol)、(1.016)咪鲜胺(prochloraz)、(1.017)丙环唑(propiconazole)、(1.018)丙硫菌唑(prothioconazole)、(1.019)啶菌噁唑(pyrisoxazole)、(1.020)螺环菌胺(spiroxamine)、(1.021)戊唑醇(tebuconazole)、(1.022)四氟醚唑(tetraconazole)、(1.023)三唑醇(triadimenol)、(1.024)十三吗啉(tridemorph)、(1.025)灭菌唑(triticonazole)、(1.026)(1R,2S,5S)-5-(4-氯苄基)-2-(氯甲基)-2-甲基-1-(1H-1,2,4-三唑-1-基甲基)环戊醇、(1.027)(1S,2R,5R)-5-(4-氯苄基)-2-(氯甲基)-2-甲基-1-(1H-1,2,4-三唑-1-基甲基)环戊醇、(1.028)(2R)-2-(1-氯环丙基)-4-[(1R)-2,2-二氯环丙基]-1-(1H-1,2,4-三唑-1-基)丁-2-醇、(1.029)(2R)-2-(1-氯环丙基)-4-[(1S)-2,2-二氯环丙基]-1-(1H-1,2,4-三唑-1-基)丁-2-醇、(1.030)(2R)-2-[4-(4-氯苯氧基)-2-(三氟甲基)苯基]-1-(1H-1,2,4-三唑-1-基)丙-2-醇、(1.031)(2S)-2-(1-氯环丙基)-4-[(1R)-2,2-二氯环丙基]-1-(1H-1,2,4-三唑-1-基)丁-2-醇、(1.032)(2S)-2-(1-氯环丙基)-4-[(1S)-2,2-二氯环丙基]-1-(1H-1,2,4-三唑-1-基)丁-2-醇、(1.033)(2S)-2-[4-(4-氯苯氧基)-2-(三氟甲基)苯基]-1-(1H-1,2,4-三唑-1-基)丙-2-醇、(1.034)(R)-[3-(4-氯-2-氟苯基)-5-(2,4-二氟苯基)-1,2-噁唑-4-基](吡啶-3-基)甲醇、(1.035)(S)-[3-(4-氯-2-氟苯基)-5-(2,4-二氟苯基)-1,2-噁唑-4-基](吡啶-3-基)甲醇、(1.036)[3-(4-氯-2-氟苯基)-5-(2,4-二氟苯基)-1,2-噁唑-4-基](吡啶-3-基)甲醇、(1.037)1-({(2R,4S)-2-[2-氯-4-(4-氯苯氧基)苯基]-4-甲基-1,3-二氧戊环-2-基}甲基)-1H-1,2,4-三唑、(1.038)1-({(2S,4S)-2-[2-氯-4-(4-氯苯氧基)苯基]-4-甲基-1,3-二氧戊环-2-基}甲基)-1H-1,2,4-三唑、(1.039)1-{[3-(2-氯苯基)-2-(2,4-二氟苯基)环氧乙烷-2-基]甲基}-1H-1,2,4-三唑-5-基硫氰酸酯、(1.040)1-{[rel(2R,3R)-3-(2-氯苯基)-2-(2,4-二氟苯基)环氧乙烷-2-基]甲基}-1H-1,2,4-三唑-5-基硫氰酸酯、(1.041)1-{[rel(2R,3S)-3-(2-氯苯基)-2-(2,4-二氟苯基)环氧乙烷-2-基]甲基}-1H-1,2,4-三唑-5-基硫氰酸酯、(1.042)2-[(2R,4R,5R)-1-(2,4-二氯苯基)-5-羟基-2,6,6-三甲基庚-4-基]-2,4-二氢-3H-1,2,4-三唑-3-硫酮、(1.043)2-[(2R,4R,5S)-1-(2,4-二氯苯基)-5-羟基-2,6,6-三甲基庚-4-基]-2,4-二氢-3H-1,2,4-三唑-3-硫酮、(1.044)2-[(2R,4S,5R)-1-(2,4-二氯苯基)-5-羟基-2,6,6-三甲基庚-4-基]-2,4-二氢-3H-1,2,4-三唑-3-硫酮、(1.045)2-[(2R,4S,5S)-1-(2,4-二氯苯基)-5-羟基-2,6,6-三甲基庚-4-基]-2,4-二氢-3H-1,2,4-三唑-3-硫酮、(1.046)2-[(2S,4R,5R)-1-(2,4-二氯苯基)-5-羟基-2,6,6-三甲基庚-4-基]-2,4-二氢-3H-1,2,4-三唑-3-硫酮、(1.047)2-[(2S,4R,5S)-1-(2,4-二氯苯基)-5-羟基-2,6,6-三甲基庚-4-基]-2,4-二氢-3H-1,2,4-三唑-3-硫酮、(1.048)2-[(2S,4S,5R)-1-(2,4-二氯苯基)-5-羟基-2,6,6-三甲基庚-4-基]-2,4-二氢-3H-1,2,4-三唑-3-硫酮、(1.049)2-[(2S,4S,5S)-1-(2,4-二氯苯基)-5-羟基-2,6,6-三甲基庚-4-基]-2,4-二氢-3H-1,2,4-三唑-3-硫酮、(1.050)2-[1-(2,4-二氯苯基)-5-羟基-2,6,6-三甲基庚-4-基]-2,4-二氢-3H-1,2,4-三唑-3-硫酮、(1.051)2-[2-氯-4-(2,4-二氯苯氧基)苯基]-1-(1H-1,2,4-三唑-1-基)丙-2-醇、(1.052)2-[2-氯-4-(4-氯苯氧基)苯基]-1-(1H-1,2,4-三唑-1-基)丁-2-醇、(1.053)2-[4-(4-氯苯氧基)-2-(三氟甲基)苯基]-1-(1H-1,2,4-三唑-1-基)丁-2-醇、(1.054)2-[4-(4-氯苯氧基)-2-(三氟甲基)苯基]-1-(1H-1,2,4-三唑-1-基)戊-2-醇、(1.055)氯氟醚菌唑(Mefentrifluconazole)、(1.056)2-{[3-(2-氯苯基)-2-(2,4-二氟苯基)环氧乙烷-2-基]甲基}-2,4-二氢-3H-1,2,4-三唑-3-硫酮、(1.057)2-{[rel(2R,3R)-3-(2-氯苯基)-2-(2,4-二氟苯基)环氧乙烷-2-基]甲基}-2,4-二氢-3H-1,2,4-三唑-3-硫酮、(1.058)2-{[rel(2R,3S)-3-(2-氯苯基)-2-(2,4-二氟苯基)环氧乙烷-2-基]甲基}-2,4-二氢-3H-1,2,4-三唑-3-硫酮、(1.059)5-(4-氯苄基)-2-(氯甲基)-2-甲基-1-(1H-1,2,4-三唑-1-基甲基)环戊醇、(1.060)5-(烯丙基硫烷基)-1-{[3-(2-氯苯基)-2-(2,4-二氟苯基)环氧乙烷-2-基]甲基}-1H-1,2,4-三唑、(1.061)5-(烯丙基硫烷基)-1-{[rel(2R,3R)-3-(2-氯苯基)-2-(2,4-二氟苯基)环氧乙烷-2-基]甲基}-1H-1,2,4-三唑、(1.062)5-(烯丙基硫烷基)-1-{[rel(2R,3S)-3-(2-氯苯基)-2-(2,4-二氟苯基)环氧乙烷-2-基]甲基}-1H-1,2,4-三唑、(1.063)N'-(2,5-二甲基-4-{[3-(1,1,2,2-四氟乙氧基)苯基]硫烷基}苯基)-N-乙基-N-甲基酰亚氨基甲酰胺、(1.064)N'-(2,5-二甲基-4-{[3-(2,2,2-三氟乙氧基)苯基]硫烷基}苯基)-N-乙基-N-甲基酰亚氨基甲酰胺、(1.065)N'-(2,5-二甲基-4-{[3-(2,2,3,3-四氟丙氧基)苯基]硫烷基}苯基)-N-乙基-N-甲基酰亚氨基甲酰胺、(1.066)N'-(2,5-二甲基-4-{[3-(五氟乙氧基)苯基]硫烷基}苯基)-N-乙基-N-甲基酰亚氨基甲酰胺、(1.067)N'-(2,5-二甲基-4-{3-[(1,1,2,2-四氟乙基)硫烷基]苯氧基}苯基)-N-乙基-N-甲基酰亚氨基甲酰胺、(1.068)N'-(2,5-二甲基-4-{3-[(2,2,2-三氟乙基)硫烷基]苯氧基}苯基)-N-乙基-N-甲基酰亚氨基甲酰胺、(1.069)N'-(2,5-二甲基-4-{3-[(2,2,3,3-四氟丙基)硫烷基]苯氧基}苯基)-N-乙基-N-甲基酰亚氨基甲酰胺、(1.070)N'-(2,5-二甲基-4-{3-[(五氟乙基)硫烷基]苯氧基}苯基)-N-乙基-N-甲基酰亚氨基甲酰胺、(1.071)N'-(2,5-二甲基-4-苯氧基苯基)-N-乙基-N-甲基酰亚氨基甲酰胺、(1.072)N'-(4-{[3-(二氟甲氧基)苯基]硫烷基}-2,5-二甲基苯基)-N-乙基-N-甲基酰亚氨基甲酰胺、(1.073)N'-(4-{3-[(二氟甲基)硫烷基]苯氧基}-2,5-二甲基苯基)-N-乙基-N-甲基酰亚氨基甲酰胺、(1.074)N'-[5-溴-6-(2,3-二氢-1H-茚-2-基氧基)-2-甲基吡啶-3-基]-N-乙基-N-甲基酰亚氨基甲酰胺、(1.075)N'-{4-[(4,5-二氯-1,3-噻唑-2-基)氧基]-2,5-二甲基苯基}-N-乙基-N-甲基酰亚氨基甲酰胺、(1.076)N'-{5-溴-6-[(1R)-1-(3,5-二氟苯基)乙氧基]-2-甲基吡啶-3-基}-N-乙基-N-甲基酰亚氨基甲酰胺、(1.077)N'-{5-溴-6-[(1S)-1-(3,5-二氟苯基)乙氧基]-2-甲基吡啶-3-基}-N-乙基-N-甲基酰亚氨基甲酰胺、(1.078)N'-{5-溴-6-[(顺式-4-异丙基环己基)氧基]-2-甲基吡啶-3-基}-N-乙基-N-甲基酰亚氨基甲酰胺、(1.079)N'-{5-溴-6-[(反式-4-异丙基环己基)氧基]-2-甲基吡啶-3-基}-N-乙基-N-甲基酰亚氨基甲酰胺、(1.080)N'-{5-溴-6-[1-(3,5-二氟苯基)乙氧基]-2-甲基吡啶-3-基}-N-乙基-N-甲基酰亚氨基甲酰胺、(1.081)Ipfentrifluconazole。
2)呼吸链复合物I或II抑制剂,例如(2.001)苯并烯氟菌唑(benzovindiflupyr)、(2.002)联苯吡菌胺(bixafen)、(2.003)啶酰菌胺(boscalid)、(2.004)萎锈灵(carboxin)、(2.005)氟吡菌酰胺(fluopyram)、(2.006)氟酰胺(flutolanil)、(2.007)氟唑菌酰胺(fluxapyroxad)、(2.008)呋吡菌胺(furametpyr)、(2.009)异丙噻菌胺(Isofetamid)、(2.010)吡唑萘菌胺(isopyrazam)(反式差向异构对映体1R,4S,9S)、(2.011)吡唑萘菌胺(反式差向异构对映体1S,4R,9R)、(2.012)吡唑萘菌胺(反式差向异构外消旋体1RS,4SR,9SR)、(2.013)吡唑萘菌胺(顺式差向异构外消旋体1RS,4SR,9RS与反式差向异构外消旋体1RS,4SR,9SR的混合物)、(2.014)吡唑萘菌胺(顺式差向异构对映体1R,4S,9R)、(2.015)吡唑萘菌胺(顺式差向异构对映体1S,4R,9S)、(2.016)吡唑萘菌胺(顺式差向异构外消旋体1RS,4SR,9RS)、(2.017)戊苯吡菌胺(penflufen)、(2.018)吡噻菌胺(penthiopyrad)、(2.019)氟唑菌酰羟胺(pydiflumetofen)、(2.020)pyraziflumid、(2.021)氟唑环菌胺(sedaxane)、(2.022)1,3-二甲基-N-(1,1,3-三甲基-2,3-二氢-1H-茚-4-基)-1H-吡唑-4-甲酰胺、(2.023)1,3-二甲基-N-[(3R)-1,1,3-三甲基-2,3-二氢-1H-茚-4-基]-1H-吡唑-4-甲酰胺、(2.024)1,3-二甲基-N-[(3S)-1,1,3-三甲基-2,3-二氢-1H-茚-4-基]-1H-吡唑-4-甲酰胺、(2.025)1-甲基-3-(三氟甲基)-N-[2'-(三氟甲基)联苯-2-基]-1H-吡唑-4-甲酰胺、(2.026)2-氟-6-(三氟甲基)-N-(1,1,3-三甲基-2,3-二氢-1H-茚-4-基)苯甲酰胺、(2.027)3-(二氟甲基)-1-甲基-N-(1,1,3-三甲基-2,3-二氢-1H-茚-4-基)-1H-吡唑-4-甲酰胺、(2.028)3-(二氟甲基)-1-甲基-N-[(3R)-1,1,3-三甲基-2,3-二氢-1H-茚-4-基]-1H-吡唑-4-甲酰胺、(2.029)3-(二氟甲基)-1-甲基-N-[(3S)-1,1,3-三甲基-2,3-二氢-1H-茚-4-基]-1H-吡唑-4-甲酰胺、(2.030)fluindapyr、(2.031)3-(二氟甲基)-N-[(3R)-7-氟-1,1,3-三甲基-2,3-二氢-1H-茚-4-基]-1-甲基-1H-吡唑-4-甲酰胺、(2.032)3-(二氟甲基)-N-[(3S)-7-氟-1,1,3-三甲基-2,3-二氢-1H-茚-4-基]-1-甲基-1H-吡唑-4-甲酰胺、(2.033)5,8-二氟-N-[2-(2-氟-4-{[4-(三氟甲基)吡啶-2-基]氧基}苯基)乙基]喹唑啉-4-胺、(2.034)N-(2-环戊基-5-氟苄基)-N-环丙基-3-(二氟甲基)-5-氟-1-甲基-1H-吡唑-4-甲酰胺、(2.035)N-(2-叔丁基-5-甲基苄基)-N-环丙基-3-(二氟甲基)-5-氟-1-甲基-1H-吡唑-4-甲酰胺、(2.036)N-(2-叔丁基苄基)-N-环丙基-3-(二氟甲基)-5-氟-1-甲基-1H-吡唑-4-甲酰胺、(2.037)N-(5-氯-2-乙基苄基)-N-环丙基-3-(二氟甲基)-5-氟-1-甲基-1H-吡唑-4-甲酰胺、(2.038)isoflucypram、(2.039)N-[(1R,4S)-9-(二氯亚甲基)-1,2,3,4-四氢-1,4-桥亚甲基萘(methanonaphthalen)-5-基]-3-(二氟甲基)-1-甲基-1H-吡唑-4-甲酰胺、(2.040)N-[(1S,4R)-9-(二氯亚甲基)-1,2,3,4-四氢-1,4-桥亚甲基萘-5-基]-3-(二氟甲基)-1-甲基-1H-吡唑-4-甲酰胺、(2.041)N-[1-(2,4-二氯苯基)-1-甲氧基丙-2-基]-3-(二氟甲基)-1-甲基-1H-吡唑-4-甲酰胺、(2.042)N-[2-氯-6-(三氟甲基)苄基]-N-环丙基-3-(二氟甲基)-5-氟-1-甲基-1H-吡唑-4-甲酰胺、(2.043)N-[3-氯-2-氟-6-(三氟甲基)苄基]-N-环丙基-3-(二氟甲基)-5-氟-1-甲基-1H-吡唑-4-甲酰胺、(2.044)N-[5-氯-2-(三氟甲基)苄基]-N-环丙基-3-(二氟甲基)-5-氟-1-甲基-1H-吡唑-4-甲酰胺、(2.045)N-环丙基-3-(二氟甲基)-5-氟-1-甲基-N-[5-甲基-2-(三氟甲基)苄基]-1H-吡唑-4-甲酰胺、(2.046)N-环丙基-3-(二氟甲基)-5-氟-N-(2-氟-6-异丙基苄基)-1-甲基-1H-吡唑-4-甲酰胺、(2.047)N-环丙基-3-(二氟甲基)-5-氟-N-(2-异丙基-5-甲基苄基)-1-甲基-1H-吡唑-4-甲酰胺、(2.048)N-环丙基-3-(二氟甲基)-5-氟-N-(2-异丙基苄基)-1-甲基-1H-吡唑-4-硫代甲酰胺、(2.049)N-环丙基-3-(二氟甲基)-5-氟-N-(2-异丙基苄基)-1-甲基-1H-吡唑-4-甲酰胺、(2.050)N-环丙基-3-(二氟甲基)-5-氟-N-(5-氟-2-异丙基苄基)-1-甲基-1H-吡唑-4-甲酰胺、(2.051)N-环丙基-3-(二氟甲基)-N-(2-乙基-4,5-二甲基苄基)-5-氟-1-甲基-1H-吡唑-4-甲酰胺、(2.052)N-环丙基-3-(二氟甲基)-N-(2-乙基-5-氟苄基)-5-氟-1-甲基-1H-吡唑-4-甲酰胺、(2.053)N-环丙基-3-(二氟甲基)-N-(2-乙基-5-甲基苄基)-5-氟-1-甲基-1H-吡唑-4-甲酰胺、(2.054)N-环丙基-N-(2-环丙基-5-氟苄基)-3-(二氟甲基)-5-氟-1-甲基-1H-吡唑-4-甲酰胺、(2.055)N-环丙基-N-(2-环丙基-5-甲基苄基)-3-(二氟甲基)-5-氟-1-甲基-1H-吡唑-4-甲酰胺、(2.056)N-环丙基-N-(2-环丙基苄基)-3-(二氟甲基)-5-氟-1-甲基-1H-吡唑-4-甲酰胺、(2.057)pyrapropoyne。
3)呼吸链复合物III抑制剂,例如(3.001)唑嘧菌胺(ametoctradin)、(3.002)安美速(amisulbrom)、(3.003)嘧菌酯(azoxystrobin)、(3.004)甲香菌酯(coumethoxystrobin)、(3.005)丁香菌酯(coumoxystrobin)、(3.006)氰霜唑(cyazofamid)、(3.007)醚菌胺(dimoxystrobin)、(3.008)烯肟菌酯(enoxastrobin)、(3.009)噁唑菌酮(famoxadone)、(3.010)咪唑菌酮(fenamidone)、(3.011)氟菌螨酯(flufenoxystrobin)、(3.012)氟嘧菌酯(fluoxastrobin)、(3.013)醚菌酯(kresoxim-methyl)、(3.014)苯氧菌胺(metominostrobin)、(3.015)肟醚菌胺(orysastrobin)、(3.016)啶氧菌酯(picoxystrobin)、(3.017)唑菌胺酯(pyraclostrobin)、(3.018)唑胺菌酯(pyrametostrobin)、(3.019)唑菌酯(pyraoxystrobin)、(3.020)肟菌酯(trifloxystrobin)、(3.021)(2E)-2-{2-[({[(1E)-1-(3-{[(E)-1-氟-2-苯基乙烯基]氧基}苯基)亚乙基]氨基}氧基)甲基]苯基}-2-(甲氧基亚氨基)-N-甲基乙酰胺、(3.022)(2E,3Z)-5-{[1-(4-氯苯基)-1H-吡唑-3-基]氧基}-2-(甲氧基亚氨基)-N,3-二甲基戊-3-烯酰胺、(3.023)(2R)-2-{2-[(2,5-二甲基苯氧基)甲基]苯基}-2-甲氧基-N-甲基乙酰胺、(3.024)(2S)-2-{2-[(2,5-二甲基苯氧基)甲基]苯基}-2-甲氧基-N-甲基乙酰胺、(3.025)(3S,6S,7R,8R)-8-苄基-3-[({3-[(异丁酰氧基)甲氧基]-4-甲氧基吡啶-2-基}羰基)氨基]-6-甲基-4,9-二氧代-1,5-二氧杂环壬烷-7-基2-甲基丙酸酯、(3.026)mandestrobin、(3.027)N-(3-乙基-3,5,5-三甲基环己基)-3-甲酰氨基-2-羟基苯甲酰胺、(3.028)(2E,3Z)-5-{[1-(4-氯-2-氟苯基)-1H-吡唑-3-基]氧基}-2-(甲氧基亚氨基)-N,3-二甲基戊-3-烯酰胺、(3.029){5-[3-(2,4-二甲基苯基)-1H-吡唑-1-基]-2-甲基苄基}氨基甲酸甲酯、(3.030)metyltetraprole、(3.031)florylpicoxamid。
4)有丝分裂和细胞分裂抑制剂,例如(4.001)多菌灵(carbendazim)、(4.002)乙霉威(diethofencarb)、(4.003)噻唑菌胺(ethaboxam)、(4.004)氟吡菌胺(fluopicolide)、(4.005)戊菌隆(pencycuron)、(4.006)噻菌灵(thiabendazole)、(4.007)甲基硫菌灵(thiophanate-methyl)、(4.008)苯酰菌胺(zoxamide)、(4.009)3-氯-4-(2,6-二氟苯基)-6-甲基-5-苯基哒嗪、(4.010)3-氯-5-(4-氯苯基)-4-(2,6-二氟苯基)-6-甲基哒嗪、(4.011)3-氯-5-(6-氯吡啶-3-基)-6-甲基-4-(2,4,6-三氟苯基)哒嗪、(4.012)4-(2-溴-4-氟苯基)-N-(2,6-二氟苯基)-1,3-二甲基-1H-吡唑-5-胺、(4.013)4-(2-溴-4-氟苯基)-N-(2-溴-6-氟苯基)-1,3-二甲基-1H-吡唑-5-胺、(4.014)4-(2-溴-4-氟苯基)-N-(2-溴苯基)-1,3-二甲基-1H-吡唑-5-胺、(4.015)4-(2-溴-4-氟苯基)-N-(2-氯-6-氟苯基)-1,3-二甲基-1H-吡唑-5-胺、(4.016)4-(2-溴-4-氟苯基)-N-(2-氯苯基)-1,3-二甲基-1H-吡唑-5-胺、(4.017)4-(2-溴-4-氟苯基)-N-(2-氟苯基)-1,3-二甲基-1H-吡唑-5-胺、(4.018)4-(2-氯-4-氟苯基)-N-(2,6-二氟苯基)-1,3-二甲基-1H-吡唑-5-胺、(4.019)4-(2-氯-4-氟苯基)-N-(2-氯-6-氟苯基)-1,3-二甲基-1H-吡唑-5-胺、(4.020)4-(2-氯-4-氟苯基)-N-(2-氯苯基)-1,3-二甲基-1H-吡唑-5-胺、(4.021)4-(2-氯-4-氟苯基)-N-(2-氟苯基)-1,3-二甲基-1H-吡唑-5-胺、(4.022)4-(4-氯苯基)-5-(2,6-二氟苯基)-3,6-二甲基哒嗪、(4.023)N-(2-溴-6-氟苯基)-4-(2-氯-4-氟苯基)-1,3-二甲基-1H-吡唑-5-胺、(4.024)N-(2-溴苯基)-4-(2-氯-4-氟苯基)-1,3-二甲基-1H-吡唑-5-胺、(4.025)N-(4-氯-2,6-二氟苯基)-4-(2-氯-4-氟苯基)-1,3-二甲基-1H-吡唑-5-胺。
5)具有多位点活性的化合物,例如(5.001)波尔多混合剂(bordeaux mixture)、(5.002)敌菌丹(captafol)、(5.003)克菌丹(captan)、(5.004)百菌清(chlorthalonil)、(5.005)氢氧化铜、(5.006)环烷酸铜、(5.007)氧化铜、(5.008)氯氧化铜、(5.009)硫酸铜(2+)、(5.010)二氰蒽醌(dithianon)、(5.011)多果定(dodine)、(5.012)灭菌丹(folpet)、(5.013)代森锰锌(mancozeb)、(5.014)代森锰(maneb)、(5.015)代森联(metiram)、(5.016)代森联锌(metiram zinc)、(5.017)羟基喹啉铜(oxine-copper)、(5.018)甲基代森锌(propineb)、(5.019)硫和包括多硫化钙的硫制剂、(5.020)福美双(thiram)、(5.021)代森锌(zineb)、(5.022)福美锌(ziram)、(5.023)6-乙基-5,7-二氧代-6,7-二氢-5H-吡咯并[3',4':5,6][1,4]二噻英并(dithiino)[2,3-c][1,2]噻唑-3-甲腈。
6)引发宿主防御的化合物,例如(6.001)苯并噻二唑(acibenzolar-S-methyl)、(6.002)异噻菌胺(isotianil)、(6.003)烯丙苯噻唑(probenazole)、(6.004)噻酰菌胺(tiadinil)。
7)氨基酸和/或蛋白质生物合成抑制剂,例如(7.001)嘧菌环胺(cyprodinil)、(7.002)春雷霉素(kasugamycin)、(7.003)春雷霉素盐酸盐水合物(kasugamycinhydrochloride hydrate)、(7.004)氧四环素(oxytetracycline)、(7.005)嘧霉胺(pyrimethanil)、(7.006)3-(5-氟-3,3,4,4-四甲基-3,4-二氢异喹啉-1-基)喹啉。
8)ATP产生抑制剂,例如(8.001)硅噻菌胺(silthiofam)。
9)细胞壁合成抑制剂,例如(9.001)苯噻菌胺(benthiavalicarb)、(9.002)烯酰吗啉(dimethomorph)、(9.003)氟吗啉(flumorph)、(9.004)异丙菌胺(iprovalicarb)、(9.005)双炔酰菌胺(mandipropamid)、(9.006)吡吗啉(pyrimorph)、(9.007)霜霉灭(valifenalate)、(9.008)(2E)-3-(4-叔丁基苯基)-3-(2-氯吡啶-4-基)-1-(吗啉-4-基)丙-2-烯-1-酮、(9.009)(2Z)-3-(4-叔丁基苯基)-3-(2-氯吡啶-4-基)-1-(吗啉-4-基)丙-2-烯-1-酮。
10)脂质和膜合成抑制剂,例如(10.001)霜霉威(propamocarb)、(10.002)霜霉威盐酸盐(propamocarb hydrochloride)、(10.003)甲基立枯磷(tolclofos-methyl)。
11)黑色素生物合成抑制剂,例如(11.001)三环唑(tricyclazole)、(11.002)2,2,2-三氟乙基{3-甲基-1-[(4-甲基苯甲酰基)氨基]丁-2-基}氨基甲酸酯。
12)核酸合成抑制剂,例如(12.001)苯霜灵(benalaxyl)、(12.002)精苯霜灵(benalaxyl-M)(kiralaxyl)、(12.003)甲霜灵(metalaxyl)、(12.004)高效甲霜灵(metalaxyl-M)(精甲霜灵(mefenoxam))。
13)信号转导抑制剂,例如(13.001)咯菌腈(fludioxonil)、(13.002)异菌脲(iprodione)、(13.003)腐霉利(procymidone)、(13.004)丙氧喹啉(proquinazid)、(13.005)喹氧灵(quinoxyfen)、(13.006)乙烯菌核利(vinclozolin)。
14)起解偶联剂作用的化合物,例如(14.001)氟啶胺(fluazinam)、(14.002)消螨多(meptyldinocap)。
15)其他化合物,例如(15.001)脱落酸(abscisic acid)、(15.002)苯噻硫氰(benthiazole)、(15.003)bethoxazin、(15.004)卡巴西霉素(capsimycin)、(15.005)香芹酮(carvone)、(15.006)灭螨猛(chinomethionat)、(15.007)硫杂灵(cufraneb)、(15.008)环氟菌胺(cyflufenamid)、(15.009)霜脲氰(cymoxanil)、(15.010)环丙磺酰胺(cyprosulfamide)、(15.011)flutianil、(15.012)三乙膦酸铝(fosetyl-aluminium)、(15.013)三乙膦酸钙(fosetyl-calcium)、(15.014)三乙膦酸钠(fosetyl-sodium)、(15.015)异硫氰酸甲酯(methyl isothiocyanate)、(15.016)苯菌酮(metrafenon)、(15.017)米多霉素(mildiomycin)、(15.018)纳他霉素(natamycin)、(15.019)福美镍(nickel dimethyldithiocarbamate)、(15.020)酞菌酯(nitrothal-isopropyl)、(15.021)oxamocarb、(15.022)氟噻唑吡乙酮(oxathiapiprolin)、(15.023)奥施康定(oxyfenthiin)、(15.024)五氯苯酚及盐、(15.025)亚磷酸及其盐、(15.026)霜霉威乙膦酸盐(propamocarb-fosetylate)、(15.027)pyriofenone(氯酚酮(chlazafenone))、(15.028)异丁乙氧喹啉(tebufloquin)、(15.029)叶枯酞(tecloftalam)、(15.030)甲磺菌胺(tolnifanide)、(15.031)1-(4-{4-[(5R)-5-(2,6-二氟苯基)-4,5-二氢-1,2-噁唑-3-基]-1,3-噻唑-2-基}哌啶-1-基)-2-[5-甲基-3-(三氟甲基)-1H-吡唑-1-基]乙酮、(15.032)1-(4-{4-[(5S)-5-(2,6-二氟苯基)-4,5-二氢-1,2-噁唑-3-基]-1,3-噻唑-2-基}哌啶-1-基)-2-[5-甲基-3-(三氟甲基)-1H-吡唑-1-基]乙酮、(15.033)2-(6-苄基吡啶-2-基)喹唑啉、(15.034)dipymetitrone、(15.035)2-[3,5-双(二氟甲基)-1H-吡唑-1-基]-1-[4-(4-{5-[2-丙-2-炔-1-基氧基)苯基]-4,5-二氢-1,2-噁唑-3-基}-1,3-噻唑-2-基)哌啶-1-基]乙酮、(15.036)2-[3,5-双(二氟甲基)-1H-吡唑-1-基]-1-[4-(4-{5-[2-氯-6-(丙-2-炔-1-基氧基)苯基]-4,5-二氢-1,2-噁唑-3-基}-1,3-噻唑-2-基)哌啶-1-基]乙酮、(15.037)2-[3,5-双(二氟甲基)-1H-吡唑-1-基]-1-[4-(4-{5-[2-氟-6-(丙-2-炔-1-基氧基)苯基]-4,5-二氢-1,2-噁唑-3-基}-1,3-噻唑-2-基)哌啶-1-基]乙酮、(15.038)2-[6-(3-氟-4-甲氧基苯基)-5-甲基吡啶-2-基]喹唑啉、(15.039)2-{(5R)-3-[2-(1-{[3,5-双(二氟甲基)-1H-吡唑-1-基]乙酰基}哌啶-4-基)-1,3-噻唑-4-基]-4,5-二氢-1,2-噁唑-5-基}-3-氯苯基甲磺酸酯、(15.040)2-{(5S)-3-[2-(1-{[3,5-双(二氟甲基)-1H-吡唑-1-基]乙酰基}哌啶-4-基)-1,3-噻唑-4-基]-4,5-二氢-1,2-噁唑-5-基}-3-氯苯基甲磺酸酯、(15.041)Ipflufenoquin、(15.042)2-{2-氟-6-[(8-氟-2-甲基喹啉-3-基)氧基]苯基}丙-2-醇、(15.043)2-{3-[2-(1-{[3,5-双(二氟甲基)-1H-吡唑-1-基]乙酰基}哌啶-4-基)-1,3-噻唑-4-基]-4,5-二氢-1,2-噁唑-5-基}-3-氯苯基甲磺酸酯、(15.044)2-{3-[2-(1-{[3,5-双(二氟甲基)-1H-吡唑-1-基]乙酰基}哌啶-4-基)-1,3-噻唑-4-基]-4,5-二氢-1,2-噁唑-5-基}苯基甲磺酸酯、(15.045)2-苯基苯酚及盐、(15.046)3-(4,4,5-三氟-3,3-二甲基-3,4-二氢异喹啉-1-基)喹啉、(15.047)quinofumelin、(15.048)4-氨基-5-氟嘧啶-2-醇(互变异构形式:4-氨基-5-氟嘧啶-2(1H)-酮)、(15.049)4-氧代-4-[(2-苯乙基)氨基]丁酸、(15.050)5-氨基-1,3,4-噻二唑-2-硫醇、(15.051)5-氯-N'-苯基-N'-(丙-2-炔-1-基)噻吩-2-磺酰肼、(15.052)5-氟-2-[(4-氟苄基)氧基]嘧啶-4-胺、(15.053)5-氟-2-[(4-甲基苄基)氧基]嘧啶-4-胺、(15.054)9-氟-2,2-二甲基-5-(喹啉-3-基)-2,3-二氢-1,4-苯并氧杂吖庚因(benzoxazepine)、(15.055)丁-3-炔-1-基{6-[({[(Z)-(1-甲基-1H-四唑-5-基)(苯基)亚甲基]氨基}氧基)甲基]吡啶-2-基}氨基甲酸酯、(15.056)(2Z)-3-氨基-2-氰基-3-苯基丙烯酸乙酯、(15.057)吩嗪-1-甲酸、(15.058)3,4,5-三羟基苯甲酸丙酯、(15.059)喹啉-8-醇、(15.060)喹啉-8-醇硫酸酯(2:1)、(15.061){6-[({[(1-甲基-1H-四唑-5-基)(苯基)亚甲基]氨基}氧基)甲基]吡啶-2-基}氨基甲酸叔丁酯、(15.062)5-氟-4-亚氨基-3-甲基-1-[(4-甲基苯基)磺酰基]-3,4-二氢嘧啶-2(1H)-酮、(15.063)aminopyrifen。
本文如上所述的所有指定的(1)至(15)类混配物都可以游离化合物的形式和/或——如果其官能团有此能力的话——其农业上可接受的盐的形式存在。
可用于防治植物病原体的生物防治剂为生物杀真菌剂。此类生物杀真菌剂的实例可以是本文中上文所述那些中的任一种。
通过下文的实验实施例将更全面地理解本发明的各个方面。
下文中记载的所有方法或操作均通过示例给出,并且对应于在可用于实现相同结果的各种方法中做出的选择。这种选择对结果的质量没有影响,因此,本领域技术人员可使用任何合适的方法来实现相同的结果。特别地,并且除非在实施例中另有说明,否则所使用的所有重组DNA技术均根据在Sambrook and Russel(2001,Molecular cloning:Alaboratory manual,第三版,Cold Spring Harbor Laboratory Press,NY)、Ausubel等人,(1994,Current Protocols in Molecular Biology,Current protocols,USA,第1卷和第2卷)和Brown(1998,Molecular Biology LabFax,第二版,Academic Press,UK)中记载的标准方案进行。用于植物分子生物学的标准材料和方法记载于Croy R.D.D.(1993,PlantMolecular Biology LabFax,BIOS Scientific Publications Ltd(UK)和BlackwellScientific Publications(UK))中。用于PCR(聚合酶链反应)的标准材料和方法也记载于Dieffenbach and Dveksler(1995,PCR Primer:A laboratory manual,Cold SpringHarbor Laboratory Press,NY)和McPherson等人,(2000,PCR-Basics:From backgroundto bench,第一版,Springer Verlag,Germany)中。
实施例
实施例1:短小芽胞杆菌菌株QST2808在田间条件下对葡萄白粉病葡萄白粉菌的作用
田间试验在选定的葡萄园田地中和在被葡萄白粉病葡萄白粉菌自然侵染的条件下进行。如国际PCT专利申请WO2017/009251中所述,使用对葡萄白粉菌特异性的qPCR检测工具监测试验田地中白粉病的自然发生和发展。
测试在种植有两个不同的葡萄树品种的葡萄园中两个不同的地点进行,两个品种均对白粉病同样敏感:
地点1:Goult,法国东南部。葡萄树品种:Roussanne。
地点2:Cuxac d’Aude,法国南部。葡萄树品种:Chardonnay。
为进行测试,将短小芽胞杆菌菌株QST2808用于处理方案中,所述方案还含有其他杀真菌产品。
使用硫基产品代替短小芽胞杆菌菌株QST2808进行比较处理。
对照组没有接受处理。
处理方案如下:
应用另外的比较处理方案(“Less QST”方案),以QST2808作为该方案的前三次处理,但缺少第一次或前三次处理,并以硫作为该方案的后三次处理,如下图所示。
1商业产品Luna Sensation含有杀真菌剂氟吡菌酰胺作为活性成分。
2商业产品Rocca含有杀真菌剂苯醚甲环唑和环氟菌胺作为活性成分。
在所述两个地点上,在葡萄园的相同发育阶段进行处理。
通过两种不同的方法评估病害的发展。
首先,使用国际PCT专利申请WO2017/009251中记载的qPCR工具,在第一次处理时在叶上分子评估葡萄白粉菌的侵染。此时,没有观察到该病害的可见症状。
第二,在该方案的最后一次处理后3至4周,在完全发育的串上目测评估该病害症状的发展。
通过在相同实验地块的几个植株上取样约一百串、计数被感染的串的数量并评估被感染的串面积的百分比,来进行对葡萄串的症状的视觉观察。
根据下式,症状的视觉观察结果表示为处理方案相对于未处理的对照组的功效的百分比:
((对照组的症状–处理组的症状)/对照组的症状)x 100
qPCR检测
qPCR测量结果显示,在Goult地点,葡萄白粉菌在处理方案开始时已经很好地存在(十分之九的测试叶显示出可检测水平的真菌),而在Cuxac d’Aude地点,其存在少得多(在处理方案开始时没有被感染的叶,在第一次处理后十分之三被感染的叶,在第二次处理后二十分之一被感染的叶)。因此,在没有可观察到的可见症状的情况下,在处理开始时的侵染状况在两个实验地点之间是不同的。
目测观察葡萄串上的症状
在处理方案结束后数周,在完全发育的串上进行观察葡萄串的症状。这些观察在两个地点上进行。
在Goult地点的结果在表1中:
处理类型 | QST2808 | 硫 | Less QST<sup>(1)</sup> |
%功效 | 97.4 | 99.1 | 99.1 |
(1)缺少使用QST2808的前三次处理
这些观察的结果表明,使用QST2808的前三次处理对于所述三次处理方案观察到的良好功效水平没有贡献。
在Cuxac d’Aude地点的结果在表2中:
处理类型 | QST2808 | 硫 | Less QST<sup>(2)</sup> | Less QST<sup>(3)</sup> |
%功效 | 94.9 | 87.8 | 96.8 | 76.7 |
(2)仅缺少使用QST2808的第一次处理
(3)缺少使用QST2808的前三次处理
这些观察的结果表明,使用QST2808的早期处理确实对观察到的处理方案的良好功效水平具有有效贡献。当这些开花前早期处理中的多于一个被除去时,处理方案的总体功效降低。
总之,两个地点葡萄串上的结果与处理方案开始时的qPCR信息结合表明,如果一旦在病原体已经在葡萄田地中很好地存在后进行这样的早期QST2808处理,则早期QST2808处理不能有助于控制葡萄白粉菌的发育。另一方面,这些结果表明,如果在轻微检测到病原体时(即当田地侵染处于潜伏期开始时)进行这样的早期QST2808处理,则早期QST2808处理确实有助于控制葡萄白粉菌的发育。
实施例2:含有COS和OGA的生物防治剂在田地条件下对葡萄白粉病葡萄白粉菌的作用
在与实施例1中进行的实验非常相同的条件、相同的位置和相同的时间下,通过用含有壳聚糖(COS)和寡聚半乳糖醛酸苷(OGA)的混合物的另一种生物防治剂处理替代使用短小芽胞杆菌菌株QST2808的处理来重复相同的实验。
目测观察葡萄串上的症状
在处理方案结束后数周,在完全发育的串上进行观察葡萄串的症状。这些观察在两个地点上进行。
在Goult地点的结果在表3中:
在Cuxac d’Aude地点的结果在表4中:
Claims (10)
1.使用生物防治剂防治具有长潜伏期的植物真菌病原体侵染植物的方法,其特征在于,当植物真菌病原体的感染处于潜伏期开始时,将有效量的生物防治剂施用于植物上。
2.根据权利要求1所述的方法,其特征在于,所述植物为作物植物。
3.根据权利要求1或2中任一项所述的方法,其特征在于,所述作物植物位于作物田地中。
4.使用生物防治剂防治具有长潜伏期的植物真菌病原体侵染植物的方法,其包括以下步骤:
(a)在可观察到可见症状之前使用检测植物真菌病原体的手段确定植物真菌病原体对植物的感染阶段;
(b)当步骤(a)的测定显示植物真菌病原体的感染处于潜伏期开始时,将有效量的生物防治剂施用于植物上。
5.使用生物防治剂防治具有长潜伏期的植物真菌病原体侵染作物田地的方法,其包括以下步骤:
(a)通过测量与植物真菌病原体的发育相关的某些参数来确定作物田地是否处于被植物真菌病原体感染的风险中,然后使用这些测量所得信息来计算作物田地中这种发展的可能性;
(b)如果步骤(a)的测定发现作物田地处于风险中,则在可观察到可见症状之前使用检测植物真菌病原体的手段确定植物真菌病原体对田地的感染阶段;
(c)当步骤(a)的测定显示植物真菌病原体的感染处于潜伏期开始时,将有效量的生物防治剂施用于田地上。
6.根据权利要求5所述的方法,其特征在于,步骤(a)的测定使用包括一个或多个旨在计算给定田地的感染风险的计算机程序的数字装置进行,其中由所测量参数得到的信息由这种数字装置用于进行这样的计算。
7.根据权利要求1至6中任一项所述的方法,其特征在于,所述具有长潜伏期的植物真菌病原体是真菌葡萄白粉病葡萄白粉菌,并且所述植物或作物为葡萄树。
8.根据权利要求1至7中任一项所述的方法,其特征在于,所述生物防治剂包含短小芽胞杆菌菌株QST2808。
9.根据权利要求1至8中任一项所述的方法,其特征在于,确定植物真菌病原体的感染处于潜伏期开始是使用对所考虑的植物真菌病原体特异性的qPCR检测手段进行的。
10.根据权利要求1至9中任一项所述的方法,其特征在于,将所述生物防治剂作为处理方案的一部分施用。
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PE20211635A1 (es) | 2021-08-24 |
US20210282404A1 (en) | 2021-09-16 |
CO2021000103A2 (es) | 2021-01-18 |
BR112020025276A2 (pt) | 2021-03-09 |
CL2021000113A1 (es) | 2021-09-03 |
AU2019303964A1 (en) | 2021-03-04 |
EP3823447A1 (en) | 2021-05-26 |
WO2020016193A1 (en) | 2020-01-23 |
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