不同熏蒸剂对草莓连作土壤养分和微生物群落的影响

【背景】草莓连年栽培导致土传病害问题突出,施用熏蒸剂进行土壤消毒因效果显著得以广泛应用。但不同熏蒸剂对土壤病原微生物的影响存在较大差异,同时对非靶标微生物和土壤理化性质也会有不同程度的影响。【目的】明确不同熏蒸剂对草莓连作土壤养分和土壤细菌、真菌群落结构的影响,为合理选择熏蒸剂提供科学依据。【方法】以连作土壤为材料设置5个处理：未熏蒸、石灰氮熏蒸、石灰熏蒸、棉隆熏蒸、威百亩熏蒸,测定熏蒸处理后土壤养分含量;采用PacBio测序平台分析土壤微生物多样性的变化。【结果】石灰氮和威百亩处理均增加了碱解氮含量,降低了有机质、有效磷和速效钾含量;棉隆处理土壤中各养分含量均增加;石灰处理除有机质含量增加外,碱解氮、有效磷和速效钾含量均降低;棉隆、石灰和威百亩处理显著降低pH值。5个处理草莓连作土壤样本中获得了1 164个细菌OTU和444个真菌OTU。细菌多样性和丰富度分析发现,4种熏蒸剂处理均增加了土壤细菌群落的丰富度,石灰氮、石灰和威百亩处理增加了土壤细菌菌落的多样性。4种熏蒸剂处理真菌菌落的丰富度低于对照;石灰、棉隆处理真菌菌落的多样性高于对照和石灰氮、威百亩处理低于对照,但差异不显著。在物种组成分析中,从门水平看,变形菌门(Proteobacteria)和芽单胞菌门(Gemmatimonadetes)为优势细菌门;与对照相比,石灰氮、石灰、棉隆处理变形菌门相对丰度增高,威百亩处理相对丰度降低。4种处理均降低了芽单胞菌门的相对丰度。其他门类中,4种处理均增加了浮霉菌门(Planctomycetota)、疣微菌门(Verrucomicrobia)的相对丰度。优势细菌群落分析表明土壤熏蒸减少了芽单胞菌属(Gemmatimonas)、藤黄单胞菌属(Luteimonas)、中慢生根瘤菌属(Mesorhizobium)等细菌的相对丰度,增加了噬几丁质菌属(Chitinophaga)、苍白杆菌属(Ochrobactrum)的相对丰度。子囊菌门(Ascomycota)为优势真菌,石灰氮、石灰、棉隆、威百亩4种处理均增加了子囊菌门的相对丰度。另外还检测到引起草莓根部土传病害的枝孢属(Cladosporium)和镰刀菌属(Fusarium)病菌,熏蒸处理后均降低了枝孢属和镰刀菌属的相对丰度,其中枝孢属在石灰氮、石灰、棉隆、威百亩处理中分别降低了1.35%、1.11%、0.90%和1.31%,镰刀菌属分别降低了0.71%、0.85%、0.19%和0.65%,但差异不显著。4种土壤熏蒸剂均增加了有益真菌毛壳菌属(Chaetomium)的相对丰度。【结论】采用熏蒸剂处理连作土壤改变了微生物群落构成,减少或灭杀土壤中的大部分致病菌属,起到有效防治草莓土传病害的作用,但不能灭杀所有病菌,而且对有益菌和土壤理化性质有不同程度的影响,因此处理后补充有益微生物非常关键。根据对病原菌的灭杀效果,石灰氮、威百亩处理的效果优于其他处理,可作为轮换施用的熏蒸剂,本试验条件下,棉隆是一种弱的处理剂。     

甲酸乙酯对松材线虫的熏蒸效果

【背景】在溴甲烷面临禁用的情形下,探讨新熏蒸剂甲酸乙酯对松材线虫的处理效果,可以科学地评估甲酸乙酯的使用前景。【方法】采用松材线虫分离液和带疫松木段,设置甲酸乙酯5个剂量梯度、5个处理温度和5个处理时间,测定其对松材线虫的毒力及CT值。【结果】在25℃下处理3、6、12、24、48h,甲酸乙酯对松材线虫的Lc50分别为2．63、1．60、0．99、0．41、0．20mg·L-1。温度对毒力有显著影响,在10～29℃,随温度升高,甲酸乙酯对松材线虫的毒力降低,19和29℃下,1．85mg·L-1甲酸乙酯处理松材线虫的死亡率分别为63％和100％。甲酸乙酯熏蒸12h内,能完全杀灭木段中的松材线虫。在23℃下处理6和12h,松材线虫死亡率达到99％时,甲酸乙酯的cr值分别为453．94和424．14mg·h-1·L-1。【结论与意义】甲酸乙酯可用于松材线虫的检疫处理。     

松墨天牛在秦巴林区不同寄主上的危害规律

松墨天牛Monochamus alternatus是次期性蛀干害虫,也是松材线虫病的主要传播媒介,廓清该害虫在秦巴林区不同寄主上的危害程度和发生规律,对于控制松材线虫病的扩散蔓延具有重要意义.本研究对秦巴林区遭受松材线虫病严重危害的3种主要松树(油松Pinus tabuliformis、华山松P.armandii和马尾松P.massoniana)进行了冬季疫木野外解析,并在统计不同胸径、不同高度疫木树干上松墨天牛侵入孔和幼虫数量的基础上,采用聚集度指标分析方法,对该害虫在不同寄主上的危害差异进行了比较研究.结果表明,松墨天牛在3种寄主上的危害程度具有显著差异,以华山松受害最重,其次分别为马尾松和油松;越冬幼虫在油松上主要危害皮下1～2 cm的边材;在华山松和马尾松上危害比较多样,以髓心部位受害最重,其后依次为心材、边材和树皮;在油松和华山松上,松墨天牛主要在树干7m以下部分危害,而在马尾松上主要危害树干7m以上部分,且虫口数量均与寄主胸径呈显著正相关.松墨天牛侵入孔和越冬幼虫在不同寄主树干上均呈聚集性分布.本研究揭示了松墨天牛在秦巴林区3种主要寄主上的危害规律,对于进一步开展大尺度地理范围松墨天牛的寄主选择性及危害规律研究提供了基础信息,也为秦巴林区松墨天牛的有效防治和松材线虫病的蔓延控制提供了新的重要信息.     

基于多光谱卫星影像与机器学习算法的松材线虫病受害林分识别研究

松材线虫病(Pine Wilt Disease, PWD)被称为“松树癌症”,具有高传染率和高死亡率,对我国森林资源构成了严重的威胁,对我国的经济、社会和生态造成了重大损失。及时发现并清理疫木是遏制松材线虫病蔓延的有效手段,精准监测疫木是防控松材线虫病的前提,但是现阶段缺少大面积识别松材线虫病疫木的技术方法。本文旨在探索哨兵-2号与Landsat-8遥感卫星影像对受害松林的识别能力,采用随机森林(Random Forest, RF)、支持向量机(Support Vector Machine, SVM)、决策树(Decision Tree, DT)和极端梯度提升(Extreme Gradient Boosting, XGBoost)等4种机器学习算法建立了松材线虫病监测模型。结果表明：基于哨兵-2号影像数据建立的监测模型对受害松林的识别准确率高于Landsat-8遥感卫星影像,其中基于10 m分辨率的影像数据建立的监测模型识别准确率最高,随机森林、决策树、支持向量机和极端梯度提升等算法建立模型的准确率分别达到了79.3%、76.2%、78.7%和78.9%。在3种不同的影像数据集中,RF...     

几种替代杀线剂对甘薯茎线虫的毒力与活性

采用熏蒸法和浸渍法测定了4种土壤熏蒸杀线剂和6种非熏蒸杀线剂对甘薯茎线虫的毒力,在此基础上以各药剂的致死中浓度剂量处理线虫,设置不同时间处理,观察了药剂对线虫的活性动态变化.结果表明： 熏蒸杀线剂棉隆、1,3-二氯丙烯、威百亩、氯化苦对甘薯茎线虫的LC₅₀依次为0.49、0.89、0.91、3.60 mg·L^-1,非熏蒸杀线剂甲维盐、阿维菌素、灭线磷、噻唑膦、涕灭威、丁硫克百威对甘薯茎线虫的LC₅₀依次为31.2、48.1、224.3、288.4、632.3、823.9 mg·L^-1.致死中浓度处理线虫后,各药剂校正死亡率随处理时间延长有不同幅度升高,棉隆、1,3-二氯丙烯、阿维菌素和甲维盐对线虫有较高的持续抑制效果,处理48 h脱离药剂处理后校正死亡率没有降低,而传统杀线剂品种灭线磷和涕灭威处理均出现线虫明显复苏的现象.表明棉隆、1,3-二氯丙烯、阿维菌素和甲维盐可以替代传统杀线剂应用于甘薯茎线虫的防治.     

温度对松褐天牛传递松材线虫数量的影响

松材线虫病是一种重要的检疫性病害,松褐天牛是传播松材线虫的主要媒介昆虫.本实验研究了温度对松褐天牛传递松材线虫数量的影响.设置22℃、28℃和34℃ 3个温度梯度,结果表明松褐天牛成虫的寿命随温度升高而缩短,其中雌虫寿命短于雄虫,且羽化进度不太一致,多数松褐天牛携带的松材线虫量在1000～9999条之间.通过对松褐天牛取食传入松枝的松材线虫量的追踪记录发现,随着温度升高,传播线虫的高峰时段随之提前.34℃高温下松褐天牛的传递率最低,表明高温对松材线虫病的发生具有抑制作用.     

一种新杀虫剂对储粮害虫和作物害虫的杀虫效果

测定以植物提取物异硫氰酸酯为主要成分而复配的宏劲杀虫剂对玉米象Sitophilus zeamais(Motschulsky)、赤拟谷盗Tribolium castaneum(Herbst)和谷蠹Rhizopertha dominica(Fabricius)3种重要储粮害虫的熏蒸效果及其对菜青虫Pieris rapae(L.)、红蜘蛛Tetranychus cinnabarinus(Boisduval)的杀虫效果。结果表明,宏劲杀虫剂48h对玉米象、赤拟谷盗和谷蠹的LC50分别是0.016,0.009和0.009μL/mL,均明显低于常规熏蒸剂磷化铝的要求处理剂量;对菜青虫和红蜘蛛的LC50分别是0.0159和1.3738mg/mL,其防效均优于对照药剂3%阿菊乳油和3%新型水分散性颗粒剂。     

化学通讯在松材线虫侵染和扩散中的作用

松材线虫为外来入侵种 ,由其引起的松材线虫病正在我国迅速扩散蔓延 ,造成我国部分地区松林资源的毁灭性破坏。松材线虫病的发生和流行与媒介天牛、寄主植物、共生真菌和细菌密切相关 ,松材线虫 -墨天牛 -松树 -共生微生物之间存在着广泛的化学联系 ,它们通过化学互作 ,调控松材线虫的行为 ,影响松材线虫的侵染和扩散     

云杉花墨天牛成虫对四种寄主松树的取食偏好性

【目的】近年来,松材线虫病扩散到我国的中温带地区,云杉花墨天牛Monochamus saltuarius成为该地区松材线虫的新媒介昆虫。由于中温带地区松材线虫危害松树种类较多,本研究对云杉花墨天牛成虫对4种寄主松树的取食偏好性进行测定,为明确云杉花墨天牛成虫补充营养特性,进而为中温带地区松材线虫病和云杉花墨天牛监测与防治工作提供理论依据。【方法】2020年5月云杉花墨天牛羽化前,在辽宁省抚顺市大伙房实验林场采集被云杉花墨天牛侵入的红松Pinus koraiensis疫木,锯成1 m长木段,两端蜡封,置于养虫笼内,每天收集初羽化的云杉花墨天牛成虫。养虫笼内四角分别放置寄主植物红松、油松P. tabulaeformis、樟子松P. sylvestris var. mongolica和长白落叶松Larix olgensis 3年生树苗,将同一天羽化的5对雌雄成虫配对放入养虫笼中央,调察其在不同寄主松树上的日均取食频次和在寄主松树各部位的日均取食量。【结果】云杉花墨天牛成虫主要停留在油松上,日均取食频次为7.05±3.87次/d,其次是红松、长白落叶松上,停留在樟子松上的频次最少,日均取食频次仅有1.02±0.81次/d;各种寄主松树上雌雄成虫间的日均取食频次差异显著。云杉花墨天牛成虫在4种松树上的日均取食量存在显著差异,其中对油松的日均取食量最高,为129.14±50.23 mm²,占取食总量的62.89%;其次是对红松、长白落叶松,对樟子松的日均取食量最低,为9.87±11.02 mm²,仅占取食总量的4.81%。另外,云杉花墨天牛成虫在同种松树植物的不同部位上取食量差异显著,在4种松树上均主要取食嫩枝。【结论】从云杉花墨天牛成虫补充营养期间在不同松树上取食频次和取食量分析,云杉花墨天牛对油松的选择性明显高于其他3种松树。     

磷化铝处理家白蚁障碍巢的野外应用

磷化铝是国家粮食仓库６０年代至今一直用来消灭储粮害虫的常备熏蒸杀虫剂。在使用中磷化铝吸收空气中的水份而分解产生磷化氢（ＰＨ３）。但目前用磷化铝来直接熏蒸毒杀白蚁的报道不多,特别是用磷化铝在野外的家白蚁巢上熏蒸毒杀尚未见报道。本文报道了用磷化铝的熏蒸毒杀能力消灭了多窝家白蚁CoptotermesformosanusShiraki,从７０年代至今,先后在宜都市、宜昌市、宜昌县、长阳县、远安县等地用磷化铝熏蒸了一些难以解决的障碍巢,所处理之处,都达到了彻底消灭的效果。为了总结经验,叙述如下。１野外自然障碍巢障碍巢是指那些在坟墓…     

Ribosomal DNA haplotype distribution of Bursaphelenchus xylophilus in Kyushu and Okinawa islands, Japan

Ribosomal DNA region sequences (partial 18S, 28S and complete ITS1, 5.8S, and ITS2) of the pinewood nematode (Bursaphelenchus xylophilus) were obtained from DNA extracted directly from wood pieces collected from wilted pine trees throughout the Kyushu and Okinawa islands, Japan. Either a 2569bp or 2573bp sequence was obtained from 88 of 143 samples. Together with the 45 rDNA sequences of pinewood nematode isolates previously reported, there were eight single nucleotide polymorphisms and two indels of two bases. Based on these mutations, nine haplotypes were estimated. The haplotype frequencies differed among regions in Kyushu island (northwest, northeast and center, southeast, and southwest), and the distribution was consistent with the invasion and spreading routes of the pinewood nematode previously estimated from past records of pine wilt and wood importation. There was no significant difference in haplotype frequencies among the collection sites on Okinawa island.     

Scanning Electron Microscopy of Pine Seedling Wood Tissue Sections Inoculated with the Pinewood Nematode Bursaphelenchus xylophilus Previously Prepared for Light Microscopy

Scanning electron microscopy (SEM) was applied to paraffin-embedded wood sections to study the histopathology of pine seedlings inoculated with the pinewood nematode (PWN), Bursaphelenchus xylophilus. The sections, which had been previously prepared and observed by light microscopy (LM) on glass slides, were originally obtained from experiments in which pine seedlings had been inoculated with PWN. The cover glass was removed by soaking the glass slide in xylene for 3 to 5 days. The glass slides were cut into small pieces so that each piece contained one wood section. Each piece of the glass slide was attached with double adhesive tape to an aluminum stub. The specimens were sputter-coated with gold and examined with a scanning electron microscope (JEOL-JSM 5200). Compared to LM (as documented in previous reports) SEM provided greater depth of focus and resolution of the damaged wood tissues, nematodes and associated bacteria. SEM made it possible to observe the relationship between bacterial distribution and nematode distribution in wood tissues. SEM observations also suggested the possibility of documenting the death of ray cells and other parenchyma cells in relation to disease development. Finally, the current study of PWN in pine seedlings demonstrated that glass slides prepared for LM observations more than 25 years earlier could be successfully processed for examination by SEM.     

Bacteria associated with the pinewood nematode <Emphasis Type="Italic">Bursaphelenchus xylophilus</Emphasis> collected in Portugal

In this study, we report on the bacterial community associated with the pinewood nematode Bursaphelenchus xylophilus from symptomatic pine wilted trees, as well as from long-term preserved B. xylophilus laboratory collection specimens, emphasizing the close bacteria–nematode associations that may contribute to pine wilt disease development.     

Transmission of the Pinewood Nematode, Bursaphelenchus xylophilus,to Slash Pine Trees and Log Bolts by a Cerambycid Beetle,Monochamus titillator, in Florida

Field-collected adults of the southern pine sawyer, Monochamus titillator (F.) (Coleoptera: Cerambycidae), naturally infested with fourth-stage juveniles (dauerlarvae) of the pinewood nematode, Bursaphelenchus xylophilus (Steiner and Buhrer, 1934) Nickle, 1970, were maturation fed on excised shoots of typical slash pine, Pinus elliottii Engelm. var elliottii, for 21 days. During August 1981, a male and female adult beetle were held in a sleeve cage placed on the terminal of a side branch of each of seven replicate, healthy 10-year-old slash pine trees. All seven branch terminals showed evidence of beetle feeding on the bark after 1 week, and pinewood nematodes were present in wood samples taken near these feeding sites. Four of the seven trees showed wilt symptoms in 4-6 weeks and died about 9 weeks after beetle feeding. Pinewood nematodes were recovered from the roots and trunks of the dead trees. Each of seven replicate slash pine log bolts was enclosed in a jar with a pair of the same beetles used in the sleeve cages. After 1 week, wood underlying beetle oviposition sites in the bark of all replicate log bolts was infested with the pinewood nematode.     

Nematicidal Activity of Plant Essential Oils and Components From Ajowan (Trachyspermum ammi), Allspice (Pimenta dioica) and Litsea (Litsea cubeba) Essential Oils Against Pine Wood Nematode (Bursaphelenchus Xylophilus)

Commercial plant essential oils from 26 plant species were tested for their nematicidal activities against the pinewood nematode, Bursaphelenchus xylophilus. Good nematicidal activity against B. xylophilus was achieved with essential oils of ajowan (Trachyspermum ammi), allspice (Pimenta dioica) and litsea (Litsea cubeba). Analysis by gas chromatography-mass spectrometry led to identification of 12, 6 and 16 major compounds from ajowan, allspice and litsea oils, respectively. These compounds from three plant essential oils were tested individually for their nematicidal activities against the pinewood nematode. LC₅₀ values of geranial, isoeugenol, methyl isoeugenol, eugenol, methyl eugenol and neral against pine wood nematodes were 0.120, 0.200, 0.210, 0.480, 0.517 and 0.525 mg/ml, respectively. The essential oils described herein merit further study as potential nematicides against the pinewood nematode.     

Pathogenicity of Bursaphelenchus xylophilus on Pines in Minnesota and Wisconsin

The pinewood nematode, Bursaphelenchus xylophilus, was inoculated into established native jack and red pines (Pinus banksiana and P. resinosa) and exotic Austrian pine (P. nigra) in Minnesota and Wisconsin forests during summer 1981. The nematode isolates did not kill established nonstressed pine trees growing in the forest. However, the same nematode isolates killed pine seedlings under greenhouse conditions. Girdling the main stem of some trees to induce stress resulted in the death of the majority of inoculated and noninoculated branches of Austrian and jack pines, but no branch death was observed on red pine. Greater numbers of nematodes were extracted from branches of inoculated, girdled trees than from nongirdled trees. The mean number of nematodes extracted from branches of inoculated, nongirdled trees was 0.3 - 14 nematodes per gram of wood.     

Draft Genome Sequence of Pseudomonas sp. Strain M47T1, Carried by Bursaphelenchus xylophilus Isolated from Pinus pinaster

The draft genome sequence of Pseudomonas sp. strain M47T1, carried by the Bursaphelenchus xylophilus pinewood nematode, the causative agent of pine wilt disease, is presented. In Pseudomonas sp. strain M47T1, genes that make this a plant growth-promoting bacterium, as well as genes potentially involved in nematotoxicity, were identified.     

Draft genome sequence of Serratia sp. strain M24T3, isolated from pinewood disease nematode Bursaphelenchus xylophilus

Here we report the draft genome sequence of Serratia sp. strain M24T3, which is associated with pinewood nematode Bursaphelenchus xylophilus, the causative agent of pine wilt disease. Serratia sp. strain M24T3 has been identified as a bionematocide for B. xylophilus in vitro, and multiple genes potentially involved in virulence and nematotoxity were identified.     

Maturation feeding and transmission of Bursaphelenchus xylophilus (Nematoda: Parasitaphelenchidae) by Monochamus alternatus (Coleoptera: Cerambycidae) inoculated with Beauveria bassiana (Deuteromycotina: Hyphomycetes)

We examined the amount of maturation feeding and transmission of pinewood nematodes, Bursaphelenchus xylophilus (Steiner et Buhrer) Nickle (Nematoda: Parasitaphelenchidae), to healthy pine (Pinus spp.) trees by pine sawyer Monochamus alternatus Hope (Coleoptera: Cerambycidae) adults infected with Beauveria bassiana (Balsamo) Vuill. (Deuteromycotina: Hyphomycetes). Inoculated beetles fed less than noninoculated beetles, probably because feeding by inoculated beetles began to decrease at about 4 d postinoculation and inoculated beetles ceased to feed for several days before their death. In inoculated beetles carrying >1,000 nematodes, some beetles died before nematode departure. The remaining heavily nematode-infested beetles lived until the beginning of nematode departure, but they had stopped feeding, preventing the nematodes from entering pine twigs. We suggest that microbial control of pine sawyer adults by B. bassiana may be effective in preventing transmission of pine wilt disease to healthy pine trees.