余甘子采后软腐病拮抗菌的筛选、鉴定及其防效

【背景】由青霉菌引起的软腐病是余甘子采后贮藏期的主要病害,造成了经济损失严重。【目的】筛选对余甘子软腐病菌具有良好拮抗效果的放线菌菌株,并初步评价拮抗菌无菌发酵液的防治效果。【方法】以PenicilliumchoerospondiatisDQ23为指示菌,采用琼脂块法结合生长速率法进行拮抗菌株的筛选,并采用形态学特征、生理生化特性结合16S rRNA基因序列进行分类鉴定,并采用浸果法初步测定其无菌发酵液对余甘子软腐病发生及果实贮藏品质的影响。【结果】菌株SC-15对青霉P.choerospondiatis DQ23有较强的拮抗作用,拮抗圈直径为18.70 mm,20%无菌发酵液对软腐病菌的抑制率可达87.8%。根据菌株形态学和生理生化特性,结合16SrRNA基因序列分析将菌株SC-15鉴定为维及尼链霉菌(Streptomycesvirginae)。菌株SC-15无菌发酵液能降低余甘子的腐烂率和腐烂指数,延缓可溶性固形物和可滴定酸的降低,其中75%发酵液浸果效果最好,贮藏8d后,果实腐烂率为53.3%,果实腐烂指数为35.2%,可溶性固形物为4.8%,可滴定酸为6.25mg/g。【结论】菌株SC-15可有效防治余甘子采后青霉软腐病的发生,具有一定的应用价值。     

库尔勒香梨黑斑病菌拮抗菌筛选及其抑菌机理

【背景】由互隔交链孢(Alternaria alternata)引起的黑斑病是库尔勒香梨采后贮期的主要病害之一,严重影响果实品质,造成巨大的经济损失。【目的】筛选对梨黑斑病菌具有高拮抗活性的菌株并探究其可能的抑菌机制,为香梨采后保鲜提供理论依据。【方法】从库尔勒香梨园健康植株根际采集土壤样品,采用稀释涂布法分离细菌并进行纯化,通过平板对峙法和琼脂打孔法筛选对梨黑斑病菌具有显著拮抗作用的菌株,结合形态学观察和16S rRNA基因序列分析进行初步鉴定;通过平板对峙法对拮抗菌抑菌谱进行测定;采用特异性平板定性检测拮抗菌产酶活性,采用3,5-二硝基水杨酸比色法法及福林-酚法定量酶活;通过孢子萌发抑制试验和扫描电镜观察,测定拮抗菌对梨黑斑病菌的拮抗作用。【结果】筛选得到2株对梨黑斑病菌有较强抑制活性的菌株JE53和JE56,经分析鉴定为多粘类芽孢杆菌(Paenibacillus polymyxa)及枯草芽孢杆菌(Bacillus subtilis),其发酵上清液对梨黑斑病菌的抑菌圈直径分别为22.49 mm和22.40 mm。抑菌谱测定结果表明,菌株JE53和JE56对马铃薯黑痣病菌等9种植物病原菌均有不同程度的抑制作用。胞外酶测定结果显示这2株菌可产生4种胞外酶;其发酵上清液能够显著抑制病原菌孢子萌发,抑制率分别为70.30%和88.87%;扫描电镜发现这2株菌可导致病原菌菌丝表面粗糙、扭曲变形。【结论】JE53和JE56对梨黑斑病菌有较好的抑制效果,具有进一步开发和应用的潜力。     

一株扩展青霉拮抗菌的分离、鉴定及抑菌活性物质

【目的】筛选有效抑制扩展青霉(Penicillium expansum)的拮抗菌,并鉴定其所产抑菌物质的主要种类及相对含量。【方法】从苹果表面分离到拮抗扩展青霉的菌株BA-16,经形态学、生理生化及16S r RNA基因序列分析对该菌进行鉴定;根据已知脂肽类抗生素合成相关基因序列设计3对特异性引物对菌株BA-16进行检测,对PCR产物克隆、测序和BLAST分析,采用酸沉淀法从菌株发酵液中制备出抑菌物质粗提液,对活性粗提物进行HPLC和MALDI-TOF-MS分析。【结果】经鉴定,BA-16被鉴定为解淀粉芽孢杆菌(Bacillus amyloliquefaciens),所得PCR产物经测序和BLAST分析,证实BA-16带有sfp和fen B基因。HPLC和MS结果显示菌株发酵液中含有Fengycin和Surfactin两种脂肽类产物,Fengycin是拮抗扩展青霉的主要因素。【结论】本研究对于苹果采后青霉病的生物防治具有良好的应用开发前景。     

芽孢杆菌FJAT-55034的鉴定、生长特性及对梨轮纹病菌的抑菌活性

【背景】由葡萄座腔菌(Botryosphaeria dothidea)引起的梨轮纹病是梨栽培生产中的主要病害之一,造成巨大的经济损失。【目的】对梨轮纹病菌具有较高拮抗活性的菌株FJAT-55034进行鉴定、生长特性及抑菌活性评价,为梨轮纹病的生物防治提供菌种资源。【方法】采用形态学观察、生理生化测定和16S rRNA基因序列分析进行拮抗菌株FJAT-55034的鉴定;通过生长曲线、培养温度和pH测定,研究该菌株的生长特性。采用抑菌圈法测定该菌株的抑菌谱;采用共培养、显微镜观察和果实回接,测定菌株FJAT-55034对梨轮纹病菌生长的抑制作用。【结果】拮抗菌株FJAT-55034鉴定为贝莱斯芽孢杆菌(Bacillusvelezensis)。该菌株在20-50℃(最适温度30℃)和pH 5.0-9.0 (最适pH值7.0)均能够生长,NaCl添加量为0-5%时,该菌株均能够较好地生长。菌株FJAT-55034对6种果树不同病原真菌均具有不同程度的抑制作用,抑菌圈直径范围为19.8-29.1 mm;与梨轮纹病菌共培养后,菌株FJAT-55034对菌丝生长的抑制率为77.2%。菌株FJAT-5...     

一株拮抗黄单胞菌的贝莱斯芽孢杆菌的分离和鉴定

【目的】为了筛选防治水稻条斑病(bacterial leaf streak,BLS)的生防细菌。【方法】以水稻条斑病菌(Xanthomonas oryzae pv. oryzicola,Xoc)的模式菌株RS105为靶标菌,采用平板稀释和抑菌圈法,从空心菜根际土壤中筛选到一株对RS105具有拮抗作用的细菌菌株504。通过形态学、生理生化特征以及16SrDNA和gyrA序列分析对菌株504进行了鉴定。利用牛津杯法测定504对植物病原黄单胞菌的拮抗活性及其无菌发酵液拮抗活性的稳定性。通过PCR扩增预测504编码合成脂肽类和聚酮类化合物的合成相关基因。采用苗期水稻注射接菌法来评价水稻组织中504对Xoc的拮抗活性。【结果】菌株鉴定结果表明504为贝莱斯芽孢杆菌,命名为Bacillusvelezensis504。抑菌实验显示,B.velezensis504对黄单胞菌属的细菌具有较好的抑菌活性,对水稻白叶枯病菌(X. oryzae pv. oryzae,Xoo)的拮抗效果最显著。基因预测结果显示,B. velezensis 504含有fenA、dhbA、sfrA、bmyA、beaS、dfnA及bacA等编码脂肽类和聚酮糖类抑菌化合物的基因簇。其无菌发酵液的活性物质耐高温和蛋白酶降解,但不耐强酸、强碱,在pH值为5.5–8.9时仍具有稳定的拮抗活性。在高感水稻品种原丰早上,B. velezensis 504对Xoc在水稻叶片中引起的水渍症状具有显著的抑制作用。【结论】B. velezensis 504能够特异性拮抗黄单胞菌,在黄单胞菌引起的细菌性病害的生物防治中将具有较大的应用潜力。     

桑树内生拮抗菌的分离鉴定及其对桑断枝烂叶病的生防初探

【目的】本研究从健康桑树茎中分离筛选对桑断枝烂叶病菌具显著拮抗作用的内生细菌,为该病生物防治奠定研究基础。【方法】采用组织培养法分离桑树内生菌,抑菌圈法和平板对峙法筛选抑菌活性稳定的内生拮抗菌;根据形态学、生理生化特征检测和基于16SrDNA、gyrA和gyrB基因的系统发育分析对拮抗菌进行菌种鉴定;利用抑菌圈法测定拮抗菌株活性发酵液热稳定性,菌丝生长速率法检测活性发酵液抑菌谱;并通过观察拮抗菌对桑断枝烂叶病菌BoeremiaexiguaGXH1菌株生长及菌丝形态的影响,扩增抑菌活性物质合成关键基因,以及采用酸沉淀法提取拮抗菌株脂肽类化合物并进行高效液相色谱串联质谱分析(LC-MS),初步探究可能的抑菌机制。【结果】从健康桑树茎中共分离获得17株桑树内生细菌,并从中筛选获得一株对桑断枝烂叶病菌B.exiguaGXH1有稳定拮抗作用的桑树内生细菌NPJ13菌株。该菌株形态学、生理生化特征与芽孢杆菌属一致,基于16SrDNA、gyrA和gyrB基因序列的系统发育分析结果显示该菌株与贝莱斯芽孢杆菌(Bacillusvelezensis)的亲缘关系最近,且处于系统发育树的最小分枝,故将NPJ13菌株鉴定为贝莱斯芽孢杆菌,命名为B. velezensis NPJ13。NPJ13菌株对灰霉病菌SWU5、核地杖菌SXSG-5、核盘菌PZ-2及烟草疫霉SWU20等12种病原真菌具有不同程度的拮抗作用,其活性发酵液具有较好的热稳定性。NPJ13菌株会导致桑断枝烂叶病菌GXH1菌丝发生扭曲、膨大、透明度增加、断裂等畸变现象;基因检测结果显示NPJ13菌株基因组中具有PKSI、NRPS、Sfp、ItuD、Srfc等5种抑菌活性物质合成关键基因,LC-MS检测结果表明菌株NPJ13脂肽类粗提物中含有表面活性素和伊枯草菌素。【结论】本研究分离筛选获得一株对桑断枝烂叶病菌具有显著拮抗作用的桑树内生细菌B. velezensis NPJ13菌株,为桑断枝烂叶病的生物防治提供了候选菌株。     

芽孢杆菌SK007的分类鉴定及其拮抗植物病原菌的功能分析

【背景】农业生产中,发掘和利用具有生防功能的微生物资源是保障粮食安全和提高作物产量的重要举措。【目的】明确土壤中芽孢杆菌SK007的分类地位,验证其对多种植物病原菌的拮抗作用,挖掘潜在的生防功能。【方法】通过16SrRNA基因和基因组分析方法确定分离菌株SK007的分类地位;采用平板对峙法研究该菌株对番茄灰霉病菌、白菜黑斑病菌、烟草赤星病菌、小麦赤霉病菌、马铃薯干腐病菌等植物病原菌的拮抗作用;采用AntiSMASH分析和预测菌株SK007的抗生素相关基因。【结果】基于16SrRNA基因、全基因组序列、平均核苷酸一致性和DNA同源性分析,结果表明菌株SK007属于Bacillus velezensis,并且具有产生脂肽类抗生素和聚酮类抗生素的基因,对多种植物病原真菌有较强的抗性。此外,菌株SK007基因组中抗生素基因簇数目较多,丰富度高。【结论】芽孢杆菌SK007在拮抗植物病原菌方面有许多优良性状,具有促进作物抗病和增产的潜力。     

草珊瑚炭疽病拮抗细菌的鉴定及其抑菌机理

【背景】草珊瑚炭疽病发生严重,目前尚未有植物内生细菌对该病原菌生物防治的研究报道。【目的】筛选对肿节风炭疽病Colletotrichumdematium具有拮抗作用的内生细菌,并对其抑菌机理进行研究。【方法】采用平板稀释法从广西不同地区采集健康肿节风植株的不同组织分离、纯化获得内生细菌。【结果】平板对峙试验结果表明,来自茎的RJ-4和JJ-5对草珊瑚炭疽病具有较强的拮抗作用,其中拮抗作用最强的菌株是RJ-4,其抑制率达到84.10%。抗菌谱测定结果表明,RJ-4、JJ-5对供试的14种病原真菌均有明显的拮抗效果,拮抗作用最强的是RJ-4,平均抑制率达到95.30%;抑菌机理研究结果表明,该菌株可以分泌蛋白酶和纤维素酶以及几丁质酶,破坏病菌菌丝,抑制病菌生长;含有拮抗细菌的发酵液对病原菌菌丝抑制明显,菌丝扭曲、断裂且分枝缠绕、菌丝颜色加深等。通过形态学特性和16S rRNA基因鉴定,RJ-4菌株为解淀粉芽孢杆菌(Bacillus amyloliquefaciens)。【结论】RJ-4菌株具有抑菌抗病功能,并能产生多种抗菌活性物质,这可为新型抗菌物质提供资源。     

核桃黑斑病拮抗放线菌WMF106的筛选、鉴定及防效

【背景】核桃黑斑病是由2种病原菌引起的细菌性病害,目前缺乏有效的生物防治方法。【目的】从核桃树根际土壤中筛选对核桃黑斑病病原菌具有拮抗效果的放线菌菌株,为该病害生防菌剂的开发提供基础。【方法】采用稀释涂布法分离放线菌,并以病原菌野油菜黄单胞菌(Xanthomonas campestris pv. campestris)和成团泛菌(Pantoea agglomerans)作为指示菌,利用平板对峙法和改良牛津杯法筛选具有高拮抗活性的菌株,通过形态学特征、生理生化特性和16SrRNA基因序列分析确定其分类地位,并测定其无菌发酵液的抗菌谱和室内防效。【结果】筛选到一株对野油菜黄单胞菌和成团泛菌均有较强拮抗作用的放线菌菌株WMF106,该菌株对2种病原菌的抑菌圈直径分别为2.38 cm和1.82 cm,无菌发酵液对2种病原菌的抑菌圈直径分别为1.75 cm和1.55 cm。根据菌株形态学、生理生化特性及16SrRNA基因序列分析,将菌株WMF106鉴定为暗蓝色链霉菌(Streptomyces caeruleatus)。该菌株对尖孢镰刀菌、腐皮镰孢菌、辣椒刺盘孢菌、灰葡萄孢菌、胶孢炭疽菌5种植物病原菌及大肠杆菌、金黄色葡萄球菌、铜绿假单胞菌、白色念珠菌、黑曲霉5种指示菌均有抑制作用,抗菌性能广谱高效,其无菌发酵液原液对离体叶片上由野油菜黄单胞菌和成团泛菌造成的核桃黑斑病防效分别为77.44%和58.33%。【结论】菌株WMF106可作为防治核桃黑斑病的生防材料,具有良好的开发价值和应用前景。     

一株蜂粮源拮抗细菌的分离鉴定及其抑菌物质特性

【目的】为发掘和利用蜂粮中拮抗菌资源,对分离获得的拮抗细菌菌株PC2进行分类鉴定,并测定其发酵液抑菌物质基本特性。【方法】采用改良牛津杯双层平板法测定菌株发酵液抑菌谱及温度、p H、紫外线和蛋白酶对其抑菌活性稳定性的影响,菌株鉴定结合形态学、生理生化特征和16S r RNA基因序列分析,硫酸铵沉淀法和盐酸沉淀有机溶剂提取法进行抑菌活性物质的初步分离。【结果】从3种蜂粮中分离筛选得到17株拮抗菌株,其中1株细菌PC2以马铃薯葡萄糖液体培养基发酵制备的无菌发酵液对7种供试菌株具有较强抑制作用,经形态、生理生化特征及16S r RNA基因序列分析,将其初步鉴定为解淀粉芽胞杆菌(Bacillus amyloliquefaciens)。菌株发酵液抑菌活性对温度、酸和紫外线具有较强的稳定性,对蛋白酶K、胃蛋白酶、碱性蛋白酶处理敏感。菌株发酵液存在抑菌蛋白和脂肽类物质。【结论】菌株PC2在食品保鲜和农业生防中具有潜在的开发应用价值。     

The Role of Chemical Cues in Host and Mate Location in the Pear Psylla Cacopsylla bidens (Homoptera: Psyllidae)

Chemical communication was shown to play a role in the pear psylla, Cacopsylla bidens. Electrophysiological (EAG) and behavioral responses were investigated in males and females pear psylla . Males were found to be attracted to females, and especially to those on host plants, but not to males, uninfested host plants, or plants infested with conspecific larvae. On the other hand, females were not attracted to males or females but displayed some attraction to host plants. Furthermore, females showed a preference for uninfested pear versus plants infested with conspecific larvae. The antennae of males gave highest electroantenographic response to volatiles from pears infested with females but not males, while females, responded also toward the volatiles of pear alone. These results indicate that females of C. bidens emit sex pheromones that are attractive to the males and suggest that, host volatiles may play a role in host selection by pear psylla females.     

A light microscopical investigation of amoebal and plasmodial mitosis in the MyxomyceteEchinostelium minutum

Summary Light microscopical observations on mitosis in living material of the amoebal and plasmodial phases of the MyxomyceteEchinostelium minutum de Bary (orderEchinosteliales) are reported for the first time. The uninucleate amoebal cell undergoes centric, open spindle mitosis whereas the multinucleate plasmodium exhibits acentric, closed spindle mitosis.     

库尔勒香梨上苹果蠹蛾发生规律及其性信息素迷向防控效果

【目的】苹果蠹蛾是我国进境检疫性有害生物。库尔勒香梨是苹果蠹蛾在新疆果树种植区的主要寄主。研究苹果蠹蛾性信息素迷向防控技术在新疆库尔勒香梨产区的应用效果,可为该害虫的绿色防控提供参考。【方法】通过持续监测与调查,掌握苹果蠹蛾在库尔勒香梨上的发生规律和危害特点,并通过定期调查诱蛾量和蛀果率,比较微胶囊迷向剂、迷向丝、常规化学农药对苹果蠹蛾的防治效果。【结果】库尔勒香梨产区苹果蠹蛾一年发生2~3代,以老熟幼虫越冬,且死亡率较低。4月中旬越冬代羽化出土,5月开始为害;第1代成虫高峰期为7月上中旬,第2代成虫高峰期为8月下旬至9月中旬。微胶囊迷向剂、迷向丝、常规化学农药处理区每周的平均诱蛾量分别为2.85、2.98、4.95头·台~(-1),平均蛀果率分别为2.25%、0.8%、0.99%,与空白对照区的差异均达极显著水平。【结论】使用性信息迷向剂能够降低苹果蠹蛾第1代、第2代幼虫及成虫的数量。     

The probing behavior of the summer-form pear psylla

Stylet insertion sites on the leaf surface and leaf cell types fed upon by nymphs and adults of the pear psylla, Psylla pyricola Foerster (Homoptera: Psyllidae) were identified using histological techniques and electronic monitoring of probing and feeding activities. Neither the nymphs nor the adult ingested predominantly from the phloem of Pyrus communis cv. Bartlett. We showed that the pear psylla ingests from all leaf cell types, but that xylem, phloem and bundle sheath cells are more acceptable for ingestion than non-vascular tissues. The possible sensory mechanisms underlying selection of stylet insertion sites on the leaf surface and acceptance of various cell types for ingestion are discussed.

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Résumé Les lieux d'insertion des stylets sur la surface de la feuille et les types cellulaires consommés par les larves et les adultes de Psylla pyricola Foerster ont été précisés par étude histologique et par enregistrement électronique du comportement de piqûre et d'alimentation. Ni les larves, ni les adultes n'ont absorbé de préférence le phloème de Pyrus communis de la variété Bartlett. Nous avons constaté que le psylle du poirier ingère le contenu de tous les types cellulaires, mais que le xylème, le phloème et autre cellules des faisceaux libéro-ligneux étaient préferés aux tissus non vasculaires. La discussion a porte sur les méchanismes sensoriels éventuellement responsables de la sélection des lieux d'insertion des stylets à la surface de la feuille et l'acceptation des différents types cellulaires lors de l'ingestion.

     

Conserving natural enemies with flowering plants: Estimating floral attractiveness to parasitic Hymenoptera and attraction’s relationship to flower and plant morphology

Flowering plants in agricultural landscapes can provide ecological services, such as nectar-provision for adult parasitic Hymenoptera. Various flowering native, introduced/established and cultivated potted plants were used to bait interception traps along the wooded margins of fields planted seasonally with either feed-corn or rye. Depending on circumstances, controls consisted of traps baited with the same species of plant without flowers, a pot/area without plants, or both. In most cases pots were rotated among trap-sites. Of the 19 plant species tested, 10 captured significantly more summed ichneumonoids and chalcidoids, seven more Braconidae, two more Ichneumonidae and six more Chalcidoidea than controls. Among Braconidae, traps baited with certain plants captured significantly more individuals of specific subfamilies. “Attractive” and “unattractive” plant species tended to cluster in a principal components vector space constructed from plant morphological characteristics (flower width, flower depth, flower density and plant height). Flower width and plant floral-area (flower width² * flower density) were the variables that most often explained the variance in capture of the different parasitoid taxa. Our study identified particular plants that could be incorporated into regional conservation biological control programs to benefit parasitoid wasps In addition, the results indicate that morphological characteristics might help identify further suitable plant candidates for agricultural landscape modification.     

Micropropagation ofPyrus andCydonia and their responses to Fe-limiting conditions

Appropriate micropropagation regimes were determined for fourPyrus species (P. amygdaliformis Vill.,P. betulaefolia Bunge,P. calleryana Dene., andP. communis L.) andCydonia oblonga L. Shoot multiplication was optimal at 10 or 20M N⁶-benzyladenine and high light intensity (135E m^–2 s^-1). Root formation of thePyrus species was stimulated by exposure of shoots to high levels (10 or 32M) of-indolebutyric acid (IBA) for 7 days or a dip in 10 mM IBA for 15 s, followed by a passage on auxin-free medium.-Naphthaleneacetic acid was more effective than IBA in stimulating rooting ofC. oblonga. The effects of Fe-limiting conditions in vitro were determined by comparing the responses of shoots and rooted plantlets to medium containing FeEDTA or FeSO₄, with or without bicarbonate. Symptoms of Fe deficiency were genotype-dependent and most severe in the presence of FeSO₄ and bicarbonate. Chlorosis was pronounced inCydonia, absent fromP. amygdaliformis andP. communis, and intermediate inP. betulaefolia andP. calleryana, indicating a good correlation between in vitro and field responses. Similar responses were obtained with rooted and unrooted shoots. These findings suggest that in vitro cultures may be used for studies of Fe-chlorosis as well as screening for tolerant rootstocks.     

油菜素内酯与美极梅奇酵母复配对葡萄灰霉病的抑制作用

【背景】生防酵母具有繁殖速度快、抗逆性强和不产生抗生素等特点,但其对病害的防控效果易受环境影响,油菜素内酯(brassinosteroid,BR)能调控植物生长发育与逆境响应平衡,可有效抑制葡萄果实灰霉病的发生。【目的】本研究旨在明确BR与生防酵母复配对葡萄果实灰霉病的抑制效果及作用机制,为新型生防制剂的研制和应用提供理论依据。【方法】采用美极梅奇酵母P01C004 (Y)、酵母和BR (YBR)、酵母和BR抑制剂(YBZ)处理“红地球”葡萄果实,处理6 h后人工接种灰霉菌孢子悬液。灰霉菌接种7 d后评价BR与生防酵母复配对灰霉菌的防治效果,并检测其抗氧化酶活性和13种酚类物质含量,利用qRT-PCR检测不同处理对葡萄BR、白藜芦醇和抗病相关基因表达水平的影响。【结果】7 d时,与Y处理相比,YBR对葡萄果实灰霉病防治效率提高了23.64%,显著提高了“红地球”葡萄果实中PPO酶活。YBR显著提高了2 d时果实中绿原酸、原儿茶酸、咖啡酸、表儿茶素和芹菜素等酚类物质含量,激活了果实内多种酚类物质的快速合成。YBR在48 h时激活了果实BR信号转导途径,显著上调VvBZR1和VvPR1基因的表达水平,更好地维持植物免疫反应,提高果实对病原菌的防御力。【结论】BR与美极梅奇酵母复配能触发果实多种抗病防御机制,提高葡萄果实灰霉病的防治效率,在采后病害防控方面表现出良好的应用前景。     

The predators' complex of pear-feeding psyllids in unsprayed wild pear trees in greece

Four insect species producing honeydew, exploited by bees, were found on wild pear trees,Pyrus amygdaliformis Vill.. Three of them were Psyllids,Cacopsylla (Hepatopsylla) notata (Flor),Cacopsylla (Thamnopsylla) pyrisuga (Förster), Cacopsylla (Hepatopsylla) pyri (L.), and one was Aphid,Dysaphis pyri D.d.F. The 1 rst one was the most abundant. Although these species are producing honeydew from spring to autumn, the bees are foraging on it only during September and October, and not every year. A total of 18 predatory species were counted. Five of them,Anthocoris nemoralis F.,Atractotomus mali Mey.,Chrysoperla carnea (Stephens),Anisochrysa flavifrons (Brauer) andAnisochrysa prasina (Burmeister) were bred in laboratory on the Psyllids. It seems that the predators of theseHomoptera may be able to cause a decrease of their population level. Three predators,Anthocoris nemoralis F.,Chrysoperla carnea (Stephens) andCoccinella septempuctata L., were very common, and the last 2 were present almost all the year round.     

Polymorphism of <Emphasis Type="Italic">SFBB</Emphasis><Superscript>−γ</Superscript> and its use for <Emphasis Type="Italic">S</Emphasis> genotyping in Japanese pear (<Emphasis Type="Italic">Pyrus pyrifolia</Emphasis>)

Japanese pear (Pyrus pyrifolia) exhibits the S-RNase-based gametophytic self-incompatibility where the pollen-part determinant, pollen S, had long remained elusive. Recent identification of S locus F-box brothers (SFBB) in Japanese pear and apple suggested that the multiple F-box genes are the pollen S candidates as they exhibited pollen specific expression, S haplotype-specific polymorphisms and linkage to the S locus. In Japanese pear, three SFBBs were identified from a single S haplotype, and they were more homologous to other haplotype genes of the same group (i.e., α-, β- and γ-groups). In this study, we isolated new seven PpSFBB ^−γ genes from different S genotypes of Japanese pear. These genes showed S haplotype-specific polymorphisms, however, sequence similarities among them were very high. Based on the sequence polymorphisms of the PpSFBB ^−γ genes, we developed a CAPS/dCAPS system for S genotyping of the Japanese pear cultivars. This new S genotyping system was found to not only be able to discriminate the S ¹–S ⁹, but also be suitable for identification of the mutant S ^4sm haplotype for the breeding of self-compatible cultivars, and detection of new S haplotypes such as S ^k.     

Studies on turfgrass snow mold caused byTyphula ishikariensis. II. Microscopical observation of infected bentgrass leaves

Light and transmission electron microscopy revealed thatTyphula ishikariensis penetrated into bentgrass leaves either through cuticles or stomata either by single hyphae or infection cushions formed on host surfaces. Time course study on infected leaves showed that penetration through stomatal subsidiary cells and their adjacent cells seemed to occur earlier than that through epidermal cells located farther from stomata. More than 30% of epidermal cells were infected by 10 days after inoculation. When hyphae penetrated through an intact cuticle of epidermal cells, they seemed to dissolve host cell walls enzymatically at penetration sites. Physical pressure also seemed to be involved in penetration.