Characterization of a gamma-3 Proteobacteria Responsible for the Syndrome "Basses Richesses" of Sugar Beet Transmitted by Pentastiridius sp. (Hemiptera, Cixiidae)

ABSTRACT The disease syndrome "basses richesses" (SBR) has affected sugar beet crops in Burgundy (France) since 1991. It mainly is associated with an uncultivable phloem-restricted bacterium-like organism (BLO) called SBR BLO. Transmission tests showed that field-collected Pentastiridius sp. (Hemiptera, Cixiidae) were able to transmit the SBR BLO to sugar beet. In the present work, sequences of a 1,507-bp 16S ribosomal (r)DNA fragment of SBR BLO were amplified from DNA extracts of SBR-affected field sugar beet plants, of field-collected Pentastiridius sp. plant-hoppers, and of Pentastiridiussp.-exposed sugar beet seedlings that expressed SBR symptoms. The sequences showed total identity, confirming the role of SBR BLO in the etiology of SBR and the vector role of Pentastiridius sp. Our surveys on SBR-affected sugar beet plants and Pentastiridius sp. planthoppers collected in different fields and different years suggest that a unique BLO was involved in SBR. Furthermore, comparison of 16S rDNA sequences permitted the identification of the SBR BLO as a new plant-pathogenic gamma-3 proteobacteria different from 'Candidatus Phlomobacter fragariae,' another BLO responsible for marginal chlorosis disease of strawberry in France. Phylogenetic analysis revealed a close relationship between the SBR bacterium and several bacteria described as endosymbionts of hemipteran insects.     

A Combination of Baiting and Different PCR Formats, Including Measurement of Real-Time Quantitative Fluorescence, For the Detection of Phytophthora fragariae in Strawberry Plants

Phytophthora fragariae, the cause of strawberry red stele disease, is a quarantine pathogen in Europe. Detecting low levels of infection requires sensitive and specific methods. In the past, Dutch and English inspection services have used bait plants to test strawberry propagation stocks destined for export. Increasingly though, PCR is being incorporated into these testing procedures in an effort to increase sensitivity and speed. Various combinations of baiting and PCR assays were compared with existing testing procedures. Water and root samples from the bait test were screened by nested PCR and the PCR amplicon was detected by several methods, including fluorescent labelled probes (TaqMan and Molecular Beacon). PCR amplification was monitored in real-time and semi-quantitative detection was possible. Because PCR reactions are sensitive to inhibitors present in extracted DNA samples, an internal control containing the primer sequences specific for P. fragariae was developed to avoid false negatives.     

Evaluation of antisera raised against Phytophthora fragariae for detecting the red core disease of strawberries by enzyme-linked immunosorbent assay (ELISA)

Antiserum was raised against pooled mycelial suspensions from five isolates (designated Pf 1, Pf 2, Pf 3, Pf 10 and Pf 11) representing five physiologic races of Phytophthora fragariae. In enzyme-linked immunosorbent assay (ELISA), this antiserum detected homologous soluble antigens at protein concentrations as low as 2 ng/ml.
Fungal antigens could also be detected in extracts of strawberry plants infected with P. fragariae. Root extracts prepared from the alpine strawberry Fragaria vesca and F. ananassa cv. Cambridge Favourite infected with any of the five isolates studied produced strong reactions in ELISA. In F. vesca , ELISA-positive material was detectable 6-8 days after inoculation before macroscopic symptoms appeared. The cultivar Red Gauntlet, which is resistant to Pf 1, 2 and 3 but susceptible to Pf 10 and 11, reflected this differential response in ELISA; the absorbance produced by extracts of plants infected with virulent isolates was significantly higher than that obtained with the corresponding extracts of plants inoculated with a virulent isolates. The recently introduced cultivars Hapil, Ostara and Providence were found to be susceptible to all isolates in this study: the corresponding root extracts were also positive in ELISA.
The antiserum also detected P cactorum infections. Nevertheless, the ELISA test described should prove valuable in screening certified strawberry stocks.     

咸宁市水稻二化螟与稻飞虱发生特点及原因分析

2004年咸宁市水稻种植面积14．67万hm^2,水稻二化螟发生面积9．33万hm^2次,是1990年以来发生最重的一年,稻飞虱发生面积8．67万hm^2,是1990年以来第2个重发生年(仅次于1991年)。由于全市植保战线及时预报,当好参谋,积极指导防治,有效地控制了二化螟、稻飞虱等病虫的为害。     

Bacterial leaf blight of strawberry (Fragaria (x) ananassa) caused by a pathovar of Xanthomonas arboricola, not similar to Xanthomonas fragariae Kennedy & King. Description of the causal organism as Xanthomonas arboricola pv. fragariae (pv. nov., comb. nov.)

A new bacterial disease of strawberry is described. This disease, called bacterial leaf blight of strawberry, is characterized by dry, brown necrotic leaf spots and large brown V-shaped lesions along the leaf margin, midrib and major veins. Symptoms are different from angular leaf spot of strawberry caused by the bacterium Xanthomonas fragariae . Strains of the bacterial leaf blight pathogen were characterized in a polyphasic approach by biochemical tests, fatty acid analysis, protein electrophoresis, serology, PCR, pigment analysis, ice-nucleation activity, AFLP analysis, DNA:DNA hybridization, pathogenicity and host range tests, and compared with a number of reference strains of X. fragariae and other Xanthomonas species. Bacterial leaf blight strains formed a homogeneous group in all tests, completely different from X. fragariae . They were the only strains causing leaf blight of strawberry upon artificial inoculation into strawberry. Fatty acid and protein electrophoretic analysis showed that the strains belong to the phenon X. campestris ( sensu latu , including pathovars now classified as belonging to X. arboricola ). AFLP analysis and DNA:DNA hybridization further clarified their taxonomic position as belonging to X. arboricola. The name X. arboricola pv. fragariae is proposed for the bacterium causing leaf blight of strawberry with strain PD2780 (LMG 19145) as pathovar type strain. Criteria for routine identification are given and the taxonomic status is discussed.     

Biochemical Characterization of Carboxylesterase in the Small Brown Planthopper Laodelphax striatellus (Fallén)

Several forms of carboxylesterase were observed in a malathion-resistant small brown planthopper, Laodelphax striatellus Fallén, by isoelectrofocusing. To study the mechanisms of increased esterase activity, esterases were purified and their biochemical characteristics were investigated in five active esterase isozymes of two resistant strains. These esterases have polymorphic characteristics and their molecular weights ranged from 66 to 70 kDa, due to variations in glycosylation. The pI values of these esterases ranged from 5.3 to 4.7. These esterases were immunologically related and NH₂-terminal amino acids were identical in all isozymes regardless of pI or molecular weight. No differences have been found in kinetic parameters (K_m and V_max) to α-naphthylacetate and specific activity toward α-naphthylacetate and malathion as a substrate in all isozymes. Resistant individuals showed high ali- and malathion carboxylesterase activities and these enhancements were caused by quantitative differences of carboxylesterases with several different pI.     

褐飞虱两个酚氧化酶原基因的特性及细菌诱导后的表达分析

为探讨褐飞虱Nilaparvata lugens（Stål）酚氧化酶原基因的发育表达模式及生物学功能,利用转录组数据和RACE方法获得两条褐飞虱酚氧化酶原基因NlPPO1（GenBank No.：ALE32751）和NlPPO2（GenBank No.：ALE32752）全长序列,褐飞虱NlPPO1全长为2316 bp,CDS编码区为2073 bp,编码690氨基酸;NlPPO2全长为2738 bp,CDS编码区为2100 bp,编码699氨基酸。通过实时荧光定量PCR方法检测NlPPO的发育表达模式及注射细菌诱导后NlPPO的表达量变化。发育表达模式结果显示,NlPPO1和NlPPO2在肠中都有较高的表达水平,而NlPPO1在表皮细胞表达量最低,NlPPO2在脂肪体表达量最低;在5龄幼虫阶段NlPPO1随时间增加表达量增加,NlPPO2表达量则维持相对较高水平;在成虫阶段NlPPO1表达量维持在较低水平,NlPPO2随时间增加表达量降低。注射大肠杆菌诱导NlPPO结果表明,诱导24 h后,NlPPO1表达量显著升高,NlPPO2表达量变化差异不显著;注射枯草芽胞杆菌诱导NlPPO结果表明,诱导24 h后,NlPPO1和NlPPO2表达量都显著升高。研究结果显示褐飞虱NlPPO1和NlPPO2的发育表达模式存在差异,对不同菌诱导免疫应答也存在差异,暗示二者具有不同的生物学功能。该结果为进一步探索酚氧化酶原在褐飞虱对病原菌的免疫响应机制奠定基础。     

褐飞虱clip丝氨酸蛋白酶基因NlCSP4的克隆及功能分析

Clip丝氨酸蛋白酶（clip-domainserineprotease,CSP）是广泛存在于昆虫体内的一类结构保守、功能多样的蛋白质水解酶类,在昆虫生长、发育和免疫等诸多生理过程中发挥重要功能。本研究克隆并鉴定获得一个褐飞虱CSP编码基因NlCSP4(GenBank登录号：OK018340）,其cDNA序列全长1911bp,编码636个氨基酸,预测蛋白质分子量为69.29kD,等电点为9.02。NlCSP4蛋白N端包含一段由25个氨基酸残基组成的信号肽和一段由44个氨基酸残基组成的clip结构域,C端具有CSP蛋白家族典型的Tryp＿SPc结构域,且其中包含特异性保守的His-Asp-Ser催化中心三元件。系统发育分析表明NlCSP4与其他同属半翅目昆虫的CSP在NJ进化树上聚为一支,而与鳞翅目昆虫CSP亲缘关系较远。qRT-PCR分析显示,NlCSP4基因表达具有明显的时空特异性,其在雌成虫中的表达量最高,且高龄若虫期（4龄和5龄）表达量显著高于卵期和1～3龄若虫期;NlCSP4基因在褐飞虱雌成虫脂肪体、肠道和卵巢中均有表达,且在脂肪体中表达量最高;金黄色葡萄球菌（革兰氏阳性细菌）...     

Effect of Timing of Application of the Biological Control Agent Microsphaeropsis ochracea on the Production and Ejection Pattern of Ascospores by Venturia inaequalis

ABSTRACT Field and in vitro trials were conducted to establish the influence of the biological control agent Microsphaeropsis ochracea on the ejection pattern of ascospores by Venturia inaequalis and on apple scab development, and to establish the best timing of application. The ejection pattern of ascospores was similar on leaves sprayed with M. ochracea and on untreated leaves. Fall application of M. ochracea combined with a delayed-fungicide program was evaluated in orchards with intermediate and high scab risk. For both orchards, it was possible to delay the first three and two infection periods in 1998 and 1999, respectively, without causing significant increase or unacceptable leaf and fruit scab incidence. To evaluate the best timing of application, sterile leaf disks were inoculated with V. inaequalis and then with M. ochracea 0, 2, 4, 6, 8, 10, 12, 14, and 16 weeks later. After incubation under optimal conditions for pseudothecia development, the number of ascospores was counted. Similarly, M. ochracea was sprayed on scabbed leaves on seven occasions from August to November 1999 and 2000. Leaves were overwintered on the orchard floor and ascospore production was evaluated the following spring. Ascospore production was reduced by 97 to 100% on leaf disks inoculated with M. ochracea less than 6 weeks after inoculation with V. inaequalis, but ascospore production increased with increasing period of time when M. ochracea was applied 8 to 16 weeks after the inoculation with V. inaequalis. In the orchard, the greatest reduction in production of ascospores (94 to 96% in 2000 and 99% in 2001) occurred on leaves sprayed with M. ochracea in August. The production of ascospores was reduced by 61 to 84% in 2000 and 93% in 2001 on leaves sprayed with M. ochracea in September, reduced by 64 to 86% in 2000 and 74 to 89% in 2001 on leaves sprayed in October, and reduced by 54 and 67% in 2000 and 2001, respectively, on leaves sprayed in November. It was concluded that M. ochracea should be applied in August or September and that ascospore maturation models and delayed-fungicide program could be used in orchards treated with this biological control agent.     

中国西北地区气候变化与同期气象场异常的关系

利用奇异值分解法（SVD）分析中国西北地区降水和气温异常与同期高空环流和下垫面温度异常的关系。研究表明：西北地区各季降水与同期北印度洋海区、西太平洋海面温度都有较好的遥相关性;西北地区降水与同期欧亚对流层的中下层的纬向和经向环流强度的异常有明显的关系;西北地区气温异常与欧亚大陆同期中高纬向和经向环流的异常有关,冬季和秋季气温异常主要与纬向环流的异常有关,春季气温异常与经向环流异常有关。     

本土寄生蜂对移殖天敌昆虫泽兰实蝇的寄生

泽兰实蝇（Procecidochares utilis Stone）是我国入侵杂草紫茎泽兰的专食性天敌昆虫。为摸清本土寄生蜂对泽兰实蝇的寄生情况，在云南昆明市选取林下和开阔地等两种生境，分别采用对角线五点取样法与邻接格子取样法，调查了紫茎泽兰的生长及感染虫瘿情况；通过饲养与解剖虫瘿，观察了寄生泽兰实蝇的本土寄生蜂种类及其寄生率。结果表明，共有6种寄生蜂从虫瘿中羽化，对虫瘿的寄生率达70％以上；其中大部分虫瘿被3种寄生蜂寄生。虽然在开阔地生境中紫茎泽兰株高和每株枝条数显著大于林下生境，植株和枝条感虫（瘿）率也显著高于林下，但寄生蜂对虫瘿的总寄生率、各种寄生蜂的寄生率在两种生境之间差异不显著。解剖表明，每虫瘿羽化出的寄生蜂数随虫瘿横径的增大而显著增多。     

Molecular identification and characterization of ‘Candidatus Phytoplasma asteris’-related strain associated with pepper (Capsicum annuum L.) yellow crinkle disease in China

Journal of General Plant Pathology - Pepper yellow crinkle disease was first reported in China. PCR amplification using primer pairs specific for 16S rRNA, groEL, tuf or secA gene of phytoplasma...     

Transmission of ‘Candidatus Phytoplasma pyri’ by naturally infected Cacopsylla pyri to peach,an approach to the epidemiology of peach yellow leaf roll (PYLR) in Spain

Peach orchards in the northeast of Spain were severely affected in 2012 by a previously unreported disease in this area. The symptoms included early reddening, leaf curling, decline, abnormal fruits, and in some cases death of the peach trees. All the infected peach samples were positive for ‘Candidatus Phytoplasma pyri’, but none were infected by the ‘Ca. Phytoplasma prunorum’. In this work, potential vectors able to transmit ‘Ca. Phytoplasma pyri’ from pear to peach and between peach trees were studied and their infective potential was analysed at different times of the year. Transmission trials of the phytoplasma with potential vectors to an artificial feeding medium for insects and to healthy peach trees were conducted. Additionally, isolated phytoplasmas were genetically characterized to determine which isolates were able to infect peach trees. Results showed that the only insect species captured inside peach plots that was a carrier of the ‘Ca. Phytoplasma pyri’ phytoplasma was Cacopsylla pyri. Other insect species captured and known to be phytoplasma transmitters were present in very low numbers, and were not infected with ‘Ca. Phytoplasma pyri’ phytoplasma. A total of 1928 individuals of C. pyri were captured in the peach orchards, of which around 49% were phytoplasma carriers. All the peach trees exposed to C. pyri in 2014, and 65% in 2015, were infected by ‘Ca. Phytoplasma pyri’ 1 year after exposure, showing that this species is able to transmit the phytoplasma to peach. Molecular characterization showed that some genotypes are preferentially determined in peach.     

Influence of Environmental Conditions on Spore Production and Budding in Taphrina deformans, the Causal Agent of Peach Leaf Curl

ABSTRACT Environment-controlled studies were carried out to determine the growth of Taphrina deformans under different conditions of temperature, humidity, and nutrient availability similar to those found on plant surfaces during the peach-growing season. Both ascospores and blastospores were able to bud at all temperatures tested (5 to 37 degrees C), with the optimum at 14 and 21 degrees C, respectively. Temperature <20 degrees C favored ascospore production and release, with the optimum at 10 degrees C. Budding was approximately two-and-a-half times higher in a film of water than on a dry substrate, with 100% relative humidity and blastospores also maintained a certain budding ability at lower humidity levels (minimum tested = 47%). Both spore types did not bud after approximately 50 to 55 h in the absence of external nutrients. In the presence of a periodically renewed carbon source, such as simple sugars, at concentrations that typically are present on peach plant surfaces, the fungus maintained its budding capability over time. Results showed that T. deformans is able to bud profusely under a wide range of environmental conditions that occur on peach tree surfaces. This work supports the hypothesis that T. deformans is a part of the normal epiphytic mycoflora of peach trees throughout the entire growing season.     

Explaining patterns of species dominance in the shrub steppe systems of the Junggar Basin （China） and Great Basin （USA）

Natural scientists have long recognized that regions with similar climate tend to have similar vegetation.Preliminary observations suggest that shrub steppe communities of the western US and western China may be two such regions with similar annual precipitation,temperature,land use,and vegetation.These cold dry shrub steppes have traditionally been grazed.Despite these similarities,patterns of species dominance are different.Annual species that are rare in China become dominant when introduced to the United States.The objective of this study was to investigate how climate,land use and community structure may explain these patterns of species dominance.Community structure and grazing intensity were measured at 5 sites in each region.This information was combined with a broader review of the literature describing the history of grazing in both basins.Climate was analyzed based on a spatially-gridded,interpolated weather time series(monthly records) and climatological summary(1961-1990 mean conditions) data set from the Climate Research Unit.We found that differences in summer precipitation and winter minimum temperature,land use intensity,and shrub size may all contribute to the dominance of annual species in the Great Basin,particularly Bromus tectorum.In particular,previous work indicates that summer precipitation and winter temperature drive the distribution of Bromus tectorum in the Great Basin.As a result,sites with wet summers and cold springs,similar to the Chinese sites,would not be expected to be dominated by Bromus tectorum.A history of more intense grazing of the Chinese sites,as described in the literature,also is likely to decrease fire frequency,and decreases litter and shrub dominance,all of which have been demonstrated to be important in Bromus tectorum establishment and ultimate dominance.Further research is necessary to determine if other annuals that follow the same pattern of scarcity in the Junggar Basin and dominance in the Great Basin are responding to the same influences.     

Altica cyanea (Col: Chrysomelidae) for the biological control of Ludwigia prostrata (Onagraceae) in China

Abstract

Altica cyanea (Weber) damages Ludwigia prostrata Roxb in China. It is multivoltine, having up to six generations a year. The host range of the beetle was examined in the field and in the laboratory, and it was demonstrated that A. cyanea occurs on and attacks the main host L. prostrata and another weed, Rotala indica Koehne. Its ability to damage L. prostrata was studied in field cages in Nanchang. A. cyanea was shown to damage these caged plants heavily and to be a promising biological agent for this weed.     

中国霸王属植物在世界该属中的系统发育地位与形态性状进化研究

通过PCR扩增和测序,得到了中国霸王属种类的rbc L和trn L-F基因片段序列数据,同时在网络数据库获得了南非、纳米比亚、澳大利亚和中亚地区的霸王属种类的rbc L和trn L-F片段,构建系统发育树,并且对霸王属系统发育信号和31个形态性状进行分析。结果显示:1中国霸王属种类聚为一大类,与南非和纳米比亚霸王属形成姐妹分支;2霸王属植物的生命周期、叶排列、叶形状及株高等8个性状具有明显的系统发育信号,其次,中国霸王属植物的萼片长度、萼片宽度、果实长度、茎性质和叶片结构等10个性状与国外霸王属有显著的差异,这与中国霸王属在该属系统发育树中形成单源分支相吻合。