AFLP markers in the detection of Scirpus × mariqueter (CYPERACEAE) hybrid in China

Scirpus × mariqueter Tang & F.T. Wang (CYPERACEAE) is a pioneer plant of the tidal zone of estuaries of East Asia. Amplified fragment length polymorphisms (AFLP) were used to infer the relationship between S. × mariqueter and its putative progenitors, Scirpus planiculmis F. Schmidt and Scirpus triqueter L. The genetic distance between S. × mariqueter and S. triqueter was much higher than that between S. × mariqueter and S. planiculmis, and samples of S. × mariqueter and those of S. planiculmis did not form a monophyletic group, while the S. triqueter group formed a distinct monophyletic group. However, we also detected an apparent clustering of individuals of S. × mariqueter and S. planiculmis separately. Our results disprove the hybrid origin of S. × mariqueter and rather suggest that it is more closely related to S. planiculmis.     

海三棱藨草及其近缘种基因组大小的测定

基因组大小是物种基因组的重要特征,通常用DNA C值来衡量,能够用于快速判断基因组倍性,并为分类学与进化生物学提供重要依据。海三棱藨草(Scirpus mariqueter)是长江口和杭州湾具有重要生态意义的标志性物种,被认为是扁秆藨草(S. planiculmis)和藨草(S. triqueter)的杂交种,因染色体小而难以准确确定倍性。近年来,部分研究者指出该物种的分类和命名存在疑点。该研究通过基因组Survey分析检测海三棱藨草样本CJ1的基因组特征,测序深度约为120 ×,并以绿豆(Vigna radiata)为参考标准,利用流式细胞术测定了海三棱藨草及其同域近缘种扁秆藨草和藨草以及海三棱藨草和扁秆藨草的杂交F1共13个样本的DNA C值和相对倍性。结果表明:(1)基因组Survey分析测得CJ1的基因组大小为244.12 Mbp,杂合率为0.68%,重复序列比例为42.38%,GC含量为37.25%。(2)流式细胞术测得来自不同区域的海三棱藨草各样本的基因组倍性相同,1C值在234.87 ～ 242.5 Mbp之间,其中CJ1的基因组大小与基因组Survey检测结果高度一致。(3)扁秆藨草的1C值在251.77 ～ 264.13 Mbp之间,藨草1C值为537.33 Mbp。根据上述基因组大小,认为海三棱藨草不可能是这两者的杂交种。该研究补充了海三棱藨草及其近缘种的基因组特征,为后续全基因组测序奠定基础,同时也否定了海三棱藨草起源于扁杆藨草和藨草杂交的假说。     

莫莫格扁秆藨草恢复湿地土壤种子库对长芒稗入侵的响应

湿地恢复过程中,时常有外来种或本地杂草入侵。土壤种子库作为未来植被的潜在种源,对湿地恢复效果具有重要的指示意义。在莫莫格国家级自然保护区,以恢复白鹤栖息地(扁秆藨草(Scirpus planiculmis)沼泽)为目的,进行了退化湿地的水文恢复;但退化湿地恢复2a后,一年生杂草长芒稗(Echinochloa caudata)在大部分区域成为建群种。以长芒稗入侵湿地和扁秆藨草自然湿地为研究对象,对比分析了长芒稗和扁秆藨草的土壤种子库及生长结实特征。结果表明,在自然湿地扁秆藨草种子库规模是长芒稗的18.42倍,而在恢复湿地长芒稗种子库大小是扁秆藨草的5.04倍。与自然湿地相比,扁秆藨草种子库密度在入侵湿地明显减少,但仍保留了一定量具有活力的种子(664.32±105.98)粒/m~2,这与研究区扁秆藨草较高的种子生产力(9210.4±1513.4)粒/m~2及种子较强的浮力(FP50=39.7d)有关,说明扁秆藨草具备通过种子库或水传播恢复的潜力。长芒稗土壤种子库密度在入侵湿地高达(3345.9±520.3)粒/m~2,明显高于自然湿地种子库规模(P0.01),说明恢复湿地受长芒稗入侵影响严重,这与长芒稗较高的种子生产力(7621.4±376.25)粒/m~2及较弱的种子浮力(FP0=5d)有关,同时也表明长芒稗通过水传播扩散的能力较弱。另外,研究区长芒稗平均高度超过1m,且盖度较大,不仅阻碍扁秆藨草种子的水播,也降低了到达地表的光照水平,从而抑制扁秆藨草更新。因此,在莫莫格受长芒稗入侵湿地,于开花结实前收获长芒稗地上植物体及凋落物应是限制长芒稗扩展、同时促进扁秆藨草恢复的有效措施。     

长江口九段沙湿地海三棱藨草生物量分配特征及其影响因子

海三棱藨草(Scirpus mariqueter)为长江河口盐沼湿地先锋植物。以长江口九段沙湿地为主要研究区域,在江亚南沙、上沙、中下沙等不同区域设置固定站点,进行植被和环境因子的取样调研,分析研究了海三棱藨草的生物量分配特征及其主要影响因子,以期为海三棱藨草的培育与恢复、盐沼湿地生态系统的保护与利用提供科学依据。结果表明:(1)不同站点海三棱藨草的茎基高度、球茎生物量占比、根茎与球茎之比均不存在显著差异(P0.05),而植株密度、植株高度、根茎生物量占比、果实生物量占比、茎叶生物量占比、须根生物量占比、果实与根茎之比、果实与球茎之比在不同站点间存在显著差异(P0.05);(2)不同站点的沉积物电导率、盐度、容积密度(5—10、25—30 cm土层)、总碳含量(10—15、20—25、25—30 cm土层)、总氮含量(5—10 cm土层)存在显著差异(P0.05);(3)海三棱藨草的果实与根茎的生物量之比和沉积物5—10、25—30 cm总碳含量,0—5、5—10 cm总氮含量,0—5 cm容积密度,茎基高度6种因子的组合存在极显著相关(P0.01);而其他生物量分配指标与本文涉及到的环境因子、植物表形参数均不存在显著相关性(P0.05)。海三棱藨草生物量分配特征是其自身生长特征与环境因子综合作用的结果。在今后的研究中需拓展生境因子涵盖范围,深入分析多因子综合作用对海三棱藨草生物量分配特征的影响。     

潮间带盐沼植物种子及实生苗漂浮起动临界剪切应力研究

盐沼是分布在海陆过渡区域,以大型草本植物为优势种的潮间带生态系统。在潮间带生态系统中,盐沼植物的种子和实生苗在潮流作用下的漂浮起动是盐沼植物自然扩散的重要前提,决定了盐沼植物能否实现有效扩散和长距离传播。然而,目前缺乏有关盐沼植物繁殖体在复杂潮滩环境下漂浮起动扩散过程的定量研究。以长江口典型盐沼先锋物种海三棱藨草(Scirpus mariqueter)为研究对象,应用U-GEMS微观侵蚀系统,测定了在不同沉积物底质条件下,海三棱藨草种子及不同萌发阶段实生苗漂浮起动时的临界剪切应力,定量分析不同萌发阶段和沉积物底质对海三棱藨草种子和实生苗漂浮起动的影响。研究结果表明：(1)不同种子萌发阶段和沉积物底质对海三棱藨草种子和实生苗漂浮起动的临界剪切应力均有极显著影响(P<0.01);(2)随着萌发阶段的进展,海三棱藨草实生苗漂浮起动所需的临界剪切应力逐渐减小,因而随潮流漂浮起动进而扩散的机会也相应增大;(3)相比于淤泥质潮滩底质,海三棱藨草种子和实生苗在粉砂质潮滩更易于漂浮起动。研究结果丰富了盐沼植物在潮间带自然扩散过程和机理的研究,也为今后长江口及其他区域开展高效、低成本、以种子为修复...     

Molecular identification of intestinal microflora in Takifugu niphobles

We investigated the intestinal microflora of coastal fish including Takifugu niphobles using both culture techniques and library cloning. As a result, the numbers of bacteria appeared on agar media were 1.0 × 10⁴ to 1.4 × 10⁹ CFU/g (colony forming units/gram), whereas those of total bacteria stained with 4′,6-diamidino-2-phenylindole were 4.7 × 10¹⁰ to 1.9 × 10¹¹ cells/gram, irrespective of different fish species. In addition, the culture technique showed that the intestinal microflora in all specimens was mainly composed of the genus Vibrio. In contrast, the direct count method showed that spirochaetes with length of 2.5-4.5 μm were present in the intestinal contents of T. niphobles at high densities, whereas such bacteria could not be detected in those of other fish species. Library cloning yielded the sequences of 16S rRNA genes that were divided into seven taxonomic categories of bacteria including Actinobacteria, Bacilli, Clostridia, Gammaproteobacteria, Mollicutes, Spirochaetes and an unclassified bacterial group. These results demonstrate that the molecular diversity of the intestinal bacteria in T. niphobles based on the clone library method reflects the direct observation by fluorescence microscopy to some extent.     

Interactions between Prorocentrum donghaiense Lu and Scrippsiella trochoidea (Stein) Loeblich III under laboratory culture

Interactions between Prorocentrum donghaiense Lu and Scrippsiella trochoidea (Stein) Loeblich III, two species of causative bloom dinoflagellates in China, were investigated using bi-algal cultures under controlled laboratory conditions. The growth of P. donghaiense and S. trochoidea were significantly suppressed when the initial cell densities were set at 1.9 × 10⁴ cells mL⁻¹ or 1.9 × 10⁵ cells mL⁻¹ for P. donghaiense and 1.0 × 10⁴ cells mL⁻¹ for S. trochoidea when the initial size/density ratio was 1:1 or 10:1, respectively, but no out-competement was observed in either bi-algal culture by the end. The simultaneous assay on the culture filtrate showed that P. donghaiense filtrate prepared at a lower initial density (1.9 × 10⁴ cells mL⁻¹) stimulated the co-cultured S. trochoidea at a density of 1.0 × 10⁴ cells mL⁻¹, but filtrate at a higher density (1.9 × 10⁵ cells mL⁻¹) depressed its growth. Differently, the filtrate of S. trochoidea at a density of 1.0 × 10⁴ cells mL⁻¹ significantly suppressed the growth of P. donghaiense at a density of 1.9 × 10⁴ cells mL⁻¹, but had little stimulatory effect on P. donghaiense at a density of 1.9 × 10⁵ cells mL⁻¹compared to the control (P > 0.05). It is likely that these two species of microalgae interact with each other mainly by releasing allelochemical substance(s) into the culture medium, and a direct cell-to-cell contact was not necessary for their mutual interaction. We then quantify their interactions in the bi-algal culture by using a mathematical model. The estimated parameters from the model showed that the inhibition exerted by S. trochoidea on P. donghaiense was about 43 and 24 times stronger than the inhibitory effect that P. donghaiense exerted on S. trochoidea when the initial size/density were 1:1 and 10:1, respectively. S. trochoidea seemed to have a survival strategy that was superior to P. donghaiense in the bi-algal culture under controlled laboratory conditions. We also observed a closely positive relationship between the initial cell density and its effect on the co-cultured microalga by measuring the fluorenscence: filtrate prepared from higher initial cell density had stronger interference on the co-cultured microalga. Moreover, pre-treated under different temperature conditions (30 °C, 60 °C and 100 °C) would significantly changed the effect of culture filtrate on the co-cultured microalga. Result inferred that P. donghaiense or S. trochoidea would release allelochemicals into the bi-algal culture medium and the allelochemicals might be a mixture with temperature-sensitive components in it.     

Walking pattern of Trichogramma nubilale Ertle & Davis (Hymenoptera; Trichogrammatidae) on various surfaces

The leaf surfaces on which mass-reared Trichogramma spp. are released for augmentative biological control may be an important factor effecting the success of these releases. We observed and recorded the walking pattern of Trichogramma nubilale females on leaves of Zea mays, Canna × generalis (Canna lilly), Silphium perfoliatum (cup plant), Abutilon theophrasti (velvetleaf), Schizachyruim scoparium (little bluestem), a smooth and a fuzzy polyester material, and waxed paper. For each surface type, a total of 10 mated, 1–2 days old, naive and fed females were observed and their walking path was traced. Females walked fastest on waxed paper and leaves of S. scoparium and slowest on leaves of A. theophrasti and the fuzzy material. Turning rates were high on Canna × generalis and waxed paper. In general, walking speed was negatively affected by the presence and density of trichomes and possibly the lack of leaf veins. The effect of surface structures, such as trichomes is likely to scale to the body size of the searching parasitoid. Parasitoids can walk over short trichomes, but short directionally pointed trichomes (as on S. scoparium) can guide the search paths in certain directions. The effects of long trichomes may depend on trichome density relative to the parasitoid body length. When trichome density is on the same order of magnitude as 1/L² (Z. mays), wasps will stand between trichomes, and will frequently run into trichomes, which will retard walking speeds and increase turning.     

Biological control of brown rot (Moniliana laxa Ehr.) on apricot (Prunus armeniaca L. cv. Hacıhaliloğlu) by Bacillus, Burkholdria, and Pseudomonas application under in vitro and in vivo conditions

In 2002 and 2003, a study was conducted to determine the effect of bacterial strains, Burkholdria OSU 7, Bacillus OSU 142, and Pseudomonas BA 8, on biological control of brown rot disease (Monilinia laxa Ehr.) on apricot cv. Hacıhaliloğlu in Malatya province of Turkey. Apricot orchard at full blooming stage was inoculated with conidial suspension (1 × 10⁶ spores/ml) of M. laxa Ehr. After inoculation, two apricot trees for each application were treated with each of the three biological control agents (Burkholdria gladii OSU 7, Bacillus subtilis OSU 142, and Pseudomonas putida BA 8) by spraying (1 × 10⁹ cfu/ml) on inoculated branches. Disease incidence was evaluated for untreated (control 1) and four different treatment groups including commercial disease management (control 2, positive control: 3% Bourdox in fall, 50% Cupper at pink flower, 30 g/100 l Corus at first blooming, and 300 g/100 l Captan at last blooming stage) and treatments including each of the three bacterial strains (OSU 7, OSU 142, and BA 8). The results showed that disease incidence for negative control (control 1) was 9.94, which was significantly higher than disease incidence for commercial application (2.57%) or bacterial treatments (2.82–5.00%) in the first year. In 2003, the lowest disease incidence observed in OSU 7 treatment (6.80%), while disease incidence rate for positive control and negative control were 9.45% and 28.46%, respectively. This result may suggest that OSU 7 has potential to be used as biopesticide for effective management of brown rot disease on apricot.     

Occurrence and toxicity of Amphidinium carterae Hulburt in the North Arabian Sea

The occurrence and toxicity of Amphidinium carterae Hulburt is hereby reported for the first time from the North Arabian Sea on the coast of Pakistan. The concentrations of 1.2 × 10⁴ cells ml⁻¹ were found in intertidal pools that were also inhabited by the brown macroalga Sargassum wightii. Both wild and cultured A. carterae cells were tested for ciguatera toxicity through exposure to brine shrimp nauplii (Artemia salina) and albino mice. Although the brine shrimp did not appear to be affected mortalities in mice ranged between 13 and 16% at doses of 7.2 × 10⁴ and 2.5 × 10⁵ cells ml⁻¹, respectively. When mice were affected pharmacological effects such as muscle contraction in lower back area, increased respiration, immobility and paralysis in hind limbs were observed for 2 h. These effects appeared to be reversible and gradually disappeared within 24 h.     

Local competitive effects of introduced Spartina alterniflora on Scirpus mariqueter at Dongtan of Chongming Island, the Yangtze River estuary and their potential ecological consequences

Dongtan of Chongming Island, Shanghai, China is a wetland of international importance, in which dominant vegetation is Scirpus mariqueter community that supports a great diversity of bird species. Spartina alterniflora, native to the eastern and gulf coasts of the USA, was intentionally introduced to the tidelands of Dongtan in May 2001. Field work were conducted at Dongtan from March 2002 to May 2003 to determine how introduced S. alterniflora affects S. mariqueter population dynamics. The results obtained here show that S. alterniflorahad strong competitive effects on S. mariqueter, and that the introduction of S. alterniflora to S. mariqueter community resulted in a significant decreases of S. mariqueter's abundance, coverage, seed and fresh corm output after two growing seasons. A concomitant consequence might be the decline in bird diversity. It is suggested that further introduction of S. alterniflora should be avoided to conserve the wetlands and their associated birds. A plan for controlling further spread of existing S. alterniflora populations in the Yangtze River estuary is urgently needed.     

Distribution by influence factors of pyrene removal in chemical enhancers assisted microbial phytoremediation of Scirpus triqueter in co-contaminated soils

Abstract

The rehabilitation of soil co-contaminated by heavy metals and polycyclic aromatic hydrocarbons (PAHs) has become a serious global issue. Chemical enhancers and strains are often used to remove PAHs from contaminated soil. In this paper, the effects of chemical enhancers, strain HD-1, and Scirpus triqueter in removing pyrene from co-contaminated soil are studied. In the pot experiment, chemical enhancers and HD-1 were added to the co-contaminated soil. On the 60^th day, the plants and soil were taken out for measurement. The result showed that the addition of chemical enhancers and microorganisms (Group PBC) alleviated the inhibition effect of plants on pollution. The accumulation of pyrene in plants of Group PC (chemical enhancers) and Group PBC (chemical enhancers and HD-1) were much higher than those in other groups. Plant enrichment was not the major way to remove pyrene from soil (less than 0.3%). Compared with the contributions of chemical enhancers, HD-1, and Scirpus triqueter, HD-1 had stronger effects on the removal of pyrene (17.23–22.80%). This study indicates that the combination of chemical enhancers, HD-1, and Scirpus triqueter constituted a beneficial composite system, in which the three elements interacted with each other and ultimately achieved the goal of removing pyrene from co-contaminated soil.     

Interactions among the entomopathogenic fungus, Beauveria bassiana (Ascomycota: Hypocreales), the parasitoid, Aphidius matricariae (Hymenoptera: Braconidae), and its host, Myzus persicae (Homoptera: Aphididae)

The effect of the entomopathogenic fungus Beauveria bassiana on the biological characteristics and life table of Aphidius matricariae, a parasitoid of the green peach aphid, Myzus persicae, was studied under laboratory conditions. Aphids were first infected with twice the LC₉₅ of B. bassiana for third-instar M. persicae (2 × 10⁸ conidia/ml). Subsequently, at different intervals they were exposed to 1-day-old mated parasitoid females for 24 h. The number of mummies produced per female and the percentage emergence of the F1 generation differed significantly as a function of the time interval between application of the fungus and exposure to the parasitoid. The interference of B. bassiana on parasitoid development was also studied by first exposing the aphid hosts to the parasitoid for 24 h and subsequently applying B. bassiana. The number of mummies produced by a female A. matricariae varied from 11.8 to 24.8 and was significantly different when the aphids were first exposed to the parasitoids and then treated with B. bassiana 24, 48, 72, and 96 h after exposure. There were no significantly different effects of B. bassiana on net reproductive rate (R₀), mean generation time (T), intrinsic rate (r_m) and the finite rate of increase (λ) of A. matricariae as a result of development in hosts exposed to low or high conidial concentrations (1 × 10², 2 × 10⁸ conidia/ml). The parasitoids developed in infected hosts had lower r_m, λ, T and DT (doubling time) values compared with those that developed in uninfected hosts but no differences were observed in R₀ values. With proper timing, A. matricariae and B. bassiana can be used in combination in the successful biological control of M. persicae.     

基于胁迫梯度假说和互惠理论的海三棱藨草种群恢复技术

滨海湿地生态修复已成为阻止海岸带生态系统退化、保护生物多样性以及提供生态服务的关键措施。以长江口原生盐沼植物海三棱藨草(Scirpus mariqueter)为研究对象,选取崇明东滩新生滩涂湿地为研究区域,通过沿潮滩高程梯度的海三棱藨草植株斑块的移植实验,探究胁迫梯度假说和互惠理论(即种内的正相互作用)对长江口海三棱藨草种群恢复的指导意义。研究结果显示:(1)在一定的胁迫梯度范围内(潮滩高程2.0 m以上),增大种植斑块可以促进海三棱藨草的种内正相互作用,显著提高种植斑块的存活率和植株密度(P0.05);(2)潮滩水文动力沉积条件与潮滩高程梯度密切相关(P0.05),水文动力沉积作用对海三棱藨草定居和生长的胁迫随高程梯度下降而增强。潮滩高程2.0 m以下处强烈的水文动力条件干扰限制了生物-物理因素的正反馈作用。滨海湿地盐沼植被修复工作的成功率可以通过改进种植方式,增强种内的正相互作用得到极大的提高。研究可为开展大规模滨海湿地盐沼植被修复工程和提高生态修复效率提供科学依据和技术支持。     

盐度及种间相互作用对海三棱藨草、互花米草萌发及生长的影响

盐度和种间作用是影响湿地植物群落构建的关键因子。然而,已有研究主要集中于植物成体阶段,我们对生活史早期更新阶段的种间相互作用了解十分有限。崇明东滩国家级自然保护区是位于长江口的重要湿地,外来入侵植物互花米草对优势土著物种海三棱藨草的竞争排斥对当地生态系统造成了严重的负面影响。通过受控实验探讨了盐度及种间作用对海三棱藨草和互花米草种子萌发及生长的影响,以深入了解更新过程在盐沼湿地植物群落构建中的作用。结果表明,在培养皿中盐度对海三棱藨草的萌发有显著抑制作用,互花米草的萌发率受盐度影响不显著但萌发进程被延迟。混种处理对两者的萌发存在一定促进效应,且其作用强度受到盐度的调控。海三棱藨草与互花米草种子在萌发阶段的相互促进并非是通过化感作用实现的,可能是由于萌发过程对盐分的吸收减弱了盐胁迫的影响。盆栽条件下,两物种混种时的萌发及生长表现（高度、地上生物量）较单种时有所下降,但差异不显著。种间竞争受环境胁迫程度及生活史阶段的影响,竞争作用在胁迫较弱的淡水环境及生活史后期更强。盐沼湿地植物群落在形成早期受到盐度等环境因子的影响较大,不同植物对盐胁迫的响应是影响种群建成的主导因素,后期种间竞争的重要性不断增加,最终决定了植物群落的整体格局。     

High toxicity of the novel bloom-forming species Chattonella ovata (Raphidophyceae) to cultured fish

A toxicological study of an axenic cell line of novel species Chattonella ovata Y. Hara et Chihara (Raphidophyceae) revealed that cultured species of sea bream (Pagrus major), horse mackerel (Trachurus japonicus), and yellowtail (Seriola quinqueradiata) were killed by 4.1–6.8 × 10³, 5.4 × 10³, and 2.8 × 10³ cells/mL, respectively. The sensitivity of the gill lamellae to C. ovata differed among the fish species tested. This finding revealed that C. ovata was highly toxic to the cultured fish. Histological examination showed that edema and hyperplasia of the secondary gill lamellae of red sea bream and horse mackerel occurred when exposed to, or killed by C. ovata, whereas severe damage in the gill lamellae was not observed in yellowtail. Chattonella produced high amounts of superoxide anion radicals and hydrogen peroxide, possibly responsible for the fish death observed. Based on the results of this study and occurrence of a red tide by this organism in China in 2001, we consider this organism to be one of the harmful algae in coastal waters. This is the first report demonstrating that C. ovata is highly toxic to fish, and that it produces superoxide and hydrogen peroxide.     

Chaetosphaeria tortuosa, the newly discovered teleomorph of Menispora tortuosa, with a key to known Menispora species

Chaetosphaeria tortuosa is described as the newly discovered teleomorph of Menispora tortuosa, based on specimens from Canada and the Czech Republic, and single spore isolations from both morphs. The fungus produces superficial, more or less globose, papillate, dark brown to black smooth perithecia (200–)220–250 × (220–)230–260 μm. The asci are unitunicate, 8-spored, cylindrical-fusiform, (110–)120–133(–145) × 12–14 with a distinct apical, nonamyloid annulus 1–1.5 μm high, 3.5–4 μm wide. The ascospores are fusiform, 19–24 × 5–6 μm, hyaline, 3-septate, smooth, and 2-seriate in the ascus. The morphology of the teleomorph and anamorph are similar to that of C. ovoidea (anamorph: M. glauca), differing in dimensions of asci and ascospores, and in the disposition and morphology of the phialides of the anamorphs. The generic concept and phylogeny of Menispora is briefly discussed, and a key to the 11 species currently accepted in the genus is provided.     

Selection of Beauveria bassiana isolates for control of the whiteflies Bemisia tabaci and Trialeurodes vaporariorum on the basis of their virulence, thermal requirements, and toxicogenic activity

As part of a 3-fold approach to select potential mycoinsecticides for whitefly control, we evaluated infectivity, thermal requirements, and toxicogenic activity of the entomopathogenic fungus Beauveria bassiana (Ascomycota: Clavicipitaceae) under laboratory conditions. Twenty-five native B. bassiana isolates and a commercially available mycoinsecticide (based on B. bassiana) were evaluated for virulence to fourth instar nymphs of sweetpotato whitefly, Bemisia tabaci, and greenhouse whitefly, Trialeurodes vaporariorum, at a concentration of 1 × 10⁷ conidia/ml. All isolates were pathogenic for both whitefly species, whereas mortality rates varied from 3 to 85%. A second series of bioassays was conducted on 10 selected isolates using four 10-fold concentrations ranging from 1 × 10⁵ to 1 × 10⁸ conidia/ml. Median lethal concentrations (LC₅₀) of the four most virulent isolates varied from 1.1 × 10⁵ to 6.2 × 10⁶ conidia/ml and average survival time (AST) of treated nymphs from 5.9 to 7.4 days. T. vaporariorum were significantly more susceptible to all B. bassiana isolates than B. tabaci. The thermal biology of the eight most virulent isolates to both whitefly species was investigated at six temperatures (10–35 °C). The colony radial growth rate was estimated from the slope of the linear regression of colony radius on time and data were then fitted to a modified generalized β function that accounted for 90.5–99.3% of the data variance. Optimum temperatures for extension rate ranged from 23.1 to 27.1 °C, whereas maximum temperatures for fungal growth varied from 31.8 to 36.6 °C. On the basis of their virulence and thermal requirements, three isolates showed promise as candidates for whitefly management in Mediterranean greenhouses. Whilst in vitro production of macromolecular compounds toxic to Galleria mellonella larvae was not a requisite for virulence, ASTs of larvae injected with Sephadex G-25 fractions from candidate isolates ranged from 1.4 to 3.7 days compared with 5–6 days for non-toxic G-25 fractions. In addition, proteinase K treatment significantly reduced their toxic activity suggesting that they were proteins and revealing the potential of these isolates to be further improved through biotechnology to kill the pest more quickly.     

Evaluation of the Entomopathogenic Fungi Beauveria bassiana and Paecilomyces fumosoroseus for Microbial Control of the Silverleaf Whitefly, Bemisia argentifolii

Collaborative research was conducted at the USDA-ARS Subtropical Agricultural Research Center in southern Texas to assess the microbial control potential of Beauveria bassiana and Paecilomyces fumosoroseus against Bemisia whiteflies. Laboratory assays demonstrated the capacity of both pathogens to infect Bemisia argentifolii nymphs on excised hibiscus leaves incubated at relative humidities as low as 25% at 23 ± 2°C (ca. 35% infection by B. bassiana and P. fumosoroseus resulted from applications of 0.6–1.4 × 10³ conidia/mm² of leaf surface). In small-scale field trials using portable air-assist sprayers, applications at a high rate of 5 × 10¹³ conidia in 180 liters water/ha produced conidial densities of ca. 1–2.5 × 10³ conidia/mm² on the lower surfaces of cucurbit leaves. Multiple applications of one isolate of P. fumosoroseus and four isolates of B. bassiana made at this rate at 4- to 5-day intervals provided >90% control of large (third- and fourth-instar) nymphs on cucumbers and cantaloupe melons. The same rate applied at 7-day intervals also provided >90% control in zucchini squash, and a one-fourth rate (1.25 × 10¹³ conidia/ha) applied at 4- to 5-day intervals reduced numbers of large nymphs by >85% in cantaloupe melons. In contrast to the high efficacy of the fungal applications against nymphs, effects against adult whiteflies were minimal. The results indicated that both B. bassiana and P. fumosoroseus have strong potential for microbial control of nymphal whiteflies infesting cucurbit crops.     

A new species of Cladophialophora (hyphomycetes) from boreal and montane bryophytes

During a survey of bryophilous fungi from boreal and montane habitats in central Alberta, a hitherto undescribed species of Cladophialophora was recovered from Polytrichum juniperinum, Aulacomnium palustre, and Sphagnum fuscum. On potato dextrose agar (PDA) colonies grew slowly, attaining a diameter of 25 mm after 30 d, were dark grey, velvety, radially sulcate, and convolute and cracked at the centre. Micronematous conidiophores gave rise to branched chains of small (1–2 × 8–22 μm), cylindrical to fusiform conidia with truncate, swollen scars at each end. Phylogenies built on the ITS and ribosomal SSU regions indicate the isolates form a monophyletic clade within the family Herpotrichiellaceae (Chaetothyriales) that is composed of two geographically based groups, each with 99 % within-group sequence similarity and 97–98 % between-group sequence similarity. A teleomorph has not been found but would likely be similar to species of Capronia. In vitro inoculation of the isolates onto axenically grown P. juniperinum produced no discernible host symptoms, and host penetration could not be detected using light microscopy. The production of polyphenol oxidases by the fungus and the role of other Cladophialophora species as latent endophytes and saprobes suggest that a potential role for the fungus is the degradation of the polyphenol-rich cell walls of mosses. A dichotomous key to species of the genus Cladophialophora is provided.