Parasitoid complex of alfalfa aphids in an IPM intensive crop system in northern Catalonia

Alfalfa aphids were monitored and aphid parasitism was determined in three crop-growing seasons (2007–2009) in commercial alfalfa fields in valleys close to the Pyrenees, where an intensive arable crop rotation is practised under non-tillage and an integrated pest management system. Acyrthosiphon pisum, Therioaphis trifolii, and Aphis craccivora were the aphid species colonizing alfalfa fields but their occurrence varied between years. Thirteen Aphidiinae parasitoid species and four Aphelinidae from the genus Aphelinus sp. were recorded. Aphidius ervi was the prevalent parasitoid species associated with A. pisum but Praon barbatum was also noticeable, especially during the first year. Trioxys complanatus and P. exsoletum were the only species associated with T. trifolii. Lysiphlebus fabarum was the prevalent parasitoid species associated with A. craccivora. The occurrence of Lysiphlebus testaceipes parasitizing A. craccivora on alfalfa is a new record for the study region because this parasitoid species had only been previously found on weeds. Only A. pisum and A. craccivora were parasitized by Aphelinus sp. Whereas A. abdominalis and A. semiflavus were associated with A. pisum, A. chaonia, and Aphelinus sp. nr. albipodus were associated with A. craccivora. Associations between aphid species and Aphidiinae or Aphelinidae parasitoid species emphasize that individual aphid species also possess individual parasitoid complexes (guilds) which do not interfere with each other in alfalfa. Alloxysta victrix, A. macrophadna, Alloxysta spp., Asaphes suspensus, A. vulgaris, Dendrocerus aphidum, Pachyneuron aphidis and Syrphophagus aphidivorus were the hyperparasitoids associated with aphid–parasitoid complexes. Our study showed A. ervi, T. complanatus, P. exsoletum, and L. fabarum as potential candidate species which could provide biological control of their respectively aphid hosts in alfalfa.     

PCR‐based identification of Erysiphe pulchra and Phyllactinia guttata from Cornus florida using ITS‐specific primers

The internal transcribed spacer (ITS) regions of rDNA and the intervening 5.8S rRNA gene for the powdery mildew fungi Erysiphe (sect. Microsphaera) pulchra and Phyllactinia guttata were amplified using standard polymerase chain reaction (PCR) protocols and the universal primer pairs, ITS₁ and ITS₄. PCR products for ITS were analysed by electrophoresis in a 1.5% agarose gel and sequenced. The size of the amplified ITS products (approximately 650 bp) were not sufficiently different to allow reliable differentiation of E. pulchra and P. guttata; however, their sequences were distinct. Specific primers for E. pulchra and P. guttata were developed and evaluated for use as diagnostic tools. The diagnostic band size from E. pulchra‐specific primer pair was 568 bp while the P. guttata band was 597 bp; the two primer pairs were highly specific to E. pulchra and P. guttata. Comparison of ITS sequences with information in the GenBank showed a very close similarity between sequences of E. pulchra isolates from Cornus florida in the USA and isolates collected on Cornus kousa in Japan. BLAST analysis of the sequence of the 650‐bp band from P. guttata revealed a close alignment with sequences of P. moricola (92%), P. kakicola (94%), and P. fraxini (92%). The sequence of P. guttata in C. florida also had a 98% identity with P. guttata in Calycanthus occidentalis and 94% identity with P. guttata in Corylus cornuta.     

Biomass,nutrient and pigment content of beech (Fagus sylvatica) saplings infected with Phytophthora citricola,P. cambivora,P. pseudosyringae and P. undulata

Fagus sylvatica saplings were infected with Phytophthora citricola, Phytophthora cambivora, Phytophthora pseudosyringae and Phytophthora undulata to study the influence of these root pathogens on total belowground and aboveground biomass, on the nutrient distribution within plants, on the concentration of plastid pigments, including tocopherol and on components of the xanthophyll cycle. Phytophthora citricola and P. cambivora infection significantly reduced total biomass of beech when compared with control plants and finally most of these plants died at the end of the experiment. However, beech invaded by the other two Phytophthora spp. did not differ from control plants and none of them was killed. Fine root length as well as the number of root tips of all infected beeches were reduced between 30 and 50%. The excellent growth of beech infected with P. pseudosyringae and P. undulata when compared with control plants was correlated with a strong increase of important root efficiency parameters. Phytophthora citricola and P. cambivora caused a significant reduction in nitrogen concentration of leaves in comparison with control and other infected plants, whereas this nutrient was slightly increased in fine and coarse roots. Furthermore, the phosphorus and potassium concentrations in leaves were impaired after infection with P. citricola. However, foliar concentrations of Ca and Mg were not affected by the different Phytophthora spp., whereas fine and coarse roots were significantly enriched with Ca in beech infected with P. citricola or P. cambivora. The concentrations of α‐tocopherol and xanthophyll cycle pigments were increased in plants infected by P. citricola and P. cambivora, indicating that several reactive oxygen species might be formed in leaves during infection.     

Seedborne Fusarium on Douglas-fir: Pathogenicity and seed stratification method to decrease Fusarium contamination

Twelve Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) seedlots from coastal British Columbia were assayed for seedborne Fusarium. All of the seedlots were contaminated with Fusarium. Percent of nonstratified seeds from individual seedlots harboring Fusarium ranged from 0.3% to 95.4%. Sixty-seven percent of the seedlots had Fusarium on less than 2% of the seeds. Post-stratification seedborne Fusarium levels were significantly less for running water imbibition compared to standing water imbibition. However, seedling growth at a container nursery was not consistently different for stratified seed imbibed initially in standing or running water. Fusarium disease symptoms were not observed in the nursery environment. The species of Fusarium isolated from seed were F. acuminatum, F. avenaceum, F. lateritium, F. moniliforme, F. oxysporum, F. poae and F. sambucinum. Twelve Fusarium isolates, comprising six species, were assessed for pathogenicity. Disease symptoms were observed after four weeks incubation and Fusarium isolates ranged in virulence from low to high. Fusarium oxysporum isolates were the most pathogenic.     

Development of Leptographium wageneri on root surfaces and other substrata

Conidia of Leptographium wageneri var. ponderosum, Leptographium wageneri var. pseudotsugae and Leptographium wageneri var. wageneri germinated on wounded roots of ponderosa pine (Pinus ponderosa), Douglas fir (Pseudotsuga menziesii), and pinyon (Pinus edulis), but not on unwounded roots. On wounds, germination occurred regardless of the combination of pathogen variety and plant species. When wounded roots of ponderosa pine and Douglas fir seedlings were inoculated with conidia of L. w. var. ponderosum and L. w. var. pseudotsugae, respectively, 87-100% of the seedlings became infected. When unwounded roots were inoculated with wooden blocks colonized with L. wageneri, 60-66% of the seedlings became infected. The development of L. w. var. ponderosum and L. w. var. pseudotsugae on agar was compared with their development on agar overlaid with either cellophane or Nucleopore membranes. Undulating hyphae were found commonly in and on the agar, but less frequently on the membranes. On both membranes, mycelia formed mats which were tightly appressed to the surface; mats were not formed on the agar. On both agar and the membranes, two or more hyphae often formed a strand. When L. w. var. ponderosum mycelia were allowed to colonize the surface of an unwounded pine root, strands, undulating hyphae and poorly formed mats were seen.     

两种兰科植物CaM及CML基因家族全基因组分析

[目的]探讨Ca M和CML基因家族在兰科植物不同组织和与菌根真菌建立共生关系过程中的潜在功能。[方法]依据已发表的拟南芥Ca M和CML基因家族蛋白序列,利用生物信息学方法对小兰屿蝴蝶兰和铁皮石斛全基因组进行搜索,通过多种生物软件或在线工具对Ca M和CML基因家族蛋白序列进行筛选及确定、蛋白结构分析、系统进化分析及结构域预测;利用转录组数据绘制heatmap图,对小兰屿蝴蝶兰不同组织(花、叶、茎、根)及石斛种子与美胞胶膜菌共生条件下Ca M和CML基因家族的表达情况进行分析。[结果]在小兰屿蝴蝶兰和铁皮石斛全基因组中均预测到4个Ca M蛋白和54个CML蛋白;小兰屿蝴蝶兰Ca M及CML家族基因中39个基因没有内含子,19个基因具有内含子;铁皮石斛Ca M及CML基因家族中有41个基因没有内含子,17个基因具有内含子;系统进化分析将小兰屿蝴蝶兰和铁皮石斛Ca M和CML蛋白家族各分为10个亚家族;小兰屿蝴蝶兰中9个基因在叶中的表达相对于花、茎、根为上调表达,2个基因在叶中的表达相对于花、茎、根却为下调表达; 3个基因在花中的表达相对于叶、茎、根为上调表达; 3个基因在花和叶中的表达相对于茎和根为上调表达;铁皮石斛中4个基因在与美胞胶膜菌共生萌发文库中的表达相对于非共生文库为上调表达; 4个基因在共生萌发文库中的表达相对于非共生文库为下调表达。[结论]Ca M及CML家族基因在兰科植物不同组织具有重要作用,且参与铁皮石斛种子与美孢胶膜菌共生萌发的生物学调控过程。     

Cronartium flaccidum produces uredinia and telia on Melampyrum nemorosum and on Finnish Vincetoxicum hirundinaria

Uredinia and telia were produced on Melampyrum nemorosum through inoculations with Cronartium flaccidum aeciospores collected from northern Finland, proving that M. nemorosum serves as the telial host of C. flaccidum. Melampyrum nemorosum plants were either grown from seeds collected from several geographical provenances or transplanted from natural forest stands. They were inoculated with several geographical aeciospore sources of C. flaccidum in a greenhouse. Vincetoxicum hirundinaria, M. pratense and M. sylvaticum were used as positive controls. Detached leaves of Melampyrum spp. and V. hirundinaria were also inoculated but uredinia and telia formed only on V. hirundinaria. Most of the aeciospore sources from northern Finland produced abundant uredinia and telia both on M. nemorosum from two geographical locations and on V. hirundinaria in the greenhouse. Statistically significant differences were observed in the dimensions of artificially produced telia, teliospores and urediniospores among the tested alternate hosts and spore sources. The results suggest that M. nemorosum is susceptible to C. flaccidum and therefore may play a role in southern Fennoscandia as an alternate host but it has potentially a greater role in central and eastern Europe, where M. nemorosum occurs commonly.     

Community of Aphyllophorales and Root Rot in Stumps of Picea abies on Clear-felled Forest Sites in Lithuania

The community of Aphyllophorales fungi in stumps of Picea abies (L.) Karst. and the occurrence of root rot caused by Heterobasidion annosum were investigated at 38 clear-felled sites in Lithuania. Fruit-bodies were recorded on 36.0% of the 3924 examined stumps. The most common were Bjerkandera adusta, Phlebiopsis gigantea, Fomitopsis pinicola, Gloeophyllum sepiarium, Stereum sanguinolentum, Trametes zonata and H. annosum. All species occurred in stumps both with and without established root rot, but B. adusta, F. pinicola, G. sepiarium, T. zonata and Trichaptum abietinum occurred less often in stumps containing root rot, while H. annosum was encountered more often. Greater numbers of fruit-bodies were found on stumps cut in summer than on those cut in winter. The fruiting became most abundant 3-4 yrs after the trees were felled. The abundance of nearly all Aphyllophorales correlated positively with the stump diameter, and on larger stumps, fruit-bodies of several different fungi were more common. The number of stumps containing H. annosum root rot in different sites varied from 9.8% to 68.8%, and was 27.6% on average. The incidence of root rot correlated neither with the age nor with the density of the felled stand, but it correlated negatively with the proportion of deciduous trees within a stand.     

Effectiveness of some biocontrol and chemical treatments against Heterobasidion annosum on Norway spruce stumps

The biocontrol agents Trichoderma harzianum, Verticillium bulbillosum, Hypholoma fasciculare, Phanerochaete velutina, Vuilleminiia comedens together with the chemical treatments propiconazole and the culture filtrate of V. bulbillosum were tested on Norway spruce stumps during 2 years in order to evaluate their effectiveness against Heterobasidion annosum. The results were compared, in 1 year, with the effectiveness of Phlebiopsis gigantea. All the treatments reduced H. annosum infections. The best results were obtained with P. velutina, propiconazole and with culture filtrates and conidial suspensions of V. bulbillosum.     

Effects of associated fungi Sclerophoma pythiophila and Cenangium ferruginosum on Gremmeniella abietina dieback in Spain

The interactions between Gremmeniella abietina and either Sclerophoma pythiophila or Cenangium ferruginosum, fungi frequently isolated from diseased twigs along with G. abietina, were studied under laboratory (dual cultures) and greenhouse conditions (double‐inoculations). Virulence of each species was also evaluated in greenhouse experiments by means of single‐inoculations. In vitro interactions were assessed on Petri dishes containing malt agar with pine needle extract, and greenhouse experiments were performed on 1‐year‐old Pinus halepensis seedlings. In vitro growth of G. abietina was inhibited by both fungi when grown in dual culture. In single‐inoculations, G. abietina caused the greatest necrosis length on P. halepensis seedlings, followed by S. pythiophila, whereas C. ferruginosum did not cause significant necrosis. In double‐inoculations, C. ferruginosum was able to reduce the length of necrosis caused by G. abietina on the P. halepensis seedlings. In contrast, necrosis length was greater in seedlings inoculated with both S. pythiophila and G. abietina than in those inoculated with G. abietina alone. Therefore, S. pythiophila seems to play a role in disease expression caused by G. abietina on P. halepensis in Spain.     

白蜡虫热激蛋白序列分析

[目的]了解白蜡虫泌蜡高峰期所表达的热激蛋白及其序列特征。[方法]从白蜡虫转录组数据库筛选获得hsp10、hsp20、hsp40、hsp60、hsp70、hsp90基因序列,进行序列联配和进化树分析,并对hsp40、hsp60、hsp90基因在不同温度下mRNA相对表达量情况进行荧光定量PCR检测。[结果]从白蜡虫转录组数据库中共筛选出32条hsp基因序列,序列分析发现它们与目前已报道的序列具有很高一致性,大部分含有保守区域和特征序列,能够与同目昆虫基因聚为一支。hsp40、hsp60、hsp90基因均能被温度诱导。[结论]在白蜡虫泌蜡高峰期共鉴定到32个热激蛋白非重复序列基因(unigene):hsp10 unigene 3个,hsp20 unigene 1个,hsp40 unigene 11个,hsp60 unigene 1个,hsp70 unigene 13个,hsp90 unigene 3个。     

Fungi associated with the fir bark beetle Cryphalus piceae in Poland

The aim of this study was to identify fungi associated with Cryphalus piceae on European silver fir (Abies alba) in Poland and to test the pathogenicity of selected isolates. Fungi were isolated from five populations of overwintered adults and their galleries. A great diversity of taxa was associated with C. piceae. In total, 2487 isolates, including 58 species distributed in 25 genera, were obtained. The two most frequently isolated fungi, an undescribed species of the genus Geosmithia and Ophiostoma piceae, appeared to be specifically associated with C. piceae, whereas Pesotum fragrans, Pesotum sp. and Sporothrix sp. were sporadically associated. Two‐year‐old seedlings of silver fir were wound‐inoculated with three species of fungi (Geosmithia sp., O. piceae and Pesotum sp.) recovered from C. piceae. Only Pesotum sp. showed pathogenic ability, but we do not consider it to be an important pathogen of A. alba.     

<Emphasis Type="Italic">Ophiostoma</Emphasis> species associated with bark beetles infesting three <Emphasis Type="Italic">Abies</Emphasis> species in Nikko,Japan

Ophiostoma species were isolated from bark beetles and Abies mariesii, A. veitchii and A. homolepis attacked by the beetles in Nikko, Tochigi, central Honshu, Japan. One to two Ophiostoma species were frequently isolated from each species of bark beetle. Ophiostoma subalpinum was the most common associate of Cryphalus montanus. Ophiostoma sp. B as well as O. subalpinum was a common fungus associated with Polygraphus proximus. Ophiostoma europhioides was isolated from Dryocoetes hectographus and D. autographus as one of the common associates. Ophiostoma sp. J and Ophiostoma sp. S were frequently isolated from D. autographus and D. striatus, respectively. These fungi seem to have specific relationships with particular bark beetles. Ophiostoma sp. B, Ophiostoma sp. J and Ophiostoma sp. S have unique morphological characteristics and appear to be new species. Five trees of A. veitchii, approximately 43 years old, were inoculated with five Ophiostoma species to assess the relative virulence of the fungi. Ophiostoma subalpinum, Ophiostoma sp. B, and O. europhioides had relatively higher virulence than the other species studied.     

Association of seed‐borne Fusarium species on Pinus ponderosa with germination and seedling viability in Argentina

Pinus ponderosa seedlots from eight seed orchards in Western Patagonia, Argentina, were evaluated for the presence of Fusarium species, and isolates obtained were subjected to pathogenicity tests. The following species were isolated: Fusariumoxysporum, Fusarium equiseti, Fusarium verticillioides, Fusarium incarnatum, Fusarium acuminatum, Fusarium graminearum and Fusarium proliferatum. With the exception of F. proliferatum and F. graminearum all Fusarium species were recovered from P. ponderosa seedlings with symptoms of damping‐off. Five Fusarium species were re‐isolated from necrotic roots of seedlings showing chlorosis (F. oxysporum, F. equiseti, F. incarnatum, F. acuminatum and F. verticillioides). High damping‐off losses occurred in Fusarium‐contaminated seed of P. ponderosa and low disease incidence occurred on less contaminated seed. The role of seed‐borne inoculum in disease spread within forest nurseries is discussed.     

黄河小浪底库区山地栓皮栎人工林土壤呼吸的季节动态

[目的]分离并量化土壤自养呼吸和异养呼吸,探讨各自贡献率及其随季节变化的动态特征。[方法]采用壕沟法和气体红外分析法,研究黄河小浪底库区山地栓皮栎人工林土壤总呼吸、自养呼吸和异养呼吸速率的季节动态变化、贡献率和环境影响因子。[结果]表明:栓皮栎人工林总土壤呼吸、自养呼吸和异养呼吸均呈夏季速率高、冬季速率低。栓皮栎土壤总呼吸、自养呼吸及异养呼吸速率与5 cm土壤温度均呈极显著指数相关,温度敏感性系数Q_(10)值大小为自养呼吸(3.40)异养呼吸(2.90)土壤总呼吸(2.45);栓皮栎土壤总呼吸、自养呼吸、异养呼吸速率与0 10 cm土壤体积含水量均显著线性相关;土壤总呼吸、自养呼吸速率与0 10 cm土壤电导率显著相关。土壤总呼吸和异养呼吸的温度敏感系数Q_(10)值均在冬季最大,夏秋季最小;而自养呼吸的Q_(10)值则呈相反的变化趋势。栓皮栎人工林自养呼吸和异养呼吸对土壤总呼吸的月贡献率为13.23%37.33%和62.67%86.76%,且自养呼吸的贡献率与土壤温度的季节变化规律相似。土壤总呼吸、异养呼吸与自养呼吸的CO2年通量分别为1 616.41、1 199.39、417.02 g·m~(-2)·a~(-1)。[结论]经过区分与定量化土壤总呼吸及其组分,确定异养呼吸为本研究区栓皮栎人工林土壤总呼吸的主要组分,作用于异养呼吸的生物与非生物因子均能显著影响整个森林生态系统表层CO_2总排放通量的大小,进一步为该研究区森林生态系统碳循环与能量流动的进一步量化研究提供参考。     

The culturable leaf mycobiome of Viscum album subsp. austriacum

In Poland's pine forests, mistletoe (Viscum album subsp. austriacum) abundance has increased over the past 20 years. This mistletoe infestation has decreased the growth and vigour of Scots pine trees. In this study, we surveyed the culturable fungi from healthy and diseased V. album subsp. austriacum leaves from two stands in Poland. In total, 63 distinct species were identified, 99.8% of which belonged to the phylum Ascomycota. The community compositions of fungi in mistletoe leaves were similar in healthy and diseased leaves as Alternaria alternata, Aureobasidium pullulans, Botryosphaeria visci, Fusarium paeoniae and Microsphaeropsis olivacea were consistently found in leaves of all symptom types. The most frequently isolated fungus from asymptomatic leaves was M. olivacea, followed by A. alternata, A. pullulans and Hypoxylon rubiginosum. In comparison, the most frequently isolated fungi from leaves with rusty-brown necrotic spots were (in decreasing order) M. olivacea, B. visci, F. paeoniae and A. alternata, while the most frequently isolated fungi from leaves with black or dark brown spots were M. olivacea, A. alternata, A. pullulans, Epicoccum layuense and F. paeoniae. This study was the first comprehensive report showing that certain fungal species may be pathogens of V. album subsp. austriacum in Poland. This study was also the first report of F. paeoniae, F. juglandicola, Diaporthe vacuae and Heterotruncatella spartii from V. album, and the first report of D. vacuae and H. spartii in Poland.     

Studies on free amino acid contents in the aeciospores of Endocronartium pini and Cronartium flaccidum

Aeciospores of Endocronartium pini (Peridermium pini) and Cronartium flaccidum were analysed with regard to free amino acid content. As a rule the quantity in Endocronartium pini is greater than in Cronartium flaccidum. Twenty four amino acids were identified. Regarding thirteen amino acids, the quantity in Endocronartium was greater than in Cronartium; three amino acids were greater only in one year; two amino acids were about the same in the aeciospores of both species, and six occurred only in one analysis or as a trace. The beetle, Lagria hirta, has been observed to eat the acciospores of Endocronartium pini.     

Insecticidal and repellent activities of medicinal plant extracts against the sweet potato whitefly, Bemisia tabaci (Hom.: Aleyrodidae) and its parasitoid Eretmocerusmundus (Hym.: Aphelinidae)

Toxicity and repellent activities of aqueous extracts of nine medicinal plants were evaluated on different life stages of the sweet potato whitefly, Bemisia tabaci. Tomato plants infested with whiteflies were dipped in 10% (wt/wt) of each plant extract for toxicity evaluation. Repellency was evaluated in a choice experiment with detached tomato leaves. All extracts evaluated were relatively ineffective against the adult stage. Extracts of Ruta chalepensis, Peganum harmala and Alkanna strigosa were effective in reducing the numbers of B. tabaci immatures similar to the reduction observed in the imidacloprid treatment. These three extracts were not detrimental B. tabaci parasitoid, Eretmocerus mundus. In addition, the plant extracts Urtica pilulifera and T. capita were repellent to B. tabaci adults. These results indicate that the extracts from the plants R. chalepensis, P. harmala and A. strigosa could act as a potential source for natural product developed for B. tabaci management.     

Associations between Tomicus destruens and Leptographium spp. in Pinus pinea and P. pinaster stands in Tuscany,central Italy

The association between Tomicus destruens and fungi of the genus Leptographium was studied in Pinus pinea and P. pinaster forests in Tuscany, central Italy. Fungi were isolated from adult beetles and from pine tissues from infested trees. On average, Leptographium spp. were associated with 18% of beetles in breeding galleries, 35% of emergent brood beetles and 18% of beetles undergoing maturation feeding in pine twigs. The fungal species most frequently identified were Leptographium wingfieldii and L. lundbergii while L. guttulatum and L. serpens were also found.     

Armillaria novae‐zelandiae and other basidiomycete wood decay fungi in New Zealand Pinus radiata thinning stumps

Two studies were undertaken in a young New Zealand Pinus radiata stand to find a biological control agent for armillaria root disease caused by Armillaria novae‐zelandiae. Fresh thinning stumps were inoculated with saprophytic basidiomycete species as wooden dowel cultures or as homogenized aqueous mycelial suspensions, while all stumps were also inoculated with dowel cultures of A. novae‐zelandiae. After a period of between one and two years, no differences were found among test fungi in the percentage of Armillaria species isolated from stumps. However, inoculation of Stereum sanguinolentum as homogenate, and of Phlebiopsis gigantea, Sistotrema brinkmannii, Hypholoma acutum and Rigidoporus concrescens as dowel cultures, significantly increased their isolation incidence in comparison with stumps not treated by these species. Phlebiopsis gigantea, Resinicium bicolor and R. concrescens were also isolated with greater frequency nearer to their dowel inoculation points. Results suggest that P. gigantea and S. sanguinolentum should be included in further testing. However, a future trial is likely to be more effective if evaluation is directed towards the portion of the stump below the soil surface, particularly the root system.