Dithiothreitol decreases in vitro activity of ADP-glucose pyrophosphorylase from leaves of apple (Malus domestica Borkh.) and many other plant species

Inclusion of dithiothreitol (DTT) in the extraction buffer and pre-incubation of apple leaf ADP-glucose pyrophosphorylase (AGPase) with DTT resulted in a decrease in AGPase activity whether the assay was performed in the presence or absence of 3-phosphoglycerate (PGA). When PGA was included in the pre-incubation mixture or when pre-incubation of AGPase with PGA was followed by DTT, the latter did not cause any decrease in AGPase activity. However, once AGPase was decreased by DTT, subsequent incubation of the enzyme with PGA did not reverse the decrease. Pre-incubation of AGPase from leaves of Arabidopsis thaliana, sorghum, soybean, tobacco, spinach, wheat, barley, tomato and potato, and tubers of potato with DTT, generally caused a decrease in AGPase activity when assayed in the presence of PGA. When assayed in the absence of PGA, however, a diverse response of AGPase was observed among species to pre-incubation with DTT. The activity of AGPase from potato tubers was increased by DTT; the activity of AGPase from both potato and tomato leaves was not affected by DTT; the activity of AGPase from leaves of other species was decreased by DTT. It is concluded that DTT decreases in vitro activity of AGPase from leaves of apple and many other plant species such that DTT should not be routinely included in the extraction or assay mixture of leaf AGPase.     

Fermentation of cashew apple juice to produce high added value products

The use of agriculture substrates in industrial biotechnological processes has been increasing because of its low cost. Cashew apples are considered an agriculture low cost product in the Brazilian Northeast because the cashew cultivation is done mainly to produce cashew nuts. About 90% of the cashew apples production is lost in the field after removing the nut. In this work, the use of clarified cashew apple juice as substrate for microbial cultivation was investigated. The results showed that cashew apple juice is a good source of reducing sugars and can be used to grow Leuconostoc mesenteroides to produce high added value products such as dextran, lactic acid, mannitol and oligosaccharides.     

Isolation and pathogenicity of some South African entomopathogenic fungi (Ascomycota) against eggs and larvae of Cydia pomonella (Lepidoptera: Tortricidae)

Virulent entomopathogenic fungal strains against Cydia pomonella (Lepidoptera: Tortricidae) were isolated and identified in the Western Cape Province of South Africa. Thirty-nine isolates belonging to six species were obtained using the insect bait method. Generally, Metarhizium robertsii (Ascomycota: Hypocreales) was the most frequently encountered species representing 51% of the total number of isolates collected from the soil samples. This is the first report of M. robertsii from southern Africa. Mortality data from an immersion bioassay indicated that the 39 fungal isolates were pathogenic against fifth instar larvae of C. pomonella inducing 47–85% insect mortalities. Two M. robertsii strains, MTL151 and GW461, induced 85% larval mortality and were selected for further evaluations. The exposure of freshly laid eggs to wax papers that were pre-treated with fungal spores ranging from 10³ to 10⁸ spores ml^?1 of MTL151 and GW461 resulted in a significant reduction of egg hatchabilities, from 95 to 66% and 93 to 71%, respectively as spore concentration increased. First instar larval neonates were exposed to apples that were pre-sprayed topically with varied conidia suspensions (10³?10⁸ spores ml^?1). The mean percentage of participating apples with larvae in their cores/flesh significantly reduced from 53 to 10% (MTL151) and 76 to 10% (GW461) of 10 apples, and a concurrent decrease in the incidence of apple fruit rot was observed as conidia concentration increased. Up to 90% of apples treated with 1 × 10⁸ spores ml^?1 had no larvae present in their cores and this result compared favourably with the commercial pesticide Fruitfly®, containing the active ingredient cypermethrin (20 g/l) used at a recommended dose of 0.25 ml/250 ml of water.     

食块茎水鸟及水位对沙湖沉水植物冬芽分布的影响

分别于2012—2013、2013—2014年度越冬期候鸟越冬前(10月份)与越冬后(4月份)采用样方法调查沙湖沉水植物冬芽的种类、密度及生物量,分析不同水位条件的2个年度鄱阳湖碟形子湖沉水植物冬芽的分布及其对食块茎水鸟食物贡献的差异性,探讨越冬水鸟取食与水位变化对沉水植物冬芽分布的影响。结果表明:刺苦草(Vallisneria spinulosa)和罗氏轮叶黑藻(Hydrilla verticillata var.rosburghii)2种沉水植物的冬芽同域分布。2013年10月2种植物冬芽的密度与生物量均显著低于2012年同期,主要原因是鄱阳湖水位年际间变化剧烈,并对水质有显著影响:与2012年相比,2013年丰水期(4—9月)沙湖与主湖区连通的时间和日平均水深显著减小,但水体浊度显著增加,不利于沉水植物生长发育。2012—2013年度越冬水鸟迁出后2种冬芽的密度和生物量均明显下降,而2013—2014年度越冬期水鸟迁出后与迁入前相比两种植物冬芽的密度和生物量均无显著变化,很可能与食块茎水鸟的取食活动和高水位对食物可利用性的负面影响有密切关系。湖泊剧烈的水位变化导致越冬水鸟的食源具有年际波动的特征,而食块茎水鸟对鄱阳湖子湖的食物利用率受越冬季冬芽丰富度和食物可及性(accessibility)的共同影响。研究结果对鄱阳湖乃至长江中下游流域沉水植被恢复、越冬水鸟保护以及生态系统功能评估具有指导意义。     

The severity of wheat diseases increases when plants and pathogens are acclimatized to elevated carbon dioxide

Wheat diseases present a constant and evolving threat to food security. We have little understanding as to how increased atmospheric carbon dioxide levels will affect wheat diseases and thus the security of grain supply. Atmospheric CO₂ exceeded the 400 ppmv benchmark in 2013 and is predicted to double or even treble by the end of the century. This study investigated the impact of both pathogen and wheat acclimation to elevated CO₂ on the development of Fusarium head blight (FHB) and Septoria tritici blotch (STB) disease of wheat. Here, plants and pathogens were cultivated under either 390 or 780 ppmv CO₂ for a period (two wheat generations, multiple pathogen subcultures) prior to standard disease trials. Acclimation of pathogens and the wheat cultivar Remus to elevated CO₂ increased the severity of both STB and FHB diseases, relative to ambient conditions. The effect of CO₂ on disease development was greater for FHB than for STB. The highest FHB disease levels and associated yield losses were recorded for elevated CO₂‐acclimated pathogen on elevated CO₂‐acclimated wheat. When similar FHB experiments were conducted using the disease‐resistant cultivar CM82036, pathogen acclimation significantly enhanced disease levels and yield loss under elevated CO₂ conditions, thereby indicating a reduction in the effectiveness of the defence pathways innate to this wheat cultivar. We conclude that acclimation to elevated CO₂ over the coming decades will have a significant influence on the outcome of plant–pathogen interactions and the durability of disease resistance.     

Culture-dependent and culture-independent assessment of bacteria in the apple phyllosphere

Aims: Bacterial communities in the apple phyllosphere were examined quantitatively and qualitatively by applying culture‐dependent and culture‐independent methods. Methods and Results: Populations estimated by viewing cells stained with 4′,6‐diamidino‐2‐phenylindole generally were at least 100–1000 times greater than populations estimated by culturing on tryptic soy agar (TSA). Of the 44 operational taxonomic units (OTUs; cut‐off threshold of 97%) detected in total, five bacterial orders containing 23 OTUs were identified by culturing on TSA, whereas nine orders containing 33 OTUs were identified by 16S rRNA gene cloning of DNA extracted from apple leaf surfaces. Twelve of the 44 OTUs were shared between cultured isolates and 16S rRNA gene clones and included the orders Burkholderiales, Pseudomonadales, Rhizobiales and Sphingomonadales. Three OTUs within the genus Sphingomonas accounted for 40% of isolates and 68% of clones. The Actinomycetales were found only among isolates, whereas the Bacteroidales, Enterobacteriales, Myxococales and Sphingobacteriales were represented in the 16S rRNA gene clone libraries but were absent among isolates. Conclusions: Culture‐independent methods revealed greater numbers and greater richness of bacteria on apple leaves than found by culturing. Significance and Impact of the Study: This is the first study to directly compare culture‐dependent and independent approaches for assessing bacterial communities in the phyllosphere. The biases introduced by different methods will have a significant impact on studies related to phyllosphere ecology, biological control of plant diseases, reservoirs of antibiotic resistance genes and food safety.     

Interactions between 'Candidatus Phytoplasma mali' and the apple endophyte Epicoccum nigrum in Catharanthus roseus plants

Aims: We investigated the ultrastructural and molecular interactions between ‘Candidatus Phytoplasma mali’ and the apple endophyte Epicoccum nigrum in the experimental host Catharanthus roseus to determine whether inoculation of endophyte could trigger defence reactions in the host. Methods and Results: Apple proliferation (AP) symptom severity was evaluated in AP‐grafted plants that were treated by E. nigrum and compared with untreated controls. Phytoplasma concentration was quantified by real‐time PCR in treated and untreated plants. Ultrastructural observations revealed that in endophyte‐treated periwinkles, modifications to phytoplasmas, such as irregular shape and cytoplasm confined to the periphery of the cell, and plant cytological changes, such as abundant callose depositions and P‐protein aggregations in the sieve elements, occurred. AP‐grafted plants that were treated by the endophyte (E. nigrum) showed a reduction in symptom severity; in particular, flowers appeared normal in shape and size, when compared with uninfected controls. Real‐time PCR indicated that phytoplasma concentration in AP‐grafted plants treated with E. nigrum was about 2·8 times lower than that in untreated ones. Conclusions: These results demonstrated that the inoculation with E. nigrum influenced phytoplasma infection in C. roseus plants; plant ultrastructural modifications allowed us to hypothesize an enhancing host defence response. Significance and Impact of the Study: At present, curative protocols against this phytoplasma are not available. Alternative approaches are thus required to reduce disease spread. Our study might represent a first step in the clarification of plant–phytoplasma–endophyte relationships to find possible strategies for the control of phytoplasma diseases.