Deep‐sequencing revealed Citrus bark cracking viroid (CBCVd) as a highly aggressive pathogen on hop

Newly emerging or re‐emerging diseases are a constant and significant threat to agricultural production, so prompt and accurate identification of the causative agents is required for rapid and appropriate disease management. Classical methods of pathogen detection can be successfully supplemented by next‐generation sequencing (NGS), whereby sequence analysis can help in the discovery of new or emerging diseases. In 2007, hop growers in Slovenia reported the appearance of severely stunted hop plants, a phenomenon that spread rapidly within hop gardens and among farms. Classical diagnostic methods were unable to detect a new pathogen; therefore, single step high‐throughput parallel sequencing of total RNA and small RNAs from plants with and without symptoms was employed to identify a novel pathogen. The sequences were assembled de novo and also mapped to reference genomes, resulting in identification of a novel sequence of Citrus bark cracking viroid (CBCVd) in the stunted hop plants. Furthermore, the presence of this novel pathogen on hop was confirmed by RT‐PCR analysis of 59 plants with symptoms from 15 hop gardens, representing the main outbreak locations identified by systematic disease monitoring, and small RNA Illumina sequencing of the bulked RNA sample. The high infectivity of the newly identified CBCVd was also confirmed by biolistic inoculation of two hop cultivars, which developed aggressive symptoms in controlled conditions. This study shows the feasibility of deep sequencing for the identification of causative agents of new diseases in hop and other plants.     

Multifunctionalised benzoxazinones in the systems Oryza sativa–Echinochloa crus‐galli and Triticum aestivum–Avena fatua as natural‐product‐based herbicide leads

BACKGROUND: Fifteen novel derivatives of D‐DIBOA, including aromatic ring modifications and the addition of side chains in positions C‐2 and N‐4, had previously been synthesised and their phytotoxicity on standard target species (STS) evaluated. This strategy combined steric, electronic, solubility and lipophilicity requirements to achieve the maximum phytotoxic activity. An evaluation of the bioactivity of these compounds on the systems Oryza sativa–Echinochloa crus‐galli and Triticum aestivum–Avena fatua is reported here. RESULTS: All compounds showed inhibition profiles on the two species Echinochloa crus‐galli (L.) Beauv. and Avena fatua L. The most marked effects were caused by 6F‐4Pr‐D‐DIBOA, 6F‐4Val‐D‐DIBOA, 6Cl‐4Pr‐D‐DIBOA and 6Cl‐4Val‐D‐DIBOA. The IC₅₀ values for the systems Echinochloa crus‐galli–Oryza sativa and Avena fatua–Triticum aestivum for all compounds were compared. The compound that showed the greatest selectivity for the system Echinochloa crus‐galli–Oryza sativa was 8Cl‐4Pr‐D‐DIBOA, which was 15 times more selective than the commercial herbicide propanil (Cotanil‐35). With regard to the system Avena fatua–Triticum aestivum, the compounds that showed the highest selectivities were 8Cl‐4Val‐D‐DIBOA and 6F‐4Pr‐D‐DIBOA. The results obtained for 6F‐4Pr‐D‐DIBOA are of great interest because of the high phytotoxicity to Avena fatua (IC₅₀ = 6 µM , r² = 0.9616). CONCLUSION: The in vitro phytotoxicity profiles and selectivities shown by the compounds described here make them candidates for higher‐level studies. 8Cl‐4Pr‐D‐DIBOA for the system Echinochloa crus‐galli‐Oryza sativa and 6F‐4Pr‐D‐DIBOA for Avena fatua‐Triticum aestivum were the most interesting compounds. Copyright © 2010 Society of Chemical Industry     

The dynamics of bacterial community in a polyculture aquaculture system of Penaeus chinensis,Rhopilema esculenta and Sinonovacula constricta

Polyculture with Chinese shrimp Penaeus chinensis, jellyfish Rhopilema esculenta and razor clam Sinonovacula constricta (S.J.R) is a common aquaculture system in Dandong, China. Investigation on the composition and function of microbial communities and their relationship with environmental factors in aquaculture systems have crucial implications. Therefore, the sequencing of 16S rRNA gene was applied to profile the bacterial community composition of water and sediment in S.J.R polyculture system in the present study. Proteobacteria was the dominant phylum in S.J.R polyculture ponds following with Actinobacteria and Bacteroidetes in water; in contrast, more diverse phyla were detected in sediments. Water temperature, pH and dissolved oxygen were determined the bacterial community composition in the S.J.R polyculture systems. Specially, Planktomarina, PS1 clade, Candidatus Aquiluna and Fibrobacteres, which can resist the phytoplankton blooms or degrade the cellulose, were more abundant in water at the early stage of cultivation in the S.J.R polyculture system. Moreover, the functions of bacterial communities in water were mainly associated with cell communication mediated, while the functions related to basal metabolism were enriched in sediments. The findings of this study allowed us to better understand the bacterial community in the S.J.R polyculture system and accumulated data for the improvement of cultural management.     

Design and validation of a RT‐qPCR procedure for diagnosis and quantification of most types of infectious pancreatic necrosis virus using a single pair of degenerated primers

Infectious pancreatic necrosis virus (IPNV) is an important virus which affects the salmonid aquaculture industry worldwide; therefore, it is important to develop rapid and reliable methods of diagnosis to detect the disease at early stages. Nowadays, RT‐qPCR is replacing other methods because it provides additional information on the viral load, which is important to have a better understanding of the virus replication level and of the stage of the infection and its risk level. The main problem stems from the high diversity of this virus, which can compromise the reliability of the diagnosis. In this study, we have designed an RT‐qPCR procedure for diagnosis and quantification of IPNV based on a single pair of primers targeted to segment B. The procedure has been validated, in vitro and in vivo, testing two different types of standards against seven reference strains and 23 field isolates from different types. The procedure is reliable for the detection of any type, with a detection limit of 31 TCID₅₀ mL^?1, 50 pfu mL^?1 or 66 RNA copies mL^?1, depending on the standard. All the standard curves showed high reliability (R² > 0.95). The results support the high reliability of this new procedure for the diagnosis and quantification of IPNV.     

Midsummer copper sprays have no effect on ‘delicious’ apple fruit color grade or typiness

Recent reports suggested that multiple midsummer copper (Cu) sprays could improve the proportion of apple fruit (Malus × domestica Borkh.) being packed in the reddest market color grades. We conducted a three‐year field experiment in a low‐Cu orchard with a history of poor fruit coloring. We applied multiple mid‐summer sprays of Cu sulfate and basic Cu sulfate in 1994 and 1995 to ‘Bisbee Delicious’ and ‘Oregon Spur II Delicious’ apple trees. Plant tissue nutrient levels, fruit color grade, and fruit typiness were evaluated in the years of application and the following year. The Cu sprays increased leaf Cu in the years of application; however, leaf Cu in the Cu‐sprayed plots returned to or very near to background concentrations in the year after sprays were terminated. The Cu treatments did not increase the proportion of fruit in the reddest market color grades or influence five fruit typiness indices of either apple cultivar. Although midsummer Cu sprays may not enhance apple fruit red color or typiness, they may be useful for increasing overall tree Cu status, particularly for long‐season cultivars which are harvested too late for postharvest sprays to be effective.