Performance of real‐time PCR and immunofluorescence for the detection of Clavibacter michiganensis subsp. sepedonicus and Ralstonia solanacearum in potato tubers in routine testing

This paper describes a comparison study of test methods and supports the use of real‐time polymerase chain reaction (PCR) for the detection of Clavibacter michiganensis subsp. sepedonicus and Ralstonia solanacearum in potato tubers in routine testing. These 2 bacteria are quarantine organisms under European Union (EU) regulatory control and testing for (latent) infections of these bacteria in seed potatoes is mandatory. Real‐time PCR tests were performed on 276 routine potato tuber samples, including samples infected with either C. michiganensis subsp. sepedonicus or R. solanacearum, and the performance of these real‐time PCR tests was compared with that of immunofluorescence (IF). Real‐time PCR tests, using different primer sets and extraction and PCR protocols, proved to be sensitive and specific for the detection of C. michiganensis subsp. sepedonicus and R. solanacearum in potato tubers in routine testing, and performed at least as well as IF. Real‐time PCR is a good addition to the detection protocols as laid down in EU regulations (EU Council Directives 2006/56/EC and 2006/63/EC).     

The role of soils in the regional carbon budget of pine forests in Karelia

The carbon pool and its fluxes were studied in Karelian pine forests of automorphic sites for the characterization of the regional carbon cycle. On the permanent sample plots arranged in pine stands (dominant forest types) of the middle and northern taiga, the structure and dynamics of the phytomass, as well as the composition of the soil organic matter, were investigated. The contribution of different components of the forest ecosystems to the regional carbon budget was studied. The carbon pools in the soils of the middle and northern taiga subzones were 330–440 × 10² and 440–550 × 10² kg/ha, respectively. The share of soil carbon in the total carbon pool of the forest biogeocenoses studied amounted to 46–35 and 33–24%, respectively. The period of considerable carbon fixation in the pine ecosystem (nearly 20 × 10² kg/ha/yr) turned out to be limited to the middle-aged stands. In the mature and overmature pine forests, only 4 × 10² kg of C/ha/yr were fixed. The main part of carbon in the soil (65–75%) was contained in the illuvial horizons, although its content there was less that 1% as compared to 30–50% in the forest litters. On industrial harvesting of wood, the main reserves of soil carbon are not spent; only the fund of mobile carbon in organic horizons of poor soils is exhausted or carbon accumulates intensely due to the development of continuous plant cover on fertile soils.     

Genetic differentiation between fake abalone and genuine Haliotis species using the forensically informative nucleotide sequencing (FINS) method

Abalones ( Haliotis species) are a popular delicacy and commonly preserved in dried form either whole or in slices or small pieces for consumption in Asian countries. Driven by the huge profit from trading abalones, dishonest traders may substitute other molluscan species for processed abalone, of which the morphological characteristics are frequently lost in the processed form. For protection of consumer rights and law enforcement against fraud, there is a need for an effective methodology to differentiate between fake and genuine abalone. This paper describes a method (validated according to the international forensic guidelines provided by SWGDAM) for the identification of fake abalone species using forensically informative nucleotide sequence (FINS) analysis. A study of the local market revealed that many claimed "abalone slice" samples on sale are not genuine. The fake abalone samples were found to be either volutids of the genus Cymbium (93%) or the muricid Concholepas concholepas (7%). This is the first report of Cymbium species being used for the preparation and sale as "abalone" in dried sliced form in Hong Kong.     

Enhancement of protective immunity in blue gourami,Trichogaster trichopterus (Pallas), against Aeromonas hydrophila and Vibrioanguillarum by A. hydrophila major adhesin

Blue gourami, Trichogaster trichopterus (Pallas), were intraperitoneally immunized with major adhesin, a 43 kDa OMP protein isolated from fish Aeromonas hydrophila, in the presence of Freund's complete adjuvant (FCA). Three weeks later, a booster injection of adhesin without FCA was administered. Control group fish were similarly treated with phosphate‐buffered saline (PBS) and FCA. Results showed that anti‐adhesin serum obtained from fish after booster immunization exhibited very strong ability in agglutinating bacterial cells. Although this antiserum had no bactericidal effect, it could significantly inhibit serologically different strains of A. hydrophila from invading EPC (Epithelioma papillosum of carp) cells in vitro. In addition, the proliferative response of head kidney leucocytes of these immune fish was significantly increased as compared to that of the control. The results also showed that the major adhesin could provide significant protective immunity to fish against the challenge by homologous and heterologous strains of A. hydrophila and one virulent strain of Vibrio anguillarum.     

Patterns of root respiration rates and morphological traits in 13 tree species in a tropical forest

The root systems of forest trees are composed of different diameters and heterogeneous physiological traits. However, the pattern of root respiration rates from finer and coarser roots across various tropical species remains unknown. To clarify how respiration is related to the morphological traits of roots, we evaluated specific root respiration and its relationships to mean root diameter (D) of various diameter and root tissue density (RTD; root mass per unit root volume; gcm(-3)) and specific root length (SRL; root length per unit root mass; mg(-1)) of the fine roots among and within 14 trees of 13 species from a primary tropical rainforest in the Pasoh Forest Reserve in Peninsular Malaysia. Coarse root (2-269mm) respiration rates increased with decreasing D, resulting in significant relationships between root respiration and diameter across species. A model based on a radial gradient of respiration rates of coarse roots simulated the exponential decrease in respiration with diameter. The respiration rate of fine roots (<2mm) was much higher and more variable than those of larger diameter roots. For fine roots, the mean respiration rates for each species increased with decreasing D. The respiration rates of fine roots declined markedly with increasing RTD and increased with increasing SRL, which explained a significant portion of the variation in the respiration among the 14 trees from 13 species examined. Our results indicate that coarse root respiration in tree species follows a basic relationship with D across species and that most of the variation in fine root respiration among species is explained by D, RTD and SRL. We found that the relationship between root respiration and morphological traits provides a quantitative basis for separating fine roots from coarse roots and that the pattern holds across different species.     

Spatiotemporal dynamics and modelling support the case for area-wide management of citrus greasy spot in a Brazilian smallholder farming region

Citrus greasy spot (CGS), caused by Zasmidium citri, induces premature defoliation and yield loss in Citrus spp. The epidemiology of CGS is well understood in high humidity areas, but remains unaddressed in Brazil, despite differing climatic conditions and disease management practices. The spatiotemporal dynamics of CGS were characterized in the Recôncavo of Bahia (Brazil) at four hierarchical levels (quadrant, plant, grove, and region). A survey conducted in 19 municipalities found the disease to be present throughout the region with an incidence of 100% in groves and plants, and higher than 70% on leaves. Index of dispersion (D) values suggest the spatial pattern of units with symptoms lies between random and regular. This was confirmed by the parameters of the binary power law for plants and their quadrants (log[A] < 0 and b < 1). No consistent differences were observed in the disease incidence at different plant heights. We introduce a compartmental model synthesizing CGS epidemiology. The collected data allow such a model to be parameterized, albeit with some ambiguity over the proportion of new infections that result from inoculum produced within the grove versus external sources of infection. By extending the model to include two populations of growers—those who control and those who do not—coupled by airborne inoculum, we investigate likely performance of cultural controls accessible to citrus growers in northeastern Brazil. The results show that control via removal of fallen leaves can be very effective. However, successful control is likely to require area-wide strategies in which a large proportion of growers actively manage disease.     

寒地水稻优质稻米抗稻瘟病性鉴定研究

通过人工接种和自然感病两种方法,共鉴定寒地水稻新品种(系)292份(次),鉴定出高抗新品种(系)22份,适合寒地稻区种植的优质、抗病、农艺性状优良的新品种(系)7份,扩大和充实了抗病基因库。     

Passive transfer of protective immunity against ichthyophthiriasis from vaccinated mother to fry in tilapias, Oreochromis aureus

Tilapias, Oreochromis aureus, were vaccinated twice with live tomites of Ichthyophthirius multifiliis in physiological saline 1 month before spawning (test group). A control group was similarly treated but with physiological saline only. Fertilized eggs were collected from the mouths of both groups and incubated until the young reached the free-swimming stage. Free-swimming fry were also collected immediately after they were expelled from the mother's mouth in other fish of the two groups. The fry were exposed to infective tomites of I. multifiliis and thereafter returned to their holding tanks. All fry obtained from the controls without mouth-brooding died after tomite exposure. However, fry with mouth-brooding before tomite exposure showed 37.3% survival. On the other hand, fry obtained from vaccinated broodstock without mouth-brooding exhibited 78.4% survival which increased to 95.3% with mouth-brooding. The protective immunity is correlated with the titres of anti-I. multifiliis antibodies in the soluble extracts of fry tissues and the mother's plasma. The results clearly indicate that protective immunity against ichthyophthiriasis “ich” in tilapia fry can not only be derived directly from the mother via eggs but also be acquired indirectly from the mouth cavity during the brooding period. The results thus suggest that proper vaccination of mothers before spawning would effectively protect fry against a protozoan disease.     

Genomic Sequence and Experimental Tractability of a New Decapod Shrimp Model,Neocaridina denticulata

The speciose Crustacea is the largest subphylum of arthropods on the planet after the Insecta. To date, however, the only publically available sequenced crustacean genome is that of the water flea, Daphnia pulex, a member of the Branchiopoda. While Daphnia is a well-established ecotoxicological model, previous study showed that one-third of genes contained in its genome are lineage-specific and could not be identified in any other metazoan genomes. To better understand the genomic evolution of crustaceans and arthropods, we have sequenced the genome of a novel shrimp model, Neocaridina denticulata, and tested its experimental malleability. A library of 170-bp nominal fragment size was constructed from DNA of a starved single adult and sequenced using the Illumina HiSeq2000 platform. Core eukaryotic genes, the mitochondrial genome, developmental patterning genes (such as Hox) and microRNA processing pathway genes are all present in this animal, suggesting it has not undergone massive genomic loss. Comparison with the published genome of Daphnia pulex has allowed us to reveal 3750 genes that are indeed specific to the lineage containing malacostracans and branchiopods, rather than Daphnia-specific (E-value: 10⁻⁶). We also show the experimental tractability of N. denticulata, which, together with the genomic resources presented here, make it an ideal model for a wide range of further aquacultural, developmental, ecotoxicological, food safety, genetic, hormonal, physiological and reproductive research, allowing better understanding of the evolution of crustaceans and other arthropods.