Comparison of diagnostics techniques in an outbreak of infectious laryngotracheitis from meat chickens

Various diagnostics techniques were compared for their ability to detect infectious laryngotracheitis (ILT) during an outbreak in chickens aged between 4 and 21 wk. Gross lesions ranged from excess mucus to accumulation of fibrinonecrotic exudate in the larynx and trachea. Syncytial cells with intranuclear inclusion bodies were found in sinus, conjunctiva, larynx, trachea, lung, and air sac. Virus isolation in chicken embryos was attempted in every case. Negative-stain electron microscopy detected herpesvirus in only 6% of the cases. Yet, isolation of ILT virus in the chorioallantoic membrane was presumed by histology in >20% of the samples and confirmed by fluorescent antibody (FA) in 35% of the embryos inoculated with conjunctivas or tracheas from affected birds. Overall, results from histology and FA tests were highly correlated. FA test has the advantage over histology of being diagnostically specific for ILT virus. Polymerase chain reaction was the most sensitive test and detected the viral DNA even in cases where histology and FA were negative. ILT virus DNA was quantified by real-time polymerase chain reaction (Re-Ti ILTV). Histologic and FA results from larynx and trachea were negative if the concentration of the viral DNA was < or =4 of log10. A viral DNA concentration higher than log10 4, as determined by Re-Ti ILTV, was required for clinical ILT to be manifested.     

Improving Drought Tolerance of Rice by Designed QTL Pyramiding

Drought is the most important factor limiting rice yields in the rainfed areas of Asia. To overcome the problem, we developed a new strategy ＇designed QTL pyramiding＇ to more efficiently develop drought tolerant （DT） rice cultivars. In the first phase of this strategy, we developed large numbers of introgression lines （ILs） in elite backgrounds using backcross （BC） breeding, each of which carries multiple genomic segments for improved DT from a known donor. Then, we genotyped all ILs with SSR markers to track the genomewide pattern of introgression in the DT ILs of 16 BC populations derived from crosses between 2 recipients （IR64 and Teqing） and four different donors. X^2 tests and linkage disequilibrium analyses of introgression in the DT ILs revealed significant frequency shifts at many loci across the genome and non-random associations at the multilocus level as a result of selection for DT in the ILs, which led us to the discovery of putative genetic networks underlying DT in the ILs. The networks each containing all DT loci detected in ILs from a BC population showed some interesting features.     

Cloning and Sequence Analysis of Lipoxygenase Gene cDNA from Cucumber Fruit （Cucumis sativus L.）

Lipoxygenases are nonheme-iron-containing dioxygenases that catalyze the hydroperoxidation of unsatrated fatty acids containing a cis, cis-1,4-pentadiene structure producing hydroperoxy acids with conjugated dienes. LOX activity has been found in a wide range of plants. Typical substrates for LOX in plants are linoleic acid and linilenic acid fatty acids. The function of various LOXs in plants is unknown, but their participation in all stages of plant growth and development has been suggested （Hildebrand, 1989; Siedow, 1991）. Some of the physiological processes in whicn lipoxygenses have been implicated include wounding （Saravitz and Siedow, 1996）, pathogen attack （Melan et al., 1993）, seed germination （Kato et al., 1992）, fruit ripening （Ferrie et al., 1994）, plant senescence （Paliyath and Droillard, 1992）. The study on the role of lipoxygenase in ripening and senescence fruit focused on tomato and strawberry. Cloning LOX gene of cucumber fruit will make us further understand the molecularaction mode of this enzyme during fxuit ripening and senescence. In this paper we isolated the partial nucleotide sequences of cucumber fiuit lipoxygenase gene and discuss the characterization of it.     

TILLING and Ecotilling the Plant Kingdom

The modem crop scientist has a large amount of available nucleotide sequence information to identify genes of potential agronomic importance. Using reverse genetic approaches, specific genes can be disrupted, and hypotheses regarding gene function directly tested in vivo. Although a number of reverse genetic methods have been introduced, many are limited in application because they are organism-specific, expensive, transgenic, or only transiently disrupt gene function. However, traditional mutagenesis using chemical mutagens has been widely used as a forward genetics strategy to create many new crop plant varieties at relatively low cost. Mutagens such as ethyl methanesulphonate （EMS） cause stable point mutations and thus produce an allelic series of truncation and missense changes that can provide a range of phenotypes （Greene et al., 2003）. TILLING （targeting induced local lesions IN genomes） is a high-throughput reverse genetic strategy that combines traditional mutagenesis and SNP discovery methods （Colbert et al., 2001; McCallum et al., 2000）. To identify mutations, target regions of-l.5 kb are amplified with fluorescently labeled gene specific primers. Heteroduplexes are then formed between wild-type and mutant strands, mismatches are cleaved by incubation with a single-strand specific nuclease,     

Mapping QTLs for Yield Stability in Durum Wheat Grown under Different Water Regimes

Among the most important Mediterranean annual crops, durum wheat is widely grown in drought-prone areas. Therefore, improving water-use efficiency （WUE） of durum wheat represents a major breeding goal. IDu-WUE （Improving Durum wheat for Water Use Efficiency and yield stability through physiological and molecular approaches） is a collaborative project among public and private research centres in Italy, Spain and WANA （West Asia and North Africa） countries （Morocco, Tunisia, Syria and Lebanon） funded by the European Union aimed at investigating the genetic variation for WUE and yield stability in durum wheat grown in Mediterranean droughtprone areas. During the first year of the project, a number of morpho-physiological traits （e.g. early vigour, flowering time, leaf rolling, number of fertile tillers, etc.）, WUE, WUE-related traits （e.g. carbon isotope discrimination, canopy temperature, chlorophyll fluorescence, etc.）, yield and its components have been investigated in a RIL population （249 lines） and a collection of ca. 190 durum wheat accessions characterized by a high level of linkage disequilibrium （Maccaferri et al., 2005）,     

Microsatellite Marker Based Assessment of Genetic Diversity among Cultivars, Landraces and Wild Relatives in Rice

India being the primary center of origin for rice had a very large treasure of local land races, most of which are out of cultivation today. The exact genetic potential and their differences from commercial varieties and the magnitude of heterogeneity still present in them are not well catalogued. Hence the need to characterize the available land races has become imminent in the modem day concept of crop improvement （Rezai and Frey, 1990）. The present study addresses the utility of SSR markers in divulging the genetic relationships at molecular level among the major component factions office gennplasm viz., local cultivars, land races and wild species collected from a wide range of agro-geographical regions of Indian subcontinent.     

Genetic Diversity of Peanut Core Collection from US Using Simple Sequence Repeats

Groundnut is a member of genus Arachis and the crop is divided into two subspecies and six botanical varieties based on morphological characteristics （Krapovickas and Gregory, 1994）. The International Crops Research Institute for the Semi-Arid Tropics （ICRISAT） holds more than 14 000 groundnut accessions from which a core collection of 1 704 have been developed （Upadhyaya et al., 2003）. Holdbrook et al. （1993） developed a groundnut core collection of 831 accessions from a total of 7 432 US groundnut accessions based on morphological characteristics. The large number and variability of accessions in GenBanks create problems in knowing which germplasm to select for breeding purpose. Only a few established cultivars and elite breeding lines have been utilised in breeding programmes. A core collection is a fraction of accessions from the entire collection which represent most of the available genetic diversity of the species. A core collection can extensively be evaluated and information derived from them can be applied to the whole collection. Identification of DNA markers associated with the botanical varieties of groundnut would be useful in genotyping, germplasm management and evolutionary studies.     

氮肥施用对柑橘果园土壤有机碳矿化的影响

土壤有机碳矿化是土壤向大气释放CO2的最大净输出途径,其与植被的净初级生产力的差值是判断生态系统碳“源”或“汇”的关键。采用室内模拟试验,在10、20、30℃ 3个温度条件下,研究施肥施用对柑橘果园土壤有机碳矿化的影响。结果表明：3种温度处理下,各施氮处理土壤有机碳矿化速率都表现为培养前期快速下降,培养后期保持相对稳定的趋势。在整个培养过程中,3种温度条件下各施氮处理的土壤CO2累积排放量为1328.25~2219.42 mg/kg,100 mg/kg（N4）处理土壤有机碳矿化量最大,CK处理最低,100 mg/kg（N4）和80 mg/kg（N3）2个高氮处理显著高于低氮50 mg/kg（N2）、30 mg/kg（N1）处理。土壤有机碳矿化速率随温度升高而增长,不同的土壤施氮条件下土壤有机碳矿化的温度敏感性不同,N2处理土壤有机碳矿化的温度敏感性最低,N4处理最高。柑橘果园土壤有机碳矿化受高施氮量影响较大,低施氮影响不明显。随着施氮量的增加土壤有机碳矿化的温度敏感性增加,氮肥施用和温度的共同作用可能使柑橘林向大气中排放的CO2增加。     

Construction of Genetic Linkage Map of Bread Wheat （Triticum aestivum L.） Using an Intervarietal Cross and QTL Map for Spike Related Traits

Most often a genetic linkage map is prepared using populations obtained from two highly diverse genotypes. However, the markers from such a map may not be useful in a breeding program as these markers may not be polymorphic among the varieties used in breeding. For the past nine years, intraspecific maps have been gaining importance and such maps based on Swiss （PaiUard et al., 2003）, Japanese （Suenaga et al., 2005）, Australian （Chaimcrs et al., 2001） wheat varieties arc available. A map based on Indian wheat varieties however has not been reported. We constructed a genetic linkage map based on a cross between two Indian bread wheat （Triticum aestivum L.） varieties, Sonalika and Kalyansona. One hundred and fifty F2 individuals were analyzed for arbitrarilyprimed polymerase Chain reaction （AP-PCR）, random amplified polymorphic DNA （RAPD）, inter simple sequence repeats （ISSR）, Sequence Tagged Microsatelhte Sites （STMS）, Amplified Fragment Length Polymorphism （AFLP） markers, seed storage proteins and known genes. A linkage map was constructed consisting of 236 markers and spanning a distance of 3 639 cM with 1 211.2 cM for A genome, 1 669.2 cM for B genome, 192.4 cM for D genome and 566.2 cM for unassigned groups,