抗寒茶树品种的筛选研究

为了获得抗寒茶树品种或资源,提高茶园防冻减灾的能力,分别采用田间自然鉴定法、叶片解剖结构和电导法对24个茶树品种和优质资源的抗寒性进行了鉴定。结果表明,‘浙农113’、‘中茶102’、‘翠峰’等茶树品种在以上抗寒性鉴定方法的评价结果中均表现为较强抗寒性,属于抗寒茶树品种。不同鉴定方法评价茶树抗寒性的结果有一定的差异,综合鉴定比较的结果较单一鉴定方法的结果更可靠,更符合生产实际。     

行距和穴内插秧密度对寒地粳稻产量和品质性状的影响

为了明确寒地粳稻优质高产栽培的最佳行距和穴内插秧密度,以优质粳稻品系‘东农09-51’为试验材料,采用随机区组试验设计,研究行距和穴内插秧密度及其互作效应对寒地粳稻产量和品质的影响。结果表明,行距对产量及其构成因素、加工品质和蒸煮食味品质性状及RVA特性（除了回冷恢复值）都有显著或极显著的影响,穴内插秧密度对产量及其构成因素、加工品质和蒸煮食味品质性状及RVA特性（除了最低粘度和最终粘度）有显著或极显著的影响,二者互作效应对产量及其构成因素、加工品质和蒸煮食味品质性状及RVA特性都有显著或极显著的影响。其中处理A2B2的产量为最高,为9775 kg/hm2,但其蒸煮食味品质并不是最佳的,较A3B2和A2B3处理稍差,这一点与RVA谱特性分析结果是一致的。从高产和优质的协调以及高效角度考虑,首先要充分利用行距和穴内插秧密度的互作效应,行株距30 cm×10 cm、穴内插秧4棵/穴更有利于提高寒地粳稻产量和改善品质。     

基于多重串联式PCR的基因碟片技术检测转基因作物

【目的】建立基于多重串联式PCR（multiplexed tandem PCR,MT-PCR）的基因碟片技术,并使之应用于转基因作物的大量、快速和稳定地检测。【方法】以转基因作物研究中常用的调控序列NOS终止子、FMV35S启动子及外源基因NPTⅡ、Cry1Ab、Cry1Ab/Ac、CP4-EPSPS、PAT和玉米内源基因IVR、棉花内源基因sad1、菜籽粕内源基因PEP为检测对象,针对每个基因设计内外2对引物,先进行一次循环数较少（10-20 cycles）的高通量多重PCR,以便在均匀地扩增各基因和调控序列的同时避免引物之间的竞争,然后利用巢式荧光定量PCR检测各个基因和调控序列,最后根据扩增曲线和熔融曲线分析结果。【结果】该方法能够快速（＜2 h）、高通量、准确地（＞0.000292 ng）检测出棉花、玉米、大米、菜籽粕中的多种转基因成分,可以分辨出3种转基因物种,适合大批量检测。【结论】该方法适合转基因作物的高通量、定量检测,具有较好的应用前景。     

Development of real-time PCR systems based on SYBR? Green I and TaqMan? technologies for specific quantitative detection of Phoma tracheiphila in infected Citrus

Real-time PCR assays based on SYBR? Green I and TaqMan? technologies were developed for in planta detection and quantification of Phoma tracheiphila, the mitosporic fungus causing ‘mal secco’ disease on citrus. Primers and a hybridization probe were designed on the basis of the internal transcribed spacer (ITS) region of the nuclear rRNA genes. The real-time PCR assays were compared with a classic isolation method in two separate experiments carried out on 6 and 24 month-old sour orange seedlings, artificially inoculated with a conidial suspension of the pathogen. Both technologies made it possible to follow the progression of infection by P. tracheiphila, enabling detection and quantification of the target fungus prior to the development of symptoms. The detection limit was 10 copies of the cloned target sequence and 15 pg of genomic DNA extracted from fungal spores. The values of the cycle threshold (Ct) were linearly correlated with the concentration of the target DNA, indicating that the method is suitable as a qualitative and quantitative assay. The presence of non-target fungal DNA had no effect on the specificity of the assay, but resulted in a 10-fold reduction of sensitivity. Total inhibition of the reaction occurred when conidia of the target pathogen were mixed with an organic soil substrate before extracting DNA using the standard protocol, while an alternative purification kit resulted in a significant decrease in sensitivity. Compared to classic methods, real-time PCR proved faster and easier to perform and showed a higher sensitivity. These results suggest that real-time PCR, based on both chemistries, has a great potential for early diagnosis of ‘mal secco’ disease and for quantitative estimation of fungal growth within host tissue.     

Are Acartia tonsa cold‐stored eggs a suitable food source for the marine ornamental species Amphiprion polymnus? A feeding study

Copepods are the natural food items for marine fish larvae but are still difficult to be cultured on a continuous basis. Therefore, suitable storage techniques for copepod subitaneous eggs are of great interest. Cold‐stored copepod embryos still develop and retain a physiological activity during the cold storage period. As a consequence, their nutritional quality may change over the time of storage with these changes possibly affecting both eggs viability and larval fish survival and development. In this study, the clownfish, Amphiprion polymnus, was used as an experimental model to compare, for the first time, the effects of diets based on enriched rotifers and Artemia (control), Acartia tonsa copepods of a continuous culture and A. tonsa originated from 6 months cold‐stored eggs. The effects of the different diets were tested through morphometric, molecular and biochemical approach. This study demonstrated that after a 6 months cold storage period, the quality of copepods obtained from those eggs was suboptimal for A. polymnus larval rearing. In fact, larvae fed those copepods showed lower growth and survival performances respect to the other experimental groups.     

Development of a multiplex polymerase chain reaction to detect five common Gram-negative bacteria of aquatic animals

A multiplex polymerase chain reaction (m-PCR) technique was developed as a rapid and accurate diagnostic tool for identifying five major Gram-negative bacilli -Vibrio vulnificus, V. parahaemolyticus, Aeromonas hydrophila, Chryseobacterium meningosepticum and Edwardsiella tarda- that cause major diseases in cultured aquatic animals in Taiwan. The expected amplicons for V. vulnificus, V. parahaemolyticus, A. hydrophila, C. meningosepticum and E. tarda were 410, 368, 685, 180 and 230bp, respectively. The assay was shown to be specific for the target pathogens. The sensitivities of detection were estimated to be 20.5fg～200pg of genomic DNA or 10(2) ～10(4) colony-forming units (cfu) of bacterial isolates when adopted as PCR templates. The m-PCR was capable of simultaneously amplifying target fragments from bacterial genome DNA mixed with the DNA extracted from viscera and tissues taken from fish without affecting the performance of the method.     

Diversity of Flavobacterium psychrophilum and the potential use of its phages for protection against bacterial cold water disease in salmonids

Flavobacterium psychrophilum causes rainbow trout fry syndrome (RTFS) and cold water disease (CWD) in salmonid aquaculture. We report characterization of F. psychrophilum strains and their bacteriophages isolated in Chilean salmonid aquaculture. Results suggest that under laboratory conditions phages can decrease mortality of salmonids from infection by their F. psychrophilum host strain. Twelve F. psychrophilum isolates were characterized, with DNA restriction patterns showing low diversity between strains despite their being obtained from different salmonid production sites and from different tissues. We isolated 15 bacteriophages able to infect some of the F. psychrophilum isolates and characterized six of them in detail. DNA genome sizes were close to 50 Kbp and corresponded to the Siphoviridae and Podoviridae families. One isolate, 6H, probably contains lipids as an essential virion component, based on its chloroform sensitivity and low buoyant density in CsCl. Each phage isolate rarely infected F. psychrophilum strains other than the strain used for its enrichment and isolation. Some bacteriophages could decrease mortality from intraperitoneal injection of its host strain when added together with the bacteria in a ratio of 10 plaque-forming units per colony-forming unit. While we recognize the artificial laboratory conditions used for these protection assays, this work is the first to demonstrate that phages might be able protect salmonids from RTFS or CWD.     

小胸鳖甲各龄期幼虫过冷却点的测定

[目的]了解新疆荒漠甲虫准噶尔小胸鳖甲幼虫耐寒性的发育阶段性变化.[方法]对新疆荒漠甲虫准噶尔小胸鳖甲各龄期幼虫的过冷却点进行测定.[结果]同一虫期个体间的过冷却点出现不同程度的变异,但均服从正态分布.不同虫期的过冷却点差异显著,其中1龄的过冷却点最低(-19.4～-24.1℃),且个体差异不大.老熟幼虫的过冷却点个体差异较大,其中7龄幼虫中过冷却点在个体间差异达到15.7℃(-5.6～-21.3℃).[结论]老龄幼虫最有可能是该虫在荒漠环境地区越冬的虫态.     

基于灰色神经网络与马尔科夫链的城市需水量组合预测

【目的】针对城市需水量预测系统具有非线性和随机波动性的特点,建立基于马尔科夫链修正的组合灰色神经网络预测模型,以提高模型的预测精度。【方法】比较分析灰色GM(1,1)模型、BP神经网络模型以及二者线性组合的灰色神经网络预测模型的预测效果,建立基于马尔科夫链修正的组合灰色神经网络预测模型,并以榆林市2000-2009年的用水量实际数据为研究对象,通过实例比较分析模型的检验预测精度。【结果】经马尔科夫链修正处理后,建立的基于马尔科夫链修正的灰色神经网络组合模型的预测精度更高,预测误差的绝对值均小于4%,且均方差σ为1.00,小于组合灰色神经网络模型与GM(1,1)模型、BP神经网络模型预测误差值的均方差。【结论】基于马尔科夫链修正的组合灰色神经网络需水量预测模型,对城市需水量的预测优于灰色神经网络及各单项预测模型,不仅预测精度高,而且能同时反映出数据序列发展变化的总体趋势和系统各状态之间的内在规律,适合描述随机波动性较大的预测问题。