A New Disease of Cherry Plum Tree with Yellow Leaf Symptoms Associated with a Novel Phytoplasma in the Aster Yellows Group

A novel phytoplasma was detected in a cherry plum（Prunus cerasifera Ehrh） tree that mainly showed yellow leaf symptom. The tree was growing in an orchard located in Yangling District, Shaanxi Province, China. The leaves started as chlorotic and yellowing along leaf minor veins and leaf tips. Chlorosis rapidly developed to inter-veinal areas with the whole leaf becoming pale yellow in about 1-4 wk. Large numbers of phytoplasma-like bodies（PLBs） were seen under transmission electron microscopy. The majority of the PLBs was spherical or elliptical vesicles, with diameters in range of 0.1-0.6 μm, and distributed in the phloem cells of the infected tissues. A 1 246-bp 16 S ribosomal RNA（rRNA） gene fragment was amplified from DNA samples extracted from the yellow leaf tissues using two phytoplasma universal primer pairs R16mF2/R16mR1 and R16F2n/R16R2. Phylogenetic analysis using the 16 S rRNA gene sequence suggested that the phytoplasma associated with the yellow leaf symptoms belongs to a novel subclade in the aster yellows（AY） group（16SrI group）. Virtual and actual restriction fragment length polymorphism（RFLP） analysis of the 16 S rRNA gene fragment revealed that the phytoplasma was distinguishable from all existing 19 subgroups in the AY group（16SrI） by four restriction sites, Hinf I, Mse I, Sau3 A I and Taq I. The similarity coefficients of comparing the RFLP pattern of the 16 S rRNA gene fragment of this phytoplasma to each of the 19 reported subgroups ranged from 0.73 to 0.87, which indicates the phytoplasma associated with the cherry plum yellow leaf（CPYL） symptoms is probably a distinct and novel subgroup lineage in the AY group（16SrI）. In addition, the novel phytoplasma was experimentally transmitted to periwinkle（Catharanthus roseus） plants from the tree with CPYL symptoms and then back to a healthy 1-yr-old cherry plum tree via dodder（Cuscuta odorata） connections.     

Histological and computed tomographic evaluation of a parasitic conjoined twin in hybrid catfish (Ictalurus punctatus [Rafinesque] X Ictalurus furcatus [Lesueur])

There is growing use of hybrid catfish (Ictalurus punctatus ♀ X Ictalurus furcatus ♂) in commercial aquaculture to utilize hybrid vigour to improve production A conjoined twin specimen found during the course of production studies by the United States Department of Agriculture Catfish Genetic Research Unit (USDA‐CGRU) was submitted to the Aquatic Research and Diagnostic Laboratory (ARDL). After preliminary inspection, it was transported to Mississippi State University, College of Veterinary Medicine for further evaluation. The specimen was examined using both computed radiography and computed tomography antemortem. Following humane euthanasia, the specimen was examined both grossly and histologically. Tissues from both fish were also submitted for genetic analysis to determine whether twins were derived from the same egg. This report records the presentation and examination of a pair of conjoined hybrid catfish (I. punctatus X Ictalurus furcatus).     

京海黄鸡 GnRHR 基因克隆、生物信息学及组织表达分析

旨在根据GenBank 上公布的原鸡GnRHR基因序列设计2对引物,采用RT-PCR方法,从京海黄鸡肌肉组织中克隆GnRHR基因的编码区序列,并使用多种生物软件和在线工具对目的序列进行生物信息学分析,分析GnRHR基因与其他物种的同源性、蛋白质的理化性质、蛋白质序列的跨膜区、亚细胞定位、亲水性、潜在的磷酸化位点、保守结构域及该基因编码蛋白的二级结构、三级结构等。同时为GnRHR基因和β-actin基因各设计1对引物,采用RT-qPCR的方法研究GnRHR基因在京海黄鸡12个组织和器官中的表达情况。最终成功克隆了京海黄鸡GnRHR基因完整的编码区序列和部分侧翼序列,Blast结果显示,京海黄鸡GnRHR基因与原鸡、番鸭、藏羚羊、野猪、马、小鼠、大鼠、家兔、绵羊、人、牛、黑猩猩和斑马鱼的同源性分别为99．7％,86．7％,55．7％,54．6％,52％,51．6％,50．8％,50％,49．9％,49.6％,49．4％,47．4％和39．3％,并成功构建系统发育树。蛋白质结构分析显示,GnRHR蛋白分子量为45．432 kDa,理论等电点为9．55,包含20种氨基酸,其中,亮氨酸含量最高,占13．8％,赖氨酸含量最低,占0．7％;不稳定系数为65．35,显示该蛋白不稳定;脂溶指数为95．54,总平均疏水指数为0．312,为非水溶性蛋白;该蛋白不属于分泌型蛋白,主要存在于胞膜上,没有信号肽结构,存在16个潜在磷酸化位点和11个糖基化位点;保守结构域分析显示存在2个低复杂度序列和7段跨膜结构,跨膜分析显示该蛋白为7次跨膜蛋白,二级结构预测结果显示,在GnRHR二级结构中,α-螺旋占29．83％,β-折叠占17．18％,无规则卷曲占52．98％,GnRHR蛋白三级结构为复杂的7次跨膜螺旋结构,且为单链蛋白。组织表达分析显示,GnRHR基因主要在垂体中高表达,表达量显著高于其他组织,在其他11个组织和器官中表达量较低。     

核苷酸对大黄鱼生长性能、肠道形态和抗氧化能力的影响

为探究饲料中添加核苷酸对大黄鱼生长性能、肠道形态和抗氧化能力的影响,实验选取初始体质量为(7.71±0.02)g的大黄鱼幼鱼为研究对象,设置2个对照组,即高鱼粉对照组FM(鱼粉含量为45.0%)和低鱼粉对照组N0(鱼粉含量为31.5%),在低鱼粉对照组基础上分别添加0、75、150、300、600和1 000 mg/kg的核苷酸,共配制出7种等氮等脂饲料,在海水浮式网箱中进行为期63 d的养殖实验。结果表明:饲料中添加核苷酸对大黄鱼的存活率、饲料效率、摄食率、鱼体组成和形体指标没有显著影响(P0.05);但增重率随着核苷酸添加量的增加呈先升高后下降趋势,在添加量为300~600 mg/kg时显著高于其他处理组(P0.05)。随着核苷酸添加量的增加,血清中超氧化物歧化酶(SOD)和总抗氧化力(T-AOC)的活力呈先升高后下降的趋势,并在300 mg/kg时显著高于低鱼粉对照组(P0.05)。核苷酸对大黄鱼肠道肌肉层厚度和微绒毛高度有显著影响(P0.05),分别在添加量为150和300mg/kg时出现最大值;而对肠道褶皱高度无显著影响(P0.05)。结果表明,饲料中添加核苷酸能促进大黄鱼生长,改善肠道形态结构和增强机体的抗氧化能力。以增重率为指标,利用折线模型计算得到大黄鱼幼鱼饲料中核苷酸的适宜添加量为194.91 mg/kg。     

小黄鱼生物学特征与资源数量的演变

通过分析20世纪50年代以来小黄鱼(Larimichthys polyactis)资源动态和生物学特征演变,阐述其生物学特征与种群数量的变动机理,揭示种群兴衰的关键因素,评估小黄鱼的渔业风险。结果显示,小黄鱼资源动态由上世纪50年代的丰盛期演变至80年代的衰退低谷期、转而又发展到90年代初期的恢复期乃至此后的稳定丰厚期;年龄结构由50年代的10龄以上缩短到80年代的1~5龄、并继续缩小至90年代以来的1~3龄,世代周期缩短;以1龄鱼为初届性成熟年龄比例由50年代的少量(5%)扩大到80年代的中等(40%)、进一步演变到21世纪初以来的大量(74%~100%);个体绝对繁殖力从60年代初至90年代初显著提高,90年代以来无显著变化;生长参数K随资源丰厚与衰退而相应地减缓与加速,具有可逆的表型特征。小黄鱼资源数量和生物学性状的变动机理表明过度捕捞可导致开发性鱼类种群衰退,但鱼类种群为抗御过度捕捞而诱导生命史特征演变,并在适宜的渔业管理措施下有望实现鱼类种群恢复;性成熟提前和繁殖力提高是种群恢复的关键因素,针对性成熟和繁殖力的演变进化程度已接近极限或达最大潜力,如不控制捕捞强度,小黄鱼渔业面临着崩溃的风险。     

The effect of feeding diets containing deoxynivalenol contaminated corn on channel catfish (Ictalurus punctatus) challenged with Edwardsiella ictaluri

Channel catfish were fed practical corn‐soybean meal diets for 10 weeks that contained various weighed amounts of ground, dried field corn contaminated with 20 mg deoxynivalenol (DON) kg⁻¹. Weighed amounts of DON corn were blended with weighed amounts of ground, clean corn that contained no DON (0 mg kg⁻¹) to yield five diets that had 0, 2.5, 5.0, 7.5 and 10.0 mg DON kg⁻¹ of diet. Results show that catfish fed diets that contained DON for 7 weeks did not experience lower weight gains or poorer feed conversion ratios that were significantly (P > 0.05) different from control‐fed fish. Mortality of catfish during the 21‐day post‐challenge period indicate that catfish fed diets containing DON‐contaminated corn that provided at least 5.0 mg DON kg⁻¹ of diet had significantly (P < 0.05) lower mortality than catfish fed the control diet or the diet that provided 2.5 mg DON kg⁻¹ of diet. The presence of DON‐contaminated corn in the experimental diets did not significantly (P > 0.05) alter fish body weight gains and appeared to provide a protective effect for channel catfish challenged with the pathogenic bacterium Edwardsiella ictaluri.     

Effects of dietary arginine supplementation on growth,protein optimization and innate immune response of channel catfish Ictalurus punctatus (Rafinesque 1818)

Juvenile channel catfish Ictalurus punctatus were fed casein–gelatin‐based semi‐purified test diets with graded amounts of arginine (ARG, 0.5%, 1%, 2% and 4% of dry diet) for a 6‐week period, to evaluate the effects of dietary ARG supplementation on performance indicators, levels of plasma ARG, ornithine, citrulline, glutamine and glutamate, as well as selected innate immune parameters. Incremental dietary ARG resulted in significant improvements in weight gain, feed efficiency and protein efficiency ratios, protein retention, protein deposition and arginine deposition. Fish fed the 4% ARG diet had significantly higher values for all performance indicators, although these values were not different from those of fish fed the 2% ARG diet with the exception of protein retention, protein deposition and arginine deposition. Conversely, fish fed the 0.5% ARG diet had significantly lower values for all analysed plasma amino acids. Similarly, both phagocyte superoxide anion production and neutrophil respiratory burst were significantly decreased in fish fed the 0.5% ARG diet. These results demonstrate that the indispensability of dietary ARG is not only related to optimized protein utilization for growth but also its supplementation to fish diets may significantly impact several aspects of the immune system, including phagocyte function and thus improve overall fish health.     

金黄口服液提取工艺的优化

为获得金黄口服液的最佳提取条件,本试验通过UV法测定不同提取条件下取得的复方口服液中盐酸小檗碱的含量,选取含量最高的提取条件作为最佳提取条件。在波长350 nm处,盐酸小檗碱在浓度为1.5625～25 μg/mL范围内,线性关系良好,回归方程为y=0.1493x+0.0583,R²=0.9996;口服液最佳提取条件为：浸泡45 min、加10倍量的水、煎煮45 min、共煎煮3次;结合实际生产和成本因素,确定中药复方水提工艺为：浸泡30 min、加10倍量的水、煎煮30 min、共煎煮3次,均接近正交方案中的最高值。     

麦类品种与种质资源对大麦黄矮病毒GAV的田间抗性鉴定

为明确不同小麦品种对大麦黄矮病毒(Barley yellow dwarf viruses, BYDV)GAV的抗性差异,采用堆测法田间人工接种鉴定了215份小麦品种及种质资源的抗病性.结果表明,不同材料对黄矮病的抗病性存在较大差异,00中13、中4、西农979、临优21号、878和H9020-17-25-6-4的病情指数分别为18.9、9.6、16.3、21.1、21.1和22.6,表现出较好的抗病性;荔高6号、偃展4110、新麦13、豫麦49、徐麦856、金穗1号、中国春和7182的病情指数分别为68.9、55.2、58.5、54.0、56.2、51.5、55.8和53.0,均高于50.0,表现为高感;其余材料病情指数在25.0~50.0之间,表现为感病.此外,红须麦、豫麦34、小冰麦和晋麦47初期表现感病,后期恢复健康,有一定的耐病性.抗病性不同的小麦品种感染GAV后,对小麦产量的影响差异很大,抗病品种如中5产量损失为5.6%,中4产量损失7.6%,耐病品种小冰麦、晋麦47和红须麦的产量损失依次为19.9%、15.2%和18.3%,而感病品种荔高6号产量损失达到47.5%.     

黄色黏虫板在3种果园对蚜虫及其天敌的诱集作用

为减少黄色黏虫板(简称黄板)对果园害虫天敌的影响,明确黄板在果园中的使用方法,于2014年5月4日至9月30日在山西省太谷县的苹果、梨、桃园中进行了黄板诱集蚜虫及其天敌的动态调查。结果表明:在3种果园黄板可以诱集8目32科的昆虫,主要类群为蚜虫、叶蝉、瓢虫、食蚜蝇、草蛉和蚜茧蜂等。在苹果园和桃园中诱蚜量呈双峰谷线状,在梨园中呈三峰状;各果园寄生性天敌的数量变化趋势与蚜虫一致且有跟随效应,但苹果园和梨园诱集的捕食性天敌均在8月下旬至8月底出现高峰期,而桃园中捕食性天敌水平一直较高。3个果园在花期至幼果期即5月4日至6月22日益害比最小,苹果园仅为1:202.8,而在其它时段的益害比均大于1:150,甚至最高可达1:19.4,表明在果园中使用黄板的害处大于益处,建议在果园中慎用黄板诱蚜。