Pathotypes and Genetic Diversity of Blumeria graminis f.sp.hordei in the Winter Barley Regions in China

Barley powdery mildew caused by Blumeria graminis f. sp. hordei (Bgh) is one of the most destructive foliar diseases of barley in the winter barley region of China. The evaluation and assessment of the virulence and diversity of Bgh populations help to determine effective sources of resistance to the pathogen. 515 isolates were collected from seven populations of Bgh on cultivated barley in seven geographically distant locations in 2006. Their virulence was determined by inoculation onto 26 differential host lines. All of the isolates belonged to 58 pathotypes and 13 of which included 81% of these isolates. The most abundant pathotype was pathotype 0047 (18.3%), the second most abundant was pathotype 0045 (11.8%) and the third most abundant was pathotype 0057 (7.8%). Most of virulent genes investigated in this study showed similar frequencies in the seven different areas. These indicate that the seven locations may be in a uniform epidemiological region and barley cultivars in these areas may have similar genetic background. Diversities within these populations and distances between these populations measured by KOIND package were different. Correlations were not found between the genetic distance and the geographical distances between different locations. This suggested that long distance spread and local epidemics existed in the major winter barley growing regions in China.     

大麦白粉菌致病型鉴定初报

用17个“Pallas”近等基因品系和3个已知抗性基因的品系作为鉴别寄主,对中国东南沿海4个地区(浙江海盐、杭州,江苏盐城和福建莆田)的220个大麦白粉菌(Blumeria graminisf.sp.hordei)单孢菌落的致病性进行鉴定。结果显示,这些菌株可划分为13个致病型,0023042(42.3%)和0023052(20.0%)为优势致病型;所有的致病型含有4-7个毒性基因,均含有毒性基因Va22,Vra和VRu2;该群体对10个分别含有Mla1,Mla3,Mla6,Mla7,Mla9,Mla12,Mla13,Mla23,Mlat和Mlg抗性基因的品系表现为低毒性,它们是抗大麦白粉病育种中可利用的抗病基因。     

陕西关中地区苹果褐斑病菌遗传多样性分析

为了揭示苹果褐斑病菌(Diplocarpon mali)的群体遗传多样性,利用ISSR分子标记技术,通过ISSR-PCR扩增,对分离自陕西省关中平原7个县(区)的64个菌株进行遗传多样性分析。结果显示,陕西关中地区苹果褐斑病菌的遗传多样性丰富,不同县(区)病菌的遗传多样性存在差异,渭南市白水县病菌的遗传多样性水平相对较高,咸阳市乾县相对较低。群体之间的遗传分化系数不同,遗传变异主要发生在群体内;不同地区群体之间存在不同程度的基因交流,其中关中中部群体的基因交流更广泛;关中东部、中部、西部地区之间的群体也存在基因交流,其中东部和西部之间群体的基因交流更频繁。NTsys聚类结果表明,种群之间的遗传亲缘关系与分离株的地理来源没有明显的相关性。     

华重楼野生种群表型变异研究

目的 综合分析华重楼Paris polyphylla var. chinensis (Franch.) Hara野生种群的表型变异规律,为其种质资源保护、药材资源开发利用和繁育改良提供理论支撑。方法 以我国分布区内7个野生华重楼地理种群为研究对象,对其17个表型性状进行系统测量,利用方差分析、变异系数分析、多重比较分析和相关性分析等多种数理统计分析方法,探讨其表型变异规律及其与地理气候因子的关系。结果 除叶片数和花萼片数外,野生华重楼其他15个表型性状在种群间和种群内有显著或极显著的差异;种群内的变异(方差分量占比为52.19%)大于种群间的变异(方差分量占比为26.39%),表型性状的平均分化系数为34.27%,种群内变异是华重楼表型变异的主要来源;各表型性状的平均变异系数为17.72%,变异幅度为6.83%~29.95%,花梗长、叶面积、花萼面积和株高的变异幅度远大于平均变异系数,叶片数、花萼片数、果球横径、果球纵径、果球横纵比和种子千粒质量的变异幅度远小于平均变异系数,表明华重楼花、叶的数量指标及果球、种子的性状稳定性大于叶片形态性状的稳定性;相关分析表明,大多数表型因子与地理气候因子之间存在显著或极显著的相关性。结论 广东清远地理种群变异最为丰富,可作为种质资源收集保存的优选对象。     

基于ISSR分析湖北省枫香资源的遗传多样性

为了给湖北省枫香资源的保护与利用提供科学依据,选取湖北省远安（YA）、红安（HA）、赤壁（CB）、竹溪（ZX）、南漳（NZ）、利川（LC）和武汉（WH）的枫香资源为研究对象,并以安徽黄山（HS）、重庆丰都（FD）、江西铜鼓（TG）和海南霸王岭（BWL）的枫香资源为对照,采用ISSR分子标记对这11个枫香群体的335份个体样品进行了遗传多样性分析。结果表明：（1）湖北省7个枫香群体的215份样本共扩增得到334个条带,其中多态性条带306条。群体内的遗传变异为84.19%,群体间遗传变异为15.81%,基因流（N_m）为2.663 3。在群体内部,HA和WH的遗传多样性最丰富,ZX的遗传多样性最低。ZX和LC的亲缘关系最近,YA和LC的亲缘关系最远。当遗传相似系数为0.94时,可将7个群体分为4个大类,NZ、LC和ZX群体聚为一类,YA和WH群体聚为一类,HA群体、CB群体分别单独聚为一类。 （2）11个枫香群体共扩增出349个条带,其中多态性条带320个,平均多态性百分比为91.69%。不同群体的多态位点百分率范围为54.29% ~ 77.14%,平均值为67.01%。WH和HA的遗传多样性最丰富,BWL的遗传多样性最低。11个群体中,18.01%的遗传变异存在于群体间,群体内的遗传变异为81.99%,基因流（N_m）为2.276 5。在群体内部,HA和WH的遗传多样性最丰富,BWL的遗传多样性最低。遗传相似度和遗传距离均显示,ZX和TG的亲缘关系最近,TG和BWL的亲缘关系最远。当遗传相似系数为0.93时,可将11个群体分为四大类：第1类包括NZ、ZX、TG和LC;第2类包括FD、HA和CB;第3类包括YA、WH和HS;第4类是BWL单独一类。 可见湖北省7个枫香群体的聚类结果与其自然地理分布大致吻合,可分为东部、中部和西部三大类;4个省外枫香群体中,除BWL单独聚成一类外,其余3个群体（TG、FD和HS）并未与湖北省内地理位置临近的群体聚为一类。     

2006年广西稻瘟病菌生理小种的组成与分布研究

利用中国统一的7个稻瘟病菌鉴别品种对2006年采自广西桂南、桂中、桂北和高寒山区4个不同稻作区28个县（市）的139个稻瘟病菌有效单孢菌株进行生理小种鉴定,结果共鉴定出7群27个生理小种,其中ZB为优势种群,出现频率高达67．62％,ZB1为优势小种,出现频率为25．18％,其次为ZB9小种,出现频率为19．42％,其他小种出现的频率较低。对不同稻作区稻瘟病菌生理小种鉴定结果表明,不同稻作区的稻瘟病菌生理小种类型有明显差异,而同一稻作区内的稻瘟病菌生理小种类型则较丰富。     

Distribution of mating types and genetic diversity induced by sexual recombination in Setosphaeria turcica in northern China

Mature ascocarps and ascospores in the heterothallic ascomycete fungus, Setosphaeria turcica, were successfully produced in Sach’s medium with barley culm as the mating stimulator after four weeks’ coincubation of two opposite mating type isolates at 25°C in darkness. A single isolate could not produce ascospores or ascocarps. The ascocarps were produced on the exposed surface and embedded parts of barley culm or in the upper layer of the medium. The asci linked themselves to ascocarp with their short handles and assembled at the bottom of the ascocarp. Many asci had four to six colorless mature ascospores with one to six septa. But asci with eight ascospores were also found. Using isolate 9914 and isolate 9961 as standard testers for mating types (MAT1 and MAT2), respectively, 94 isolates of S. turcica collected from northern China in 1999, 2003, and 2004 were grouped into three mating types: MAT1 (53 isolates), MAT2 (31 isolates) and MAT12 (10 isolates). The MAT12 isolates, which were first found in China, were compatible with not only MAT1 isolates but also MAT2 isolates. No MAT12 isolates were found in 1999, but 2 MAT12 isolates and 8 MAT12 isolates were found in 2001 and 2003, respectively. The geographic distribution of different mating types was unequal among locations. Generally the frequency of MAT1 was significantly higher than that of MAT2 and MAT12. The unequal distribution of mating types suggested a low frequency of genetic recombination. The pathogenicity of different mating type isolates was tested on the susceptible corn inbred B37 and the results revealed that the disease latency period, disease incidence, lesion area and conidia production were not significantly different among the three mating type groups. However, the pathogenicity of the progeny isolates of isolate 99-12 (MAT2, race 1) and isolate 99-15 (MAT1, race 0) was significantly different from the parent isolates, isolate 99-12 and isolate 99-15, suggesting that sexual recombination could cause significantly virulence variation in S. turcica. Random amplification of polymorphic DNA (RAPD) analysis also revealed high genotype diversity among the progeny isolates, indicating that the sexual recombination could also produce significant genetic variation in the fungal pathogen.     

山西草莓斑驳病毒cp基因克隆及遗传多样性分析

旨在研究草莓斑驳病毒（SMoV）在山西省的分布、结构变异及遗传多样性。从山西省7个主要草莓种植区随机采取159份草莓叶片进行RT-PCR病毒检测，结合在线数据库，使用MEGA5构建系统发育树，采用SDTv 1.2软件进行序列相似性分析，运用DnaSP v5.10软件分析cp基因的遗传多样性，采用RDP v.4.31 软件检测SMoV中的重组事件。RT-PCR检测后发现，从山西省各地区收集的159份草莓叶片中有65份样品呈阳性，检出率为38.46%。这些阳性样品经过分离、测序、克隆，获得16个SMoV分离物。结合在线的19个SMoV分离物系统进化树分析显示，35个SMoV分离物被分成两组，组1包含28个分离物，均来自中国；组2包含7个分离物，分别来自加拿大、日本和美国。经选择压分析和中性检验，组1和组2 SMoV分离物之间存在显著的遗传差异，且中性检验均为负值，SMoV种群呈扩张状态。序列相似性分析表明，35株分离物的核苷酸同一性为81.34%~100%，氨基酸的一致性为94.77%~100%，呈现出较高的相似性。可见：SMoV存在较高的遗传变异，负选择压力可能是导致SMoV遗传多样性的原因。     

中国奶牛牛支原体流行病学调查分析

为调查中国奶牛养殖地区牛支原体(Mycoplasma bovis)的流行情况,从2013—2019年,在中国6个奶牛养殖区域集约化牧场,通过随机采样收集犊牛鼻拭子1 878份,对样品进行M.bovis分离鉴定,并分析M.bovis阳性率时间分布,空间分布以及犊牛日龄与M.bovis阳性率的关系。结果表明:1)空间上,中国不同奶牛养殖地区,M.bovis阳性率是不同的。其中西北区(38.7%)高于华东区(35.6%)、华北区(33.6%)、华中区(30.8%)、华南区(29.9%)和西南区(27.1%)。2)时间上,2013—2015年M.bovis 阳性率平均值为47.9%,三年间M.bovis阳性率差异不显著(P>0.05)。2016—2019年,M.bovis阳性率平均值为21.1%,四年间M.bovis阳性率无显著性差异。2013—2015年M.bovis 阳性率显著高于2016—2019年M.bovis 阳性率(P<0.05)。3)季节上,夏秋季节M.bovis平均阳性率为31.3%,冬春季节M.bovis阳性率平均值为33.9%,不同季节M.bovis阳性率差异不显著(P>0.05)。4)通过线性回归分析,表明M.bovis 阳性率与犊牛日龄呈正相关。综上,2013—2019年,中国奶牛养殖地区M.bovis阳性率平均值为32.6%,M.bovis在中国呈长期流行趋势。季节并不影响M.bovis在犊牛中的流行。M.bovis阳性率与犊牛日龄呈线性正相关。本研究丰富了中国奶牛养殖地区牛支原体流行情况数据,从病原学角度为预防和临床用药提供重要的数据支撑。     

烟蚜茧蜂规模扩繁冬寄主筛选研究

为筛选出烟蚜茧蜂(Aphidius gifuensis Ashmead)规模扩繁中的最佳繁蚜冬寄主,比较了自然条件下7种不同冬寄主上烟蚜的种群数量变化,烟蚜茧蜂对各寄主上烟蚜的防控效果及僵蚜羽化率.结果表明:烟蚜在7种寄主上种群数量的自然增长从大到小依次表现为烤烟K326、芥菜、芥菜型油菜、甘蓝型油菜、萝卜、红菜薹、白菜;芥菜上烟蚜茧蜂的寄生率为70.56%,显著高于其他寄主;7种冬寄主上僵蚜羽化率均高于70%,其中芥菜最高,为97.33%,萝卜最低,为72.87%,二者差异具有统计学意义.烟蚜茧蜂均能寄生7种冬寄主植物上所饲养的烟蚜且僵蚜羽化率相对较高,但总体而言芥菜上烟蚜的种群数量增长较快,烟蚜茧蜂的寄生率和僵蚜羽化率均最高,可利用芥菜来替代烟草作为冬季繁蚜寄主,也可在春烟种植前对芥菜田释放烟蚜茧蜂使其在田间建立自然种群.     

Sensitivities of Phytophthora infestans to Metalaxyl, Cymoxanil, and Dimethomorph

The isolates of Phytophthora infestans on tomato in the Guangxi Zhuang Autonomous Region, China, were determined for the sensitivities to metalaxyl, cymoxanil and dimethomorph to give the basic information for integrating disease management. Sensitivities were tested by measuring the radial growth on agar medium amended with fungicide, compared with the floating-leaf-disk method. 239 isolates were collected from eight tomato growing areas during 2000-2006. The testing results indicated that the frequencies of sensitive, intermediate, and resistant isolates to metalaxyl were 42.26, 35.98, and 23.53%, respectively. Variations in sensitivities amongst isolates from different areas or different years were very high for metalaxyl. All isolates from Tianlin and Wuxuan were sensitive to metalaxyl, but the metalaxyl-resistant isolates predominated in Tianyang, with the frequency of 51.35%. The EC50 values of certain isolates from Tianyang were higher than 500 μg·mL^-1 and their resistance levels were over 100 000 folds. Cymoxanil has been used for nearly 10 years in Guangxi, and dimethomorph has been used for 5-6 years. However, there was no decrease in sensitivity of P. infestans populations and the sensitivities of the pathogen were nearly normally distributed. Hence, their mean ECs0 value [cymoxanil （0.1647 ±0.0255） μg·mL^-1, dimethomorph （0.0970 ± 0.0052） μg·mL^-1 could be used as the baseline sensitivities for monitoring the field resistance development. The comparison with the floating-leaf-disk method indicates that both the techniques provided equivalent results. These studies suggested that metalaxyl can be continuously applied in Tianlin, Wuxuan, and Nanning due to the resistant isolates that have not been found, while for those areas with resistant isolate, the use of metalaxyl should be reduced or alternated, and cymoxanil or dimethomorph was recommended for controlling late blight disease of tomato.     

东南沿海地区大麦白粉菌群体毒性分析

利用19个含有已知抗大麦白粉病基因的鉴别寄主,对采自福建莆田、江苏盐城、浙江海盐和杭州4个地区的213个大麦白粉病菌株,进行毒性基因鉴定和毒性频率分析.结果显示,在被调查的白粉菌群体中,尽管某些毒性基因的频率在不同地区有所不同,但大部分亚群体中毒性基因的频率在不同地区极为接近;在东南沿海4个地区大麦抗白粉病育种中,具有利用价值的抗病基因是:Mla11,Mlmw,Mlkb,Mlmlw,Mlk2,(Mla10 MlDu2)和(MlKl Mla8),无利用价值的抗病基因是:Mla8,Mlnn,Mlh,(Mlra Mla8),(MlRu2 Mla8),(MlBw Mla8),(Mlra MlDr2)和(MlLa Mla8).     

玉米内生真菌的分离与鉴定

对不同品种的玉米种子及不同地区、不同部位的玉米内生真菌进行分离和鉴定。结果显示,8个不同品种的玉米种子,内生真菌检出率为25%,不同玉米品种的种子内所含的内生真菌种类和数目都不同,最高的检出率为40%,最低的检出率为10%。从玉米种子中分离得到的92个菌株,经形态学鉴定,分别属于14个属,出现频率最高的是木霉属,为34.78%。从不同地区的玉米组织中一共分离得到内生真菌168株,其中10株未产孢的没有鉴定,其他菌株经形态学鉴定分别属于26个属,以青霉属、曲霉属、头孢霉属为主,占内生真菌分离物总数的41.07%。对形态学鉴定为白僵菌属的10个菌株进行分子鉴定,均确定为球孢白僵菌(Beauveria bassiana)。此外,球孢白僵菌作为玉米的内生真菌,在国内还是首次分离得到。     

入侵我国中南三省(区)草地贪夜蛾的单倍体型和群体遗传结构分析

【目的】揭示入侵我国中南三省(区)的草地贪夜蛾Spodoptera frugiperda种群间可能存在的遗传变异,探讨其入侵来源、入侵路线和扩散方式,为制定有效控制和阻击草地贪夜蛾的方案提供理论依据和遗传信息。【方法】利用分子标记技术从线粒体DNA和核DNA角度研究草地贪夜蛾种群的遗传多样性和种群结构。对采集自我国广东、广西和湖南3个省(区)的草地贪夜蛾,利用COI和Tpi基因片段进行了单倍体型分析,利用9个微卫星标记分析种群间的遗传结构和遗传多样性。【结果】供试样品中,除了湖南张家界群体有2个样品为COI玉米型单倍体型、1个样品为杂合型,其他群体的个体均为COI水稻型单倍体型。基于SSR标记的群体遗传分析结果表明,被检测的群体遗传多样性较巴西等国家的美洲群体低,各群体遗传分化不明显。广东东莞群体与湖南张家界群体的遗传距离最近,与广西南宁群体的遗传距离较远。【结论】遗传距离与实际距离之间不一定呈正相关,通过遗传结构的分析有助于推测草地贪夜蛾扩散过程中,天气背景场、地理隔离和人为传播各自所起的作用。入侵湖南省张家界的草地贪夜蛾出现了低频率COI玉米型和高频率的CO I水稻型单倍体型,说明虫源的非单一性;草地贪夜蛾在入侵地定殖时间较短,目前未形成明显的遗传分化。     

天山樱桃种质资源表型多样性研究

在对新疆4个天山樱桃野生居群16个表型性状比较分析的基础上,采用性状变异分析、聚类分析和主成分分析等方法研究其遗传多样性水平和遗传结构。结果表明:表型变异系数、Simpson指数和Shannonweaver指数均为裕民县野生居群最大,分别为21.15%、0.98和3.97,喀拉峻野生居群最小,分别为16.18%、0.82和2.31。16个表型性状的变异系数为8.67%~48.78%,其中单果质量、果实纵径和种核质量的变异系数较高,花瓣宽、雄蕊数目和可溶性固形物质量分数的变异系数较低;平均表型分化系数为25.77%,表明居群内变异大于居群间变异。在阈值为5.27时,4个天山樱桃野生居群被分为3大类,特克斯与喀拉峻野生居群欧式距离最小(3.75),大西沟与裕民县野生居群欧式距离最大(7.16);16个表型性状的主成分分析中,前4个主成分的累积贡献率达76.02%。裕民县野生居群的表型多样性最丰富,特克斯与喀拉峻野生居群遗传关系最近,果实纵径、果实横径、雌蕊长、1a生枝长度和花冠直径是天山樱桃进行表型观测时的重要指标。     

食源性大肠杆菌的PCR检测

为了将PCR技术更好地应用于食品安全检测,利用多重PCR对大肠杆菌uidA基因和7种毒理基因进行扩增,并对人为污染大肠杆菌的水和牛奶进行普通和实时定量PCR限度检测。结果发现,2组多重PCR反应即可完成uidA和7种毒理基因的检测;普通PCR和实时定量PCR检测限度相同,对人为污染水检测限度为2.8×10 cfu/mL, 对人为污染牛奶的检测限度为2.8×10⁴ cfu/mL。     

新疆棉花枯萎病菌的RAPD分析

将供试新疆棉花枯萎病菌32个代表菌株及3个标准菌株进行RAPD分析，筛选出14个随机引物，共产生252个RAPD分子标记，其中79.4％具有多态性。通过聚类分析将供试菌株划分为5个RAPD组，此结果与常规鉴别寄主反应法划分生理小种的结果基本一致，RAPD分析为传统方法的结果提供了分子水平的证据，从分子水平上证明新疆棉花枯萎菌群体内7号小种占绝对优势。RAPD可作为一种标记手段用于棉花枯萎病菌生理小种及遗传分化的检测。     

粉枝莓ISSR遗传多样性和果实香气成分多样性分析

为明确西藏粉枝莓的多样性和亲缘关系,利用ISSR分子标记技术和HS-SPME/GC-MS技术,对粉枝莓7个居群的遗传多样性和果实香气成分多样性进行研究。结果表明:11条引物共得到102个清晰的扩增条带,其中多态性条带99个,PPB达到97.06%,香农信息指数I为0.490 9,表明粉枝莓具有较高的遗传多样性水平。Nei’s遗传结构(GST,0.287 2)和分子变异分析(AMOVA)的结果表明大部分的遗传变异分布在居群内(71.28%)。通过遗传分化系数GST估计的基因流为1.241 1。45个粉枝莓果实共检测出28种挥发性成分,基于果实香气成分的欧式距离为0.028 6~0.144 6。利用Mantel检验结果表明粉枝莓ISSR遗传距离和果实香气成分欧式距离无显著相关性(r=-0.153 2,P=0.676 0)。采用28种香气成分对45个样本进行主成分分析,前4个主成分的累积贡献率达到74.20%,较好的反映了粉枝莓果实香气成分的组成情况。研究表明,粉枝莓存在较丰富的遗传多样性,为西藏粉枝莓资源的进一步研究和利用提供参考。     

宁夏5个野生酸枣种群遗传变异性分析

以收集到的宁夏5个野生酸枣种群及3个栽培品种的叶片和果实为材料,测定其表型性状、生理和果实品质指标,分析种群的遗传多样性。结果表明：5个野生酸枣种群的表型性状平均变异系数为4.46%,果实品质平均变异系数为3.49%,叶片生理指标平均变异系数为15.89%,说明果实的遗传性状变化较小,而叶片的变化较大;5个野生酸枣种群遗传变异性大小的顺序为中宁县牛首山种群>中卫市日照山种群>银川市贺兰山滚钟口种群>灵武市白芨滩种群>吴忠市红寺堡种群。聚类分析表明,3个栽培品种为第Ⅰ类;5个野生酸枣种群中,中卫市日照山种群与中宁县牛首山种群为第Ⅱ类;银川市贺兰山滚钟口种群与灵武市白芨滩种群为第Ⅲ类;吴忠市红寺堡种群为第Ⅳ类。     

Evaluation of the Pathotypes of Xanthomonas oryzae pv. oryzae and Preliminary Analysis of the Resistant Reactions of Main Japonica Rice in the Yunnan Plateau, China

The pathogenicity of 36 isolates of Xanthomonas oryzae pv. oryzae （Xoo）, which were collected from japonica rice varieties in the Yunnan Plateau, China, was evaluated. It was evaluated on 29 rice varieties including a set of seven varieties to identify pathogenicity, i.e., Haonuoyang, TN1, Kogyoku, Zhenzhu＇ai, IR26, Nanjing 33, and Kinmaze, which may be considered as a set of differential varieties for Xoo races from Yunnan japonica rice. The efficiency of the seven varieties was further confirmed. The results showed reversible and specific interactions between isolates and varieties. The isolates were classified into nine pathotypes from pathotyp Ⅰ to Ⅸ according to their pathogenic reactions on the seven rice varieties. The pathotype V was the epidemic, whereas pathogen Ⅶ was the most pathogenic. Most japonica varieties grown in the Yunnan Plateau were susceptible to Xoo. The rice lines IRBB21 （Xa-21）, Zhachanglong （Xa-22,, Xa- 24,）, and IR1545-339 （xa-5）, which were resistant to all the isolates tested, can be used as donors of resistant genes for bacterial blight in japonica rice breeding in the Yunnan Plateau.