2株鸭源 H11N9亚型禽流感病毒的基因遗传进化及致病性分析

为了解从湖南省洞庭湖区鸭群中分离的2株 H11N9亚型禽流感病毒变异特点、进化规律及生物学特性，本研究对2株H11N9亚型禽流感病毒的HA、NA序列进行同源性和遗传进化分析，并用2株毒株对SPF鸡进行致病性试验。结果显示，本试验分离到2株 H11N9亚型禽流感毒株的 HA裂解位点均没有多个连续的碱性氨基酸插入，属于低致病性毒株；HA基因的受体结合位点均非常保守，具有典型的禽源性特征；NA基因序列与在周边国家野鸟中分离的H11N9亚型毒株的氨基酸同源性较高；鼻腔接种SPF鸡后，均能使鸡感染并通过喉头或泄殖腔排毒，但感染的鸡均不表现明显的临床症状，并且不能使同居鸡感染排毒。     

Zoospore chemotaxis of closely related legume‐root infecting Phytophthora species towards host isoflavones

Phytophthora niederhauserii, P. pisi, P. sojae and P. vignae are closely related species that are pathogenic to various legume plants. While P. sojae and P. vignae are reported to specifically infect soybean and cowpea, respectively, P. pisi is reported to attack pea and faba bean. Phytophthora niederhauserii is considered to have a broad host range. Zoospores of some Phytophthora species are chemotactically attracted to the isoflavones that are secreted by their host plants. The focus of the current study was to determine the chemotaxic behaviour of zoospores from closely related legume‐root infecting Phytophthora species and to investigate the correlation, if any, to host preference as determined by greenhouse pathogenicity tests. The results showed that P. sojae and P. vignae were attracted to the non‐soybean isoflavone prunetin as well as to the soybean isoflavones genistein and daidzein, which is in contrast with their host specificity on soybean and cowpea, respectively. On the other hand, P. pisi and P. niederhauserii were only attracted to prunetin, previously reported to be produced by pea, but not to the isoflavones associated with the non‐host soybean. The lack of responsiveness to genistein and daidzein in P. pisi may represent a recent adaptation to the host specialization towards pea. However, the affinity of P. niederhauserii to prunetin shows that this trait can also be present in taxa not specifically associated with legume hosts.     

甘肃加工型番茄叶果真菌病原鉴定及主要病害

经田间采集、分离培养、致病性测定,鉴定出13种加工型番茄真菌病原及病害。其中早疫病(Alternaria solani)、灰霉病(Botrytis cinerea)、叶霉病(Fulvia fulva)、斑枯病(Septoria lycopersici)、白霉病(Ramularia lycopersici)为甘肃省加工型番茄主要流行性病害,是制约甘肃省加工型番茄发展的主要病害。内丝白粉病(Leveillula brassicae)、柱隔孢菌(Ramularia lycopersici)侵染加工型番茄在国内外未见报道,属新病害。黑斑病(Alternaria brassicae)、果腐病(Fusarium moniliforme)、枯萎病(Fusarium oxysporium)、斑枯病(Septoria lycopersici)、煤污病(Cercospora fuligena)侵染加工型番茄在甘肃省属首次报道。     

Sequence variation in the putative effector gene SIX8 facilitates molecular differentiation of Fusarium oxysporum f. sp. cubense

Fusarium oxysporum f. sp. cubense (Foc), causal agent of fusarium wilt of banana, is among the most destructive pathogens of banana and plantain. The development of a molecular diagnostic capable of reliably distinguishing between the various races of the pathogen is of key importance to disease management. However, attempts to distinguish isolates using the standard molecular loci typically used for fungal phylogenetics have been complicated by a poor correlation between phylogeny and pathogenicity. Among the available alternative loci are several putative effector genes, known as SIX genes, which have been successfully used to differentiate the three races of F. oxysporum f. sp. lycopersici. In this study, an international collection of Foc isolates was screened for the presence of the putative effector SIX8. Using a PCR and sequencing approach, variation in Foc‐SIX8 was identified which allowed race 4 to be differentiated from race 1 and 2 isolates, and tropical and subtropical race 4 isolates to be distinguished from one another.     

宿主湿度环境对球孢白僵菌致病力的影响

【目的】研究宿主湿度环境对球孢白僵菌致病力的影响,为采取适当防治措施以减少白僵病对蚕茧生产的危害提供参考依据。【方法】以广西象州蚕区球孢白僵菌感染家蚕获得的白僵病蚕尸体,观察不同湿度环境中白僵病蚕尸体形态的变化情况,并每隔15d进行一次分生孢子致病力测试。【结果】在25℃下的3种湿度环境（60%RH、75%RH和90%RH）中,白僵病蚕尸体在同一宿存时间内,球孢白僵菌活力与宿存湿度关系密切,在60%RH中宿存的蚕体保存较完好,60d后仍然可见淡黄白色的分生孢子,尸体质硬;随湿度增加,宿存病原活力逐渐降低。3种湿度条件下球孢白僵菌在2龄病蚕尸体中宿存30d内,对家蚕仍有很强的致病性,其半致死时间（LT50）分别为2.915、3.071和5.222d;在60%~75%RH条件下,宿存45~60d的球孢白僵菌对家蚕仍具有一定的致病力;而在90%RH条件下,宿存45d后球孢白僵菌分生孢子数量急剧下降,无法测定其致病力。【结论】球孢白僵菌在病蚕体中宿存60d内对家蚕的致死率仍然很高,因此及时清除病蚕尸体并使尸体病原性灭活,可降低白僵菌的二次感染。     

石楠拟盘多毛孢致病毒素除草作用的初步研究

石楠拟盘多毛孢(Pestalotiopsis photiniae)是草莓根腐病的优势致病菌,主要通过毒素引发病害。为明确该病菌毒素对常见几种杂草的影响,本试验利用离体叶片和种子根芽伸长法研究了该病菌粗毒素对几种杂草的致病作用。结果表明:该粗毒素对供试的13种杂草叶片均有不同程度的致病作用,致病强度与杂草种类及粗毒素浓度有关,其中车前、马齿苋、马唐、反枝苋和灰菜5种杂草受害最重,经较低浓度(0.036mg/mL)毒素处理后1d即开始出现褐色斑点,第3天病斑面积达到最大(50%以上);该粗毒素(0.036mg/mL)对车前等5种杂草种子萌发、幼根和幼芽伸长也均有显著的抑制作用,且随毒素浓度升高抑制作用增强,抑制率均在53%以上,但对小麦和大豆种子萌发及根芽伸长均无明显的抑制作用;粗毒素经紫外线照射后,对车前和蒲公英的除草活性在自然光照下比黑暗下明显增强。这些结果揭示石楠拟盘多毛孢粗毒素有一定的除草潜力。     

Incomplete systemic movement of Xanthomonas campestris pv. musacearum and the occurrence of latent infections in xanthomonas wilt‐infected banana mats

Management of banana xanthomonas wilt (XW) (caused by Xanthomonas campestris pv. musacearum, Xcm) has been impeded by poor adoption of control options that are complex, cumbersome and costly. To improve XW management, this study investigated Xcm survival and latent infections in subsequent generations, survival of latently infected planting materials (suckers), incidence of latent infections in symptomless plants in mats having diseased plants, and XW status across farms and markets in districts previously devastated but currently endemic. On‐station experiments were protected from new infections. Latent bacteria at low levels were detected in up to 20% of the third generation suckers, with a significant (P < 0·05) reduction (43–20%) in subsequent generations. Only 3–6% of latently infected suckers succumbed to XW. Incidence of Xcm in symptomless suckers from farmers' fields (with up to 70% incidence) was low (3%) while it increased (8–25%) with disease severity in mats in controlled experiments. In the surveyed districts, incidence had significantly declined with yields observed to have recovered relative to earlier reports, although latent infections remained high. This study provides evidence that if new infections are prevented, fields with high XW incidence can be rejuvenated. It showed incomplete systemic movement of Xcm in mats coupled to a gradual decline of bacterial load in subsequent generations to levels that cannot initiate disease. These studies explain the current successes in farms practising single diseased plant removal instead of whole mat rouging, and gives hope to farmers lacking access to clean planting material.     

一株GPV的分离鉴定及致病性研究

为了确定导致吉林省某地区10日龄雏鹅发病的病原，从病死雏鹅的肝脏中分离到一株病毒，根据病毒的电镜照片，动物回归试验及PCR鉴定，确定为鹅细小病毒。PCR产物测序后经BLAST比对显示，分离毒株与NCBI收录的GPVVP3基因同源性均在92％以上，与GPV／CH／HLJ02／08VP3基因的同源性最高，达99％。致病性研究表明，分离株的ELD50为10^-6.5／0．2mL，毒力较强，且该毒株能被GPV标准血清所中和。     

桉树青枯病病原菌的分子鉴定及其致病性测定

对华南4个地区的桉树青枯病病原菌进行了分子鉴定和致病性测试研究,以期为有效预防其发生提供依据.分别对来源于不同区域的11株桉树青枯病病原菌进行分子鉴定和致病力强弱测试比较,11株病原菌与Ralstonia solanacearum序列形成支持率为86％分枝,表明其病原菌16S rRNA基因序列与Ralstonia solanacearum具有高度同源性.以桉树无性系DH32-29组培苗为材料,采用伤根法对11个菌株进行致病力测试,不同菌株间在致病性上差异显著(P＜0.01或P＜0.05),以发病高峰期、高峰期发病率和平均发病率进行K-均值聚类分析(K=3),结果表明来源于广东、广西、海南的各菌株间致病性存在差异,HY01、HY02、DA01、HP04菌株与HP05分为1类,DA02和DA03为第2类,而YJ01、HP01、HP02和HP03为第3类.对4个不同地区(采样点)的青枯病菌株平均发病率进行方差分析,结果发现来源广东阳江菌株致病性显著地低于其他来源菌株.不同地理来源菌株之间在致病性上无明显相关性,同一地区内同时存在着致病力强弱菌株,HP03的生物型4-1菌株与其他生物型3菌株在致病性亦无明显的差别.     

水稻白叶枯病菌avrBs3/PthA家族基因的随机缺失及突变体PXO99Δavr功能的研究

利用来源于水稻白叶枯菌株JXOⅢ的无毒基因avrXa3构建的同源序列突变转化单元,同源重组得到PXO99~A菌株的1个缺失突变体PXO99△avr。PXO99~A及其突变体基因组DNA以内切酶BamHⅠ完全消化后进行同源检测,发现突变体PXO99△avr中有3条杂交带消失,有2条带信号减弱。进一步以相同探针检测PXO99~A的基因文库,限制性酶切分析和Southern杂交归类,共获得13个不同的阳性克隆。众多avrBs3/PthA基因在基因组中单独存在,或2个和2个以上串联排列。经比对,克隆p5所含条带与突变体缺失的5条带大小相同。因此认为随机交换产生的突变体中有5个基因被敲除。将此文库克隆(p5)分别导入PXO99~A和突变体PXO99△avr中,苗期接种试验表明:p5互补突变体能使其毒性恢复接近PXO99~A的致病表型,推测此克隆正是突变体缺失的基因。同时也表明:缺失的5个串联基因的综合作用主要表现为毒性功能。