针叶树病原菌异担子菌属的物种多样性及相关检疫建议

异担子菌属Heterobasidion种类是北半球针叶树最严重的森林病原菌,在世界范围内能侵染27种针叶树,对欧洲和北美洲的经营林已造成重大损失。基于传统形态学研究,认为异担子菌属有2个种,即多年异担子菌H. annosum和岛生异担子菌H. insulare,然而单孢交配实验研究证明2个种为复合种。分子系统发育分析研究表明：异担子菌属包括15个种,其中5个种为森林病原菌[冷杉异担子菌H. abietinum、多年异担子菌（狭义）H. annosum s. str.、变孔异担子菌H. irregulare、西方异担子菌H. occidentale和小孔异担子菌H. parviporum];10个种为腐生菌（淀粉孢异担子菌H. amyloideum、南洋杉异担子菌H. araucariae、阿曼德异担子菌H. armandii、南方异担子菌H. australe、岛生异担子菌H. insulare、林芝异担子菌H. linzhiense、东方异担子菌H. orientale、拟岛生异担子菌H. subinsulare、拟小孔异担子菌H. subparviporum和西藏异担子菌H. tibeticum）。多年异担子菌（狭义）H. annosum s. str.、小孔异担子菌H. parviporum和冷杉异担子菌H. abietinum分布于欧洲,分别是松属、云杉属和冷杉属林木的病原菌。变孔异担子菌H. irregulare和西方异担子菌H. occidentale分布于北美洲,前者侵染松属和柏属树木,后者侵染冷杉属、铁杉属、黄杉属和巨杉属树木。虽然广义的多年异担子菌H. annosum sensu lato曾在我国报道,但基于目前研究结果表明该种为拟小孔异担子菌H. subparviporum。目前世界上最具侵染力的5种病原菌即狭义的多年异担子菌H. annosum s. str.、小孔异担子菌H. parviporum、冷杉异担子菌H. abietinum、变孔异担子菌H. irregulare和西方异担子菌H. occidentale还未在我国发现,也未列为对外检疫对象,因此建议将其列为进境植物检疫性有害生物。RNA聚合酶II大亚基序列（RPB1）在异担子菌属种类鉴定中敏感性和特异性最高,研究证明该分子方法可应用于鉴别不同异担子菌种类的有效基因标记,在海关部门进行原木和木质产品的检疫中可运用该分子标记进行林木病原异担子菌的检测。     

人工栽培蛹虫草的病原真菌

蛹虫草规模化栽培过程中,真菌病害普遍发生且危害严重。本研究对人工栽培蛹虫草中真菌病害进行调研,对病原真菌进行分离、纯化、鉴定及致病性检验,并分析病害发生的特点。结果发现引起蛹虫草病害的病原真菌主要有虫草生齿梗孢、产扁虫菌素单端孢、镰刀菌、裂褶菌、哈茨木霉、淡紫拟青霉、稻绿核菌、粉红枝穗霉、卵孢单端孢、扩展青霉、黄曲霉和黑曲霉。其中虫草生齿梗孢为引起蛹虫草侵染性病害的主要病原真菌。虫草生齿梗孢、产扁虫菌素单端孢、镰刀菌、裂褶菌和哈茨木霉主要为害蛹虫草子实体;淡紫拟青霉、稻绿核菌、粉红枝穗霉、卵孢单端孢、扩展青霉、黄曲霉和黑曲霉主要为害栽培料与蛹虫草菌丝体。镰刀菌、裂褶菌、哈茨木霉、淡紫拟青霉、稻绿核菌和粉红枝穗霉为引起蛹虫草病害的首次报道。本研究为蛹虫草病害防控奠定基础,以促进产业健康发展。     

西藏种子植物分布新资料

本文报道了分布于西藏的4个中国新记录种, 即须弥四带芹(Tetrataenium wallichii)、巴洛特栎(Quercus baloot)、尼泊尔姜味草(Clinopodium nepalense)和喜马拉雅耳稃草(Garnotia emodi); 以及24个西藏新记录种, 即翼叶棱子芹(Pleurospermum decurrens)、线叶球兰(Hoya linearis)、尼泊尔菊三七(Gynura nepalensis)、一枝黄花(Solidago decurrens)、拟游藤卫矛(Euonymus vaganoides)、云南木鳖(Momordica subangulata subsp. renigera)、马㼎儿(Zehneria japonica)、攀援胡颓子(Elaeagnus sarmentosa)、四棱猪屎豆(Crotalaria tetragona)、大叶千斤拔(Flemingia macrophylla)、毛喉鞘蕊花(Coleus forskohlii)、独龙木姜子(Litsea taronensis)、长序大苞兰(Bulbophyllum purpureofuscum)、球花石豆兰(B. repens)、黄兰(Cephalantheropsis obcordata)、双角厚唇兰(Dendrobium forrestii)、密花兰(Diglyphosa latifolia)、云南盆距兰(Gastrochilus yunnanensis)、扁茎羊耳蒜(Liparis assamica)、耳唇兰(Otochilus porrectus)、疏花无叶莲(Petrosavia sakuraii)、长序羊角藤(Gynochthodes lacunosa)、黑蒴(Alectra avensis)和赤杨叶(Alniphyllum fortunei), 其中无叶莲科为西藏新记录科, 一枝黄花属(Solidago)、马㼎儿属(Zehneria)、千斤拔属(Flemingia)、鞘蕊花属(Coleus)、密花兰属(Diglyphosa)、无叶莲属(Petrosavia)、羊角藤属(Gynochthodes)、黑蒴属(Alectra)和赤杨叶属(Alniphyllum)等9个属为西藏新记录属。凭证标本均存放于中国科学院昆明植物研究所标本馆(KUN)。     

沙地樟子松带状混交林土壤碳氮磷化学计量特征

探究不同沙地樟子松混交林土壤碳氮磷化学计量变化规律,为其混交经营提供科学依据.以沙地樟子松纯林为对照,在沙地樟子松×榆树及沙地樟子松×怀槐带状混交林中,沿沙地樟子松和伴生树种两个方向在距离混交中心0、1、2、3和4 m处的不同土层采集土样,分析土壤有机C、全N、全P、速效N、速效P含量及其计量比的特征.结果表明: 沙地樟子松混交林土壤有机C、全N和速效N含量高于纯林, 沙地樟子松与榆树混交主要增加了深层土壤有机C和全N含量,以及C/N和C/P, 沙地樟子松与怀槐混交提高了土壤N含量, 降低了P含量.随着远离混交林中心,沙地樟子松×榆树混交林中沙地樟子松林带的土壤C/N先升高后降低,全P和速效P含量下降,N/P增加;榆树林带土壤C/N下降,速效P含量先升高后降低.沙地樟子松×怀槐混交林土壤全N含量在沙地樟子松林带先降低后升高,在怀槐林带先升高后降低.沙地樟子松混交林提高了土壤C、N储量,沙地樟子松应与榆树行间混交,与怀槐间隔2行混交.     

中国小菇属十个新记录种

报道了小菇科小菇属真菌10个中国新记录种,香菌组：橙盖小菇Mycena aurantiidisca、黄白小菇Mycena flavoalba、粉黄小菇Mycena floridula;棘刺组：异刺小菇Mycena heteracantha;纤柄组：碱味小菇Mycena amygdalina;脆足组：粉被小菇Mycena zephirus;冬生组：绣线菊小菇Mycena speirea、冬生小菇Mycena hiemalis;小菇组：绒柄小菇Mycena flos-nivium,分别来自吉林等11个省份、自治区。提供了每个物种的形态描述和线条图,以及与相近种的讨论。共计90条自测及下载ITS序列,在采用贝叶斯法和最大似然法构建的小菇属系统发育树中,新记录种均得到分子数据支持。凭证标本存放于吉林农业大学菌物标本馆（HMJAU）。     

Identical digeneans in coral reef fishes from French Polynesia and the Great Barrier Reef (Australia) demonstrated by morphology and molecules

Three coral reef fish species, Zanclus cornutus, Chaetodon vagabundus and Naso lituratus, were collected in French Polynesia and on the Great Barrier Reef, Queensland. These fish species were each infected by one morphologically similar digenean species in both localities; Schistorchis zancli Hanson, 1953 was found in Zanclus cornutus, Preptetos laguncula Bray and Cribb, 1996 in Naso lituratus and Neohypocreadium dorsoporum Machida and Uchida, 1987 in Chaetodon vagabundus. In addition, on the Great Barrier Reef P. laguncula was also found in Naso unicornis and N. dorsoporum in Chaetodon ephippium and Chaetodon flavirostris. Morphometric differences between the species from the two sites were only slight. Sequences from the second internal transcribed spacer of the ribosomal DNA of each worm revealed total homology or negligible divergence between samples from hosts caught in French Polynesia and on the Great Barrier Reef. These results show that across more than 6000 km these digeneans are similar in morphology and genotype. Some species of fishes and molluscs are considered to have distributions that encompass the entire tropical Indo-West Pacific. These findings suggest that at least some of their parasites have similarly broad distributions.     

中国鬼伞类真菌的分类

鬼伞类真菌涉及蘑菇科的鬼伞属Coprinus以及小脆柄菇科的小鬼伞属Coprinellus、拟鬼伞属Coprinopsis、近地伞属Parasola、刺毛鬼伞属Tulosesus和心孢鬼伞属Narcissea。基于标本研究和文献记载,确认我国该类真菌57种：小鬼伞属9种,刺毛鬼伞属10种,心孢鬼伞属3种,拟鬼伞属25种,近地伞属8种,鬼伞属2种。本文记载1个新组合——速亡型心孢鬼伞Narcissea ephemeroides,中国新记录种11个：黄鳞小鬼伞Coprinellus ellisii、甜味小鬼伞Coprinellus saccharinus、锐突拟鬼伞Coprinopsis acuminata、非洲雪白拟鬼伞Coprinopsis afronivea、美丽拟鬼伞Coprinopsis bellula、钟孢拟鬼伞Coprinopsis mitraespora、麻醉拟鬼伞Coprinopsis narcotica、厚壁拟鬼伞Coprinopsis pachyderma、近雪白拟鬼伞Coprinopsis pseudonivea、施罗特近地伞Parasola schroeteri和刺毛近地伞Parasola setulosa。编制了中国鬼伞类真菌分种检索表,对新组合和中国新记录种进行形态学描述,并提供线条图。结合ITS和LSU序列片段,采用了最大似然法和贝叶斯分析法建立系统发育树,反映各类群之间的系统演化关系。     

Pathogenicity of Heterorhabditis bacteriophora, Steinernema glaseri, and S. scarabaei (Rhabditida: Heterorhabditidae, Steinernematidae) Against 12 White Grub Species (Coleoptera: Scarabaeidae)

We compared the pathogenicity of the entomopathogenic nematodes Heterorhabditis bacteriophora, Steinernema glaseri, and S. scarabaei against third instars of 12 white grub species. The Japanese beetle (Popillia japonica) was highly susceptible to all nematode species. Oriental beetle [Exomala (=Anomala) orientalis], European chafer (Rhizotrogus majalis), Asiatic garden beetle (Maladera castanea), and the May/June beetles Phyllophaga crinita, Ph. congrua, and Ph. (Subgenus Phytalus) georgiana were highly susceptible to S. scarabaei but had mediocre to low susceptibility to H. bacteriophora and S. glaseri. The black turfgrass ataenius (Ataenius spretulus) was very susceptible to H. bacteriophora but had mediocre susceptibility to S. glaseri and S. scarabaei. Northern (Cyclocephala borealis) and southern masked chafer (C. lurida) had mediocre and southwestern masked chafer (C. pasadenae) and green June beetle (Cotinis nitida) had low susceptibility to all nematode species.     

两种乳菇菌丝生长的最适培养基与菌根合成

通过测量菌丝直径和干重对松乳菇（rcb-74）和靓丽乳菇（rll-107,rmsh-118）进行最适菌丝生长培养基的筛选。结果表明所有菌株均在改良的biotin-aneurine-folic acid（BAF）培养基上表现出较大的菌丝直径以及最大菌丝干重。用改良的BAF培养基制备乳菇液体菌丝,接种云南松、马尾松和油松,并在13-30d后发现rcb-74与云南松和油松形成菌根,rll-107与马尾松形成菌根,rmsh-118与云南松形成菌根。     

大围山樱属植物群落结构及物种多样性

研究大围山樱属植物群落结构特征和物种多样性,为野生樱属植物保护和利用提供科学依据.结果 表明:大围山樱属植物有钟花樱桃群落、尾叶樱桃群落、华中樱桃群落和雪落樱桃群落,生活型结构以高位芽植物为主,植物区系组成以泛热带植物成分为主.物种多样性为灌木层＞乔木层,尾叶樱桃群落＞华中樱桃群落＞钟花樱桃群落＞雪落樱桃群落.种群以中...     

毛白杨茎干不同方位夜间液流变化规律及其主要影响因子

为明确毛白杨(Populus tomentosa)不同方位夜间蒸腾量(Nt)及茎干充水量(Sr)等夜间液流活动的规律, 探究不同方位Nt和Sr的主要影响因子, 该研究使用热扩散的方法监测了宽窄行模式下栽植的毛白杨茎干不同方位夜间液流, 并用图像法区分Nt和Sr。使用自动气象站和机械式张力计监测太阳总辐射(R_s, kW·m^-2)、空气温度(T_a, ℃)、空气相对湿度(RH, %)、风速(v, m·s^-1)、土壤水势(ψ, kPa)等环境因子。通过比较各方位的Nt和Sr等液流活动的大小情况及其与环境因子之间的相关性得到方位间夜间液流的差异性以及各方位夜间液流的主要影响因子。结果显示: 宽行距位于东侧的样树西方位的Nt和Sr均最大, 其中西方位的Sr显著大于其他3个方位; 北方位的Nt显著小于其他3个方位; 其他方位间的Nt和Sr无显著差异; 各方位夜间茎干充水量占夜间液流量的比例(Sr/Q)无显著差异。宽行距位于西侧的样树西方位的Nt和Sr亦均最大, 其中西方位的Sr显著大于东方位和南方位; 南方位的Nt最小, 显著小于西方位和北方位, 其他方位间的Nt和Sr无显著差异; 南方位的Sr/Q显著大于其他3个方位。各方位的Nt和Sr均与水汽压亏缺(VPD)有显著的正相关关系, 部分方位Nt和Sr与T_a和RH有显著相关关系, 没有任何方位Nt和Sr与v和ψ有显著相关关系。Nt和Sr方位间的差异(Nt_CV、Sr_CV)与VPD、T_a、RH、v和ψ均无显著相关关系。此外, Sr受白天的液流活动的影响显著。综上所述, 毛白杨不同方位Nt和Sr等液流活动具有较大的差异, 且西方位是优势方位; VPD是影响各方位Nt和Sr的主要气象因子。     

Expression patterns of hedgehog, wingless, and decapentaplegic during gut formation of Gryllus bimaculatus (cricket).

We observed expression patterns of hedgehog (hh), wingless (wg), and decapentaplegic (dpp) during gut development of Gryllus bimaculatus (the cricket), a typical hemimetabolous insect, and compared with those observed in Drosophila, a typical holometabolous insect. Gryllus hh(Gbhh) and Gbwg are expressed in both foregut and hindgut, while Gbdpp is expressed only in the hindgut: at the boundaries between the small and large intestine, and between the large intestine and rectum. Although the expression patterns of Gbhh and Gbwg are essentially comparable to those observed in Drosophila, the expression pattern of Gbdpp differs from those of the Drosophila dpp.     

贵州梨芽枯间座壳菌属Diaporthe的种类

为明确引起贵州梨芽枯病的间座壳菌种类,本研究进行了病害调查、病芽采集和病菌分离及纯化,根据其菌落特征与ITS序列鉴定,共获得94个间座壳属Diaporthe菌株,从中选取40个代表菌株进行多基因(ITS、TEF、CAL、HIS和TUB)系统发育分析和形态学观察,结果显示它们分属于间座壳属的8个种,包括核桃间座壳菌D. biguttulata、榆树间座壳菌D. eres、椭圆间座壳菌D. ellipsoidea、梭状间座壳菌D. fusicola、香港间座壳菌D. hongkongensis、塞纳间座壳菌D. sennae、大豆间座壳菌D. sojae和蜜柑间座壳菌D. unshiuensis,其中D. ellipsoidea、D. biguttulata和D. sennae为侵染梨的首次报道。玉露香梨离体枝条接种结果显示,8种间座壳的代表菌株均可产生与田间相同的症状,分离比最高的D. eres致病力较强,表明它们均为梨芽枯病的病原菌,其中D. eres为病原优势种。本研究首次报道了引起梨芽枯的间座壳菌种类组成,为该病的发生规律及防控技术研究提供了理论依据。     

Azimuthal variation in nighttime sap flow and its mainly influence factors of Populus tomentosa

为明确毛白杨(Populus tomentosa)不同方位夜间蒸腾量(Nt)及茎干充水量(Sr)等夜间液流活动的规律, 探究不同方位Nt和Sr的主要影响因子, 该研究使用热扩散的方法监测了宽窄行模式下栽植的毛白杨茎干不同方位夜间液流, 并用图像法区分Nt和Sr。使用自动气象站和机械式张力计监测太阳总辐射(R_s, kW·m^-2)、空气温度(T_a, ℃)、空气相对湿度(RH, %)、风速(v, m·s^-1)、土壤水势(ψ, kPa)等环境因子。通过比较各方位的Nt和Sr等液流活动的大小情况及其与环境因子之间的相关性得到方位间夜间液流的差异性以及各方位夜间液流的主要影响因子。结果显示: 宽行距位于东侧的样树西方位的Nt和Sr均最大, 其中西方位的Sr显著大于其他3个方位; 北方位的Nt显著小于其他3个方位; 其他方位间的Nt和Sr无显著差异; 各方位夜间茎干充水量占夜间液流量的比例(Sr/Q)无显著差异。宽行距位于西侧的样树西方位的Nt和Sr亦均最大, 其中西方位的Sr显著大于东方位和南方位; 南方位的Nt最小, 显著小于西方位和北方位, 其他方位间的Nt和Sr无显著差异; 南方位的Sr/Q显著大于其他3个方位。各方位的Nt和Sr均与水汽压亏缺(VPD)有显著的正相关关系, 部分方位Nt和Sr与T_a和RH有显著相关关系, 没有任何方位Nt和Sr与v和ψ有显著相关关系。Nt和Sr方位间的差异(Nt_CV、Sr_CV)与VPD、T_a、RH、v和ψ均无显著相关关系。此外, Sr受白天的液流活动的影响显著。综上所述, 毛白杨不同方位Nt和Sr等液流活动具有较大的差异, 且西方位是优势方位; VPD是影响各方位Nt和Sr的主要气象因子。     

Differentiation among three species of bovine Thelazia (Nematoda: Thelaziidae) by polymerase chain reaction-restriction fragment length polymorphism of the first internal transcribed spacer ITS-1 (rDNA).

Thelazia gulosa, Thelazia rhodesi and Thelazia skrjabini are nematodes transmitted by some species of Musca (Diptera: Muscidae) which cause ocular infestations in bovines. Differences in the rDNA of these species were determined by a PCR using different sets of relatively conserved oligonucleotide primers. PCR on the first internal transcribed spacer (ITS-1) revealed differences in size in Thelazia species (437 bp for T. gulosa, 370 bp for T. rhodesi and 506 bp for T. skrjabini) while the DNA control of Musca spp. was not amplified. The ITS-1 amplicons of the three species were sequenced and then analysed. The GC contents ranged from 26 to 36% and the level of differences in the nucleotide sequences of ITS-1 was lower between T. skrjabini and T. gulosa (39%) than the latter and T. rhodesi (49–56%). Restriction fragment length polymorphism (RFLP) of ITS-1 amplicons was also carried out and the restriction profiles compared. Clear genetic differences among the three Thelazia examined were demonstrated by using the enzymes HpaII, CpoI and SspI. This PCR–RFLP for the delineation of T. gulosa, T. rhodesi and T. skrjabini offers prospects as a molecular epidemiological tool to study parasite transmission patterns and prevalence.     

基于多基因分析对我国红曲霉属Monascus真菌的系统发育研究

红曲霉属Monascus具有重要的经济和生态价值,但对该属的物种识别和系统发育学研究有限。本研究首先对红曲霉属红色组内物种开展ITS和LSU序列的测定并综合分析,结合GenBank中相关物种序列及菌株的形态学特征,探讨红曲霉属的系统发育关系,鉴定出红色组3个种,弗罗里达组7个种。其次采用红曲霉色素表型控制基因簇部分基因pksKS序列进行分析,以期解决ITS和LSU等基因序列分析无法有效区分红色红曲与紫色红曲不同形态种的问题。通过分子克隆测序与直接测序结果的比较,以及对3个血红红曲菌株单核苷酸多态性的分析,解析了血红红曲种内遗传差异,首次在红曲霉属真菌中发现疑似红色红曲血红自然杂交种。最后综合ITS、LSU、pksKS序列和形态学分析的结果,统一了红曲霉属内物种的系统发育关系,确认红色组包括3个种：红色红曲M. ruber、紫色红曲M. purpureus、血红红曲M. sanguineus;发现3个变种：红色红曲发白变种M. ruber var. albidulus、紫色红曲橙色变种M. purpureus var. aurantiacus及紫色红曲火红色变种M. purpureus var. rutilus;弗罗里达组包括7个种：弗罗里达红曲M. floridanus、苍白红曲M. pallens、新月红曲M. lunisporas、阿根廷红曲M. argentinensis、累西腓红曲M. recifensis、黄色红曲M. flavipigmentosum、蜂蜜红曲M. mellicola。另外,发现1个疑似杂交种：红色红曲血红杂种M. ruber × sanguineus。结果表明,红曲霉属具有丰富的物种多样性,而且通过多基因和形态学分析可以将相近种区分开来。     

入侵豚草与本地植物竞争对丛枝菌根真菌多样性的影响

外来植物入侵常引起入侵地丛枝菌根真菌（arbuscular mycorrhizal fungi,AMF）多样性的变化,但AMF多样性的变化与入侵域本地植物种类相关性目前尚不清楚。本研究采用野外同质园试验研究豚草与不同本地植物竞争时对AMF多样性的影响。本试验共设入侵豚草单种、本地植物狗尾草Setaria viridis单种、藜 Chenopodium album单种、黄香草木犀Melilotus officinalis单种、豚草和狗尾草混种、豚草和藜混种、豚草和黄香草木犀混种以及裸土（无任何植物生长）对照8个处理。采集根区土样后分离、鉴定AMF,分析植株生物量和AMF物种多样性变化间的关系。结果表明,对豚草而言,单种处理中生物量均高于混种处理;豚草分别与3种本地植物竞争时,与黄香草木犀混种中豚草竞争能力最强,而与藜混种中竞争能力最弱。豚草分别与3种本地植物混种时土壤中AMF种丰度和香农-威纳指数分别显著高于豚草单种和本地植物单种,其中豚草与藜混种中AMF的种丰度和香农-威纳指数最高,与狗尾草混种中AMF的种丰度和香农-威纳指数最低,而豚草分别与3种本地植物混种处理中孢子密度显著低于本地植物单种（豚草与黄香草木犀混种除外）。豚草单种、狗尾草单种、藜单种、豚草分别与狗尾草和藜混种4个处理中AMF优势种均为网状球囊霉Glomus reticulatum和缩隔球囊霉Septoglomus constrictum;黄香草木犀单种中优势种是网状球囊霉、缩隔球囊霉和根内根孢囊霉Rhizophagus Intraradices,豚草与黄香草木犀混种中优势种是网状球囊霉和根内根孢囊霉;狗尾草单种、豚草与狗尾草混种和豚草与藜混种3个处理的特有种分别是木薯根孢囊霉Rhizoglomus manihotis、近明球囊霉Claroideoglomus claroideum和美丽盾巨孢囊霉Scutellospora calospora。狗尾草单种生物量与孢子密度呈显著正相关,豚草与狗尾草混种中豚草的生物量与均匀度呈显著负相关,豚草与黄香草木犀混种中豚草的生物量与孢子密度呈显著正相关,与香农威纳指数呈显著负相关。综上可知,豚草与本地植物竞争对AMF多样性的影响具有异质性,这与本地植物种类密切相关。     

Anatomy of the stimulative sequences flanking the ARS consensus sequence of chromosome VI in Saccharomyces cerevisiae

We have analyzed the relationship between autonomously replicating sequence (ARS) structure and function for three ARS (ARS605, ARS607 and ARS609) from chromosome VI of Saccharomyces cerevisiae by systematic XhoI-linker mutation in the ARS consensus sequence (ACS) and flanking sequences. All mutations that encroached upon the ACS destroyed ARS activity. DNA sequences stimulative for ARS function were identified on either side of the ACS of ARS605 and only on the 3'-side of the ACS of ARS607. In ARS609, however, no such stimulative sequences were observed. Base substitutions complementary to the wild-type sequence of those stimulative regions, in ARS605 and ARS607, that did not change the AG of unwinding nor affected ARS activity suggests that these regions have, at least, a function as DNA-unwinding elements (DUE). ARS605, ARS607 and ARS609 DNA are of low AG value and showed hypersensitivity to single-strand-specific nuclease when inserted in negatively supercoiled plasmid. Linker mutations inhibitory for ARS activity (5L11 and 7L14) also caused significant changes in local nucleotide (nt) sensitivity within the ACS and its adjoining regions. Complementary base substitutions, however, did not affect these changes in local nt sensitivity. These results imply that the stimulative regions flanking the ACS are necessary to produce an optimum conformation around the ACS which may be important for full ARS activity.     

Genetic diversity and phylogenetic relationships of bacteria belonging to the Ochrobactrum-Brucella group by recA and 16S rRNA gene-based comparative sequence analysis

The genetic diversity and phylogenetic interrelationships among 106 Ochrobactrum strains (O. anthropi: 72, O. intermedium: 22, O. tritici: 5, O. oryzae: 2, O. grignonense: 2, O. gallinifaecis: 1, O. lupini: 2), the type strains of the eight Brucella species and other closely related taxa were studied by recA and rrs gene (16S rRNA) comparative sequence analysis. Both markers correctly delineated the various Ochrobactrum species; however, resolution at the subspecies level was considerably higher in the recA gene-based approach. Phylogenetic analyses using neighbor-joining, parsimony, and maximum likelihood algorithms generated trees with similar topologies but the overall branching order, and also the order of the subclades, were not stable in either assay, which could be explained by generally high recA and rrs sequence similarities. Ochrobactrum and Pseudochrobactrum formed separate clades distinct from other Alphaproteobacteria with Bartonella, Agrobacterium, and Rhizobium as the closest relatives. O. gallinifaecis was the most distinct member, when compared to the type species O. anthropi, with rrs and recA similarities of 96.2% and 81.4%. Brucella species were indistinguishable, exhibiting high rrs and recA gene similarities of 98.6% and 85.5% compared with Ochrobactrum intermedium. At the protein level, all RecA sequences among the various Ochrobactrum species and between Ochrobactrum and Brucella were highly similar with only a few amino acid substitutions. O. anthropi and O. tritici were indistinguishable by means of their RecA proteins. A set of initially biochemically classified strains did not cluster within their assigned species and they either grouped within other known species or grouped as potential novel Ochrobactrum species. In further investigations, these strains were reclassified and described as novel species. In summary, Ochrobactrum is a highly diverse genus comprising several novel species. We recommend recA- in addition to rrs gene-analysis for correct species allocation and subtyping of novel Ochrobactrum isolates.     

The chorion genes of the medfly. II. DNA sequence evolution of the autosomal chorion genes s18, s15, s19 and s16 in Diptera

We present a total of approximately 15 kb of DNA sequences, encompassing four chorion genes Ccs18, Ccs15, Ccs19, Cc16 and their flanking DNA in the medfly C. capitata. Comparison of coding regions, introns and intergenic sequences in five Dipteran species, D. melanogaster, D. subobscura, D. virilis, D. grimshawi and C. capitata documented an extensive divergence in introns and coding regions, but few well conserved elements in the proximal 5′ flanking regions in all species. These elements are related to conserved regulatory features of three of the genes, including tissue- and temporal regulation. In the fourth, gene s15, significant alterations in the 5′ flanking region may be responsible for its changed temporal regulation in C. capitata. One long intergenic sequence, located in the distal 5′ flanking region of gene s18, is homologous to ACE3, a major amplification control element and contains an 80-bp A/T-rich sequence, known to stimulate strong binding of the origin recognition complex (ORC) in D. melanogaster. Analysis of the nucleotide composition of all chorion genes in C. capitata and D. melanogaster showed that C. capitata exhibit less biased representation of synonymous codons than does D. melanogaster.