利用16S rDNA基因序列进行害竹飞虱(半翅目:飞虱科)的分子鉴定

采用DNA测序技术测定了中国飞虱科5属10种15个地理种群的线粒体16S rDNA基因长约480 bp片段的序列,分析了其序列组成及变异特征.所获序列中,A+T约占77.5％,其中,190个核苷酸位点发生了变异(约占39.6％).利用MEGA 4.0软件计算遗传距离,外群与内群的遗传距离平均值(0.317)最高,内群各属间和各属内遗传距离平均值(分别为0.209和0.132)次之,而种内不同地理种群间的遗传距离平均值(0.013)最低.构建了10种飞虱科昆虫的MP和NJ分子系统树,结果显示:各属飞虱基本上形成单系群;各种飞虱各自不同地理种群均聚为一支,构成一单系群,可以很好的区分不同种类的飞虱.上述结果与形态学研究结果非常吻合.     

中国广义金合欢属植物亲缘关系的分子分析

通过ITS-rpl16序列和mat K序列的分子分析,对中国现有金合欢属的各亚属间的亲缘关系进行了研究,结果表明:我国现有广义金合欢属的14个种大致被聚为相思亚属、皮刺亚属和金合欢亚属3大类,原产中国的3种皮刺亚属的种聚成1支,表明印度金合欢和羽叶金合欢关系更近,二者再与儿茶形成姐妹类群。基于选自美洲、非洲、澳大利亚和中国乡土种(印度金合欢、羽叶金合欢和儿茶)共40个金合欢属种和1个外类群的mat K序列建立的系统发育树表明:皮刺亚属和相思亚属的亲缘性关系比与金合欢亚属的亲缘关系更近,相思亚属和金合欢亚属均是单系类群,而皮刺亚属属于并系类群;中国的几个乡土种都与非洲一些种的亲缘关系较近,聚为1支。     

铁皮石斛内生细菌群落结构分析

采集人工种植2年的铁皮石斛,将根、茎和叶研磨为粉末,采用NA培养基分离纯化其内生细菌,酶切去除冗余,利用16S rRNA基因序列同源性分析,对铁皮石斛根、茎和叶中分离得到的49株内生细菌进行分离鉴定。结果表明,获得的49株内生细菌在根、茎、叶中分别有24株、15株和10株,分别属于芽孢杆菌属的5个种、假单孢杆菌属的1个种和沙雷氏菌属的1个种。分离出的内生细菌表现出一定程度的细菌区系相似性,也表现出细菌区系的器官特异性。铁皮石斛内生细菌在石斛根部分布的属、种数量明显比茎、叶多,土壤可能是铁皮石斛内生细菌的主要来源;铁皮石斛不同部位内生细菌群落结构存在一定差异,群落种数根＞茎＞叶。研究获得荧光假单胞杆菌20株,占总内生细菌数量的40.82％,判断其可能为优势菌株。     

多疣狭口蛙的两性异形

测量和比较采自昆明东北郊的131只成年多疣狭口蛙K a lou la verrucosa标本(72♂59♀),同时野外观察雌雄蛙的伏卧姿势、运动方式和繁殖行为.成年雌雄蛙的体长、吻长、鼓膜直径、内突长、前臂宽、后肢全长和足长7个形态特征差异显著.对与体长相关的前臂宽、后肢全长和足长进一步回归分析表明,雄蛙前臂宽随体长的增长速率显著大于雌蛙,雌蛙后肢全长和足长随体长的增长速率显著大于雄蛙.雄蛙的前臂宽、后肢全长和足长均大于相同体长雌蛙的前臂宽、后肢全长和足长.繁殖期雄蛙前肢内侧及胸腹部布满分泌物,雌蛙无.雄蛙为2/3蹼,雌蛙约为1/3蹼.雄蛙具单咽外声囊,雌蛙无.雄蛙肛部疣粒大而光滑,雌蛙肛部疣粒细长而尖.     

唐菖蒲2种主要病毒的多重RT-PCR检测

根据黄瓜花叶病毒(cucumber mosaic virus,CMV)和烟草花叶病毒(tobacco mosaic virus,TMV)外壳蛋白基因序列分别设计特异性引物对,运用单重RT-PCR检测到病毒并筛选出适合多重RT-PCR的引物对。以唐菖蒲18S rRNA为内参照,建立同时检测CMV和TMV的多重RT-PCR方法。此方法可从带病样品中扩增出CMV(629bp)和TMV(423bp)2条特异片段。测序结果表明:CMV扩增产物与其他植物分离物核苷酸的同源性为93%～97%,TMV扩增产物与其他分离物核苷酸的同源性可达99%。     

绣线菊丛枝病病原的分子鉴定

用植原体16S rRNA基因的通用引物,对表现黄化、丛枝、顶枯等症状的绣线菊DNA进行PCR扩增,得到了约1.2kb的特异性片段.测序结果证明其病原为植原体.RFLP和序列分析表明:该分离物属于翠菊黄化组的16SrⅠ -B亚组.与GenBank中其他植原体分离物序列比较,发现该分离物与16SrⅠ -B亚组中的西方翠菊黄化植原体(SAY)同源性高达99.6%.     

鸭嘴竹叶蛙

正鸭嘴竹叶蛙(Odorrananasuta),蛙科臭蛙属,是费梁等2001年发表的蛙类新种,模式产地位于海南省琼中县五指山。鸭嘴竹叶蛙为中国特有种,除海南有分布外,广西也有该物种的主要分布区。国际自然保护联盟(IUCN)把鸭嘴竹叶蛙列为易危级别。形态特征鸭嘴竹叶蛙雄性体长55—65毫米,雌性体长70—80毫米。背面颜色多为暗褐色、绿色或褐绿色;头侧,颞部为红棕色;唇缘至颌腺,腹侧和股后部均为浅棕黄色,密布褐色细斑;四肢分布褐黑色横纹3-5     

福建省蕨类植物区系分析

对福建省蕨类植物的区系组成和地理成分进行了分析,其结果表明:(1)福建省共有蕨类植物46科、107属、396种,优势科为鳞毛蕨科、金星蕨科、水龙骨科、蹄盖蕨科、铁角蕨科和凤尾蕨科等11个科,优势属为鳞毛蕨属、毛蕨属和凤尾蕨属等;(2)从蕨类植物科、属分布区类型所占比例来看,该地区区系性质以热带成分为主;而从种的地理成分来看,则表现为热带成分与东亚成分的综合体,以中国特有种、中国-日本分布、热带亚洲最为丰富,反映了东亚与热带亚洲的蕨类植物区系的统一性及紧密联系;(3)区系起源古老。     

浙江建德蕨类植物区系研究

依据2008-2016年野外线路调查数据以及相关文献资料,运用植物区系地理学方法,对建德市的蕨类植物进行区系组成和分布区类型的分析,并将其与周边地区和省区的植物区系进行比较。结果表明:(1)建德市现有蕨类植物34科60属103种(含变种和变型),优势科为鳞毛蕨科Dryopteridaceae,水龙骨科Polypodiaceae;优势属为鳞毛蕨属Dryopteris;(2)根据现代地理分布并参照吴征镒对中国种子植物属的分布区类型的划分,建德市蕨类植物科的分布区类型有5个类型,属的分布区类型有8个类型,均以世界分布和泛热带分布占优势,表明建德市蕨类植物的热带起源;(3)科、属特有现象不明显,中国特有种占较高比例,石灰岩地区分布的种类比较特殊,并发育了地区特有种;(4)地史、化石证据表明,建德市蕨类植物区系起源古老,联系广泛,特别是与印度、中南半岛、日本等地联系十分紧密。     

黑龙江省发展林蛙产业的初步探讨

中国林蛙(Rana chensinensis),俗称田鸡,又叫哈士蟆,属于两栖纲无尾目蛙科蛙属.据《中药大辞典》记载, 哈士蟆包括2个亚种,即中国林蛙和黑龙江林蛙,主产于黑龙江、辽宁、吉林3省,属国家保护的有重要经济、科学研究价值的两栖动物.它不仅在保护森林方面有很大的作用,而且是一种名贵的滋补保健食品,是食药两用的珍贵蛙种,具有明显的润肺养阴、补肾益精、补脑益智等功能.     

银杏内生细菌XZNUM033的鉴定及其抗杨树变色真菌活性物质的理化性质

银杏内生细菌XZNUM 033对杨树变色真菌—可可球二孢菌具有很强的抑制活性,通过对该菌株培养液一些理化性质的测定,结果表明:培养液在pH值为7.0时,其抗可可球二孢菌活性最强,培养液对温度、光照、紫外线都具有较好的稳定性。根据银杏内生细菌XZNUM 033的菌落、菌体形态、革兰氏染色、芽孢染色、生理生化特性以及16S rRNA序列分析,初步把银杏内生细菌XZNUM 033定为解淀粉芽孢杆菌。     

Molecular identification of a phytoplasma associated with Sophora Root yellows

A phytoplasma infecting Sophora Root (Sophora alopecuroides) was detected and identified in Alar, Xinjiang Uygur Autonomous Region of China. Typical phytoplasma bodies were observed in sieve tubes of the diseased plants by transmission electron microscopy. A partial 16S rRNA gene and ribosomal protein (rp) genes containing rpl22 (rplV) and rps3 (rpsC) were amplified by direct and nested PCR. Based on the sequence similarity of the 16S rRNA and rp genes with accompanying phylogenetic analyses, the phytoplasma associated with Sophora Root yellows belongs to the 16SrI group (aster yellows group). Virtual RFLP analysis of these 16S rRNA and rp gene sequences showed distinct differences from those of reference phytoplasma strains representing previously described subgroups of the 16SrI group. Moreover, the similarity coefficient (0.92) of the RFLP profile of this phytoplasma was less than the threshold similarity coefficient (0.97) required for subgroup classification. Thus, the phytoplasma isolate of Sophora Root plants, designated as ‘SoRY’, represents a new subgroup. Furthermore, this is the first report of phytoplasma disease associated with Sophora Root plants.     

Phylogeographic variation among isolates of the Sirococcus conigenus P group

In this study the phylogeographic variation among isolates of the Sirococcus conigenus P group and the phylogenetic relationships of S. conigenus with Sirococcus clavigignenti‐juglandacearum and other species previously placed in the genus Sirococcus were investigated. A collection of 33 isolates originating from Picea, Pinus and Larix in Europe, North America and Bhutan were characterized by sequence analyses of the internal transcribed spacer (ITS) region (including ITS1, 5.8S ribosomal DNA, ITS2) of the nuclear rDNA and a portion of the β‐tubulin gene. In phylogenetic analyses most isolates from pine, spruce and larch formed a distinct clade, representing the P group of S. conigenus, which was separated from the T group of this pathogen. Four isolates from Picea in Europe and Canada formed a third clade within S. conigenus and these isolates are referred to as the S group. The P group consisted of five distinct ITS haplotypes, which partly differed in their optimum growth temperature and their growth rates at 25°C on malt extract agar. Nested clade analysis resolved the five haplotypes into three distinct clades and revealed significant genetic/geographic associations for some of the haplotypes. Parsimony analysis of the small subunit (18S) ribosomal DNA sequences confirmed the phylogenetic affinities between S. conigenus and S. clavigignenti‐juglandacearum. In contrast, Godronia cassandrae and Hormococcus conorum, which formerly had been placed in the genus Sirococcus, were found to be only distantly related to S. conigenus and S. clavigignenti‐juglandacearum.     

Epitypification,characterization and phylogenetic positioning of Pseudobeltrania cedrelae,the causal agent of pseudobeltrania spot on Cedrela fissilis

The fungus Pseudobeltrania cedrelae, the type species of the genus, is the causal agent of an important leaf spot in seedlings and adult plant of cedar (Cedrela fissilis). Due to the contradictory phylogenetic position of the genus Pseudobeltrania, epitypification of P. cedrelae was carried out based on a culture obtained from the same locality and host of the original type. Samples were collected, and 10 isolates of P. cedrelae associated with lesions on cedar leaflets were obtained. For morphological characterization, conidia, conidiophores, conidiogeneous cells, conidiogeneous loci and basal cells were taken both from the fungus obtained from leaf lesions and from that obtained in slide culture. Mycelial growth rates and sporulation were evaluated in six different culture media. For molecular phylogeny, maximum parsimony analyses were performed from the ITS and 28S sequences of the isolates. Both the morphological characteristics of the fungal structures obtained from symptomatic leaves and the slide culture technique presented variations. In foliar lesions, isolates presented the same morphological characteristics as the type material. Mycelial growth rate and sporulation of P. cedrelae were greatest on malt extract agar and V8 juice agar. Pseudobeltrania cedrelae was pathogenic when inoculated into healthy cedar plants. According to the phylogenetic tree, isolates grouped in the same clade, but in a distinct clade of Pseudobeltrania ocoteae. The results suggested that P. ocoteae belongs to the genus Hemibeltrania. This paper presents new information on P. cedrelae that contributes to clarifying the phylogeny of the Beltraniaceae.     

Phylogenetic and pathogenicity evaluation of the marasmioid fungus Marasmius palmivorus causing fruit bunch rot disease of oil palm

Marasmius palmivorus is a marasmioid fungal species that exhibits parasitic behaviour, although most marasmioids are rarely parasitic. The fungus has been reported to cause fruit bunch rot disease of oil palm and coconut, but only a few studies on its pathogenic behaviour are available, particularly on oil palm. Hence, there is a need to assess the ability of the fungus to act as a pathogen and to study its molecular evolution and taxonomy. Nine isolates of M. palmivorus were successfully isolated from basidiocarps and diseased fruitlets of oil palm collected from oil palm plantations and were morphologically characterized on potato dextrose agar (PDA) followed by molecular identification based on nucleotide sequence alignments of internal transcribed spacer (ITS) regions of ribosomal RNA (rRNA) gene clusters with sequences from GenBank. Koch's Postulates confirmed that M. palmivorus could infect oil palm fruitlets with symptoms similar to those observed for bunch rot disease. Molecular phylogenetic studies using nucleotide sequences of ITS and the nuclear ribosomal large subunit (nLSU) showed that isolates sharing the same phenotypic characteristics of Marasmius palmivorus and Marasmiellus palmivorus are monophyletic and share a common ancestor. The fungus has also been shown to be more closely related to the genus Marasmius than Marasmiellus; therefore, we support retention of the taxon name of the pathogen causing bunch rot disease of oil palm as Marasmius palmivorus.     

Identification of a phytoplasma associated with Syringa reticulata witches' broom disease in China

During the summer of 2018, one Syringa reticulata plant showing witches' broom and small leaves was observed in Beijing, China. Molecular diagnostic tools and electron microscopic cell observation were used to detect the possible pathogen of this disease. As a result, the phytoplasma in the symptomatic S. reticulata tree was confirmed by amplifying the 16S rRNA gene using the phytoplasma‐specific universal primer pair R16mF2/R16mR1 and observation with transmission electron microscopy. The rp and tuf genes of the phytoplasma were also cloned and sequenced as the 16S rRNA gene. Sequence and phylogenetic analyses of the 16S rRNA, rp and tuf genes indicated that the phytoplasma associated with S. reticulata witches' broom (SrWB) disease belonged to the 16SrV‐B subgroup, and it was closely related to the 16SrV‐B subgroup phytoplasma strain jujube witches' broom, which causes serious disease of jujube fruit trees in China. This study shows the S. reticulata tree as a new host of a phytoplasma belonging to the 16SrV‐B subgroup in China.     

Arambarria the pathogen involved in canker rot of Eucalyptus,native trees wood rots and grapevine diseases in the Southern Hemisphere

Arambarria (Hymenochaetales, Basidiomycota) is a monotypic genus recently described to accommodate specimens from the Patagonian forests of Argentina wrongly assigned in the past to Inocutis jamaicensis. On the basis of a wide sampling of strains and phylogenetic analysis inferred from combined sequences including the nuc rDNA ITS1‐5.8‐ITS2 region, 28S rDNA D1‐D2 domains and partial sequences of translation elongation factor 1‐a (tef1‐α), we demonstrate that this genus is associated with an important canker rot of eucalypt plantations in Uruguay, to wood rots of many native and exotic hosts, and to “hoja de malvón” and chlorotic leafroll of grapevines diseases in Central Chile, Central Argentina and Uruguay, formerly assigned to I. jamaicensis and/or Fomitiporella sp. The combined phylogenetic analysis showed the existence of three closely related clades that corresponded to (1) the Pampas of Uruguay and Argentina (“uruguay” clade), (2) the Monte, Chaco Serrano and Yungas forests of Argentina (“cognata” clade) and (3) the Patagonian Andes forests and Chilean Province (“destruens” clade). Lack of morphological differences between taxa from the three clades, their occurrence in both native and exotic hosts, previous results showing interfertility between isolates from Uruguay and Argentina, and the lack of full support in the concatenated ITS + 28S + tef1‐α analysis, prevents us to distinguish and describe three different taxa; the proper name of the taxon being Arambarria cognata comb. nov. A fourth, distinctly separated clade corresponded to South African strains isolated from vineyards representing an undescribed taxon associated with Esca grapevine disease in that country. Arambarria is shown to be unrelated to Inocutis, with which it was confused in the past and, so far, remains restricted to the Northern Hemisphere in America (Mexico, Jamaica and the USA).     

An integrative study of Sakia sisanganensis n. sp. (Rhabditida: Tylenchidae) from Sisangan forest,Iran, and new morphological observations for the genus

Further data on the morphology (the lip region characters) and phylogeny of the genus Sakia are presented. The new observations were based on scanning electron microscopy (SEM) and light microscopy (LM). A new species, Sakia sisanganensis n. sp., was recovered from rotten wood of a dead beech tree (Fagus orientalis) in northern Iran, herein described and illustrated based on an integrative approach, that is morphological, morphometric and molecular characters. The new species is characterized by a combination of the following features: fine transverse striae and vestigial single band in the lateral field in SEM. Labial area dorso‐ventrally flattened. Oral region with two concentric hexagonal plates, the inner one apparently elevated. Amphidial openings short, slit‐like. Stylet delicate. Median bulb fusiform to spindle‐shaped with weak valvular apparatus. Spermatheca functional. Tail filiform with faintly pointed tip and males common. The new species was morphologically compared with four known species of the genus, viz., S. alii, S. arboris, S. castori and S. indica, all having indistinct lateral fields. Molecular phylogenetic analyses were performed based on the small subunit ribosomal RNA gene (SSU rDNA). In the Bayesian inference (BI), S. sisanganensis n. sp. with two isolates was strongly supported as a sister taxon of a clade harbouring S. arboris + Lelenchus species. However, in the maximum likelihood (ML) analysis, the new species formed a clade with S. arboris, thus supporting the reciprocal monophyly of the genera Sakia and Lelenchus. Accordingly, the test of monophyly was performed (using Bayes factor) and the results did not reject the monophyly of sakia (i.e., S. sisanganensis n. sp. and S. arboris as sister taxa) based on the currently available data.     

河北绣线菊属植物研究

作者在编写《河北灌木志》绣线菊属时,对采于河北境内的本属植物标本进行了分类与分布研究。现知河北原产17种3变种,其中1新种,2河北新分布记录种,1河北新分布记录变种。编制了分种检索表;指出了相近种的区别点;简略记述了适生条件及分布区域。