黑龙江省蜜环菌生物种的地理分布概况

自1999年至2002年间对黑龙江省境内几个主要林区中的蜜环菌生物种进行了调查、采集和鉴定,并对其寄主种类和子实体发生规律以及地理分布特点进行了总结分析。结果表明:黑龙江省境内至少存在5个蜜环菌生物种,即芥黄蜜环菌Armillaria sinapina,高卢蜜环菌A.gallica,黄盖蜜环菌A.luteopileata,奥氏蜜环菌A.ostoyae和中国生物种F;寄主包括10个针叶和阔叶树种;与其他几种蜜环菌生物种相比较,A.gallica是黑龙江省境内出现概率最大的一个生物种;根据采集经验发现,温度和湿度是影响子实体发生的主要因子。     

中国北方珊瑚状真菌

本文描述了采集于吉林、黑龙江省的珊瑚菌,并对其系统分类位置和生物地理分布做了记录。描述的类群分别隶属于Clavaria, Lentaria, Multiclavula, Pterula, Ramaria,和Rcrmariopsis属,其中Ranurria crmericana var. orientalis为新变种。     

大兴安岭和长白山地区蜜环菌生物种的研究

从大兴安岭和长白山采集到30号蜜环菌(Armillaria mellea)标本,选其中有代表性的10个号的担子果获得单孢株。交配试验表明每一担子果都具有双因子异宗配合系。不同子实体交配型之间交配结果表明,在大兴安岭和长白山地区蜜环菌目前至少存在5个生物种,分别称为生物种A、B、C、D和E。将这5个生物种的单孢菌种与欧洲5个蜜环菌生物种的单孢菌株进行配合,生物种B与欧洲A.gallica,生物种E与欧洲A.ostoyae亲和交配,因此将生物种B和E分别定为A.gallica和A.ostoyae。生物种A、C和D则不与任何欧洲生物种交配。     

粗毛栓菌诱变菌株SAH-12漆酶基因的克隆与序列分析

粗毛栓菌Trametes gallica诱变菌株SAH-12是通过紫外诱变选育得到的漆酶高产菌株。为了对其漆酶基因进行研究和利用,采用cDNA末端快速扩增(Rapid Amplification of cDNA Ends,RACE)技术,从T.gallica诱变菌株SAH-12分离得到漆酶基因全长cDNA Lacc1(GenBank accession No.DQ431716)及其对应的结构基因Lac1(DQ431715)。该基因属于真菌漆酶基因家族,与来自出发菌T.gallica漆酶基因lacA(AY875867)在成熟肽编码区的同源性最高(一致性为98%)。Lacc1全长1891bp,由40bp的5'-UTR、1554bp的完整ORF和297bp的3'-UTR构成,具有polyA加尾信号AATACA和59bp的polyA结构;其完整ORF可编码21个氨基酸残基组成的信号肽和496个氨基酸残基组成的成熟蛋白。在Lacc1基因的推导氨基酸序列中有4个潜在的N-糖基化位点和4个参与二硫键形成的Cys残基,且含有真菌漆酶Ⅰ、Ⅱ、Ⅲ型铜离子结合区的4个高度保守序列。结构基因Lac1全长2338bp,含10个内含子和11个外显子,各内含子长度在51bp~76bp之间,且其序列均符合5'-gt…ag-3'规则。     

粗毛栓菌诱变菌株SAH-12漆酶基因的克隆与序列分析

粗毛栓菌Trametes gallica诱变菌株SAH-12是通过紫外诱变选育得到的漆酶高产菌株。为了对其漆酶基因进行研究和利用,采用cDNA末端快速扩增(Rapid Amplification of cDNA Ends,RACE)技术,从T.gallica诱变菌株SAH-12分离得到漆酶基因全长cDNA Lacc1(GenBank accession No.DQ431716)及其对应的结构基因Lac1(DQ431715)。该基因属于真菌漆酶基因家族,与来自出发菌T.gallica漆酶基因lacA(AY875867)在成熟肽编码区的同源性最高(一致性为98%)。Lacc1全长1891bp,由40bp的5'-UTR、1554bp的完整ORF和297bp的3'-UTR构成,具有polyA加尾信号AATACA和59bp的polyA结构;其完整ORF可编码21个氨基酸残基组成的信号肽和496个氨基酸残基组成的成熟蛋白。在Lacc1基因的推导氨基酸序列中有4个潜在的N-糖基化位点和4个参与二硫键形成的Cys残基,且含有真菌漆酶Ⅰ、Ⅱ、Ⅲ型铜离子结合区的4个高度保守序列。结构基因Lac1全长2338bp,含10个内含子和11个外显子,各内含子长度在51bp~76bp之间,且其序列均符合5'-gt…ag-3'规则。     

大兴安岭和长白山地区蜜环菌生物种的研究

从大兴安岭和长白山采集到３０号蜜环菌标本,选其中有代表性的１０个号的担子果获得单孢株。交配试验表明每一担子果都具有双因子异宗配合系。不同子实体交配型之间交配结果表明,在大兴安岭和长白山地区蜜环菌目前至少存在５个生物种,分别称为生物种Ａ、Ｂ、Ｃ、Ｄ和Ｅ。将这５个生物种的单孢菌种与欧洲５个蜜环菌生物种的单孢菌株进行配合,生物种Ｂ与欧洲Ａ．ｇａｌｌｉｃａ,生物种Ｅ与欧洲Ａ．ｏｓｔｏｙａｅ亲和交配,因此将     

蜜环菌属真菌CPCC 400891中倍半萜合成酶多样性与功能研究

本论文以蜜环菌属（Armillaria mellea）真菌CPCC 400891为研究对象，基因组挖掘分析发现该菌株基因组中含有18个倍半萜合成酶编码基因，但其功能却鲜有报道。采用色氨酸营养缺陷型表达载体pYET，并以酿酒酵母（Saccharomyces cerevisiae）BJ5464为宿主菌分别表达蜜环菌来源的18个倍半萜合成酶编码基因（Arm_STS1-Arm_STS16）。结合气相色谱-质谱联用仪（gas chromatography-mass spectrometer，GC-MS）分析平台，对构建的重组菌（BJ-Arm_STS1-BJ-Arm_STS16）发酵产物进行采集分析。结果显示，蜜环菌CPCC 400891来源的12个倍半萜合成酶归属于新的类群：亚家族Ⅳ。同时，10个倍半萜合成酶在酿酒酵母中确定表达。根据美国国家标准与技术研究院（NIST）的标准参考数据库（https：//webbook.nist.gov/）检索比对，共鉴定17个倍半萜（醇）分子，包括α-雪松烯、β-雪松烯、罗汉柏烯、杜松萜烯、β-花柏烯和毕橙茄醇等。GC-MS检测分析发现，亚家族Ⅳ类群倍半萜合成酶均能合成多个（种）倍半萜分子。综上所述，本研究为后续改造并优化特定倍半萜合成酶，从而为深入开展STSs的催化机制研究奠定基础，也为从蘑菇类大型真菌中发现更多新颖的倍半萜合成酶提供依据和思考。     

担子菌门总革菌科中国新记录种紫黑点壳菌

[背景] 在陕西省的毛泡桐腐木桩上观察和采集到一株绒毛状真菌子实体,编号为HMNWAFU-CF-HS002。[目的] 为了确定该菌的分类地位,对其进行形态观察及分子鉴定。[方法] 获取宏观彩色图像并对显微结构进行测量、统计和绘图。此外,用PDA培养基分离纯化该菌的培养物,并结合最大似然法、最大简约法和贝叶斯法进行rDNA ITS分子系统学研究。[结果] HMNWAFU-CF-HS002的形态特征与Punctularia atropurpurascens高度相似。系统发育分析将HMNWAFU-CF-HS002聚在P.atropurpurascens的单系分支中。[结论] 结合形态学特征与系统发育结果,HMNWAFU-CF-HS002被鉴定紫黑点壳菌（Punctularia atropurpurascens）,为中国的新记录种。此外,毛泡桐为其新记录寄主。至此,Punctularia属下的3个种,均在中国有分布记录。     

中国蜜环菌生物种的RAPD分析

以中国蜜环菌(Armillaria mellea (vahl: Fr.) Kummer)生物种B．生物种E分别与欧洲种A. gallica 和A. ostoyae交配,用AP—PCR技术分析了中国五个生物种及欧洲2个种的代表菌株的系统发育关系。根据系统聚类分析的结果,将其分为4个群：生物种B与A．Gallia, A与C,D与E,A. ostoyae 单独为一群。这与交配试验及RAPD图谱直观分析的结果一致,最小支撑树也支持将中国生物种B鉴定为A．Gallica。证明RAPD是研究蜜环菌生物种的进化关系的有用手段。     

中国炭黑粉菌属及近似属(黑粉菌目)

本文报道我国炭黑粉菌属12种,其中新种1个,即小孢炭黑粉菌Anthracoidea microspora L. Guo;中国新记录种4个,即柄囊苔草炭黑粉菌Anthracoidea eleocharidis,林氏炭黑粉菌Anthracoidea lindebergiae(Kukkonen) Kukkonen,迷炭黑粉菌Anthracoidea misandrae Kukkonen和尼泊尔炭黑粉菌Anthracoidea nepalensis Kakishima & Ono,及核黑粉菌2种和异腥黑粉菌1种。     

Advances toward DNA-based identification and phylogeny of North American Armillaria species using elongation factor-1 alpha gene

The translation elongation factor-1 alpha (EF-1α) gene was used to examine the phylogenetic relationships among 30 previously characterized isolates representing ten North American Armillaria species: A. solidipes (=A. ostoyae), A. gemina, A. calvescens, A. sinapina, A. mellea, A. gallica, A. nabsnona, North American biological species X, A. cepistipes, and A. tabescens. The phylogenetic relationships revealed clear separation of all ten North American Armillaria species, with the exception of one A. gallica isolate that perhaps represents an unnamed cryptic species. These results indicate that the EF-1α gene could potentially serve as a diagnostic tool for distinguishing among currently recognized North American biological species of Armillaria.     

A novel Fomitiporia species associated with esca on grapevine in South Africa

Esca disease of grapevine is a complex trunk disease consisting of several symptoms, one of which, white rot, has been found to be caused by various basidiomycetes within the order Hymenochaetales. During recent surveys of esca-related pathogens in South African vineyards, several unidentified basidiomycetes were isolated from white rot occurring in diseased vines. A new Fomitiporia species, F. capensis, is described based on fruit body morphology and combined internal transcribed spacer (ITS) and large-subunit (LSU) phylogeny, where it forms a clearly delineated and well-supported clade. Morphologically, F. capensis is similar to F. punctata in that both species essentially lack setae. Fomitiporia capensis, F. punctata and F. aethiopica produce similarly sized basidiospores but differ in terms of host range, pore size and, possibly, fruit body shape. Phylogenetically, F. capensis appears to be related to F. tenuis, though morphologically, the species differ significantly in that F. tenuis has smaller pores and smaller basidiospores. Fomitiporia capensis was found to occur widely as vegetative mycelium throughout the Western Cape Province, though fruit bodies were scarce in comparison. A vineyard with fruit bodies was also found in Limpopo in the north east of the country. Fruit bodies were found growing on the underside of the cordon of living vines displaying external symptoms typically associated with esca, or general decline and dieback symptoms together with internal white rot.     

Molecular phylogeny of Armillaria from the Patagonian Andes

A number of species in the plant pathogen genus Armillaria are known from South America where they cause root rot disease on a wide variety of hosts. Knowledge pertaining to phylogenetic relationships of these species with those of other Armillaria species is almost non-existent. In addition, very few cultures representing these species are available, making DNA-based phylogenetic analyses impossible. The aim of this study was to characterise a collection of Armillaria isolates from the Patagonian Andes using DNA sequences and to determine their phylogenetic relationships with other Armillaria species. DNA sequences were obtained from the internal transcribed regions (ITS1, 5.8S and ITS4) and ribosomal large subunit (LSU) gene and used in phylogenetic analyses. Phylogenetic trees generated from the sequences separated the Armillaria isolates into four lineages. Lineages I and II represented A. novae-zelandiae and A. luteobubalina, respectively. Isolates belonging to A. novae-zelandiae from Malaysia, New Zealand, Australia and South America showed considerable intra-clade sub-structure. Lineages III and IV are probably distinct species and are most closely related to A. hinnulea and an unnamed species isolated from New Zealand and Kenya. This is the first comprehensive study of the phylogenetic relationships of Armillaria species from Patagonia and it provides a foundation for future research in this region.     

发生在云南与东盟边境的一种新害虫——热带拂粉蚧(同翅目:粉蚧科)

【背景】热带拂粉蚧属于粉蚧类昆虫,食性杂,是危害热带和亚热带水果、蔬菜和园林植物的重要害虫,主要随水果、苗木和交通工具等介质进行远距离传播。2014年5月,在我国云南与缅甸边境发现该虫危害草本植物马松子。【方法】在收集整理热带拂粉蚧生物学、地理学等信息的基础上,介绍了其主要形态特征、寄主、分布、生物学特性,并对该有害生物的危害和防治进行了综合分析。【结果】热带拂粉蚧雌成虫长椭圆形,触角8节,体长2.5~3.0 mm,宽1.5~2.0 mm,主要通过孤雌生殖繁殖后代,形态特征与双条拂粉蚧非常相似。我国的广东、广西、海南、云南、福建等热带地区适合该虫生存和危害,一旦入侵我国会对花卉、水果和蔬菜等造成危害,给相关产业带来损失。【结论与意义】热带拂粉蚧在我国有传播扩散的可能,加强检疫是防范该虫入侵的主要手段,生物防治是治理该虫的重要措施。     

Diversity and ecology of <Emphasis Type="Italic">Armillaria</Emphasis> species in virgin forests in the Ukrainian Carpathians

In this study, we investigated the diversity and ecology of Armillaria species in virgin pure beech and mixed conifer forests (15,000 ha) of the Carpathian Biosphere Reserve in Ukraine. Armillaria rhizomorphs were systematically sampled, both from the soil and from the root collar of trees (epiphytic), on 79 plots (25 × 20 m) of a 1.5 × 1.5 km grid. In both forest massifs, rhizomorphs were present in the majority of the soil samples, with an estimated dry weight of 512 kg/ha in the pure beech forests and 223 kg/ha in the mixed conifer forests. Similarly, in both forest massifs, most of the trees inspected had rhizomorphs at the root collar. Species identification based on DNA analyses showed that all five annulated European Armillaria species occur in these virgin forests, as previously observed in managed forests in central Europe. However, differences in the frequencies of the single species were observed. The predominance of the preferentially saprotrophic A. cepistipes and A. gallica (84 and 15% of the specimens, respectively) and the absence of significant pathogenic activity suggest that in these virgin forests Armillaria species are most likely to behave as saprotrophs. Forest management may increase the frequency of the pathogenic species A. ostoyae, which is rare in virgin forests.     

The phylogenetic position of an <Emphasis Type="Italic">Armillaria</Emphasis> species from Amami-Oshima,a subtropical island of Japan,based on elongation factor and ITS sequences

An undetermined Armillaria species was collected on Amami-Oshima, a subtropical island of Japan. The phylogenetic position of the Armillaria sp. was determined using sequences of the elongation factor-1α (EF-1α) gene and the internal transcribed spacer (ITS) region (ITS1-5.8S-ITS2) of ribosomal DNA (rDNA). The phylogenetic analyses based on EF-1α and ITS sequences showed that this species differs from known Japanese taxa of Armillaria. The sequences of this species and A. novae-zelandiae from Southeast Asia were contained in a strongly supported clade, which was adjacent to a well-supported sister clade containing A. novae-zelandiae from Australia and New Zealand.     

Interaction dynamics between saprobic lignicolous fungi and Armillaria in controlled environments: Exploring the potential for competitive exclusion of Armillaria on peach

Armillaria root rot, caused by Armillaria tabescens and Armillaria mellea, is a major cause of premature tree death in peach orchards in the southeastern United States. The root systems of infected trees can become entirely colonized by Armillaria, serving as an inoculum source for adjacent trees and providing massive inoculum levels in replant situations. If dead or dying trees could be colonized by an effective competitor of Armillaria before their removal, the extent of root colonization by the pathogen could be reduced, thus decreasing the threat to adjacent trees and/or subsequent plantings. Interactions between five species of saprobic lignicolous fungi (Ganoderma lucidum, Hypholoma fasciculare, Phanerochaete velutina, Schizophyllum commune, and Xylaria hypoxylon) and the two Armillaria species were examined in controlled conditions to provide proof of concept for competitive exclusion of Armillaria from peach roots. On agar-coated glass slides, all five potential antagonists induced detrimental reactions in >58% of the Armillaria hyphae observed, with the majority resulting in hyphal swelling or granulation. On poplar wood blocks, all antagonists consistently either overgrew Armillaria colonies or—in the case of S. commune—engaged in deadlock reactions; in all cases, the viability of Armillaria colonies was reduced to <30% of that of unchallenged controls. When inoculated simultaneously onto opposite ends of peach root segments, all antagonists consistently reduced growth and viability of Armillaria on and under the bark, whereby reduction of pathogen growth underneath the bark, Armillaria’s primary ecological niche, was most pronounced for G. lucidum, S. commune, and X. hypoxylon. When root segments were allowed to be colonized entirely by Armillaria before being inoculated with the antagonists, the latter were able to overgrow the pathogen on the root surface but unable to pre-empt it from underneath the bark. In summary,G. lucidum, S. commune, and X. hypoxylon caused strong hyphal and mycelial interference reactions and the most pronounced reductions in growth of Armillaria above and below the bark, indicating that they would be the most promising candidates for field-scale evaluations to restrict colonization of dead or dying peach trees by Armillaria in the orchard.     

VARIATION IN RIBOSOMAL DNA AMONG BIOLOGICAL SPECIES OF ARMILLARIA,A GENUS OF ROOT-INFECTING FUNGI

Armillaria is a genus of root-infecting fungi composed of several biological species in North America and Europe. To examine relatedness among biological species, ribosomal DNA (rDNA) from one isolate was cloned and rDNAs from 30 isolates were mapped for eight restriction enzymes. The positions of the large (26S) and small (18S) rRNA cistrons were found by Northern hybridizations of total cellular RNA with rDNA subclones and by alignment of maps with conserved restriction sites present in rRNA genes of other fungi. Nine restriction-site and two length polymorphisms were observed. Eight North American (Roman numerals) and five European (species epithets) biological species could be placed in six classes with respect to rDNA maps (rDNA class 1: I and A. ostoyae; class 2: II; class 3: A. borealis; class 4: V, IX, and X; class 5: III, VII, A. lutea, and A. cepistipes; and class 6: VI and A. mellea). Most, but not all, polymorphisms were in intergenic regions.     

Natural establishment of a parasitoid complex on Bactrocera latifrons (Diptera: Tephritidae) in Hawaii

Bactrocera latifrons (Hendel) (Diptera: Tephritidae) is the most recent of four tephritid fruit fly species accidentally introduced into Hawaii. Although parasitoids have been released against other tephritid fruit fly species and have shown partial success in Hawaii, no parasitoids were released until 2004 to suppress populations of B. latifrons. The present study was conducted to document the parasitoid complex that has naturally established against B. latifrons in Hawaii and to assess whether there is a need for improving the biological control of this species. Based on ripe turkeyberry (Solanum torvum Sw) fruit collections over three consecutive years B. latifrons was the dominant tephritid fruit fly infestating turkeyberry at all four sites surveyed, across three major islands in Hawaii. The overall percentage parasitism of B. latifrons ranged from a low of 0.8% (Hana, Maui) to a high of 8.8% (Kahaluu, Oahu). Five primary parasitoid species were recovered from individually held B. latifrons puparia: Fopius arisanus (Sonan), Psyttalia incisi (Silvestri), Diachasmimorpha longicaudata (Ashmead), D. tryoni (Cameron), and Tetrastichus giffardianus Silvestri. F. arisanus was the predominant parasitoid at three of the four sites. Low levels of parasitism suggest that there is a need to improve biological control of B. latifrons, to minimize chances of this species causing economic impacts on crop production in Hawaii. We discuss the possibility of improving biological control of B. latifrons through augmentative releases of F. arisanus or introduction and release of specific and efficient new parasitoid species.