多基因联合分析棒束孢属Isaria（Ascomycota,Cordycipitaceae）系统发育关系

对棒束孢属Isaria及近缘属物种开展5基因（nrSSU、nrLSU、tef-1α、rpb1和 rpb2）测序并联合分析,结合GenBank相关类群序列,探讨棒束孢属系统发育关系,最终获得95个菌株、58个明确分类群的2-5基因序列。利用MEGA和MrBayes软件进行多基因聚类分析,结果表明棒束孢属多系起源于虫草菌科中,分3个不同分支。A支主要由Isaria cicadae、I. teniupes、I. coleopterorum、I. fumosorosea和I. cateniannulata等组成;B支包括I. poprawkii、I. locustica、I. javanica、I. amoenerosea和I. cateniobliqua;C支仅有I. farinosa。分支间被Cordyceps militaris、C. ninchukispora、C. pruinosa等隔开。棒束孢在形态上,主要以瓶梗基部膨大、尖端变细及孢子呈链状等特征与其他类群分开,但同时也发现有棒状分生孢子梗和单孢子类型。基于节点的分歧时间预测分析,推测棒束孢属首次分化于70Mya,但棒束孢属主要物种形成却在60-55Mya,且3个分支的棒束孢物种为快速同时形成,而后大多数类群表现遗传稳定。同时发现,与Isaria Clade A较近一支有粉被玛利亚霉Mariannaea pruinosa（C. pruinosa无性型）和蛹草蚧霉Lecanicillium militaris（C. militaris无性型）;与粉棒束孢距离最近一支有Akanthomyces aculeatus（C. tuberculata无性型）和L. attenuatum（C. confragosa无性型）,是两个不同的属征分类群,且相互间遗传距离较近。根据棒束孢属及其近缘种属形态特征的复杂性推测,棒束孢属在快速物种形成中,其近缘类群存在一定程度的丢失和选择性演化。     

Cuburbitacins from the cruciferae

The distribution of cucurbitacins in seeds and growing plants of Iberis umbellata L. has been investigated. The testas of 'Giant Pink', 'Purple', 'Crimson' and 'Iceberg' (Dutch) strains of Iberis umbellata contain Cucurbitacin B with traces of D and E. Seeds of 'Iceberg' (Danish) strain were exceptional in containing cucurbitacins E and I but no B, and this strain was also distinguished by larger redder seeds and by testas which were not readily separable into two layers. Cucurbitacins are rare in other crucifers but they are present in I. gibraltarica L., I. sempervirens L. and Lepidium sativum L.     

基于多组织稳定同位素比值的热带大西洋4种鲨鱼营养生态位分化

不同代谢速率组织间的碳、氮稳定同位素比值(δ¹³C和δ¹⁵N)可以反映生物不同时间尺度的摄食信息,对探讨物种间摄食、栖息地利用和营养生态位的变化具有重要的指示作用。本研究以在大西洋热带海域兼捕的大青鲨、长鳍鲭鲨、拟锥齿鲨和尖吻鲭鲨为对象,通过测定其肌肉、肝脏和血液的δ¹³C和δ¹⁵N值,探讨4种鲨鱼营养生态位分化。结果表明: 与长鳍鲭鲨相比,尖吻鲭鲨、拟锥齿鲨和大青鲨的δ¹⁵N值相似且相对较高;大青鲨与其他鲨鱼存在摄食隔离,表现出独特的营养生态位;尖吻鲭鲨营养生态位宽幅最大,摄食食物种类和(或)栖息环境类型更多样化,其与拟锥齿鲨的营养生态位重叠度最高,说明两种鲨鱼具有潜在的资源竞争关系。尖吻鲭鲨、拟锥齿鲨和大青鲨组织间的δ¹³C、δ¹⁵N差值与其叉长均无显著相关关系,说明3种鲨鱼近期内无明显摄食变化;而长鳍鲭鲨的肝脏、血液和肌肉组织的δ¹⁵N差值与叉长显著相关,说明长鳍鲭鲨在短期内存在摄食变化。肝脏和血液的δ¹³C、δ¹⁵N值的相似性反映了两种组织整合摄食时间周期相近,其较高的代谢速率可以反映相对短时间周期的摄食信息。     

马兰的学名考证

马蔺是鸢尾科鸢尾属的植物,在我国北方甚常见,它的学名比较混乱,不同的作者引用的学名差异甚大。因此本文作一粗略考证,试图澄清,但因水平所限,讹误之处,敬请批评指正。     

中国马兜铃属和关木通属(马兜铃科)概览

马兜铃属(广义) (Aristolochia sensu lato)具有花单被、花萼管状、合蕊柱、子房下位、中轴胎座、胚珠多数、蒴果等主要特征, 广布于全世界的热带、亚热带和温带地区, 约有600种, 是马兜铃科中种类最多的属。依据Flora of China, 我国有本属植物45种, 其中33种为中国特有。近些年, 国内大量的新类群被相继报道, 特别是云南、广西两地, 而另一些类群则得到了确认、恢复、重新发表或修订。最近, 基于形态和分子证据, 关木通属(Isotrema)因其花萼管急剧弯曲; 合蕊柱3裂; 雄蕊6, 成对与合蕊柱裂片对生; 蒴果由上而下开裂等区别特征而从广义马兜铃属中被分出独立成属。本文基于大量的野外调查、标本鉴定、数码照片考证和相关文献的仔细研究, 重新梳理了中国马兜铃属和关木通属的种类及分布情况, 确认现阶段中国马兜铃属17种和关木通属58种1亚种, 并一一记述同时编制了最新的检索表。其中, 柔叶关木通(I. mollis)、线叶关木通(I. neolongifolia)的种级地位得到重新确认, 并提供了图版。探讨了优贵马兜铃(A. gentilis)、川滇马兜铃(A. chuandianensis)和纤细马兜铃(A. gracillima)的关系, 昆明关木通(I. kunmingense)与波氏关木通(A. bonatii)的关系, 以及卵叶关木通(I. ovatifolium)、葫芦叶关木通(I. cucurbitoides)、西藏关木通(I. griffithii)、过石珠(I. versicolor)、大别山关木通(I. dabieshanensis)等复合群和袋形关木通(I. saccata)等物种存在的问题。     

中国丝盖伞属裂盖组的三个新种

丝盖伞属裂盖组Inocybe sect. Rimosae sensu lato是一个分类难度较大的类群,组内的一些种类被认为是复合种。分子系统发育分析表明该组并非单系群,该组成员隶属于丝盖伞科下的2个属级分支,Inosperma分支和Pseudosperma分支。本文结合形态学特征和分子序列分析,对我国丝盖伞属裂盖组进行了分类学研究,发现3个新种：长白丝盖伞Inocybe changbaiensis、新茶褐丝盖伞I. neoumbrinella和云南丝盖伞I. yunnanensis。长白丝盖伞和新茶褐丝盖伞产自中国东北,前者生于针阔混交林下,后者生于柳树林下;云南丝盖伞产自云南省昆明市,生于阔叶树下。基于LSU序列的系统发育分析结果显示,长白丝盖伞隶属于Inosperma分支,新茶褐丝盖伞和云南丝盖伞隶属于Pseudosperma分支。本文对此进行了形态描述和特征图示,并讨论了新种与近缘种之间的异同。编制了中国丝盖伞属裂盖组已知种类的分种检索表。     

5-氮杂胞苷对桦褐孔菌多酚积累的影响

本文旨在研究DNA甲基转移酶抑制剂5-氮杂胞苷对桦褐孔菌多酚合成的调控。以液体摇瓶法培养桦褐孔菌,并在培养液中添加5-氮杂胞苷。采用荧光定量PCR测定与多酚合成相关的编码苯丙氨酸解氨酶（pal）、4-香豆酸辅酶A连接酶（4cl）和硬毛素合成酶（sps）基因表达水平,染色质免疫沉淀技术对编码硬毛素合成酶基因启动子区的组蛋白甲基化修饰进行检测,Folin-Ciocalteu法测定桦褐孔菌细胞内和发酵液中多酚的含量。结果显示,5-氮杂胞苷的添加提高了桦褐孔菌体内pal、4cl和sps基因的表达水平,改变了sps启动子区的组蛋白甲基化修饰,即降低了H3K9三甲基化修饰水平,提高了H3K4和H3K36三甲基化修饰水平,显著提高了桦褐孔菌细胞内和发酵液中多酚的积累量。5-氮杂胞苷诱导下桦褐孔菌细胞内多酚积累量达(46.6±2.8)mg/g,明显高于对照组多酚积累量(28.7±1.0)mg/g,并且胞外多酚的含量由对照组的(66.9±1.3)mg/L提高至(92.3±2.3)mg/L。此外,经5-氮杂胞苷处理后胞内多酚清除DPPH自由基、超氧阴离子和羟自由基的能力显著提高。可见,5-氮杂胞苷可以作为调节因子激发桦褐孔菌液体培养条件下多酚的合成,可以成为进一步提高桦褐孔菌液体发酵产物中多酚产量的技术手段之一。     

The anthocyanidins of some new guinea impatiens

17 strains of New Guinea Impatiens belonging to the I. schlecteri—linearifolia group and to the I. hawkeri—mooreana—nivea group were analysed for the anthocyanidin pigments. Pelargonidin predominated in the schlecteri group white malvidin was most common in the mooreana group.     

Effects of glyphosate on interspecific competition between Solidago canadensis and Imperata cylindrica

Aims The extensive use of herbicide to control invasive plants would change the relationship between alien and neighboring plants. In order to provide data for rational use of herbicide and a theoretical reference for further studies on the ecological effects of glyphosate, we explored the variation of the relationship between an invasive plant Solidago canadensis and a native plant Imperata cylindrica when they were sprayed glyphosate.
Methods A replacement series experiment was conducted from June to August 2016 in Wetland Ecosystem Research Station of Hangzhou Bay, State Forestry Administration, to examine the effects of glyphosate at seven concentration levels (0, 0.3, 0.6, 0.9, 1.2, 1.5, 1.8 mL·L^-1) on the growth and interspecific competition of S. canadensis and I. cylindrica.
Important findings (1) Glyphosate treatment significantly inhibited the growth of S. canadensis and I. cylindrica (p < 0.05). During the test, cumulative growth of height and leaf number of S. canadensis were apparently reduced with the increase of glyphosate concentration, but the leaf number of S. canadensis treated with 0.3- 1.5 mL·L^-1 glyphosate was re-growing with time, while the one treated with 1.8 mL·L^-1 was mostly dead. The withering rate of tiller and green leaf of I. cylindrica also significantly increased with the increase of glyphosate concentration, and the growth indices of this plant treated with 0.3-0.6 mL·L^-1 were also re-growing with time. (2) Glyphosate treatment significantly affected interspecific competition (p < 0.05), which diminished as the glyphosate concentration increased. (3) Interspecific competition has significant influence on the biomass allocation of S. canadensis (p < 0.05). When facing competition, S. canadensis would allocate more organic matter to root and thus increase the ratio of root to shoot. Competition only inhibited the tiller number and total biomass of I. cylindrica, but insignificantly affected its ratio of root to shoot. (4) The interaction between glyphosate treatment and S. canadensis-I. cylindrica interspecific relationship also significantly influenced the biomass of S. canadensis and I. cylindrica (p < 0.05), but insignificantly affected the root/shoot ratio of two plants. Different plants have different tolerance to glyphosate stress. Compared with native plant I. cylindrica, S. canadensis has stronger tolerance to glyphosate. Low-concentration glyphosate could decrease the competitive intensity between S. canadensis and I. cylindrica, which may disturb the structure and dynamics of plant communities.     

土赤壳属三个中国新记录种

通过形态特征观察、rDNA-ITS和histone H3序列测定分析,报道分离自西藏药用植物一把伞南星Arisaema erubescens和工布乌头Aconitum kongboense的土赤壳属3个中国新记录种：埃什特雷莫什土赤壳Ilyonectria estremocensis、假毁土赤壳Ilyonectria pseudodestructans和强壮土赤壳Ilyonectria robusta,并对其进行了形态学描述和系统发育分析。研究菌株保存于北京协和医学院药用植物研究所生物技术中心菌种库。     

浙江凤仙花属的订正——国产凤仙花属植物的研究(二)

作者对浙江的凤仙花属进行了订正, 共记载有11种, 除描述了Impatiens chekiangensis, I.platysepala, I.chlroxantha和I.tienmushanica 4新种外, 还有3个新记录, 对I.cosmia Hook.f.作了归并, 并附有分种检索表, 本文是《国产凤仙花属植物研究》的续篇.     

入侵植物五爪金龙根系深色有隔内生真菌的侵染特征

深色有隔内生真菌（DSE）广泛侵染华南地区外来入侵植物五爪金龙。显微观察发现五爪金龙根系具有大量的泡囊,其最显著的特征是大量脂质的富集和被分解成小脂质体,并通过透明或深色菌丝转移,说明了五爪金龙和DSE之间进行碳转移。富含脂质的泡囊可产生透明菌丝和自我复制,说明泡囊具有繁殖功能。富含脂质的泡囊和透明菌丝等非典型结构没有可分辨的或具透明的细胞壁,是活跃的真菌结构。透明菌丝和富含脂质的泡囊主要定殖在生理活性强的根组织中。深色有隔菌丝和微菌核是DSE的典型结构,主要定殖在相对不活跃的根组织中。DSE定殖于五爪金龙根的维管柱、皮层、表皮细胞和表面,同时延伸到土壤,在宿主植物和土壤之间形成连续的系统的真菌网络。根据DSE的多态性质、庞大的真菌网络和脂质的富集及其分布,探讨了DSE和入侵植物五爪金龙潜在的互惠共生关系。     

花群珊瑚对两种微藻的摄食

以花群珊瑚为材料,采用碳清除率测定、PCR分子营养标记和组织学观察的方法,研究花群珊瑚是否摄食亚心形扁藻和球等鞭金藻。结果表明: 花群珊瑚对球等鞭金藻的碳清除率显著高于亚心形扁藻,分别为0.44和0.11 pg·mL^-1·polyp^-1·h^-1。球等鞭金藻烯酰基载体蛋白还原酶基因片段作为分子标记在投喂组珊瑚组织中可扩增出目的片段。亚心形扁藻18S rRNA基因的引物在相应投喂组珊瑚组织中也可扩增出目的片段。组织学观察发现: 投喂组珊瑚水螅体内部隔膜较宽,隔膜与体壁的胃层都存在大量食物泡。亚心形扁藻投喂组在体壁胃层的食物泡中和口道沟处均发现未消化完全的亚心形扁藻。球等鞭金藻投喂组则在隔膜胃层和体壁胃层内的食物泡中发现所摄食金藻。因此,碳清除率、分子营养标记以及组织学观察均表明花群珊瑚对亚心形扁藻与球等鞭金藻存在摄食现象。     

Anti-microtubule activity of the traditional Chinese medicine herb Northern Ban Lan (Isatis tinctoria) leads to glucobrassicin

Traditional Chinese medicine (TCM) belongs to the most elaborate and extensive systems of plant-based healing. The herb Northern Ban Lan (Isatis tinctoria) is famous for its antiviral and anti-inflammatory activity. Although numerous components isolated from I. tinctoria have been characterized so far, their modes of action have remained unclear. Here, we show that extracts from I. tinctoria exert anti-microtubular activity. Using time-lapse microscopy in living tobacco BY-2 (Nicotiana tabacum L. cv Bright Yellow 2) cells expressing green fluorescent protein-tubulin, we use activity-guided fractionation to screen out the biologically active compounds of I. tinctoria. Among 54 fractions obtained from either leaves or roots of I. tinctoria by methanol (MeOH/H₂O 8:2), or ethyl acetate extraction, one specific methanolic root fraction was selected, because it efficiently and rapidly eliminated microtubules. By combination of further purification with ultra-high-performance liquid chromatography and high-resolution tandem mass spectrometry most of the bioactivity could be assigned to the glucosinolate compound glucobrassicin. Glucobrassicin can also affect microtubules and induce apoptosis in HeLa cells. In the light of these findings, the antiviral activity of Northern Ban Lan is discussed in the context of microtubules being hijacked by many viral pathogens for cell-to-cell spread.     

Genetic diversity and population structure of endangered Isoetes coreana in South Korea based on RAPD analysis

Isoetes coreana Chung and Choi is on the list of critically endangered species in South Korea. Using randomly amplified polymorphic DNA (RAPD) markers, we investigated the genetic diversity within and between seven local populations of I. coreana. Ten RAPD primers produced a total of 94 bands, of which 59 (62.8%) were polymorphic. A low level of genetic diversity was recognized within the populations of I. coreana: polymorphic loci (P), with values ranging from 3.4% to 33.9%, and a mean value of 15.5% being observed. Analysis of molecular variance (AMOVA) showed that genetic diversity was greater among populations (81.6%) than within populations (19.7%) or among the three regions included in this study (Han River, Youngsan River, and Nakdong River). In addition, a high degree of genetic differentiation (θ^B = 0.742) was detected among the populations. These results indicate that genetic differentiation has occurred very rapidly. However, the rate of gene flow between populations was found to be as low as 0.059, irrespective of the genetic and geographical distances between the populations, which indicates that genetic drift must have played an important role in forming the present populations of I. coreana. Because a reduction of genetic diversity as a result of genetic drift is undesirable, increasing the gene flow between populations of Korean quillwort I. coreana should be considered as a conservation strategy.     

入侵植物加拿大一枝黄花根际解钾菌多样性及解钾活性

入侵植物加拿大一枝黄花(Solidago canadensis)具有较强的钾(K)富集能力, 这可能和其对土壤微生物群落的改变有关。根际解钾菌能够将植物难以利用的矿物态钾转化为植物可以利用的可溶性钾, 而加拿大一枝黄花如何影响根际解钾菌多样性和解钾活性尚未明了。该研究以浙江省杭州湾湿地围垦区内自然生长的加拿大一枝黄花和其伴生本地植物白茅(Imperata cylindrica)为研究对象, 比较了加拿大一枝黄花和白茅体内及土壤中的钾含量水平, 钾供给水平对生物量积累的影响, 以及根际解钾菌的数量、多样性和解钾活性的差异。结果表明, 加拿大一枝黄花茎、叶中的钾含量均显著高于白茅, 分别是白茅的1.59和7.33倍; 加拿大一枝黄花和白茅的土壤全钾含量差异不显著, 速效钾含量在0-10 cm土层中差异显著、在10-20 cm土层中差异不显著。随着钾供应水平提高, 加拿大一枝黄花和白茅的生物量均显著增加。利用解钾培养基计数培养后发现, 加拿大一枝黄花根际解钾菌的数量是白茅的3.51倍。分离培养后将出现解钾圈的菌株进行鉴定, 利用解钾液体培养实验测定其解钾量, 发现从加拿大一枝黄花根际土中分离得到的15个解钾菌株中, 有9个具有高效解钾能力, 其处理液中K ⁺含量较空白对照高出85.11%-192.54%, 其中菌株H2-20解钾能力最强, 解钾量为10.657 mg·L ^-1。加拿大一枝黄花根际解钾菌解钾作用显著高于白茅。经16S rDNA鉴定发现, 加拿大一枝黄花15个根际解钾菌株分属11个属, 其中有6个属已经被报道证实具有明显解钾能力。这些结果表明加拿大一枝黄花根际解钾菌数量较为丰富, 且大多具有较高解钾活性, 可能对其钾富集具有重要贡献。     

Modelling various sculptures in the Cretaceous bivalve Inoceramus hobetsensis

The geometry of the external shell sculpture in the Late Cretaceous inoceramid bivalve Inoceramus hobetsensis Nagao & Matsumoto, 1939 was studied both empirically and theoretically. A large sample, collected from the Upper Cretaceous of Hokkaido, Japan, shows remarkably high intraspecific variation in the shell sculptural pattern. Quasi-commarginal ribs, slightly oblique to the external growth increments, occur in some specimens. These sculptures are commonly irregular in strength and spacing, and their features are successfully modelled by computer simulations when the commarginal ribs are superposed with nearly concentric divaricate rib. Computer models indicate that the divergent sculpture element, often found in other inoceramids, was present throughout the evolution of I. hobetsensis and was developing in the evolutionary lineage from I. hobetsensis nonsulcatus to I. hobetsensis hobetsensis, although it was only weakly expressed. The results also suggest that some apparently distinct sculptural patterns of I. hobetsensis are the result of minor changes in the morphogenetic program.     

Rearing Tardigrades: Results and Problems

We report our first results of attempts to rear four species of eutardigrades inhabiting different substrates, feeding on different kinds of food and characterized by different sexual conditions and modes of reproduction. Attempts were carried out to follow individual terrestrial carnivorous (Macrobiotus richtersi, M. joannae) and limnic herbivorous (Diphascon cf. scoticum; Isohypsibius monoicus) species. Carnivorous leaf litter-dwelling species were reared in small dishes containing agar as substrate and bacteriophagous nematodes as food. Five generations were obtained with the triploid thelytokous strain of M. richtersi, whereas three generations were obtained with the hermaphrodite species M. joannae. Diphascon cf. scoticum and I. monoicus were reared in small dishes containing algae as food and substrate. Several generations were obtained for both species. Males were never found in D. cf. scoticum and I. monoicus was hermaphroditic. Specimens isolated from hatchings were maintained and reproduced in both species, demonstrating parthenogenesis in the first one and self-fertilization in the latter. Consideration of the problems and on the future applications of tardigrade rearing are discussed.     

蝙蝠蛾幼虫感染粉棒束孢后的病症、组织病理变化及侵入途径

蝙蝠蛾幼虫的饲养是冬虫夏草培植的先决条件,粉棒束孢作为蝙蝠蛾幼虫饲养过程中的一种常见和主要致病菌,具有极强的致病性和流行性。结合形态学观察和组织切片技术,本文研究了蝙蝠蛾幼虫感染病原菌粉棒束孢后的症状、主要组织病理变化及粉棒束孢侵染蝙蝠蛾幼虫的途径。结果表明,用浓度为1×10⁵个/mL的粉棒束孢分生孢子悬液处理蝙蝠蛾5龄幼虫,4d后在腹部气门处有黑斑出现,气门变黑,虫体随后瘫痪死亡、体色改变、收缩化僵、菌丝层包裹、形成众多孢梗束等症状。幼虫被粉棒束孢感染后,菌丝体首先利用血淋巴进行增殖,脂肪体、肌肉、消化道、丝腺和马氏管等主要组织器官先后出现不同程度的破坏,虫体体腔被大量菌丝充斥并最终僵化。组织切片观察结果表明,粉棒束孢可以通过气门侵入蝙蝠蛾幼虫体内并造成虫体死亡,菌丝穿透气管进入血淋巴后断裂成许多长短不一的短菌丝段,目前尚未观察到菌丝直接穿透幼虫体壁和从消化道侵入的现象。本研究为蝙蝠蛾幼虫饲养过程中的病害防治提供信息参考。     

白囊耙齿菌产锰过氧化物酶条件优化及其对染料的脱色

锰过氧化物酶（manganese peroxidase,MnP）是白腐真菌降解多种异生物质的主要降解酶之一。本研究对白囊耙齿菌Irpex lacteus产MnP的酶活曲线进行监测,利用单因素和正交试验对I. lacteus产MnP的发酵条件进行优化,同时检测了I. lacteus的MnP粗酶液对5种染料的脱色效果。结果显示,I. lacteus在培养5d时MnP活性较大;I. lacteus产MnP较优的条件为：可溶性淀粉20g/L、尿素1g/L、pH 6.3、CaCl₂ 1mmol/L、FeCl₃ 1mmol/L,该条件下MnP活性达29.24U/L,与优化前MnP活性相比提高了1.25倍;I. lacteus的MnP粗酶液对5种染料均可脱色,其中对直接大红和活性红的脱色效果更为明显,脱色5d后的脱色率分别达到82%和81%。