广西葛根内生细菌的分离鉴定及其促生特性

【背景】植物内生菌长期与宿主共生,对宿主生长发育产生影响。葛根作为重要的药食两用作物,葛根内生菌的研究具有重要实践意义。【目的】对广西葛根根部内生细菌进行分离、鉴定及促植物生长特性分析,旨在了解该药食同源植物内生细菌种群结构及其促生特性,为分析内生菌群体在药食同源植物产量和品质形成的作用及其内生细菌资源的开发利用提供参考。【方法】采用6种不同的培养基从广西葛根的根瘤、根系和根愈伤组织分离内生细菌,16S rRNA基因测序和系统发育分析内生细菌的分布特征和遗传多样性,采用生理生化方法测定分离菌株的固氮活性、溶磷特性、产生嗜铁素、分泌吲哚乙酸(indole-3-aceticacid,IAA)等促生特性。【结果】从葛根根瘤、根系和根部愈伤组织中共分离得到223个菌株,16S rRNA基因测序鉴定这些菌株隶属于2门4纲10科19属,其中芽孢杆菌属、假单胞菌属、土壤杆菌属、肠杆菌属为葛根优势菌群;内生细菌数量和群落组成存在明显的组织特异性,其数量表现为根瘤>根系>根愈伤组织,但其种群多样性表现为根愈伤组织>根系>根瘤。不同培养基分离出的细菌种群丰富度有差异。从供试菌株中筛...     

从种内、种间化感作用分析入侵种薇甘菊及其近邻种的非协同进化趋势

为预测入侵植物与其邻近物种间的非协同进化趋势,分别提取薇甘菊(Mikania micrantha)及其近邻种五爪金龙(Ipomoea cairica)、葛藤(Pueraria lobata)和鸡矢藤(Paederia scandens)的叶片水提液处理幼苗,对薇甘菊及其近邻种的种内、种间化感作用进行研究。结果表明,薇甘菊对3种近邻种的化感作用呈现不同趋势(SE=0.50, 0.61,–0.16),但种内存在较强的化感促进作用(SE=0.61),说明其种内化感竞争较弱;葛藤的种内化感促进作用(SE=0.32)弱于薇甘菊,种间的化感促进作用与薇甘菊相似(SE=0.52, 0.50);五爪金龙种内化感促进作用(SE=0.06)弱于薇甘菊和葛藤(SE=0.32),种间促进作用(SE=0.24)弱于薇甘菊(SE=0.61);鸡矢藤的种内、种间化感作用均表现为抑制作用(SE=–0.18,–0.07),说明种内、种间化感竞争较强。野外调查表明4种植物分布的频度和多度均表现为薇甘菊葛藤五爪金龙鸡矢藤,这与化感竞争力结果一致。可见,种内与种间化感作用在入侵植物与近邻植物的综合竞争力中扮演着极为重要的作用,这为入侵种与近邻种之间的非协同进化趋势指明了方向。     

Species of the water-fern megaspore genus Molaspora from a Cenomanian deposit in western France: occurrence,sporoderm ultrastructure and evolutionary relationships

The partially reticulate sculpture of Molaspora aspera sp. nov., a marsileaceous megaspore from a Cenomanian deposit in western France, distinguishes it from other species of Molaspora. An acrolamella entirely surrounds and obscures a small tetrad scar, a feature that has been demonstrated hitherto within members of the genus only in M. fibrosa. It was also encountered for the first time in M. lobata, with which the new species is associated in the same French mesofossil assemblage. The ultrastructure of the sporoderm of M. aspera is similar to that of M. lobata, but differs particularly in that the inner epispore is markedly thicker and may also contain numerous large, homogeneous spherules or, alternatively, holes of comparable dimensions and only a few small spherules. It is possible that these are a response to some hostile bacterial or other activity when the developing sporoderm was partially permeable. The cavity replacing part of the epispore in one of the specimens, and in the specimen of M. lobata examined, may be a preservational feature or have served to increase buoyancy of the spore in water. Molaspora lobata is very similar to megaspores of fossil and extant Regnellidium, but M. aspera bears some resemblance to other members of extant Marsileaceae and certain species of Cretaceous Arcellites, although there are significant differences between them. This suggests that Molaspora is a heterogeneous taxon embracing megaspores produced by water ferns of more than one natural genus, of which only Regnellidium has survived to the present day.     

Taxonomic revision of the Koeleria splendens C. Presl group (Poaceae) in Italy based on morphological characters

Abstract

The Italian populations of Koeleria splendens C. Presl, critical group of the Mediterranean flora, are examined from the taxonomic and nomenclatural viewpoints. On the basis of literature data, herbarium investigations and field surveys, many morphologically well-differentiated taxa have been recognized. This group is widespread in the Italian peninsula and Sicily, where it is represented by the following units: K. splendens C. Presl, with four subspecies [subsp. splendens, subsp. grandiflora (Bertol. ex Schultes) Domin, subsp. brutia Brullo, Gangale & Uzunov, and subsp. ophiolitica subsp. nova], K. lucana sp. nova, K. subcaudata (Ascherson & Graebner) Ujhelyi, K. australis Kerner, K. callieri (Domin) Ujhelyi, and K. insubrica sp. nova. As concerns their distribution, both K. splendens and K. lucana are endemic to the Italian territory, while the other species occur also in some eastern Mediterranean countries. The taxonomic position of K. lobata, recently treated by some authors as a synonym of K. splendens, is also examined. A phylogenetic analysis was performed using morphological characters. Finally, a detailed iconography of the main morphological features and an analytical key of the Italian taxa (K. lobata included) are given.     

Genome size stability in six medicinal plant species propagated <Emphasis Type="Italic">in vitro</Emphasis>

DNA content was estimated by flow cytometry in seedlings and in vitro clones of six species: Oenothera paradoxa, Inula verbascifolia ssp. aschersoniana, Rubus chamaemorus, Solidago virgaurea, S. graminifolia and Pueraria lobata. With the exception of P. lobata, there was no difference in genome sizes between seedlings and in vitro cultured plants from any species, indicative that they maintain their genetic stability during in vitro culture. This confirms the usefulness of tissue culture for production of certified plant material to obtain herbal medicines.     

Pachyphragma andGagria (Brassicaceae) Revisited: Molecular data indicate close relationship toThlaspi S.Str.

The systematic position ofPachyphragma macrophyllum (Brassicaceae) is unsettled. This species was either treated as a monotypic section ofThlaspi s.l. or was treated as a distinct genus.Gagria lobata apparently closely related toPachyphragma, was also given generic rank. To unravel the phylogenetic relationships ofPachyphragma andGagria, ITS sequence studies were conducted using as a reference representatives of the main lineages ofThlaspi s.l. The molecular data indicate thatGagria lobata andPachyphragma macrophyllum are closely related to members ofThlaspi s.str., an assemblage of species well separated from the otherThlaspi s.l. lineages.     

The Botanophila–Epichloë association in cultivated Festuca in Oregon: evidence of simple fungivory

We investigated the Botanophila (Diptera: Anthomyiidae)–Epichloë (Ascomycetes: Clavicipitaceae) interaction in cultivated Festuca spp. (fine fescue) in Oregon in western USA. Epichloë spp. are endophytic fungi of grasses in the subfamily Pooideae. They develop a felt‐like stroma on the surface of grass culms and a dense mycelium within the culms that typically prevents seed head emergence. As a result, seed yields are suppressed, and hence the disease is known as choke. Studies of Epichloë spp. on wild grasses indicate that the fly–fungus interaction is an obligatory mutualism. During oviposition, Botanophila transfers Epichloë spermatia between stromata of opposite mating types, and the perithecia that develop after fungal fertilization serve as food for Botanophila larvae. In the current study, we surveyed 19 cultivated fields of Festuca spp. in Oregon, and observed choke caused by Epichloë festucae Leuchtmann, Schardl and Siegl in 10 of these. However, perithecia were observed in only four fields, and on only 1.0–2.6% of stromata. Perithecial development was also low, and rarely covered 50% of the stroma surface. Despite the absence or low frequency of fertilized stromata, Botanophila lobata Collin larvae were present in all choke‐infested fields. Infestation levels ranged from 2.5 to 70.7%, based on an examination of 175–200 stromata from each field. Only eight (= 2%) of the 450 stromata with B. lobata had perithecia, and the greater majority of B. lobata larvae completed development and exited from unfertilized stromata. This is the first report of the B. lobata–E. festucae association in the USA, and of B. lobata larvae developing successfully on unfertilized Epichloë stromata. The average pupal weight (0.0032 g) did not differ significantly from pupae (0.0030 g) originating from larvae that had developed on fertilized stromata of E. typhina on Dactylis glomerata in a neighboring field. This result indicates that in cultivated fine fescue fields in Oregon, B. lobata forages on E. festucae, but fly development is not dependent on the fertilized stromata of Epichloë.     

Preparation,Structural Analysis and Antioxidant Activity of Polysaccharides and Their Derivatives from Pueraria lobata

Pueraria lobata polysaccharides (PLPs) were obtained by hot water extraction using Pueraria lobata as raw material. Structural analysis revealed that PLPs may have a repetitive backbone units of →4) -α-D-Glcp (1→4-α-D-Glcp (1→. Phosphorylated Pueraria lobata polysaccharides (P-PLPs), carboxymethylated Pueraria lobata polysaccharides (CM-PLPs) and acetylated Pueraria lobata polysaccharides (Ac-PLPs) were obtained by chemical modifications of PLPs, respectively. The physicochemical properties and antioxidant activities of these four Pueraria lobata polysaccharides were studied in comparison. In particular, the clearance rate of P-PLPs exceeded 80 %, and was expected to achieve the same effect as V_c. The results showed that the effects of different chemical modifications on the antioxidant activity of PLPs varied greatly.     

Potential distribution and environmental threat of <Emphasis Type="Italic">Pueraria lobata</Emphasis>

Pueraria lobata (kudzu) is an invasive weed originating from East Asia. Local infestations have been recently observed in Switzerland and northern Italy; however, the potential for P. lobata to spread and to become abundant and damaging in the Alpine countries is not known. The aim of this study was to project the potential distribution of P. lobata under current climate in Switzerland, Austria and Slovenia and parts of northern Italy using the ecoclimatic model CLIMEX. In addition, areas at risk were identified where P. lobata may occur as a strong and aggressive competitor. This was derived from the plants’ distribution and climatic requirements in the south-eastern United States where the heaviest infestations occur. Projections show that 60.84% of the total land area of northern Italy, followed by 47.08% of Slovenia, 21.01% of Austria and only 1.97% of Switzerland are climatically suitable. P. lobata may become a troublesome weed due to very favourable climatic conditions only in some parts of northern Italy and Slovenia. In climatically suitable areas, any occurrence of the plant should be carefully observed. In infested and highly climatically suitable areas, there is a need for strategic management to prevent further spread of P. lobata.     

20-hydroxyecdysone accumulation and regulation in Ajuga lobata D. Don suspension culture

Suspension culture of Ajuga lobata D. Don cells provides a method of synthesis of the phytoecdysteroid 20-hydroxyecdysone (20E) which can regulate the molting process of larvae. We characterized the culture conditions to optimize 20E production. Growth of A. lobata D. Don cells fits the logistic equation curve with a growth cycle of 19 days. Medium conductivity was negatively correlated with dry cell weight and 20E accumulation, thus could be used to determine the optimal time for cell harvest. Continuous subculture reduced 20E synthesis, but supplementing medium with 20E precursors mevalonic (MVA), α-Pinene, and nitric oxide (NO) can significantly promote cell growth and influence 20E accumulation. Combination of α-Pinene, MVA, and SNP significantly elevated 20E accumulation, thus may synergistically enhance 20E synthesis in A. lobata D. Don. The optimal concentrations of α-Pinene, MVA, and NO donor SNP in suspension culture were 50 μL L^?1, 10 mg L^?1, and 80 μmol L^?1.     

The feeding behaviour of a sit-and wait-predator,Ranatra dispar (Heteroptera: Nepidae): optimal foraging and feeding dynamics

Summary Nitrogen and phosphorus flow in litterfall and throughfall were studied in two California Quercus species (the evergreen Q. agrifolia and deciduous Q. lobata) before, during, and after an outbreak of the California oak moth, Phryganidia californica. All of the foliage of both oak species was removed by the herbivore during the course of this outbreak. During the outbreak, total N and P flow to the ground more than doubled from Q. agrifolia and increased to a lesser extent from Q. lobata over the previous year. The composition of the litter during the outbreak year shifted so that in Q. agrifolia, almost 70% of the total N and P flow to the ground moved through frass and insect remains, while in Q. lobata, approximately 60% of the N and 40% of the P moved through frass and insect remains. Short-term leaching experiments showed that nitrogen was far more rapidly lost from Phryganidia frass than from leaf litter of either species. These results and the relative frequency of Phryganidia outbreaks suggest that this herbivore has significant effects on the nutrient cycling beneath these trees.     

The distribution of the thermally tolerant symbiont lineage (Symbiodinium clade D) in corals from Hawaii: correlations with host and the history of ocean thermal stress

Spatially intimate symbioses, such as those between scleractinian corals and unicellular algae belonging to the genus Symbiodinium, can potentially adapt to changes in the environment by altering the taxonomic composition of their endosymbiont communities. We quantified the spatial relationship between the cumulative frequency of thermal stress anomalies (TSAs) and the taxonomic composition of Symbiodinium in the corals Montipora capitata, Porites lobata, and Porites compressa across the Hawaiian archipelago. Specifically, we investigated whether thermally tolerant clade D Symbiodinium was in greater abundance in corals from sites with high frequencies of TSAs. We recovered 2305 Symbiodinium ITS2 sequences from 242 coral colonies in lagoonal reef habitats at Pearl and Hermes Atoll, French Frigate Shoals, and Kaneohe Bay, Oahu in 2007. Sequences were grouped into 26 operational taxonomic units (OTUs) with 12 OTUs associated with Montipora and 21 with Porites. Both coral genera associated with Symbiodinium in clade C, and these co‐occurred with clade D in M. capitata and clade G in P. lobata. The latter represents the first report of clade G Symbiodinium in P. lobata. In M. capitata (but not Porites spp.), there was a significant correlation between the presence of Symbiodinium in clade D and a thermal history characterized by high cumulative frequency of TSAs. The endogenous community composition of Symbiodinium and an association with clade D symbionts after long‐term thermal disturbance appear strongly dependent on the taxa of the coral host.     

Taxonomy ofNeurolaena (Asteraceae-Heliantheae)

A taxonomic treatment of the tropical genusNeurolaena (tribeHeliantheae, subtribeGalinsoginae) is rendered. Ten species are recognized: the widespread, annual, weedy,N. lobata and nine relatively rare, relictual, shrubby or small tree species, mostly restricted to southern Mexico and Central America. Distributional maps, key to species, and complete synonymy are presented. A chromosomal and chemical review is also provided.     

Description and ecological traits of a new species of Pitydiplosis (Diptera: Cecidomyiidae) that induces leaf galls on Pueraria (Fabaceae) in East Asia,with a possible diversification scenario of intraspecific groups

A gall midge that induces thick lenticular galls on leaflets of Pueraria species (Fabaceae) in Japan, mainland China, Taiwan and South Korea is described as Pitydiplosis puerariae sp. nov. (Diptera: Cecidomyiidae). Tanaostigmodes puerariae (Hymenoptera: Tanaostigmatidae), described earlier from mainland China as an inducer of the lenticular gall, is regarded to be an inquiline. Pitydiplosis puerariae is distinguishable from the only known congener, the Nearctic Pitydiplosis packardi, by the male genitalia with entire aedeagus and with hypoproct that is as long as cerci and bilobed with a U‐shaped emargination. DNA sequencing data indicate the existence of three genetically different intraspecific groups: (i) “YNT‐montana group” induces galls on Pueraria montana on the Yaeyama Islands, Japan and in northern Taiwan; (ii) “CT‐montana group” on P. montana in central Taiwan; (iii) and “JCK‐lobata group” on Pueraria lobata in mainland China, South Korea and Japan north of Okinoerabu Island. A possible diversification scenario of the three groups is hypothesized based on DNA sequencing data and geohistorical information. A distribution gap of the gall midge on five islands between Tokunoshima and Ishigaki Islands, Japan was confirmed by intensive field surveys. Ecological traits and adult behavior of Pity. puerariae are also described. Its possibility as a potential biological control agent against P. lobata seems counter‐indicated.     

Priority effects and competition by a native species inhibit an invasive species and may assist restoration

Selecting native species for restoration is often done without proper ecological background, particularly with regard to how native and invasive species interact. Here, we provide insights suggesting that such information may greatly enhance restoration success. The performance of the native vine, Pueraria lobata, and that of the invasive bitter vine, Mikania micrantha, were investigated in South China to test how priority effects (timing and rate of germination and seedling growth) and competition (phytochemical effects and competitive ability) impact invasive plant performance. We found that, in the absence of competition, the germination rate of M. micrantha, but not of P. lobata, was significantly affected by light availability. P. lobata seedlings also performed better than those of M. micrantha during early growth phases. Under competition, negative phytochemical effects of P. lobata on M. micrantha were strong and we found M. micrantha to have lower performance when grown with P. lobata compared to when grown by itself. Relative interaction indexes indicated that, under interspecific competition, P. lobata negatively affected (i.e., inhibited) M. micrantha, whereas M. micrantha positively affected (i.e., facilitated) P. lobata. Higher photosynthetic efficiency and soil nutrient utilization put P. lobata at a further advantage over M. micrantha. Field trails corroborated these experimental findings, showing little recruitment of M. micrantha in previously invaded and cleared field plots that were sown with P. lobata. Thus, P. lobata is a promising candidate for ecological restoration and for reducing impacts of M. micrantha in China. This research illustrates that careful species selection may improve restoration outcomes, a finding that may also apply to other invaded ecosystems and species.     

Parasitoids of Paratachardina lobata (Hem., Kerriidae): surveys for biological control of the invasive lobate lac scale

Paratachardina lobata is an invasive pest in southern Florida, threatening a great number of economically important and native plants. The lobate lac scale does not cause problems in its area of origin, India and Sri Lanka, presumably due to various parasitoid wasps. In an attempt to discover promising parasitoids for biological control against the invasive pest in Florida, P. lobata infesting Pongamia pinnata (Fabaceae) was collected bimonthly from August 2005 to June 2006 at 14 sites in southern India. Four parasitoids were demonstrably associated with P. lobata: a eulophid Aprostocetus bangaloricus Narendran, an encyrtid Ooencyrtus kerriae Hayat and two aphelinids, Marietta leopardina Motschulsky and an undescribed Coccophagus Westwood sp. These chalcidoid wasps were found regularly at all heavily infested sites with an average emergence number per collection period of 62.8, 31.7, 11.9 and 90.5 respectively. The mean number emerged 20–28 days after the collection date, excluding the Coccophagus sp., which occurred significantly later, on average 41 days after collection. Species emergence was examined and parasitized scales dissected. Parasitoid remains were interpreted to understand the mode of parasitism. The Coccophagus sp. was found to be a secondary parasitoid. Marietta leopardina occurred as a primary parasitoid, but only in low number and this species is also known to be hyperparasitic on chalcidoid wasps. Aprostocetus bangaloricus and O. kerriae are promising candidates for the lobate lac scale control in Florida. They are primary parasitoids of P. lobata and occurred at almost every collection site, especially where P. lobata was very abundant.     

Prey selection by coral-feeding butterflyfishes: strategies to maximize the profit

Synopsis Factors that structure preferences among food corals were examined for the obligate coral-feeding butterflyfishChaetodon multicinctus. In the field, fish show a simple repetitious pattern of foraging composed of (1) pre-encounter search for coral colonies, and (2) post-encounter inspection/orientation, bite, and consumption of polyps. Rose coral,Pocillopora meandrina, and the massive coral,Porites lobata, were taken in higher proportions than their percentage substrate cover, while finger coral,Porites compressa, was taken in lower proportion. Paired presentations of coral colonies in the lab gave similar results:Poc. meandrina was preferred overPor. lobata which was preferred overPor. compressa. Poc. meandrina tissue had the highest energy content, lowest handling time, and highest profitability. Energy content did not differ amongPorites tissues, but handling time was greater and more inspective eye movements were made while foraging on the branched finger coral,Por. compressa. Experimental manipulation of coral colony morphology indicate preferences amongPorites are most likely structured by handling costs. Predictions of a simple prey-choice foraging model are supported in theC. multicinctus system if abundance of the branched coralPor. compressa is estimated as that available to fishes rather than percentage substrate cover. The relative size and abundance of stinging nematocysts are also consistent with observed foraging patterns in the field, but await immunological confirmation. Coral-feeding butterflyfishes offer unique opportunities to test models of foraging ecology in reef fishes, and the direction of future studies is suggested.     

Genetic analysis of the Linnaean Ulva lactuca (Ulvales,Chlorophyta) holotype and related type specimens reveals name misapplications,unexpected origins,and new synonymies

Current usage of the name Ulva lactuca, the generitype of Ulva, remains uncertain. Genetic analyses were performed on the U. lactuca Linnaean holotype, the U. fasciata epitype, the U. fenestrata holotype, the U. lobata lectotype, and the U. stipitata lectotype. The U. lactuca holotype is nearly identical in rbcL sequence to the epitype of U. fasciata, a warm temperate to tropical species, rather than the cold temperate species to which the name U. lactuca has generally been applied. We hypothesize that the holotype specimen of U. lactuca came from the Indo‐Pacific rather than northern Europe. Our analyses indicate that U. fasciata and U. lobata are heterotypic synonyms of U. lactuca. Ulva fenestrata is the earliest name for northern hemisphere, cold temperate Atlantic and Pacific species, with U. stipitata a junior synonym. DNA sequencing of type specimens provides an unequivocal method for applying names to Ulva species.     

葛(豆科)叶的解剖学研究

利用光学显微镜和扫描电镜观察了葛(Pueraria lobata)叶的解剖学特征。结果表明,葛叶片的上、下表皮都只有一层表皮细胞,上表皮比下表皮厚。上、下表皮都有腺毛和非腺毛。气孔主要分布在下表皮,下表皮的气孔密度为(261±17)mm-2,上表皮只有(6±3)mm-2。叶肉由两层栅栏组织细胞和一层海绵组织细胞构成。叶肉细胞中有丰富的叶绿体。在栅栏组织和海绵组织之间有一层平行于叶脉的薄壁细胞。叶脉中含有大量的草酸钙晶体。葛叶的这些形态特征与其喜阳、耐旱的特点相适应。     

粉葛主要营养物质、异黄酮成分的积累规律及其相关性分析

为确定粉葛的最佳采收时期及各部位的应用价值，探索粉葛主要营养物质与异黄酮成分的积累规律及相关关系，该研究以广西藤县粉葛为材料，采用测试盒法、酶重量法、索氏抽提法、高效液相色谱法等生理生化技术，测定了成熟期不同部位异黄酮成分的含量及不同生长时期粉葛块根主要营养物质、异黄酮成分的含量，并分析了各营养物质与异黄酮成分积累的相互关系。结果表明：(1)在成熟期，粉葛不同部位的染料木素含量差异均不显著，但叶片的大豆苷元含量显著小于中藤，根头和藤的总异黄酮、葛根素、大豆苷的含量显著大于叶片和块根，并且根头和藤的葛根素含量均超过1.00%。(2)不同生长时期，粉葛的淀粉、多糖、粗蛋白、可溶性蛋白的积累在11月和12月达到最大，不溶性膳食纤维的积累在12月最小，可溶性膳食纤维在8月和12月的积累量最大，总异黄酮和葛根素在8月和9月的积累量最大。(3)总异黄酮、葛根素的积累与淀粉、多糖的积累呈显著负相关，总异黄酮、葛根素的积累与不溶性膳食纤维的积累呈显著正相关。综上结果认为：粉葛的根头、藤蔓含有丰富的总异黄酮、葛根素、大豆苷，具有很好的药用开发价值；粉葛药用的最佳采收期为8月和9月，粉葛食用的最佳采收期为...