The Herbivore Assemblage, Herbivory and Leaf Chemistry of the Mangrove Kandelia obovata in Two Contrasting Forests in Hong Kong

The herbivore assemblage, intensity of herbivory and factors determining herbivory levels on the mangrove Kandelia obovata (previously K. candel, Rhizophoraceae) were studied over a 13-month period at two forests with contrasting growing conditions in Hong Kong. Mai Po was part of an eutrophic embayment in the Pearl River estuary and generally offered more favourable conditions for mangrove growth, whereas Ting Kok had a rocky substratum and oceanic salinity. Twenty-four insect herbivore species were recorded on K. obovata, with lepidopteran larvae that consume leaf lamina being the dominant species. While leaf litter production was similar at the two forests, herbivory level at Mai Po (mean = 3.9% in terms of leaf area loss) was more severe than that at Ting Kok (mean = 2.3%). Peak herbivory levels were found in summer at both locations (6.5% for Mai Po and 3.8% for Ting Kok). Young leaves of K. obovata at both locations were generally preferred by the herbivores from the period of late spring to summer. Concentrations of most feeding deterrents (ash, crude fibre, and total soluble tannins) were significantly higher in both young and mature leaves at Ting Kok, whereas leaf nutrients (total nitrogen and water) were the same at the two sites. Young leaves at Ting Kok contained about 30% more tannins than their counterparts at Mai Po. Significant differences in leaf chemistry also existed between young and mature leaves at either site. The differences were concomitant with the observed patterns of leaf herbivory on K. obovata, and suggest a potential relationship between environmental quality and plant defence against herbivory.     

Effects of plant density and age on the mating system of Kandelia candel Druce (Rhizophoraceae), a viviparous mangrove species

Mating system is an important factor affecting gene flow and genetic structure of plant populations. Based on identification of chlorophyll-deficient offsprings on heterozygous individuals, selfing rates were estimated for Kandelia candel, a viviparous mangrove species. Outcrossing rates were significantly decreased when population was thinned, indicating that plant density plays an important role in plant reproductive features. Selfing rate of young population was significantly higher than that of mature population, due to pollen flow barriers in young population, created by dense leaves and twigs.     

Assessment of genetic diversity among and within Achillea species using amplified fragment length polymorphism (AFLP)

Amplified fragment length polymorphism (AFLP) markers were used to assess the genetic diversity of 57 Achillea accessions belonging to five species, A. millefolium, A. filipendulina, A. tenuifolia, A. santolina and A. biebersteinii. Nine AFLP primer combinations were used, which produced 301 polymorphic bands. In most species, a high level of genetic variation was detected among the genotypes. The Jaccard's similarity indices (J), based on AFLP profiles, were subjected to UPGMA cluster analysis. Application of Mantel's test for cophenetic correlation to the cluster analysis indicated the high fitness of the accessions to a group (r = 0.918). The dendrogram generated revealed five major groups corresponding to five species. The principle coordinate analysis (PCoA) data confirmed the results of the clustering. Among the species, A. teunifolia and A. santolina showed the greatest and the least genetic diversity, respectively. A. filipendulina accessions were acquired primarily from the same ecological regions of western Iran. Accessions belonging to A. biebersteinii originated from the Isfahan province and were separated from other species at the root of the dendrogram. The results of the clustering method, based on AFLP markers, corresponded closely with the geographical origins of the genotypes. The results of the present study could contribute to a better understanding and management of conservation and exploitation of the Achillea germplasm.     

Isolation and characterization of microsatellite loci in the red mangrove Rhizophora mangle (Rhizophoraceae) and its related species

Fourteen microsatellite markers were isolated from the red mangrove Rhizophora mangle (Rhizophoraceae), a widely distributed mangrove plant in the New World and West Africa. The range of expected heterozygosity of these markers was 0.000–0.672 in the two populations of R. mangle. Cross-species testing was examined for five other species of Rhizophora, and Kandelia candel and Bruguiera gymnorrhiza. All 14 markers could be amplified in R. samoensis, thirteen in R. racemosa, and six markers in all other species of Rhizophora. Our findings greatly increase the utility of these markers.     

Histological analysis in shoot organogenesis from hypocotyl explants of<Emphasis Type="Italic"> Kandelia candel</Emphasis> (Rhizophoraceae)

In vitro culture of hypocotyl explants from Kandelia candel, a common mangrove species, on hormone-free Murashige and Skoog (MS) medium resulted in shoot formation. Since the hypocotyls showed good potential for in vitro shoot multiplication, the process of bud primordium formation was analyzed from a histological viewpoint. A wound periderm first appeared at the top, exposed cut surface of the explants. The wound-induced meristem continued to divide giving rise to suberized cells oriented towards the cut surface. After formation of the suberized cell layers, the meristem and its inner derivatives differentiated into multilayered, uniformly packed parenchyma cells. Bud primordia differentiated from the dense cytoplasmic cells of the wound-induced meristem just beneath the suberized layer near the severed vascular bundles. Each explant produced several visible shoot buds. Furthermore, histological sections revealed that additional bud primordia were present within the explant just underneath the suberized cells and that these bud primordia appeared to be arrested in their development. The fact that additional bud primordia were present within the explant suggests that further manipulation of the explant is helpful to maximize the potential of this system.     

Population genetic structure of three tree species in the mangrove genus Ceriops (Rhizophoraceae) from the Indo West Pacific

Ceriops is a viviparous mangrove with widespread species Ceriops decandra and C. tagal, and an endemic species C. australis. Genetic diversity of the three species was screened in 30 populations collected from 23 locations in the Indo West Pacific (IWP) using Inter-simple sequence repeats (ISSR) and sequences of partial nuclear gene (G3pdh) and chloroplast DNA (trnV-trnM). At the species level, the total gene diversity (Ht) revealed by ISSRs was 0.270, 0.118, and 0.089 in C. decandra, C. tagal, and C. australis, respectively. A total of six haplotypes of G3pdh and five haplotypes of trnV-trnM were recognized among the three species. Only C. decandra was detected containing more than one haplotype from each sequence data set (four G3pdh haplotypes and three trnV-trnM haplotypes). At the population level, genetic diversity of Ceriops was relatively low inferred from ISSRs (He = 0.028, 0.023, and 0.053 in C. decandra, C. tagal, and C. australis, respectively). No haplotype diversity within population was detected from any of the three species. Cluster analysis based on ISSRs identified three major geographical groups in correspond to the East Indian Ocean (EIO), South China Sea (SCS), and North Australia (NA) in both C. decandra and C. tagal. The cladogram from DNA sequences also detected the same three geographical groups in C. decandra. Analysis of molecular variance (AMOVA) revealed that most of the total variation was accounted for by differentiation between the three major geographical regions of both C. decandra and C. tagal. The significant genetic structure may result from the geological events in these regions during the recent Pleistocene glaciations. This study also provided insights into the phylogenetics of Ceriops. Yelin Huang and Fengxiao Tan contributed equally to this work.     

Ten polymorphic microsatellite loci for the Atlantic halibut (Hippoglossus hippoglossus) and cross-species application in related species

In the present study, 10 polymorphic microsatellite DNA loci from Atlantic halibut (Hippoglossus hippoglossus) were isolated and characterized. The number of alleles for these loci ranged from 2 to 4 in tested 24 individuals. Observed and expected heterozygosities per locus varied from 0.21 to 0.70 and from 0.31 to 0.65, respectively. Most of these 10 microsatellite loci were successfully amplified and showed polymorphic in five related species. These loci will be useful for the assessment of genetic diversity and population structure of Atlantic halibut. H. Ding and C. Shao contributed equally to this work.     

红树林恢复初期莱氏锥滨螺分布动态

树栖腹足类动物是红树林底栖动物的重要组成部分,其组成和分布受到红树植被的影响。因此,在红树林生态修复中,腹足类动物的恢复依赖于植被的发育。滨螺科(Littorinidae)锥滨螺属(Mainwaringia)动物是亚洲红树林的广布种,但针对其在红树林中的分布及种群特征的研究较少,对红树林植被恢复过程中锥滨螺属动物的恢复动态更是鲜有研究。调查了厦门下潭尾湿地秋茄(Kandelia obovata)红树林恢复初期,新记录种莱氏锥滨螺(Mainwaringia leithii)种群恢复的时间动态,并分析其与植被参数的关系。研究发现,红树林恢复初期,秋茄植被生长迅速,株高、冠幅、基径和覆盖度分别由0.5年生幼林的41.3 cm、25.0 cm、1.04 cm和25%增长为8年生样地的179.2 cm、76.7 cm、5.96 cm和95%。在秋茄胚轴种植6个月左右开始记录到莱氏锥滨螺的个体,其密度和生物量在1.5 a后达到最大(分别为136个/m²和1.86 g/m²),而后逐渐降低,种植3 a后秋茄林内莱氏锥滨螺仅零星分布。除8年生的样地,莱氏锥滨...     

Isolation and characterization of microsatellite loci in the Japanese pipistrelle (Pipistrellus abramus)

We describe the first set of ten microsatellite markers isolated in Pipistrellus abramus. The number of alleles per locus ranged from 7 to 13. The observed and expected heterozygosities values ranged from 0.486 to 0.971 and from 0.752 to 0.876, respectively. Three loci revealed significant departure from Hardy–Weinberg equilibrium and no linkage disequilibrium was found between loci pairs. These informative microsatellite markers will be a powerful molecular tool for studying the population genetic structure of P. abramus, as well as other species of this genus.     

引进红树拉关木和两种乡土红树离子平衡及光合作用的比较研究

为了比较引进红树与乡土红树的耐盐性差异,该研究以引进红树植物拉关木(Laguncularia racemosa)和乡土红树植物木榄(Bruguiera gymnorrhiza)与秋茄(Kandelia obovata)幼苗作为实验材料,分析其在不同Na Cl浓度(100、200、300、400 mmol·L~(-1))处理下各器官离子浓度(Na~+、Cl~–、K~+、Ca~(2+)和Mg~(2+))和叶光合作用的变化。结果表明:(1)高盐胁迫(400 mmol·L~(-1)Na Cl,28 d)处理下,拉关木根系Na~+增幅较小,秋茄根、叶Cl~–含量增幅均高于木榄和拉关木,说明拉关木在较高的盐浓度时能限制根系对Na~+、Cl~–的吸收,减少向地上部分运输。(2)高盐胁迫均增加3种红树根、叶的K~+浓度(木榄叶K~+略有降低,差异不显著),表明3种红树均可吸收K~+,来限制Na~+对植物的伤害;同时,降低3种红树根Ca~(2+)浓度,但拉关木根Ca~(2+)下降幅度小于秋茄和木榄,说明拉关木具有更强的防止Ca~(2+)流失的能力。(3)拉关木根维持Na~+/K~+、Na~+/Ca~(2+)平衡的能力强于秋茄和木榄。(4)高盐胁迫引起秋茄与木榄光合速率均降低,而拉关木光合速率却增加了54.1%。综上所述,拉关木能限制根系对Na Cl的吸收,有效维持Na~+/K~+、Na~+/Ca~(2+)的平衡,并保持较高的光合速率,这表明拉关木与木榄和秋茄相比具有更高的耐盐性。     

Forest structure of a subtropical mangrove along a river inferred from potential tree height and biomass

The hypothesis was tested that potential tree height and biomass in mangroves decrease downstream with the tidal gradient along the Okukubi River in Okinawa Island, Japan. The mangrove stands consisted of Bruguiera gymnorrhiza (L.) Lamk. and Kandelia obovata (S., L.) Yong (Rhizophoraceae). Four sites were selected considering the distance from the mouth of the river. Soil salinity increased downstream, while soil total nitrogen content decreased. The soil redox potential did not vary along the river. Maximum gross photosynthesis and tree height for each species decreased downstream. The potential tree height (H_max) inferred from the stem diameter (D_0.1)–tree height (H) relationship ( a, h, coefficient) in each species decreased downstream. The tree density (ρ)–mean tree size () relationships ( K, α, coefficient) determined for four sites revealed that the mean tree size at any given tree density decreased downstream, which indicates the decrease of potential biomass. Furthermore, an index for biomass () was homogeneous within a site regardless of tree density, i.e. the value of α at each site did not differ significantly from 1.0 (p > 0.05). The decreases in potential tree height and biomass may be partially ascribed to the stressful environments at the downstream sites characterized by high salinity (>2.6%) and nitrogen-poor soils (<0.25 ppt) in our study area.     

Characterization of novel microsatellite loci in the great leaf-nosed bat, Hipposideros armiger and cross-amplification in other related species

Eight microsatellite loci were isolated from an enriched genomic library of the great leaf-nosed bat, Hipposideros armiger. The polymorphism of these loci was tested on a population of 48 individuals from Anhui Province, China. All loci revealed the polymorphism ranging from three to 12 alleles. The observed heterozygosity values were from 0.213 to 0.875 and expected heterozygosity from 0.232 to 0.820. No significant linkage disequilibrium was detected. Two loci significantly deviated from Hardy–Weinberg equilibrium after Bonferroni correction. In addition, successful cross-amplification also suggested that these microsatellite markers will facilitate research on the population genetics and gene flow of H. armiger and other related species.     

Fifteen novel polymorphic microsatellites in rock carp, Procypris rabaudi (Tchang), an endemic fish species in the upper reaches of the Yangtze River drainage

Fifteen new microsatellite markers were isolated in rock carp, Procypris rabaudi (Tchang), from an AC-enriched genomic library. The allele number of these loci ranged from three to 13 (average 6.87 per locus) in the tested 24 individuals. Polymorphism information content (PIC) ranged from 0.453 to 0.821 with an average of 0.719. Average observed and expected heterozygosities were 0.712 (range 0.565–0.917) and 0.772 (range 0.542–0.855), respectively. These microsatellite markers could be useful in the population genetic study of rock carp.     

An improved amplified fragment length polymorphism (AFLP) protocol for discrimination of Listeria isolates

Nine restriction enzyme combinations and 108 different primer combinations were initially tested for suitability for amplified fragment length polymorphism (AFLP) analysis of Listeria monocytogenes; the combination of HindIII and HpyCH4IV showed consistently strong signals on gels, amplified an adequate number of DNA fragments and detected polymorphism among closely related strains based on AscI macrorestriction profiles. AFLP also distinguished between L. monocytogenes, L. innocua, L. ivanovii, L. seeligeri, L. welshimeri and L. grayi species. All Listeria species showed species-specific clusters, with less than 33% similarity between different species. A total of 34 L. monocytogenes strains were characterised by using both AFLP and pulsed-field gel electrophoresis (PFGE). The results of AFLP analysis of L. monocytogenes strains were in concordance with those obtained by PFGE. Both methods identified 29 different genotypes of L. monocytogenes and had a high discrimination index (> 0.999). By combining the results of AFLP and PFGE, subtype discrimination was further improved. Numerical analysis of both AFLP and PFGE profiles yielded three genomic groups of L. monocytogenes strains. AFLP was found to be faster and less labour-intensive than PFGE. We conclude that the AFLP protocol is a highly discriminatory, reproducible and valuable tool in characterisation of Listeria strains and may also be suitable for Listeria species identification.     

Development and characterization of ten novel microsatellite markers from olive ridley sea turtle (Lepidochelys olivacea)

Olive ridleys, one of the widely distributed marine turtle species has undergone declines in recent years due to multiple anthropogenic factors warranting conservation efforts for which assessment of genetic variability in existing populations become critical. Here we describe development of ten new microsatellite markers from a short sequence repeat-enriched partial genomic DNA library, which are found to be highly informative for genetic studies. Eight of these markers when tested on 83 olive ridley turtles revealed high allelic diversity (4–27 alleles per marker), and high observed and expected heterozygosity estimates that ranged from 0.29 to 0.82 and 0.62 to 0.94, respectively. Two microsatellites were monomorphic in the tested olive ridley samples, but were found to be informative/polymorphic when tested on related marine turtle species. More importantly, nine of the new markers showed robust cross-species amplifications in three related species Dermochelys coriacea, Chelonia mydas and Eretmochelys imbricata. Thus, this study describes ten new microsatellite markers and also demonstrates their potential as efficient genetic markers in studies related to parentage analysis, population structure, phylogeography and species relationships of olive ridleys and other marine turtle species.     

Trypanosoma evansi: genetic variability detected using amplified restriction fragment length polymorphism (AFLP) and random amplified polymorphic DNA (RAPD) analysis of Kenyan isolates

We compared two methods to generate polymorphic markers to investigate the population genetics of Trypanosoma evansi; random amplified polymorphic DNA (RAPD) and amplified restriction fragment length polymorphism (AFLP) analyses. AFLP accessed many more polymorphisms than RAPD. Cluster analysis of the AFLP data showed that 12 T.evansi isolates were very similar ('type A') whereas 2 isolates differed substantially ('type B'). Type A isolates have been generally regarded as genetically identical but AFLP analysis was able to identify multiple differences between them and split the type A T. evansi isolates into two distinct clades.     

Population structure and genetic diversity of Huperzia serrata (Huperziaceae) based on amplified fragment length polymorphism (AFLP) markers

The levels and pattern of the genetic variation within and among natural populations of Huperzia serrata were investigated using amplified fragment length polymorphism markers. Seven primer combinations used in the study amplified 615 discernible bands with 532 (86.5%) being polymorphic, indicating a considerable high level of genetic diversity at the species level. AMOVA analysis revealed a low level of genetic differentiation among the ten populations. The UPGMA cluster of all samples showed that individuals from the same population occasionally failed to cluster in one distinct group. A Mantel test showed no significant correlation between genetic distance and geographical distance (r = 0.278, P = 0.891), suggesting that the gene flow was not restricted geographically. A number of factors that might affect the genetic profiles of H. serrata included clonal growth, selective effect of niche and outcrossing, as well as the effective wind-dispersal of spores.     

AFLP analysis of phylogenetic relationships among myrmecophytic species of Macaranga (Euphorbiaceae) and their allies

The palaeotropic pioneer tree genus Macaranga Thouars (Euphorbiaceae) is characterized by various types of mutualistic interactions with specific ant partners (mainly Crematogaster spp.). About 30 species are obligate ant-plants (myrmecophytes). We used amplified fragment length polymorphism (AFLP) markers to assess phylogenetic relationships among 108 Macaranga specimens from 43 species, including all available taxa from the three sections known to contain myrmecophytes. Eight primer combinations produced 426 bands that were scored as presence/absence characters. Banding patterns were analyzed phenetically, cladistically and by principal coordinates analysis. Monophyly of section Pruinosae is clearly supported. There is also good evidence for a monophyletic section Pachystemon that includes the puncticulata group. The monophyly of section Winklerianae and relationships between the three sections remain ambiguous. Section Pachystemon is subdivided into four well-supported monophyletic subclades that presumably correspond to taxonomic entities.We acknowledge the support by the Deutsche Forschungsgemeinschaft (DFG Fi606/4-1, DFG We1830/2-1, 4-1 and 4-2), which in part was granted in the frame of the DFG-SPP 1127 Radiations: origins of biological diversity. Part of the plant material was kindly supplied by Dr. H. Feldhaar (University of Würzburg), Dr. U. Moog (University of Kassel) and Dr. F. Slik (Leiden University Branch, Nationaal Herbarium Nederland). We thank the University of Malaysia (Dr. Rosli b. Hashim) and Taman Taman Sabah (Datuk Lamri Ali; Dr. J. Nais) for permits and logistic support, and EPU for permission to conduct research in Malaysia.     

Genome wide AFLP markers support cryptic species in Coniophora (Boletales)

Numerous fungal morphospecies include cryptic species that routinely are detected by sequencing a few unlinked DNA loci. However, whether the patterns observed by multi-locus sequencing are compatible with genome wide data, such as amplified fragment length polymorphisms (AFLPs), is not well known for fungi. In this study we compared the ability of three DNA loci and AFLP data to discern between cryptic fungal lineages in the three morphospecies Coniophora olivacea, Coniophora arida, and Coniophora puteana. The sequences and the AFLP data were highly congruent in delimiting the morphotaxa into multiple cryptic species. However, while the DNA sequences indicated introgression or hybridization between some of the cryptic lineages the AFLP data did not. We conclude that as few as three polymorphic DNA loci was sufficient to recognize cryptic lineages within the studied Coniophora taxa. However, based on analyses of a few (three) sequenced loci the hybridization could not easily be distinguished from incomplete lineage sorting. Hence, great caution should be taken when concluding about hybridization based on data from just a few loci.     

Taxonomy of North and Central American diploid wild potato (Solanum sect. Petota) species: AFLP data

Solanum section Petota, the potato and its wild relatives, includes about 200 wild species distributed from the southwestern United States to central Argentina and adjacent Chile, with about 30 species in North and Central America. The North/Central American region and the South American region all include diploids, tetraploids, and hexaploids. Chloroplast DNA restriction enzyme data from a prior study showed that 13 of the North/Central American species formed a clade containing only diploids, but there was low resolution within the clade. This Amplified Fragment Length Polymorphism (AFLP) study is conducted to provide additional resolution within the North/Central American diploids and complements the chloroplast results, and prior morphological results. Wagner parsimony and phenetic analyses mostly agreed with the morphological data in supporting currently recognized species except that they suggest that S. brachistotrichium and S. stenophyllidium are conspecific. Our new AFLP data, in combination with the cpDNA and morphological data, also support sister taxon relationships for the following diploid species from North and Central America: 1) S. cardiophyllum subsp. ehrenbergii and S. stenophyllidium, 2) S. tarnii and S. trifidum, 3) S. jamesii and S. pinnatisectum, 4) S. lesteri and S. polyadenium, and 5) S. clarum and S. morelliforme.This work represents partial fulfillment for the requirements of a Ph.D. degree in Plant Breeding and Genetics at the University of Wisconsin-Madison. We thank committee members Paul Berry, Michael Havey, Thomas Osborn, and Kenneth Sytsma. We also thank John Bamberg and Staff of the Unites States Potato Genebank for germplasm and locality data; Charles Nicolet and staff of the University of Wisconsin Biotechnology Center for technical help; Lynn Hummel and staff at Walnut Street Greenhouse for help in growing plants; and lab partners Brian Karas, Iris Peralta, Celeste Raker, and Sarah Stephenson for technical advice. This study was supported by CONACYT (Mexico) scholarship number 116742 granted to Sabina I. Lara-Cabrera, and the United States Department of Agriculture. Names are necessary to report data. However, the USDA neither guarantees nor warrants the standard of the product, and the use of the name by the USDA implies no approval of the product to the exclusion of others that may also be suitable.