Mapping species differences for adventitious rooting in a <Emphasis Type="Italic">Corymbia torelliana</Emphasis> × <Emphasis Type="Italic">Corymbia citriodora</Emphasis> subspecies <Emphasis Type="Italic">variegata</Emphasis> hybrid

Quantitative trait loci (QTL) detection was carried out for adventitious rooting and associated propagation traits in a second-generation outbred Corymbia torelliana × Corymbia citriodora subspecies variegata hybrid family (n = 186). The parental species of this cross are divergent in their capacity to develop roots adventitiously on stem cuttings and their propensity to form lignotubers. For the ten traits studied, there was one or two QTL detected, with some QTL explaining large amounts of phenotypic variation (e.g. 66% for one QTL for percentage rooting), suggesting that major effects influence rooting in this cross. Collocation of QTL for many strongly genetically correlated rooting traits to a single region on linkage group 12 suggested pleiotropy. A three locus model was most parsimonious for linkage group 12, however, as differences in QTL position and lower genetic correlations suggested separate loci for each of the traits of shoot production and root initiation. Species differences were thought to be the major source of phenotypic variation for some rooting rate and root quality traits because of the major QTL effects and up to 59-fold larger homospecific deviations (attributed to species differences) relative to heterospecific deviations (attributed to standing variation within species) evident at some QTL for these traits. A large homospecific/heterospecific ratio at major QTL suggested that the gene action evident in one cross may be indicative of gene action more broadly in hybrids between these species for some traits.     

Inheritance and linkage analysis of five enzyme loci in interspecific hybrids of toadlets,genus Bombina

Progeny produced from Bombina bombina, B. variegata, and field-collected interspecific hybrids have been analyzed for the inheritance of five enzyme loci, which are fixed for alternate alleles in the parental species. Lactate dehydrogenase (Ldh-1), NAD-dependent malate dehydrogenase (Mdh-1), creatine kinase (Ck), adenylate kinase (Ak), and glucosephosphate isomerase (Gpi) are all inherited in a Mendelian manner as codominant alleles at nuclear loci. Both parental alleles are equally functional in artificial F₁ hybrids (female B. bombina×male B. variegata) at each of the loci studied. No linkage between any pair of loci was observed. Discovery of this inherited biochemical variation combined with a technique for assaying individual genotypes without killing the animals makes feasible studies of hybrid population structure heretofore impossible.This work was partly supported by the Polish Academy of Sciences within the project MR-II/6.     

Isolation and characterization of ten microsatellite loci in Blue-footed (Sula nebouxii) and Peruvian Boobies (Sula variegata)

Ten microsatellite loci were isolated and characterized from Blue-footed (Sula nebouxii) and Peruvian Boobies (S. variegata). The loci were screened in 24 Blue-footed Boobies and 27 Peruvian Boobies: 8 were polymorphic in Blue-footed Boobies with between 2 and 10 alleles per locus and 9 were polymorphic in Peruvian Boobies with between 2 and 12 alleles per locus. Observed heterozygosity ranged from 0.29 to 0.84. These loci were also tested in Brown Boobies (S. leucogaster) and were variably polymorphic. These new loci are currently being used to assess population genetic structure in Blue-footed and Peruvian Boobies and will also be used to examine hybridization between the species.     

Eight new microsatellite loci for the critically endangered fire‐bellied toad Bombina bombina and their cross‐species applicability among anurans

We describe eight new microsatellite loci for the critically endangered fire‐bellied toad, Bombina bombina. Seven of them are polymorphic with two to seven alleles per locus, an expected heterozygosity between 0.41 and 0.8, and an observed heterozygosity between 0.27 and 0.7. The yield of new loci was relatively low, presumably due to mildly repetitive sequence motifs in microsatellite flanking regions. As typical for anurans, cross‐species amplification was limited (here, to congeners Bombina orientalis and Bombina variegata). Combining these new loci with those already available provides a reasonable number of loci for population studies and pedigree analysis in Bombina.     

Comparative responses of three fuel wood yielding plants to PEG-induced water stress at seedling stage

Three species of fast growing fuel wood yielding plants locally available (Acacia holosericea, Bauhinia variegata and Cassia siamea) were characterized in respect of their responses to water stress. Seedlings (25 days) of these species, exposed to two levels of water stress (−0.5 and −1.0 MPa) induced by PEG-6000 for 24 h, were analysed for relative water content (RWC) and the contents of chlorophyll, protein, soluble sugars and proline in leaves along with activities of catalase, peroxidase and superoxide dismutase (SOD). RWC was lower in stressed compared to the unstressed seedlings. However, stress-induced decline in RWC was lowest in B. variegata. Chlorophyll and protein contents declined with increasing levels of water stress, decline being least in B. variegata. Soluble sugar and proline contents increased under water stress particularly in B. variegata. The enzyme activity of catalase (EC-1.11.1.6), peroxidase (EC-1.11.1.7) and SOD (EC-1.15.1.1) decreased with increased levels of water stress. Such decline in the activity of these enzymes was least in B. variegata. Apparently, B. variegata is potentially the species most tolerant to water stress among these three fuel wood-yielding plants.     

A barcode analysis of the genus Lobophora (Dictyotales,Phaeophyceae) in the western Atlantic Ocean with four novel species and the epitypification of L. variegata (J.V. Lamouroux) E.C. Oliveira

In the western Atlantic Ocean, the brown algal genus Lobophora is currently represented by a single species, L. variegata, with a type locality designated by Lamouroux as ‘Antilles’. In this study, we used molecular-assisted alpha taxonomy (MAAT) to assess species diversity of Lobophora in Bermuda, the Florida Keys, St. Croix and Guadeloupe (Lesser Antilles). Using cox1 and cox3 sequences as barcode markers, five species of Lobophora, four of them novel, were delineated, all previously having been identified in the area as L. variegata. Our morphological and habitat studies, made possible by abundant sampling, have revealed unique characters for each of these western Atlantic species, including distinct cellular arrangements, as well as different depth ranges for certain species. Observations made from Lamouroux’s holotype of Dictyota variegata (= Lobophora variegata) allowed us to assess the anatomy of this species, which enabled us to easily align this early taxon to one of our genetic species from the western Atlantic. As the type was unavailable for genetic analysis, we selected a recent St. Croix (Virgin Is., Antilles) specimen as the epitype to support it with molecular sequence data.     

Morphological differentiation correlates with ecological but not with genetic divergence in a Gehyra gecko

Body size affects life history, the ecological niche of an organism and its interactions with other organisms. Resultantly, marked differences in body size between related organisms are often an indication of a species boundary. This is particularly evident in the Gehyra variegata species complex of geckos, which displays differential body sizes between genetically divergent species, but high levels of intraspecific morphological conservatism. We report on a Gehyra population that displays extraordinary body size differentiation in comparison with other G. variegata species. We used morphological and environmental data to show this population is phenotypically and ecologically distinct from its parapatric congener Gehyra lazelli and that morphology and ecology are significantly correlated. Contrastingly, mtDNA analysis indicates paraphyly between the two groups, and allele frequencies at six microsatellite loci show no population structure concordant with morpho‐/ecotype. These results suggest either ecological speciation or environmentally induced phenotypic polymorphism, in an otherwise morphologically conservative group.     

Nine new tetranucleotide microsatellite markers for the fire‐bellied toad (Bombina bombina)

We describe nine new polymorphic tetranucleotide microsatellite loci isolated from the fire‐bellied toad (Bombina bombina). The relative yield of new loci was higher than described in previous studies in amphibians: out of 12 loci initially evaluated, nine were polymorphic and amplifying reliably. Number of alleles ranged from four to 10 and observed heterozygosities from 0.47 to 0.91. Seven loci were polymorphic also in Bombina variegata and five in Bombina orientalis. Enrichment protocols yielding long flanking regions potentially overcome difficulties (i.e, low yield of reliable loci relative to number of clones screened) which have been reported in microsatellite development in anurans.     

Inheritance of allozyme loci inBombina: One linkage group established

Two hybridizing European species of fire-bellied toads,Bombina bombina andB. variegata, have alternate electromorphs fixed at a number of allozyme loci. Segregation of alleles at seven allozyme loci (Ldh-1, Mdh-1, Ak, Ck, Gpi, Np, andEst-) was studied in a backross progeny of an F₁ interspecific hybrid male and aB. bombina female. Mendelian inheritance of allozyme forms at all seven loci was ascertained. Except for two loci,Gpi andEst-, which were found to be tightly linked (1 cM apart), other loci showed independent segregation.     

Hybridization between the fire-bellied toads Bombina bombina and Bombina variegata in the karst regions of Slovakia and Hungary: morphological and allozyme evidence

Geographic variability and genetic interactions in the contact zone between the fire-bellied toads, Bombina bombina and B. variegata, were studied using analysis of morphological and genetic variation in sixteen samples from the Slovak Karst and Aggtelek Karst regions. Genotype frequencies at four marker loci (Ldh-1, Mdh-1, Adk, Hem) demonstrate the existence of a hybrid zone with highly variable population structures. While some samples appear to represent panmictic hybrid populations, other samples are very heterogeneous. Pure individuals of both species occurred together with hybrids at one site. Habitat segregation among genetically differentiated demes probably causes this heterogeneity of population structures. Increased frequencies of the allele Ldh-1^M, which is present in low proportion in B. bombina south of the contact zone, were found in some hybrid populations.     

Biotic interactions between Eriopis connexa and Hippodamia variegata,a native and an exotic coccinellid species associated with alfalfa fields in Chile

Human‐assisted introductions, including those in the context of biological control, are considered to be one of the most important factors of global environmental change. However, the mechanisms underlying environmental changes, such as a decrease in the relative abundance of native species, are poorly understood. Since the introduction of the ladybird beetle, Hippodamia variegata (Goeze) (Coleoptera: Coccinellidae), in Chile in the 1970s for biological control of aphids, a reduction in the relative abundance of the native ladybird beetle Eriopis connexa (Germar) has been noticed. To explore the role of cannibalism, intraguild predation (IGP) or competition as possible mechanisms that might increase dominance of H. variegata over E. connexa, several laboratory experiments were carried out. The native and the exotic species were very similar in their voracity and biotic interactions. Although H. variegata was able to maintain constant reproductive performance, E. connexa reproduction decreased at lower densities of aphids, Acyrthosiphon pisum (Harris) (Hemiptera: Aphididae). The impacts of cannibalism, IGP, and competition exerted by these species were very symmetrical in both larvae and adults. We conclude that these biotic factors may not fully explain the increase in relative abundance of H. variegata. The higher sensitivity of E. connexa to prey patch size, along with other factors such as chemical‐mediated negative interspecific interactions may be operating in alfalfa fields, changing the composition of associated coccinellid assemblages.     

Shade facilitates an invasive stem succulent in a chenopod shrubland in South Australia

Abstract The invasive stem succulent Orbea variegata (L) Haw. (Asclepiadaceae) tends to be more abundant underneath shrubs than in open areas in chenopod shrublands near Whyalla, South Australia. To assess the role of facilitation in the life cycle of O. variegata, we investigated the effect of chenopod shrubs on different life stages of the species by experimentally manipulating temperature, light, soil moisture and nutrient levels. Experimental results suggest that the reduction in light and temperature under shrubs, but not increased nutrient levels, are the main facilitative mechanisms for O. variegata. Temperatures above 30°C, which are more likely to occur on the soil surface of open areas than under shrubs, inhibited seed germination. Seedling survival at low watering frequency and the growth of established ramets were increased by 75–80% shade cloth. Ramets growing in full light contained a high concentration of anthocyanin pigments. One of the functions of these pigments is to absorb excess radiation, suggesting that O. variegata experiences radiation stress in full light. In the field O. variegata performed considerably better under Atriplex vesicaria Heward ex Benth. (Chenopodiaceae) or under 75–80% shade cloth than in full light. Monthly irrigation of 20 mm did not reduce this positive effect of the A. vesicaria canopy on O. variegata, suggesting that O. variegata is inhibited by high light intensities or temperatures, independent of water availability. In conclusion, whereas shrub canopies do not seem to be required for the establishment or survival of O. variegata, shrubs improve adult growth and can improve establishment. The possibility of exotic plants being facilitated by other plants has to be taken into account when assessing the probability and rate of invasion.     

Microsatellite DNA markers for Australian geckos

Nine highly polymorphic microsatellite markers were developed for Gehyra variegata from an enriched library screened for tetranucleotide repeats. An average of 19.44 alleles per locus and an average observed heterozygosity of 0.86 were found in 81 samples from fragmented populations in the Western Australian wheatbelt. Different gecko species exhibit varying rates of extinction in fragmented landscapes and the genetic markers were developed to test whether these difference are due to different rates of dispersal. The loci were tested for their ability to cross amplify in 12 species of Australian Gekkonidae and Pygopodidae, in an attempt to identify species which might be amenable to population genetic analysis using these microsatellite markers.     

水淹生境下秋华柳对镉污染土壤研究修复能力

以耐水淹和耐重金属的秋华柳（Salix variegata）作为试验材料,从土壤角度出发,探究秋华柳在水淹条件下对镉污染土壤的修复能力。设置无植物和种植秋华柳两个处理组,分别对两组设置两个水分处理组：正常供水组（CK）及土壤水淹组（FL）,4个镉浓度处理组：对照组（0 mg/kg）、低浓度（0.5 mg/kg）、中浓度（2 mg/kg）及高浓度（10 mg/kg）。分别于处理后的第30天和第60天对各处理组的土壤和水淹组水样进行取样。试验结果表明：（1）镉处理浓度越高,土壤中镉活性态浓度越高,生物毒性越强。（2）水淹显著降低土壤活性态Cd浓度（P < 0.05）,增大土壤修复难度。（3）种植秋华柳对30d土壤Cd全量及各形态镉浓度均无显著影响（P > 0.05）;对第60天正常供水和水淹组土壤中交换态、碳酸盐结合态的镉浓度以及60d水淹组土壤Cd全量均有显著影响（P < 0.05）。（4）秋华柳种植显著降低土壤Cd迁移系数,30d正常供水组、水淹组以及60d正常供水组和水淹组平均降幅分别为2.0%、4.12%、9.71%、9.32%。处理时间过短和试验用苗生物量小可能是秋华柳对土壤全量Cd影响不大的主要原因,但秋华柳均显著降低正常供水和水淹组土壤迁移系数且二组降幅差异不大。研究表明：水淹生境下,秋华柳对Cd污染的土壤仍有较好的修复能力。     

Taxonomy and biogeography of Australian species of the Ropalidia stigma group and R. variegata group (Hymenoptera: Vespidae)

The taxonomy of the Australian species of two Ropalidia species groups, the R. stigma group and R. variegata group, is revised and their distribution patterns are discussed. Two species in the R. stigma group (R. darwini and R. elegantula), and four species in the R. variegata group (R. flavinoda, R. gregaria, R. mutabilis and R. revolutionalis) are recognized in Australia, with a new synonymy of R. mutabilis torresiana Richards, 1978, under R. gregaria (de Saussure, 1854). Icaria torrida Smith, 1863, from Seram Island is synonymized with R. unicolor (Smith, 1859).     

Factors affecting detectability of prey DNA in the gut contents of invertebrate predators: a polymerase chain reaction-based method

This study aimed to determine factors that influence the detection of DNA of Plutella xylostella L. (Lepidoptera: Plutellidae) in the gut contents of arthropod predators when the polymerase chain reaction is used to amplify a diagnostic fragment of the gene coding for cytochrome oxidase subunit I. The effects of temperature, time since feeding, subsequent food intake, sex, weight, and species of predator on prey detectability were studied in the laboratory. Three types of predator were studied: the spider Venator spenceri Hogg. (Araneae: Lycosidae), a bug with sucking mouthparts, Nabis kinbergii (Reuter) (Heteroptera: Nabidae), and a coccinellid with chewing mouthparts, Hippodamia variegata (Goeze) (Coleoptera: Coccinellidae). In all experiments, the detectability of prey DNA was negatively correlated with time post‐feeding. The duration of detectability differed among the predator species. The time calculated for median detection success at 20 °C ranged from 49.6 h in V. spenceri to 36.1 h in N. kinbergii and 17.1 h in H. variegata. In H. variegata, but not in V. spenceri, the rate of detection decreased with increasing temperature. Subsequent food intake did not affect the detectability of DNA of P. xylostella in V. spenceri. In H. variegata, sex and weight of the predator did not influence detection of prey DNA. In addition, this study uncovered potential sources of error caused by detection of prey DNA following secondary cannibalistic and intraguild predation. The results provide essential information for the interpretation of prey detection data from field‐collected predators’ gut contents.     

Foraging responses of Coccinella septempunctata,Hippodamia variegata and Chrysoperla carnea to changing in density of two aphid species

The successful use of predators in classical biocontrol programmes needs several background laboratory investigations, one of which is the evaluation of predator behavioural responses to changes in the density of their prey. The impact effect of the density of two prey species [Myzus persicae Sulzer and Aphis craccivora Koch (Hemiptera: Aphididae)] on the predation rates of third-instar Chrysoperla carnea Stephens (Chrysopidae: Neuroptera) and fourth-instar Coccinella septempunctata L. and Hippodamia variegata Goeze (Coccinellidae: Coleoptera) larvae was studied. Although prey species, predator species, prey density, and their interactions all had significant effects on the numbers of aphids consumed, the type of functional response did not vary, remaining a type II response in all treatments. However, the type II parameters differed among predator species on the same prey species, and for each predator species on the two prey species. Chrysoperla. carnea on M. persicae and H. variegata on A. craccivora were more voracious than other predators. In the context of functional response and biological control, the release of these predators, that show inverse density-dependent mortality, has to be started in early season to build up their population on low aphid densities and attack later high aphid populations.     

Cultivar-specificity genes of the nitrogen-fixing soybean symbiont,Rhizobium fredii USDA257, also regulate nodulation of Erythrina SPP.

Rhizobium fredii USDA257 is a Gram-negative soil bacterium from China that fixes nitrogen in symbiosis with primitive soybean (Glycine max [L.] Merr.) cultivars such as Peking, but not with advanced cultivars such as McCall. Mutation of either of two bacterial loci, nolBTUVWX or nolC, removes this cultivar-specificity constraint, allowing the bacterium to nodulate advanced cultivars. We have discovered that USDA257 forms nodules on Erythrina costaricensis M. Micheli, but not on six other species of this genus, which includes trees and shrubs from the tropics. Inactivation of nolBU or nolC broadens the symbiosis to include other Erythrina species. Whereas nolBU-mutants acquire the capacity to nodulate E. fusca Loureiro, E. variegata L., and E. vespertilio Benth., nolC-mutants could nodulate only E. variegata. Nodules are determinate and often clustered in root axils near the root crown. E. variegata nodules containing a nolBU-mutant consist of well-developed inner and outer cortical layers, as well as a bacteroid zone. The cortical layers are separated from one another by a meristematic region that is associated with large darkly staining cells, and cells of the inner cortex contain numerous starch grains. Histochemical staining of E. variegata roots inoculated with a strain containing a nolB-lacZ gene fusion confirms that nolB and nolC are expressed during the initial stages of bacterial interaction with the host. R. fredii enters elongated root-hairs via infection threads, which ramify within root-hairs and are associated with meristematic activity in the adjacent underlying cortical cells.     

Relationships between early growth and Quambalaria shoot blight tolerance in Corymbia citriodora progeny trials established in Queensland, Australia

The fungal pathogen Quambalaria pitereka can cause significant damage to spotted gum (Corymbia sp.) plantations in Australia. A series of seven progeny trials, involving seed from a range-wide collection from 527 individuals within 25 native populations of Corymbia citriodora sub-species variegata, were assessed for height growth and damage from Quambalaria around 1 year after planting. Infection at this young age has been found to detrimentally impact growth, form, and wood quality for many years. Genetic variance was found to be significant at both the provenance and family level. However, selection of families within provenances should lead to greater levels of genetic gain than what can be realized from selecting among provenances as estimates of additive genetic variance were consistently greater than estimates of variance among populations. Strong relationships between height and Quambalaria shoot blight (QSB) damage assessments in these trials were evidenced by very high genetic correlations between the traits; therefore, selection for any of these traits could be used to identify more productive and QSB-tolerant populations. While both provenances and families were found to interact with the trial environments at a similar level across traits, genetic correlations indicated that rankings for growth were be less stable than rankings for QSB tolerance across environments. Genetic parameter estimates derived from general and generalized linear models were very similar and either analytical method could be used to evaluate fungal damage.     

GENETIC ANALYSIS OF A HYBRID ZONE BETWEEN THE FIRE-BELLIED TOADS,BOMBINA BOMBINA AND B. VARIEGATA,NEAR CRACOW IN SOUTHERN POLAND

The fire-bellied toads Bombina bombina and B. variegata differ extensively in biochemistry, morphology, and behavior. We use a survey of five diagnostic enzyme loci across the hybrid zone near Cracow in Southern Poland to estimate the dispersal rate, selection pressures, and numbers of loci which maintain this zone. The enzyme clines coincide closely with each other and with morphological and mitochondrial DNA clines. Although the zone lies on a broad transition between environments suitable for bombina and variegata, the close concordance of diverse characters, together with increased aberrations and mortality in hybrids, suggest that the zone is maintained largely by selection against hybrids. There are strong “linkage disequilibria” between each pair of (unlinked) enzyme loci (R? = 0.129 [2-unit support limits: 0.119–0.139]). These are probably caused by gene flow into the zone, and they give an estimate of dispersal (σ = 890 [790–940] m gen^?½). The clines are sharply stepped, with most of the change occurring within 6.15 (5.45–6.45) km, but with long tails of introgression on either side. This implies that the effective selection pressure on each enzyme marker (due largely to disequilibrium with other loci) is s* = 0.17 (0.159–0.181) at the center but that the selection acting directly on the enzyme loci is weak or zero (s_e < 0.0038). The stepped pattern implies a barrier to gene flow of 220 (48–415) km. This would substantially delay neutral introgression but would have little effect on advantageous alleles; the two taxa need not evolve independently. Strong selection is needed to maintain such a barrier: hybrid populations must have their mean fitness reduced by a factor of 0.65 (0.60–0.77). This selection must be spread over a large number of loci to account for the concordant patterns and the observed cline widths (N = 300 [80–2,000]).