Isolation and cross-amplification of the first set of polymorphic microsatellite markers of two high-Andean cushion plants

In the southern Andes mountains (27–\(39{^{\circ }}\hbox {S}\)) Azorella madreporica and Laretia acaulis, two Apiaceae cushion plant species commonly known as yaretas, conform a well-established altitudinal vegetation belt along the lower Andean zone. These species have been considered as fundamental components of several ecological dynamics within their communities; however, high-mountain ecosystems are increasingly threatened worldwide by natural and anthropogenic pressures and the southern Andes are not the exception. Recognizing that genetic information is crucial for the success of any conservation or restoration initiative in wild populations, we developed and cross-amplified 28 specifically designed microsatellite markers (14 in A. madreporica and 14 in L. acaulis), and also tested the cross amplification of 25 markers from the related species Azorella selago. In a region which is particularly vulnerable to global change trends, this new polymorphic microsatellite loci will be useful in the study of the genetic diversity of these high-mountain cushion plants, which are pivotal in the structuring of their native ecosystems.     

<Emphasis Type="Italic">Lioptilodes friasi</Emphasis> (Lepidoptera: Pterophoridae) Niche Breadth in the Chilean Mediterranean Matorral Biome: Trophic and Altitudinal Dimensions

Understanding the factors driving the diet breadth of phytophagous insects remains one of the main questions in ecological research. In this study we explored the diet breadth and plant-insect associations in the plume moth Lioptilodes friasi Vargas & Parra (Lepidoptera: Pterophoridae). This phytophagous insect was originally described in association with a single host species, Haplopappus foliosus (Asteraceae), a native shrub of the Chilean Mediterranean matorral. In order to address the breadth of host plant choice, we surveyed other Haplopappus species growing along the elevation gradient of central Chile from sea level to 2600 m. We were able to obtain L. friasi adults from five additional Haplopappus species: Haplopappus chrysantemifolius and Haplopappus decurrens from the coastal zone, Haplopappus multifolius and Haplopappus schumanii from the mid-elevation zone, and Haplopappus scrobiculatus at high elevation. Our results demonstrate that the genus-specialized endophagous herbivore L. friasi has a wider distribution and climatic tolerance than previously described. Its biogeographical range extends from the lowland coastal habitats up to the Andean subnival level. We propose that shared flower phenotypic traits such as morphology and chemical composition may have allowed the colonization of closely related Haplopappus species in central Chile, the expansion of which is limited by the harsh high elevation conditions.     

On oviposition in weevils of the genus <Emphasis Type="Italic">Larinus</Emphasis> Dej. (Coleoptera,Curculionidae)

The following actions performed by females of several Larinus Dej. species during egg laying are described: search of an appropriate place on the plant, making the hole for the egg, oviposition proper, and sealing the hole. The hole preparation takes the longest time and the greatest effort. Only one individual usually completes development in one flower head. Females of Larinus vulpes Ol. prefer larger flower heads for oviposition and occasionally lay eggs into stems. The ability of females to distinguish the flower heads with already laid eggs is discussed. Species of Larinus may be divided into two groups with “early” and “late” oviposition. The evolution of egg laying in the genus Larinus is discussed.     

Climatic niche evolution in the Andean genus <Emphasis Type="Italic">Menonvillea</Emphasis> (Cremolobeae: Brassicaceae)

The study of how climatic niches change over evolutionary time has recently attracted the interest of many researchers. Different methodologies have been employed principally to analyze the temporal dynamics of the niche and specially to test for the presence of phylogenetic niche conservatism. Menonvillea, a genus of Brassicaceae including 24 species, is distributed primarily along the Andes of Argentina and Chile, with some taxa growing in southern Patagonia and others in the Atacama Desert and the Chilean Matorral. The genus is highly diversified morphologically but also presents a remarkably wide ecological range, growing from the high Andean elevations, to the dry coastal deserts in Chile, or the Patagonia Steppe in Argentina. In this study, we used molecular phylogenies together with climatic data to study climatic niche evolution in the genus. The results show that the main climatic niche shifts in Menonvillea occurred between the sections Cuneata-Scapigera and sect. Menonvillea throughout the Mid-Late Miocene, and associated with the two main geographical distribution centers of the genus: the highlands of the central-southern Andes and the Atacama Desert-Chilean Matorral, respectively. Climatic niches in these lineages were mainly differentiated by the aridity and potential evapotranspiration, the minimum temperatures of the coldest month, and the temperature annual range and seasonality. Niche evolution in Menonvillea deviated from a Brownian motion process, with most of the climatic dimension best-fitting to an Ornstein-Uhlenbeck model of multiple adaptive peaks. Our results also indicated that higher aridity levels and lower annual temperature ranges were associated with the evolution of the annual habit, as exemplified by the distribution of sect. Menonvillea. Finally, the results suggested that climatic niche evolution in Menonvillea exhibited some degree of phylogenetic niche conservatism, fundamentally within the two main lineages (sect. Menonvillea and sects. Cuneata-Scapigera).     

The evolution of cultivated potatoes

In the first stages of potato evolution in the northern Andes, diploid cultivated species of theSolanum stenotomum complex were selected, in all probability, from wild progenitors in theS. brevicaule complex. TetraploidSolanum tuberosum ssp.andigena arose by fusion of unreduced gametes of a parent in theS. stenotomum complex with those of an unidentified wild species having actinomorphic calyces. Unreduced male gametes of several diploid species fertilized eggs of ssp. andigena leading to extensive introgression.Solanum tuberosum ssp.tuberosum probably originated from a cross between ssp.andigena as staminate parent and an unidentified wild species which contributed cytoplasmic sterility factors encoded in mitochondria and/or plastids having a distinctive type of DNA. Derivatives of this hybridization, which may have occurred in northwestern Argentina, evolved to ssp. tuberosum in southern Chile and southern Argentina. In the 1570’s ssp.andigena was imported to Europe and spread from there to become a major crop with worldwide distribution. In the 1840's it was essentially eliminated by late blight,Phytophthora infestans.Solanum tuberosum ssp.tuberosum was introduced from Chile into North America and Europe in the late 1800's, and in turn achieved a worldwide distribution, filling the vacated agricultural niche of ssp.andigena. The differences between ssp.andigena and ssp.tuberosum in South America are sufficient that the two could reasonably be considered to be separate species. Since the 1960’s the two taxa have been hybridized often in breeding programs. Neotuberosum, a northern-adapted strain of ssp. andigena, has been selected to mimic ssp.tuberosum. Substitution back-cross products have been produced that have the chromosomal genes of ssp.tuberosum combined with cytoplasmic factors of Andean species. These breeding activities are blurring the distinctions between the two subspecies throughout much of the world, though they remain distinct in their native areas in South America.     

Rate of consumption,biological parameters,and population growth capacity of <Emphasis Type="Italic">Orius laevigatus</Emphasis> fed on <Emphasis Type="Italic">Spodoptera exigua</Emphasis>

Orius laevigatus (Fieber) (Hemiptera: Anthocoridae) is already successfully used to control Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae) in protected crops. In the present work, the predatory capacity of O. laevigatus on the eggs and larvae of Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae) as well as the population growth capacity of O. laevigatus fed on eggs of the noctuid were determined. Fourth and 5th instar nymphs had a greater consumption of eggs than younger ones. Females consumed more eggs than males. Only 5th instar nymphs and adults preyed on the neonatal larvae of S. exigua. The intrinsic rate of natural increase did not differ between O. laevigatus fed with S. exigua eggs and those offered eggs of the substitute host Ephestia kuehniella (Zeller) (Lepidoptera: Pyralidae). Thus, O. laevigatus is a good candidate for the biological control of S. exigua, a cosmopolitan pest of many crops.     

Proteomics and functional analyses of <Emphasis Type="Italic">Arabidopsis</Emphasis> nitrilases involved in the defense response to microbial pathogens

Main conclusion

Proteomics and functional analyses of the Arabidopsis – Pseudomonas syringae pv. tomato interactions reveal that Arabidopsis nitrilases are required for plant defense and R gene-mediated resistant responses to microbial pathogens. A high-throughput in planta proteome screen has identified Arabidopsis nitrilase 2 (AtNIT2), which was de novo-induced by Pseudomonas syringae pv. tomato (Pst) infection. The AtNIT2, AtNIT3, and AtNIT4 genes, but not AtNIT1, were distinctly induced in Arabidopsis leaves by Pst infection. Notably, avirulent Pst DC3000 (avrRpt2) infection led to significant induction of AtNIT2 and AtNIT4 in leaves. Pst DC3000 and Pst DC3000 (avrRpt2) significantly grew well in leaves of nitrilase transgenic (nit2i-2) and mutant (nit1-1 and nit3-1) lines compared to the wild-type leaves. In contrast, NIT2 overexpression in nit2 mutants led to significantly high growth of the two Pst strains in leaves. The nitrilase transgenic and mutant lines exhibited enhanced susceptibility to Hyaloperonospora arabidopsidis infection. The nit2 mutation enhanced Pst DC3000 (avrRpt2) growth in salicylic acid (SA)-deficient NahG transgenic and sid2 and npr1 mutant lines. Infection with Pst DC3000 or Pst DC3000 (avrRpt2) induced lower levels of indole-3-acetic acid (IAA) in nit2i and nit2i NahG plants than in wild-type plants, but did not alter the IAA level in NahG transgenic plants. This suggests that Arabidopsis nitrilase 2 is involved in IAA signaling of defense and R gene-mediated resistance responses to Pst infection. Quantification of SA in these transgenic and mutant plants demonstrates that Arabidopsis nitrilase 2 is not required for SA-mediated defense response to the virulent Pst DC3000 but regulates SA-mediated resistance to the avirulent Pst DC3000 (avrRpt2). These results collectively suggest that Arabidopsis nitrilase genes are involved in plant defense and R gene-mediated resistant responses to microbial pathogens.

     

Diversification of <Emphasis Type="Italic">Caiophora</Emphasis> (Loasaceae subfam. Loasoideae) during the uplift of the Central Andes

Andean orogeny and the ecological changes that followed promoted diversification in plant and animal lineages since the Early Miocene. The angiosperm genus Caiophora (Loasaceae, subfam. Loasoideae) comprises around 50 species that are endemic to South America. These are distributed from southern Ecuador to Central Chile and Argentina. Bee pollination and distribution at low-intermediate elevations probably represent the ancestral condition in the lineage that includes Caiophora and its allied genera. The majority of Caiophora species grow at high elevations in the Andes, where some depend on vertebrate pollination. Previous studies did not resolve phylogenetic relationships within Caiophora, which precluded the dating of the origin and divergence of this group. We used markers of one nuclear (ITS) and one plastid region (trnS ^GCU -trnG ^UUC ) to solve phylogenetic relationships among 19 Caiophora species (including different accessions). We also included 10 species of the allied genera Blumenbachia and Loasa. Aosa rostrata and Xylopodia klaprothioides were used as outgroups. Phylogenetic reconstruction strongly supports the monophyly of Caiophora, and although several clades within this genus are poorly supported, our study yielded a better infra-generic resolution than previous studies. The origin of Caiophora is dated to the Early-Middle Miocene and can be related to the uplift of the Cordilleras Frontal and Principal and to Pacific marine transgressions. According to our estimations, Caiophora began to diversify during the Middle-Late Miocene and this unfolding proceeded eastwards during the Pliocene and the Pleistocene, in parallel to the uplift of different Andean mountain ranges.     

Biogeography and phenology of oviposition preference and larval performance of <Emphasis Type="Italic">Pieris virginiensis</Emphasis> butterflies on native and invasive host plants

In invaded environments, formerly reliable cues might no longer be associated with adaptive outcomes and organisms can become trapped by their evolved responses. The invasion of Alliaria petiolata (garlic mustard) into the native habitat of Pieris virginiensis (West Virginia White) is one such example. Female butterflies oviposit on the invasive plant because it is related to their preferred native host plant Cardamine diphylla (toothwort), but larvae are unable to complete development. We have studied the impact of the A. petiolata invasion on P. virginiensis butterflies in the Southeastern USA by comparing oviposition preference and larval survival on both plants in North Carolina (NC) populations without A. petiolata and West Virginia (WV) populations where A. petiolata is present. Larval survival to the 3rd instar was equally low in both populations when raised on A. petiolata. Mean oviposition preference on the two plants also did not differ between populations. However, we found a seasonal effect on preference between early and late season flights within WV populations. Late season females laid 99% of total eggs on A. petiolata while early season females utilized both host plants. Late season females were also less likely to lay eggs than early season females. This change in preference toward A. petiolata could be driven by the early senescence of C. diphylla and suggests a seasonal component to the impact of A. petiolata. Therefore, the already short flight season of P. virginiensis could become further constrained in invaded populations.     

Identification and polymorphism of pectinase genes <Emphasis Type="Italic">PGU</Emphasis> in the <Emphasis Type="Italic">Saccharomyces bayanus</Emphasis> complex

Pectinase (endo-polygalacturonase) is the key enzyme splitting plant pectin. The corresponding single gene PGU1 is documented for the yeast S. cerevisiae. On the basis of phylogenetic analysis of the PGU nucleotide sequence available in the GenBank, a family of divergent PGU genes is found in the species complex S. bayanus: S. bayanus var. uvarum, S. eubayanus, and hybrid taxon S. pastorianus. The PGU genes have different chromosome localization.     

Observations on the ecology of the endangered butterfly <Emphasis Type="Italic">Zerynthia cassandra</Emphasis> in a protected area of Northern Italy

Zerynthia cassandra is a butterfly endemic to the Italian peninsula. It was recently recognized as a sister species of Zerynthia polyxena. Z. cassandra is oligophagous as it feeds on a very restricted number of toxic plants belonging to the genus Aristolochia: Aristolochia pallida and A. rotunda. The paper reports the results of a research on the ecology of this species which was studied in the natural reserve "Stagni di Lungavilla" (Northern Italy) where the butterfly was living in a small patch of grassy vegetation bordered by a ditch at the margins of a pond. Z. cassandra develops one generation a year. Oviposition occurred in April. Eggs were mainly laid on the lower side of A. rotunda blades. The height of eggs from the ground depended on climatic trends, which affected the host plant phenology. Eggs mortality was mainly due to consumption of leaves by snails (Cepaea nemoralis) and slugs (Arion sp.). Larvae completed their development from April to May, while chrysalises were the aestivating and overwintering stage. Females did not lay eggs on host plants when they were part of the undergrowth. A decline of the butterfly due to habitat changes was recorded during the research (2001–2005). According to information coming from local butterfly watchers the butterfly used to be quite common in the natural reserve until 70’s. Its decline was caused by agricultural intensification and the natural evolution of vegetation towards woodlands, which gradually reduced the host plant density, so making the habitat unsuitable and causing the local extinction of the butterfly.     

Banding and FISH in three species of <Emphasis Type="Italic">Vernonia</Emphasis>, subsection <Emphasis Type="Italic">Macrocephalae</Emphasis> (Asteraceae,Vernonieae)

To complement our knowledge about the karyotypes of the genus Vernonia Schreb., different techniques of chromosome banding, including AgNOR, triple staining with fluorochromes CMA/DA/DAPI (CDD), and fluorescence in situ hybridization (FISH) for the 45S rDNA probe, were applied to three species of subsection Macrocephalae. Vernonia bardanoides was collected from an area of cerrado (savanna) vegetation in Itirapina, São Paulo State, Brazil, and V. linearifolia and V. tomentella were collected from areas of rocky, open altitudinal vegetation in Joaquim Felicio and Diamantina, respectively, in Minas Gerais State. All species showed two terminal CMA⁺ and NOR bands. FISH indicated two terminal 45S rDNA sites in V. linearifolia and V. tomentella, and six in V. bardanoides.     

Reproduction and development of <Emphasis Type="Italic">Pholis picta</Emphasis> (Perciformes: Pholidae) and <Emphasis Type="Italic">Opisthocentrus zonope</Emphasis> (Stichaeidae)

In the Peter the Great Bay (the Sea of Japan), reproduction and embryonic larval development of Pholis picta and Opisthocentrus zonope are investigated. Both species spawn in November–December at the water temperature decreasing from 7.0 to 0.2°C. Fecundity of Pholis picta is 494–1228 eggs, and of Opisthocentrus zonope—368–1360 eggs. Both species lay eggs in empty shells of Gray mussel. Parent fish permanently stay in the shell but do not actively care for the eggs. Morphological changes in the process of embryogenesis are compared between the related species. Prolarvae and larvae are described for identification, with special attention to pigmentation. The spawning behavior of adult fish and behavior of prolarvae are described.     

Biodiversity of aerobic methylobacteria associated with the phyllosphere of the southern Moscow region

During the summer period (15–25°C), 34 strains of methylotrophic bacteria associated with different species of herbs, shrub, and trees in Pushchino (Moscow oblast, Russia) were isolated on the medium with methanol. Predominance of pink-colored Methylobacterium strains in the phyllosphere of many plants was confirmed by microscopy, enumeration of the colonies from grass leaves, and sequencing of the 16S rRNA genes. Colorless and yellow-pigmented methylotrophs belonged to the genera Methylophilus, Methylobacillus, Hansschlegelia, Methylopila, Xanthobacter, and Paracoccus. All isolates were able to synthesize plant hormones auxins from L-tryptophan (5?50 μg/mL) and are probably plant symbionts.     

<Emphasis Type="Italic">Quinua</Emphasis> and Relatives (<Emphasis Type="Italic">Chenopodium</Emphasis> sect.<Emphasis Type="Italic">Chenopodium</Emphasis> subsect.<Emphasis Type="Italic">Celluloid</Emphasis>)

Traditionally viewed as an Andean grain crop,Chenopodium quinoa Willd. includes domesticated populations that are not Andean, and Andean populations that are not domesticated. Comparative analysis of leaf morphology and allozyme frequencies have demonstrated that Andean populations, both domesticated(quinua) and free-living(ajara), represent an exceptionally homogeneous unit that is well differentiated from allied domesticates of coastal Chile(quingua) and freeliving populations of the Argentine lowlands(C. hircinum). This pattern of relationships indicates that Andean populations represent a monophyletic crop/weed system that has possibly developed through cyclic differentiation (natural vs. human selection) and introgressive hybridization. Relative levels of variation suggest that this complex originated in the southern Andes, possibly from wild types allied withC. hircinum, with subsequent dispersal north to Colombia and south to the Chilean coast. Coastal populations were apparently isolated from post-dispersal differentiation and homogenization that occurred in the Andes. Other data point toward a center of origin in the northern Andes with secondary centers of genetic diversity subsequently developing in the southern Andes and the plains of Argentina. Comparative linkage of South American taxa, all tetraploid, with North American tetraploids of the subsection will eventually clarify this problem. While the possibility of a direct phyletic connection betweenC. quinoa and the Mexican domesticate(C. berlandieri subsp. nuttalliae,) cannot be excluded, available evidence indicates that the latter represents an autonomous lineage that is associated with the basal tetraploid, C. b. subsp.berlandieri, through var.sinuatum, whereas South American taxa show possible affinities to either var. zschackei or var.berlandieri. An extinct domesticate of eastern North America,C. b. subsp.jonesianum, represents either another instance of independent domestication, possibly from subsp. b. var.zschackei, or a northeastern outlier of subsp.nuttalliae.     

Morphological and molecular investigations of <Emphasis Type="Italic">Gagea</Emphasis> (Liliaceae) in southeastern Kazakhstan with special reference to putative altitudinal hybrid zones

The most important center of speciation in the genus Gagea is thought to be in Central Asia. Here, we focus on species diversity in southeastern Kazakhstan (around Almaty, Ili-Alatau range of the Western Tian-Shan mountains). We studied an elevational transect, reaching from lowland steppes to the alpine zone (500–2750 m a. s. l.), and carried out detailed morphological and molecular investigations for populations of Gagea spp. Nine species were identified in different altitudinal zones; one of these (Gagea almaatensis) is described as new to science. We could detect two altitudinal contact zones between closely related species: G. filiformis and G. granulosa (sect. Minimae), and G. almaatensis and G. kuraminica (sect. Gagea). Morphological and molecular investigations (ITS data and cpDNA networks) indicate ongoing hybridization of co-occurring G. filiformis into G. granulosa and putative bidirectional hybridization events between G. almaatensis and G. kuraminica.     

Chemical defences of native European coccinellid eggs against intraguild predation by the invasive Asian coccinellid, <Emphasis Type="Italic">Harmonia axyridis</Emphasis> (Pallas)

Harmonia axyridis (Pallas) is a coccinellid of Asian origin that has recently invaded substantial parts of Europe and is suspected to affect native coccinellid populations through intraguild predation and competition for food. Previous work has shown that two species from the Calvia genus appeared to be well protected against H. axyridis predation. To deepen our understanding on chemical protection of Calvia spp. and the predation risk by H. axyridis, we tested for susceptibility and palatability of Calvia spp. and H. axyridis eggs against predation by H. axyridis neonate larvae. Results show that eggs of C. quatuordecimguttata were mostly not eaten by H. axyridis, while eggs of the congeneric C. decemguttata were found to be largely unprotected against predation by the invasive coccinellid. We also observed that H. axyridis first instars successfully cannibalized on conspecific eggs. Removing the surface chemicals from C. quatuordecimguttata eggs resulted in significantly reduced protection from being preyed upon by H. axyridis, while applying these extracts onto C. decemguttata and H. axyridis eggs resulted in increased protection against H. axyridis larvae. The importance of surface chemicals in the interactions between H. axyridis and native coccinellids was confirmed by GC–MS analysis, showing a high diversity of hydrocarbons located on the surface of C. quatuordecimguttata eggs, i.e. more than twice as many when compared to C. decemguttata. Survival of H. axyridis larvae feeding on eggs of C. quatuordecimguttata, C. decemguttata or conspecific eggs, from which surface chemicals were removed by washing them with hexane, was not different from survival on unwashed eggs.     

Molecular phylogeny of pectinase genes <Emphasis Type="Italic">PGU</Emphasis> in the yeast genus <Emphasis Type="Italic">Saccharomyces</Emphasis>

Using yeast genome databases and literature data, phylogenetic analysis of pectinase PGU genes from 112 Saccharomyces strains assigned to the biological species S. arboricola, S. bayanus (var. uvarum), S. cariocanus, S. cerevisiae, S. kudriavzevii, S. mikatae, S. paradoxus, and the hybrid taxon S. pastorianus (syn. S. carlsbergensis) was carried out. A superfamily of divergent PGU genes was found. Natural interspecies transfer of the PGU gene both from S. cerevisiae to S. bayanus and from S. paradoxus to S. cerevisiae may, however, occur. Within the Saccharomyces species, identity of the PGU nucleotide sequences was 98.8–100% for S. cerevisiae, 86.1–95.7% for S. bayanus (var. uvarum), 94–98.3% for S. kudriavzevii, and 96.8–100% for S. paradoxus/S. cariocanus. For the first time, a family of polymeric PGU1b, PGU2b, PGU3b and PGU4b genes is documented for the yeast S. bayanus var. uvarum, a variety important for winemaking.     

Plant and supplementary food sources effect the development of <Emphasis Type="Italic">Dicyphus errans</Emphasis> (Hemiptera: Miridae)

Dicyphus errans (Wolff) (Hemiptera: Miridae) is an omnivorous predator of several pests attacking tomato and other vegetable crops. The nymphal development of D. errans was studied in the presence of Ephestia kuehniella Zeller (Lepidoptera: Pyralidae) eggs with or without a water source and in the presence of a leaf of cucumber, eggplant or tomato supplemented with variable food types (E. kuehniella eggs, Artemia sp. cysts, pollen or milk powder) or without the provision of any food. Water provision was found to be essential for the completion of nymphal development even when animal food was offered to predators. When nymphs foraged on leaves in the absence of any food type, development was significantly more favoured on eggplant and cucumber than on tomato. E. kuehniella eggs and Artemia sp. cysts enhanced development of D. errans in comparison to milk powder and pollen. Development and female weight were improved when the food types were offered to the nymphs on a plant leaf than when were provided together with only water. This study contributes to understanding the importance of water vs. plant feeding for the development of D. errans and reveals implications for its mass rearing and application in biological control.