Genetic variation inOrchis papilionacea (Orchidaceae) from the Central Mediterranean region: Taxonomic inferences at the intraspecific level

The genetic differentiation ofOrchis papilionacea from Italy, Corsica, Sardinia, Sicily, and Greece was analyzed by isozyme studies. No significant differences at any of the 28 loci tested were observed between samples of the putative subspeciespapilionacea from Italy,grandiflora from Sicily andheroica from Greece, or by comparing sympatricpapilionacea-like andgrandiflora-like subsamples from Sardinia and Corsica. The genetic relationships found among the populations studied reflect more their geographic origin than their taxonomic assignment;papilionacea, grandiflora, andheroica are, therefore, at least in the study area, not to be considered as subspecies, but as simple morphs, controlled by few genes (possibly two) with Mendelian inheritance. The genetic divergence observed among the samples studied is very low and consistent with that of geographic populations of a single cohesive gene pool, linked by a high gene flow (Nm = 5.9).     

Karyomorphological analyses and heterochromatin characteristics disclose phyletic relationships among 2n = 32 and 2n = 36 species ofOrchis (Orchidaceae)

The karyotypes of eight taxa ofOrchis L. with 2n = 32 and 2n = 36 have been investigated using morphometrical measurements following staining with Feulgen, Giemsa (C-banding) and the DNA specific fluorochrome Hoechst 33258. The karyotypes ofO. coriophora subsp.fragrans, andO. papilionacea proved to be the most asymmetrical, whileO. morio andO. longicornu exhibited the most symmetrical karyotypes. Using C-banding and the fluorochrome H33258 only the taxa with high asymmetry indices showed the presence of differentially stained chromatin bands. In most chromosomes heterochromatin bands were present at the telomeric position. The present results seem to indicate that the analysed species do not form a homogeneous group and further subdivisions are possible, which, in turn, do not always correlate with divisions based on morphological characters. Both karyomorphology and heterochromatin distribution coincide in indicating a possible evolutionary pathway.     

Hybriden und hybridogene Sippen ausOphrys bertolonii undO. atrata (Orchidaceae)

Flower polymorphism in 15 ItalianOphrys populations (N = 282) was analysed in detail. Statistical and graphical evaluation of data demonstrates morphological relationships and amount of hybrid character coherence.O. bertolonii s. str. andO. atrata are stable parental taxa, but occasionally form hybrids and hybrid swarms. Much evidence suggests that it was through such homogamic hybridization, involving these and other species ofOphrys, that the stabilized taxaO. bertoloniiformis andO. promontorii (ecologically different, but sympatric with the parents on Gargano), andO. benacensis (geographically separated from the parents in the Insubric region) originated. These differentiation-hybridization cycles are linked to the break-down and new establishment of incomplete prezygotic crossing barriers (e.g. different flower biology, flowering time, habitat preference).

     

Genetic variation and natural hybridization betweenOrchis laxiflora andOrchis palustris (Orchidaceae)

Genetic divergence between population samples ofOrchis laxiflora and ofO. palustris from various European locations was studied by electrophoretic analysis of 25 enzyme loci. An average genetic distance of D_Nei = 1.24 was found between the two taxa, with 12 out of 25 loci showing alternative alleles (diagnostic loci). Genetic heterogeneity was observed within bothO. laxiflora andO. palustris, when northern and southeastern populations were compared, being lower in the former taxon (D = 0.06), than in the latter (D = 0.16). Karyologically, 2n = 36 was found for bothO. laxiflora andO. palustris. O. laxiflora andO. palustris produce hybrids, described asO. ×intermedia. Genotype analysis of several sympatric samples showed the presence of hybrid zones, including F₁ hybrids and, in low proportions, recombinant classes, putatively assigned to F_n and backcrosses, as well as a few introgressed individuals of both taxa. These data indicate that hybrids are only partially fertile, with a very limited mixing up of the two parental gene pools; this is also shown by the lack of significant lowering of genetic distances when sympatric and allopatric heterospecific samples are compared. Accordingly,O. laxiflora andO. palustris form a syngameon; nevertheless they can be considered as good taxonomic species, with virtually distinct gene pools, which evolve independently. The genetic variability inO. laxiflora andO. palustris is remarkably low ( _e = 0.05 and _e = 0.02, respectively). In particular, nearly complete absence of polymorphic loci was found inO. palustris from northcentral Europe. Two hypotheses are considered to explain the low genetic variability of this endangered species.     

Populationsstatistische Analysen bestätigen die Heterogeneität vonOphrys „arachnitiformis“ (Orchidaceae)

Statistical population analyses have been carried out on so-calledOphrys arachnitiformis, partly sympatricO. sphecodes s. l. as well as on other species ofOphrys (O. holosericea, O. exaltata, O. apulica, O. cornuta, O. biscutella, O. bertolonii, O. bertoloniiformis.). Altogether 63 populations with 521 individuals were studied in regard to 54 morphological parameters. — It can be shown thatO. arachnitiformis is an extremely heterogeneous assemblage which includes taxa of most diverse origin, different degrees of stability and nomenclatorial rank. Only common trait is their very diverse relationships withO. specodes s.l.—The basionymO. arachnitiformis Gren. & Phil. refers to a colour variant ofO. sphecodes Mill. subsp.sphecodes only, and is not applicable to other segments of the assemblage.A late flowering, morphologically and biologically well isolated group of populations from southern France without any perceptible hybrid influence is established as a new species:Ophrys splendida Gölz & Reinhard. Populations from north and central Italy with distinct characters but hybrid influences fromO. holosericea/O. exaltata are calledO. tyrrhena Gölz & Reinhard.O. morisii, an endemic to Sardinia, represents an independent species of still obscure evolution. The hybrid interpretation ofO. catalaunica is statistically corroborated, but somewhat modified. For the remaining segments ofO. arachnitiformis investigated (from Castilia, Dalmatia, and the Monte Gargano) no formal taxonomic proposals are made, but some suggestions about their genesis are possible.

     

Molecular (RAPD) analysis on Italian taxa of theOphrys bertolonii aggregate (Orchidaceae)

The RAPD technique has been used for analyzing the genetic material of seven Italian populations of theOphrys bertolonii aggregate (Orchidaceae). For comparison the related speciesO. sphegodes andO. fuciflora have also been analyzed. A high genetic variability has been observed within the same populations, and on the whole onlyO. bertolonii can be safely differentiated from the other taxa of the aggregate, which are undistinguishable with this method.     

Genetic relationships between European and Siberian larch,Larix spp. (Pinaceae), studied by allozymes. Is the Polish larch a hybrid between these two species?

Thirteen populations ofLarix decidua subsp.decidua and subsp.polonica, and three populations ofL. sibirica were investigated by starch-gel electrophoresis. In the populations assayed 61 alleles at 17 loci were revealed. The allozyme data support the earlier observations about close relationships between these two larch species. Nei's genetic distances betweenL. decidua andL. sibirica were relatively small (D = 0.057), however, almost five times larger, on average, than those between populations of the same species. Results obtained in this study disagree withBobrov's hypothesis about the hybrid origin of the Polish larch and suggest a direct origin from the European larch.     

Phylogenetic relationships inSecale (Poaceae): An isozymatic study

The genetic frequencies of 9 isozyme loci have been estimated in 23 samples of 4 species ofSecale by means of starch gel electrophoresis. The populations ofS. silvestre andS. vavilovii were monomorphic and uniform within each species, those ofS. montanum andS. cereale were polymorphic for most of the isozyme loci. On the basis of isozyme patterns as well as allelic and genotypic frequencies of isozyme loci,S. silvestre can be distinguished fromS. vavilovii, and both fromS. cereale andS. montanum; but there is no clear differentiation between the two latter species. Clusters constructed from genetic distances separateS. silvestre andS. vavilovii, whereasS. cereale andS. montanum were grouped together. The isozymatic data presented here, along with cytogenetic and life habit data, agree with the generally admitted existence of 4 species inSecale, and support the relationships suggested byKhush & Stebbins (1961).     

Morphological and molecular characterization of a natural hybrid betweenOrchis laxiflora andO. morio (Orchidaceae)

A putative natural hybrid betweenOrchis laxiflora andO. morio (Orchidaceae) from southern Italy, formerly known asO. alata, was characterized both on morphological and molecular grounds in order to confirm its hybrid status and to trace its maternal lineage. The morphological characters of the putative hybrid showed intermediacy between those of the parent species, and restriction fragment length polymorphism (RFLP) analysis of a region of the nuclear ribosomal DNA confirmed its hybrid origin. Chloroplast DNA RFLP analysis indicated thatO. morio provided the maternal genome.     

Three new species and a new combination in the genusTorulomyces from soil

Seven isolates ofTorulomyces from Asian and Australian soil samples were studied in comparison with known taxa of the genus and withMonocillium indicum, the type species ofMonocillium. Three new species,Torulomyces parviverrucosus, T. laevis, andT. ovatus, are described, andT. brunneus is described as a new combination. Conidial characteristics, especially their shape and surface structure, are useful taxonomic criteria for distinguishing species ofTorulomyces. Monocillium is considered to be a distinct genus.     

Intrasexual Competition and Mating Behavior in <Emphasis Type="Italic">Ptomascopus morio</Emphasis> (Coleoptera: Silphidae Nicrophorinae)

In nicrophorine beetles, genus Nicrophorus care their larva using small vertebrate carrion, whereas genus Ptomaucopusreproduce with small vertebrate carrion but show no parental care. Aggression and sexual behavior were examined in Ptomascopus morio and Nicrophorus quadripunctatus. Nicrophorus quadripunctatus had intense female–female as well as male–male contests. In Ptomascopus morio, by contrast, female–female aggression was rarely observed. Male–male aggression (pushing, biting, male–male mounting) in Ptomascopus morio was observed when a resource for breeding was present, whether or not a female was present. The lack of female–female aggression, and male–male aggression when resources but not females are present, suggest that the mating system of Ptomascopus morio is resource defense polygyny. Large males of Ptomascopus morio were also found to exhibit mate choice, preferring large females over small females.     

Localized hybridization betweenSaxifraga aizoides andS. mutata (Saxifragaceae): Reproductive ecology ofS. ×hausmannii and implications for conservation

The roles of hybridization and of introgression as potential threats to small native populations are current topics of interest both for evolutionary and conservation biologists. Two factors are mainly involved in the threat to natural populations through hybridization: demographic swamping and genetic assimilation. At prealpine locations, twoSaxifraga species with very different growth habits,S. aizoides andS. mutata, form a hybrid,S. ×hausmannii. Investigation of meiosis and crossing experiments revealed regular pairing of the 26 chromosomes and development of viable seeds in the hybrid. Analysis of hybrid indices, GPI allozymes, germination experiments and seed sizes showed an intermediate position ofS. ×hausmannii between its parental species.Saxifraga aizoides andS. mutata were separated by only weak hybridization barriers. Introgression occurred mostly in direction ofS. aizoides, although to a small amount. There is no actual threat to small, prealpine populations ofS. aizoides andS. mutata through interspecific gene flow because hybridization is localized, resulting from different flowering phenologies of the parental species and from the narrow habitat requirements ofS. ×hausmannii, which are similar to those ofS. aizoides. The major threat to local, prealpine populations ofS. aizoides andS. mutata is the loss of landscape dynamics. Both species need open erosion slopes for successful recruitment.     

Pollination ofOphrys (Orchidaceae) in Cyprus

In the southern part of Cyprus the pollinator —Ophrys (Orchidaceae) relationships and its specifity have been investigated from the end of February until the middle of March 1986. 12Ophrys spp. were found. To date, only a single pollinator reference has been reported from this island. We found the following pollinators:Melecta tuberculata (Ophrys kotschyi),Eucera dimidiata (Ophrys flavomarginata),Eucera gaullei (Ophrys umbilicata),Eucera paulusi (Ophrys bornmuelleri),Anthophora erschowi (Ophrys elegans),Andrena torda (Ophrys sicula =O. lutea subsp.minor),Andrena cinereophila (Ophrys fusca, small-flowered),Andrena flavipes (Ophrys israelitica),Andrena morio (Ophrys iricolor andOphrys transhyrcana),Andrena bimaculata (Ophrys sphegodes aggr., probably formerly confused withO. transhyrcana). Most interestingly, it could be verified thatO. flavomarginata/O. umbilicata, O. bornmuelleri/O. levantina andO. transhyrcana/O. sphegodes aggr. (possiblyO. sintenisii) are different biospecies. This is a result of genetic isolation due to varying pollinators, and of differences in flower morphology.     

Cytology ofCrocus sativus and its allies (Iridaceae)

Populations ofC. cartwrightianus, C. hadriaticus andC. thomasii from the Balkans have 2n = 16 as has the Middle EasternC. pallasii subsp.haussknechtii. C. dispathaceus andC. pallasii subsp.pallasii have 2n = 14 whileC. pallasii subsp.turcicus has 2n = 12.C. niveus has 2n = 28 andC. sativus is an autotriploid with 2n = 24. Karyotype variation was found between populations ofC. pallasii subsp.pallasii in Central Turkey and also inC. pallasii subsp.turcicus.     

Generic delimitation of Aralia (Araliaceae)

Aralia, described by Linnaeus, remains a poorly defined genus. Many satellite genera ofAralia have been proposed, and some have been accepted without critical evaluation. These genera includeCoudenbergia, Megalopanax, Parapentapanax, andPentapanax. This study examined the historical background of the establishment of these genera and their diagnostic characters. It was found that (1)Aralia s. str. is a relatively diverse genus; (2) the separation ofPentapanax andParapentapanax fromAralia is unsubstantiated; (3)Megalopanax is synonymous withCoudenbergia; and (4) the boundaries betweenAralia and bothCoudenbergia andPentapanax intergrade. It is proposed, therefore, to mergeCoudenbergia, Parapentapanax, andPentapanax withAralia, which has nomenclatural priority.Aralia is herein defined as consisting of those species in Araliaceae with pinnate leaves, articulated rachises, five to eight-merous flowers, petals imbricate in buds, and articulated pedicels. The revised generic concept requires the following new combinations or new names:Aralia castanopsisicola, A. franchetti, A. gigantea, A. laevis, A. leschenaultii, A. parasitica, A. rex, A. subcordata, A. verticillata, andA. warmingiana.     

Pollination biology ofOpuntia polyacantha andOpuntia phaeacantha (Cactaceae) in southern Colorado

The phenology, compatibility system, and pollinator fauna ofOpuntia polyacantha Haw. andO. phaeacantha Engelm. in southern Colorado were studied and compared. The total blooming periods overlap, but the peak of blooming differs between the species withO. polyacantha blooming first and for longer. Neither species is apomictic,O. polyacantha is largely self-incompatible andO. phaeacantha is self-compatible but may not automatically fully self-pollinate. Examination of flowering structures showed that the former has larger flowers with more perianth parts and heavier (but fewer) anthers. Further the flowers of the allogamous species are visited by greater diversity (but similar abundance) of pollinators. Medium to large bees of the generaDiadasia, Lithurge, Melissodes, Bombus, Agapostemon andMegachile were found to be effective pollinators of the Opuntias studied. The two species were compared as to their floral attributes, breeding systems, and pollinators, and the possible role of competition for pollinators is discussed with respect to its role in their evolutionary paths.     

Taxonomical investigations onHelleborus viridis s. l. (Ranunculaceae) in Northern Italy

The taxon commonly namedHelleborus viridis in Lombardy (NW. Italy) differs from both, subsp.viridis and subsp.occidentalis, and is shown to fall within the variation range ofH. odorus subsp.laxus which has been reported so far for NE. Italy only. TrueH. viridis within Italy grows only in the Maritime Alps.     

Embryogeny in someCypereae

The paper presents a comparative account of embryo development inPycreus pumilus Nees.,Cyperus alternifolius Willd.,Mariscus paniceus Vahl. andRemirea maritima Abul., belonging to the tribeCypereae. Embryogeny follows the Juncus variation of the Onagrad type or conforms to Period I, Series A₂, Megarchetype IV. InPycreus andRemirea the precocious differentiation of dermatogen initials is followed by that of plerome and periblem elements. Mature embryos ofPycreus, Mariscus, andRemirea are identical withCyperus. Embryogeny and structure of mature embryo support the fusion of these taxa withCyperus. Embryogeny in SomeCyperaceae 1.     

Electrophoretic study and genetic affinity in theCampanula elatines andC. fragilis (Campanulaceae) rock-plants group from Italy and W. Jugoslavia

Variability and genetic divergence of 11 field populations of seven species of theCampanula elatines andC. fragilis rock-plants group from the Mediterranean and pre-Alpine areas have been studied by starch-gel electrophoretic techniques.Campanula isophylla, C. elatines, C. elatinoides, C. fragilis subsp.fragilis and subs.cavolinii, C. garganica were collected in Italy, whilstC. fenestrellata subsp.istriaca andC. portenschlagiana came from W. Jugoslavia. Twelve enzymatic loci for each population were genetically analysed: PGI-1 and 2, PGM-2, IDH-1 and -2, SKDH, ME, ADH, GOT-2, MPI-1 and -2, SOD-1. The genetic distances among the above mentioned entities have been calculated by Nei's index and depicted in a dendrogram.     

Frontiers in mycoplasma pathogenicity

Four different strains ofLactobacillus delbrueckii subsp.bulgaricus (Ss₁ and Yop₁₂) andStreptococcus salivarius subsp.thermophilus (Ss₂ and Yop₉) were isolated from two different yogurt sources in Argentina. In medium containing different carbon sources: lactose, fructose, sucrose or glucose plus fructose, the growth of a mixed culture (Yop₁₂+Ss₂) shows stimulation ofS. thermophilus and inhibition ofL. bulgaricus with respect to pure cultures. Both microorganisms in mixed culture grew less well on glucose plus galactose. However, in medium with glucose or galactose, both microorganisms were stimulated.