Selagineae and Manuleeae, two tribes or one? Phylogenetic studies in the Scrophulariaceae

 Based on results from a cladistic analysis of molecular characters using parsimony and Bayesian inference, the tribe Selagineae (Scrophulariaceae, sometimes Selaginaceae or Globulariaceae), is found to have arisen within Manuleeae (Scrophulariaceae). The inclusion of Selagineae into Manuleeae is therefore proposed. The result is corroborated by morphological characters, and indicates that the typical Selagineae condition with one ovule per ovary locule has arisen several times within the Manuleeae. Two former Manuleeae taxa are excluded from Manuleeae; Antherothamnus and Camptoloma. The former shows affinity with Scrophularia and Verbascum, and the latter groups with Buddleja and related genera. The controversy between parsimony and likelihood based methods are briefly discussed as is the possibility of historical bias in prior studies. Received February 9, 2001 Accepted June 1, 2001     

The varieties of Bauhinia pottsii G. Don in Thailand (Leguminosae-Caesalpinioideae)

 Bauhinia pottsii G. Don, B. subsessilis Craib, B. velutina (Wall. ex Benth.) Baker, B. mollissima (Wall.) Prain, and B. decipiens Craib were recently recognized as five varieties of Bauhinia pottsii G. Don. They grow naturally in southern Myanmar, Thailand, Indochina and the Malay Peninsula. Bauhinia pottsii var. decipiens (Craib) K. Larsen & S. S. Larsen is endemic to Thailand and known only from the type collection, so it was omitted from this study. Quantitative and qualitative morphological characters were examined in 200 specimens using multivariate and univariate analyses to determine the taxonomic relationship among the four. Some variation in qualitative characters was found between the varieties which separated them as previously defined in Flora of Thailand. Forty-three quantitative characters were subjected to cluster analysis to allow an objective classification into groups. The groups were subsequently evaluated by a canonical discriminant analysis. It was found that these characters collectively support the four varieties as defined by qualitative characters. However, the close relationship of varieties mollissima (Wall. ex Prain) K. Larsen & S. S. Larsen and velutina (Wall. ex Benth.) K. Larsen & S. S. Larsen is observed. The linear discriminant function has an overall rate of 98.5% correct classification and is useful for variety classification. Among quantitative characters, petal-claw length and ovarystalk length together with some qualitative characters are useful for key construction to separate the four varieties. Received April 24, 2001 Accepted December 11, 2001     

A reassessment of tribal affinities of Cratystylis and Haegiela (Asteraceae) based on three chloroplast DNA sequences

 The tribal affinities of Cratystylis and Haegiela were assessed using three chloroplast DNA sequences, the trnL/F spacer, the trnL intron and the matK coding region. The outgroup was represented by two species of the subfamily Barnadesioideae, whereas one to seven genera (45 species including Cratystylis and Haegiela) of the tribes of the Asteroideae [Anthemideae (6 genera), Astereae (7), Calenduleae (2), Gnaphalieae (7), Heliantheae s.l. (5), Inuleae s.str. (3), Plucheeae (3), Senecioneae (4)] and Cichorioideae, [Arctotideae (1), Cardueae (2), Lactuceae (2), Liabeae (1), Mutisieae (1) and Vernonieae (1)] comprise the ingroup. Phylogenetic analysis indicates that Cratystylis has strong support as a member of the tribe Plucheeae, whereas Haegiela is a member of Gnaphalieae. At some point in their taxonomic history, both genera have been placed in these tribes and there are good morphological and chemical characters that justify these placements. The monotypic Haegiela was once included in Epaltes (Plucheeae) and this study has confirmed the need for the separation of the two genera. The genus Cratystylis appears to be monophyletic. Received August 26, 2002; accepted September 19, 2002 Published online: February 7, 2003     

Cytogenetics of new Guizotia Cass. (Compositae) interspecific hybrids pertaining to genomic and phylogenetic affinities

 The following crosses were made using four recognized species/subspecies and a new population of Guizotia, referred to as Chelelu after the name of the locality in Ethiopia from which it was collected: G. scabra subsp. schimperi × Chelelu, Chelelu × G. scabra subsp. scabra, G. zavattarii × G. arborescens and Chelelu × G. zavattarii (all accessions with 2n=30). Plant morphology as well as mitotic and meiotic chromosome analysis confirmed the hybrid nature of the obtained progeny. At metaphase I of meiosis, the F₁ hybrid plants (2n=30) showed a mean of about 95%, 31%, 63% and 0.50% of the pollen mother cells with 15 bivalents, and a mean of about 14.95, 13.75, 14.40 and 7.86 bivalents per cell, respectively. The respective mean pollen stainability was about 67%, 19%, 31% and 2%. From the results it was concluded that Chelelu is more closely related to G. scabra subsp. schimperi than to G. scabra subsp. scabra but more to the latter than to G. zavattarii. Guizotia zavattarii and G. arborescens are closely related to each other. Based on the cytological observations made, the probable basic chromosome number for the genus is discussed. Received November 5, 1999 Accepted August 21, 2001     

Genetic differentiation and speciation in prealpine Cochlearia: Allohexaploid Cochlearia bavarica Vogt (Brassicaceae) compared to its diploid ancestor Cochlearia pyrenaica DC. in Germany and Austria

 Significant geographic partitioning of genetic variation within Cochlearia bavarica was found within populations from Allg?u and SE Bavaria (Germany) exhibiting significant genetic differentiation. It has been demonstrated that allohexaploid C. bavarica evolved via hybridization between diploid C. pyrenaica and tetraploid C. officinalis. Presently, only C. pyrenaica is distributed throughout inland Central Europe. It has been concluded that C. bavarica is of inter- or postglacial origin, and its speciation was not influenced by human activities. Isozyme analysis revealed that there is a correlation between interpopulational genetic distances and geographic distances among C. bavarica populations from both regions, and which is not the case for C. pyrenaica in Germany and Austria. Only high alpine C. excelsa is significantly differentiated among the diploid taxa analysed here. Geographically structured distribution of alleles and their frequencies in C. bavarica populations could not be explained with the distribution of these alleles in C. pyrenaica. The presented findings favour disruption of a former wider distribution area rather than migration of C. bavarica or a polytopic origin of this species. Received April 17, 2001 Accepted February 1, 2002     

Phylogenetic relationships of the genus Secale based on the characterisation of rDNA ITS sequences

 Sequence analysis of the internal transcribed spacer of the 18S-5.8S-26S rDNA (ITS-1) region was performed in order to analyse the phylogenetic relationships of eleven taxa of cultivated and wild rye species. The ITS regions were amplified using designed primers. At least ten positive clones of each taxonomic unit were sequenced and compared. Two different ITS sequences were found in three taxa: Secale sylvestre Host, Secale strictum ssp. kuprijanovii Grossh. and Secale strictum ssp. africanum Stapf. Secale sylvestre Host was the species that showed the greatest number of comparative differences in the sequences, and was the most distant of all the taxonomic units analysed. A certain degree of variation was found among all four subspecies of S. strictum analysed. S. strictum Presl ssp. strictum was most closely related to S. strictum ssp. africanum Stapf and S. strictum ssp. kuprijanovii Grossh to S. strictum ssp. anatolicum (Boiss.) Hammer. S. vavilovii showed similarities with this group of subspecies and with the S. cereale group. No differences were found between the weed forms of S. cereale and cultivated rye. Received March 8, 2002; accepted May 31, 2002 Published online: November 20, 2002 Address of the authors: Alfredo De Bustos, Nicolás Jouve (e-mail: nicolas.jouve@uah.es), Department of Cell Biology and Genetics, University of Alcalá, E-28871 Alcalá de Henares (Madrid), Spain.     

A phylogenetic analysis of Primulaceae s.l. based on internal transcribed spacer (ITS) DNA sequence data

 Phylogenetic relationships in Primulaceae were investigated by analysis of nuclear rDNA ITS sequences. Thirty-four species of Primulaceae, two of Myrsinaceae and four outgroup taxa were analyzed. In accordance to the results of recently published papers on the phylogeny of Primulaceae we found the family to be paraphyletic and resolved the positions of some genera. Our results show (a) the rather basal position of Centunculus within Lysimachieae, the genus thus being rather distantly related to Anagallis, (b) the close relationship between Lysimachia sect. Lerouxia, Anagallis, Asterolinon, and Pelletiera, (c) the well-supported monophyly of a group consisting of the four genera Hottonia, Omphalogramma, Bryocarpum, and Soldanella, and (d) the affinity of Stimpsonia to the Myrsinaceae-Lysimachieae-Ardisiandra clade. The ITS sequence data do not provide sufficient information to resolve basal relationships within the Primulaceae s.l. There is evidence against the monophyly of the large genera Primula, Androsace, and Lysimachia. In contrast to the phylogenetic reconstructions based on plastid gene sequences, Cyclamen does not appear as a member of the Myrsinaceae-Lysimachieae clade, but its position remains unclear. Revised July 10, 2002; accepted November 21, 2002 Published online: March 20, 2003     

Selfing facilitates reproductive isolation among three sympatric species of Pitcairnia (Bromeliaceae)

 The reproductive biology of three sympatric species Pitcairnia flammea, P. corcovadensis and P. albiflos (Bromeliaceae) was studied on Corcovado Mountain in the city of Rio de Janeiro, Brazil. Controlled pollination experiments were also conducted using plants transplanted to a greenhouse. The three species are self-compatible and interspecifically cross compatible. Pitcairnia corcovadensis was principally an autonomous self-pollinator, having scarce pollinator activity, anthers and stigmas at the same level, and absence of nectar. In P. flammea, many flowers opened per day, the stigma and anther were close to each other, and its nectar was regularly consumed by hummingbirds; all factors that promote geitonogamy in this species. Although fully self-compatible, P. albiflos always opened a few flowers per days showing an adaptation to cross-pollination. Observations during two flowering seasons showed that despite different times for peak flowering, blooming of these three species overlapped in April. They grow in mixed clumps that may facilitate promiscuous pollination between them but no intermediate forms were observed under natural conditions. The absence of temporal isolation, geographic isolation and isolation via post-pollination reproduction suggests that evolution toward selfing was important to avoid hybridization between these sympatric species. Received May 10, 2001 Accepted February 25, 2002     

The phylogeny of Lens (Leguminosae): new insight from ITS sequence analysis

 Lens includes L. culinaris subsp. culinaris (the cultivated lentil) and several wild species distributed from the Mediterranean region to western Asia. We compared sequence variation in the ITS region among species of Lens in an effort to end persisting uncertainty regarding the phylogeny of the genus. The parsimony analysis revealed a single minimum-length tree with a topology congruent with patterns derived by previous studies of nuclear and chloroplast DNA RFLPs. The basal and highly divergent status of the L. nigricans clade is depicted, and the progenitor-derivative relationship between L. culinaris subsp. orientalis and L. culinaris subsp. culinaris is reaffirmed. Resolution in the tree was improved by combining the ITS data set with a pre-existing set of chloroplast DNA restriction site data obtained from the same group of samples. Received May 8, 2000 Accepted October 26, 2001     

Isozyme evidence of the parental origin and possible fertility of the hybrid Potamogeton×fluitans Roth

 The hybrid Potamogeton×fluitans Roth is thought to have resulted from hybridization between P. lucens L. and P. natans L. This hybrid has only been recorded at a few locations in the British Isles. At most of these sites the number of individuals found have been quite low. The one exception to this being a population in the Moors River, Dorset and South Hampshire, England, where a substantially larger population exists. Using seven different enzyme systems, this study provided support for the putative parental origin of this hybrid. In addition, the population in the Moors River was shown to contain numerous multi-locus phenotypes suggesting that these individuals may be partially fertile and may backcross to one of its parental species, or alternatively undergo selfing or crossing to produce an F₂ generation. Received December 1, 2000 Accepted June 20, 2001     

Nectar biodiversity: a short review

 Nectaries differ in many aspects but a common feature is some kind of advantage for the plant conferred by foraging of consumers which may defend the plant from predators in the case of extrafloral nectaries, or be agents of pollination in the case of floral nectaries. This minireview is concerned mainly with floral nectaries and examines the following characteristics: position in flower; nectary structure; origin of carbohydrates, aminoacids and proteins; manner of exposure of nectar; site of nectar presentation; volume and production of nectar in time; sexual expression of flower and nectary morphology; nectar composition and floral sexual expression; variability of nectar composition; fate of nectar; energy cost of nectar production. The species of certain large families, such as Brassicaceae, Lamiaceae and Asteraceae, resemble each other in nectary organisation; other families, such as Cucurbitaceae and Ranunculaceae, have various types of organisation. A scheme is presented to illustrate factors influencing nectary and nectar biodiversity. Received July 23, 2002; accepted September 18, 2002 Published online: June 2, 2003     

Morphology and ecology of two new species of Polypleurum (Podostemaceae) from Kerala, India

 Two new species of the Asiatic genus Polypleurum (Podostemaceae – Podostemoideae), Polypleurum prostratum and Polypleurum disciforme from Kerala, India are described. They differ in the shape of the thalloid plant body from the other species of Polypleurum. A key to the species occurring in India is provided. Though sharing the habitat with some other species of the Podostemaceae, the new species inhabit special niches in the rocky riverbed. – The chemical components of the (hard) water in which they grow differ considerably from what is known from black water rivers of the New World. The results thus widen considerably our knowledge of the ecological needs of Podostemaceae. Received December 10, 2002; accepted January 10, 2003 Published online: March 31, 2003     

Floral organogenesis of Chloranthus sessilifolius, with special emphasis on the morphological nature of the androecium of Chloranthus (Chloranthaceae)

 Floral organogenesis of Chloranthus sessilifolius K. F. Wu is described. The inflorescence primordium is dome-like in the beginning and then elongates, and bract primordia initiate almost decussately. Each floral primordium, arising from the axil of a bract, soon becomes a scale-like structure, with three primordia of androecial lobes originating from its abaxial part, and the gynoecial primordium in adaxial position. As the androecial lobes become more distinct, four thecae are already in differentiation, and the gynoecial primordium appears as a shallow disc. The androecial lobes do not extend their length until the thecae approach maturity and the stigma is differentiated. The androecial lobes are united at all the stages of development, and the entire androecium falls off as a unit at the end of anthesis. Based on these results, combined with published evidence from neobotany, palaeobotany and phylogenetic studies, the morphological nature of the androecium of Chloranthus is further discussed. Our studies support the viewpoint that the androecial structure of Chloranthus may have arisen by splitting of a single stamen with 2 marginal thecae. Received May 2, 2001 Accepted December 18, 2001     

The correlation between development of atypical bisexual flowers and phylogeny in the Aroideae (Araceae)

 In the intermediate zone of the inflorescence of genera of Aroideae one can find flowers with male and female characteristics. Until now, two types of developmental sequences of atypical bisexual flowers (ABFs) have been recognized: the Philodendron type and the Cercestis type. In the Philodendron type, bisexual flowers generally consist of functional carpels and staminodes inserted on the same whorl. In the Cercestis type, the gynoecium and stamens are inserted on two different whorls. These different ontogenetic patterns represent two different pathways in the evolution of unisexual flowers in this subfamily. A molecular phylogenetic analysis of 33 genera of Araceae, based on the chloroplast trnL intron and trnL–F intergenic spacer sequences was carried out. We use this phylogenetic analysis and those published by French et al. (1995) and Mayo et al. (1997) to examine the distribution of the two types of ABFs in selected genera. Our results suggest that the two developmental patterns of ABFs in Aroideae sensu Mayo et al. (1997) do not correspond to two separate evolutionary lineages but rather are more or less consistent within clades. Although this new molecular phylogeny does not include all aroid genera, it corroborates in general, at the subfamily level, the molecular analysis of French et al. (1995) based on chloroplast DNA restriction site data and the analysis of Mayo et al. (1997) based on morphological and anatomical data. Received March 15, 2001 Accepted October 11, 2001     

The nectary as the primary site of infection by Erwinia amylovora (Burr.) Winslow et al.: a mini review

 Erwinia amylovora is the causative agent of fire blight, a bacterial disease existing as an unsolved problem in most countries where pome fruits like apple (Malus domestica) and pear (Pyrus communis) or ornamental plants of Rosaceae are grown. The primary site of colonization is the open flower. As for the establishment of the disease, the importance of various organs within the flowers is considerably different. The usual place for developing a large epiphytic population is the stigma. The actual infection will be attained by the external washing (rain, heavy dew) of bacteria from the stigma to the hypanthium. The bacteria penetrate through the openings of the nectary, so, the nectarthodes are the main entrance sites for them. Nectar is an excellent medium for growth of fire blight bacteria. Most often, however, the incidence of disease is significantly less than the percentage of colonized flowers. Little is known about the interrelationships of free moisture, nectar sugar concentration, ovary water potential, fine-structural characteristics of nectary versus the disease incidence and severity. The aim of this paper is to review the ecology and infection biology of Erwinia amylovora on floral surfaces and in floral tissues. Received July 8, 2002; accepted October 17, 2002 Published online: June 2, 2003     

Phylogenetic relationships of Mangifera species revealed by ITS sequences of nuclear ribosomal DNA and a possibility of their hybrid origin

 The phylogenetic relationships among 14 Mangifera L. species of Thailand were analyzed by comparing sequences of the internal transcribed spacer (ITS) region of nuclear ribosomal DNA (nrDNA). Parsimony and neighbor joining (NJ) analyses revealed that the common mango (M. indica L.) was closely related to M. laurina Bl., M. sylvatica Roxb., and M. oblongifolia Hook. f. Mangifera foetida Lour. and M. odorata Griff. were also related to M. indica in both parsimonious and NJ trees, although these two species are classified into a different subgenus (subgenus Limus) from the subgenus Mangifera to which M. indica belongs. ITS sequence analysis revealed that several species have nucleotide additivity (two different nucleotides at the same locus) at several sites in the ITS region. Also, M. indica had several polymorphisms among cultivars. This finding may suggest a possibility of hybrid origin of Mangifera species, although Mangifera species are all assumed to be diploid having chromosome number of 2n=2x=40. Received February 7, 2001 Accepted October 28, 2001     

Floral nectaries and nectar production in brown mustard (Brassica juncea) and white mustard (Sinapis alba) (Brassicaceae)

 The dynamics and abundance of nectar secretion as well as sugar productivity were studied in flowers of brown mustard (Brassica juncea) cv. Małopolska and white mustard (Sinapis alba) cv. Borowska. Moreover, floral nectaries were examined under LM and SEM. In both cultivars lateral and median pairs of nectaries secreted nectar. However, differences were found in morphology and activity of these pairs. The lateral nectaries produced more nectar than the median ones. Nectar secretion started at loose bud and peaked during anther dehiscence. Average amount of nectarsecreted by 100 flowers of cv. Małopolska and cv. Borowska were 119.9 mg and 134 mg. Mean concentration of nectar was 26.7% and 23.4%, respectively. One hundred flowers of cv. Małopolska and cv. Borowska secreted 28.4 mg and 24.9 mg of sugars in nectar. Estimated sugar productivity per 1 ha of crop was 65.5 kg and 71.2 kg, respectively. Received August 28, 2002; accepted November 2, 2002 Published online: June 2, 2003     

Pollination and breeding system of Eritrichium nanum (Boraginaceae)

 Flower development, pollination and breeding system of the high alpine cushion plant, Eritrichium nanum (Boraginaceae), were investigated in nine populations from the European Alps at altitudes of 2700 m–3200 m. Peak flowering period lasts longer than a month, from mid-June to the end of July. In contrast to statements in the literature that flowers are protogynous and nutlets remain in their calyx until spring we found a distinct protandry and nutlets being dispersed before mid-September. Various insects from 12 families, but mostly Diptera, frequently visited E. nanum flowers, with flies from the families Anthomyiidae and Muscidae being the predominant visitors. Under optimal conditions (max. solar radiation, min. wind force), visitation rates of 200 simultaneously observed flowers reached 32.5–46.7 insects per hour, i.e. 0.16–0.24 insects per flower per hour. However, the commonly observed Anthomyiidae and Muscidae clearly preferred the white-yellowish flowers of Saxifraga exarata and Saxifraga bryoides which are abundant at E. nanum sites and which are certainly also pollinated by species of these two fly families. The flowers of these Saxifraga species offer plenty of nectar and may compete for pollinators with E. nanum, when they are flowering in its proximity. However, various other insects like Pontia callidice and Psodos sp. (Lepidoptera) as well as Andrena sp. (Hymenoptera) and especially Eristalis tenax and closely related hoverflies showed a higher degree of flower constancy to E. nanum, often flying from a blue Eritrichium cushion to the next and hence causing outcrossing. The five fornices of E. nanum flowers which obstruct the tube containing stamens and nectar, are a feature which differs distinctly from the syndrome of fly-pollinated flowers with easily accessible nectar. Consequently pollination by flies in E. nanum seems to be caused mainly by the unfavorable ecological conditions at high altitudes, where flies are the most frequent insects. Bagging experiments showed that outcrossing and geitonogamy are the prevailing pollination modes, and autogamy, although possible, plays only a minor role. Received February 13, 2001 Accepted November 23, 2001