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[Molecular identification of Araceae-infecting viruses and their ecology].

A filamentous virus and a spherical virus were detected from partial purification with natural-infected Araceae plants. The filamentous virus was confirmed as dasheen mosaic virus (DsMV) by RNA spot hybridization (RSH) and sequence determination, while the spherical virus was identified as a new strain of cucumber mosaic virus (CMV) by enzyme-linked immunosorbent assay and sequence determination of the 3' end genes of RNA3. CMV was firstly confirmed as one of the principal viruses infecting the Araceae plants under natural condition. According to the sequence similarity of coat protein, Araceae-infecting CMV was then decided belonging to CMV subgroup I, but independent from other isolates in this subgroup. By dot-RNA hybridization to DsMV and CMV probes with total RNA extracted from leaf tissue, field samples collected from some parts of southern China were detected for their natural infection. The infection ratio of field samples infected by DsMV and CMV were 73.3% and 46.7% for samples collected from Hainan Province, 100% and 38.5% for those from Hunan Province, 93.0% and 38.5% for that from Zhejiang Province, and 100% and no infection for samples from Shanghai, respectively. The infection of viruses on Araceae plants was obviously affected by such ecological factors as geological distribution, climate and host species. The occurrence of CMV was greatly influenced by the above factors, but DsMV always existed as the most commonly distributed virus.

Amino Acid Sequence↗

Role and location of NAD malic enzyme in thermogenic tissues of Araceae.

This work was done to discover how those nonphotosynthetic tissues of the Araceae that become thermogenic release, as CO2, carbon recently fixed by phosphoenolpyruvate carboxylase. Extracts of clubs of the spadix of Arum maculatum showed no activity for phosphoenolpyruvate carboxykinase and low activities of NADP malic enzyme. NAD malic enzyme activity in the above extracts and in those of thermogenic tissues of other Araceae was appreciable. Analysis of homogenates of clubs of Typhonium giraldii by differential centrifugation and sucrose gradients showed that NAD malic enzyme was confined to mitochondria. Centrifugation of mitochondria after freezing and thawing left all the NAD malic enzyme in the supernatant. NAD malic enzyme in isolated, intact mitochondria was completely latent, and was completely protected from exogenous trypsin. The responses of this latency and protection to different concentrations of Triton X-100 suggested that none of the NAD malic enzyme was accessible from either the outside or the intermembrane space of the mitochondria. Treatment of excised clubs of A. maculatum with 2-N-butylmalonate largely prevented the development of the rapid respiration responsible for thermogenesis, and severely inhibited dark fixation of 14CO2. The conclusion is that in mature clubs of the Araceae phosphoenolpyruvate is converted to malate in the cytosol by phosphoenolpyruvate carboxylase and NAD malate dehydrogenase, and that this malate then enters the mitochondrial matrix where it is converted to pyruvate by NAD malic enzyme.

Carbon Dioxide↗

Intercontinental biogeography of subfamily Orontioideae (Symplocarpus, Lysichiton, and Orontium) of Araceae in Eastern Asia and North America.

Symplocarpus, Lysichiton, and Orontium (Orontioideae) are three of the few north temperate genera of the primarily tropical Araceae. Symplocarpus is disjunctly distributed in eastern Asia (3 spp.) and eastern North America (1 sp.); Lysichiton has an intercontinental discontinuous distribution in eastern Asia (1 sp.) and northwestern North America (1 sp.); and the monotypic Orontium is restricted to eastern North America. Phylogenetic analysis of the trnL-F and ndhF sequences supports (1) the monophyly of both Symplocarpus and Lysichiton, (2) the sister-group relationship of Symplocarpus and Lysichiton, and (3) the clade of Orontium, Symplocarpus, and Lysichiton. Although Symplocarpus shows a much wider disjunction than Lysichiton, the estimated divergence time of the former [4.49+/-1.69 or 6.88+/-4.18 million years ago (mya)] was similar to that of the latter (4.02+/-1.60 or 7.18+/-4.33 mya) based on the penalized likelihood and the Bayesian dating methods, respectively. Eastern Asia was suggested to be the ancestral area of the Symplocarpus-Lysichiton clade based on the dispersal-vicariance analysis. Our biogeographic results support independent migrations of Symplocarpus and Lysichiton across the Bering land bridge in the late Tertiary (Pliocene/late Miocene). Fossil evidence suggests Orontioideae dated back to the late Cretaceous in the temperate Northern Hemisphere (72 mya). The relative rate test shows similar substitution rates of the trnL-F sequences between the proto and the true aroids, although the latter has substantially higher species diversity. The proto Araceae perhaps suffered from a higher rate of extinction in the temperate zone associated with periods of climatic cooling in the Tertiary.

Animals↗

Araceae from the Early Cretaceous of Portugal: evidence on the emergence of monocotyledons.

A new species (Mayoa portugallica genus novum species novum) of highly characteristic inaperturate, striate fossil pollen is described from the Early Cretaceous (Barremian-Aptian) of Torres Vedras in the Western Portuguese Basin. Based on comparison with extant taxa, Mayoa is assigned to the tribe Spathiphylleae (subfamily Monsteroideae) of the extant monocotyledonous family Araceae. Recognition of Araceae in the Early Cretaceous is consistent with the position of this family and other Alismatales as the sister group to all other monocots except Acorus. The early occurrence is also consistent with the position of Spathiphylleae with respect to the bulk of aroid diversity. Mayoa occurs in the earliest fossil floras (from circa 110 to 120 million years ago) that contain angiosperm flowers, carpels, and stamens. The new fossil provides unequivocal evidence of monocots in early angiosperm assemblages that also include a variety of key "magnoliid" lineages (e.g., Chloranthaceae) but only a limited diversity of eudicots.

Araceae↗

Multiple acquisitions via horizontal transfer of a group I intron in the mitochondrial cox1 gene during evolution of the Araceae family.

A group I intron has recently been shown to have invaded mitochondrial cox1 genes by horizontal transfer many times during the broad course of angiosperm evolution. To investigate the frequency of acquisition of this intron within a more closely related group of plants, we determined its distribution and inferred its evolutionary history among 14 genera of the monocot family Araceae. Southern blot hybridizations showed that 6 of the 14 genera contain this intron in their cox1 genes. Nucleotide sequencing showed that these six introns are highly similar in sequence (97.7%-99.4% identity) and identical in length (966 nt). Phylogenetic evidence from parsimony reconstructions of intron distribution and phylogenetic analyses of intron sequences is consistent with a largely vertical history of intron transmission in the family; the simplest scenarios posit but one intron gain and two losses. Despite this, however, striking differences in lengths of exonic co-conversion tracts, coupled with the absence of co-conversion in intron-lacking taxa, indicate that the six intron-containing Araceae probably acquired their introns by at least three and quite possibly five separate horizontal transfers. The highly similar nature of these independently acquired introns implies a closely related set of donor organisms.

Base Sequence↗

The major tuber storage protein of araceae species is a lectin. Characterization and molecular cloning of the lectin from Arum maculatum L.

A new lectin was purified from tubers of Arum maculatum L. by affinity chromatography on immobilized asialofetuin. Although this lectin is also retained on mannose-Sepharose 4B, under the appropriate conditions free mannose is a poor inhibitor of its agglutination activity. Pure preparations of the Arum lectin apparently yielded a single polypeptide band of approximately 12 kD upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis. However, N-terminal sequencing of the purified protein combined with molecular cloning of the lectin have shown that the lectin is composed of two different 12-kD lectin subunits that are synthesized on a single large precursor translated from an mRNA of approximately 1400 nucleotides. Lectins with similar properties were also isolated from the Araceae species Colocasia esculenta (L.) Schott, Xanthosoma sagittifolium (L.) Schott, and Dieffenbachia sequina Schott. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration of the different Araceae lectins have shown that they are tetrameric proteins composed of lectin subunits of 12 to 14 kD. Interestingly, these lectins are the most prominent proteins in the tuber tissue. Evidence is presented that a previously described major storage protein of Colocasia tubers corresponds to the lectin.

Amino Acid Sequence↗

Programmed cell death and leaf morphogenesis in Monstera obliqua (Araceae).

The unusual perforations in the leaf blades of Monstera obliqua (Araceae) arise through programmed cell death early in leaf development. At each perforation site, a discrete subpopulation of cells undergoes programmed cell death simultaneously, while neighboring protoderm and ground meristem cells are unaffected. Nuclei of cells within the perforation site become terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL)-positive, indicating that DNA cleavage is an early event. Gel electrophoresis indicates that DNA cleavage is random and does not result in bands that represent multiples of internucleosomal units. Ultrastructural analysis of cells at the same stage reveals misshapen, densely stained nuclei with condensed chromatin, disrupted vacuoles, and condensed cytoplasm. Cell walls within the perforation site remain intact, although a small disk of dying tissue becomes detached from neighboring healthy tissues as the leaf expands and stretches the minute perforation. Exposed ground meristem cells at the rim of the perforation differentiate as epidermal cells. The cell biology of perforation formation in Monstera resembles that in the aquatic plant Aponogeton madagascariensis (Aponogetonaceae; Gunawardena et al. 2004), but the absence of cell wall degradation and the simultaneous execution of programmed cell death throughout the perforation site reflect the convergent evolution of this distinct mode of leaf morphogenesis in these distantly related plants.

Apoptosis↗

Conventional and novel modes of exine patterning in members of the Araceae--the consequence of ecological paradigm shifts?

In the family Araceae, the members of all subfamilies except Aroideae follow the conventional mode of exine formation pattern, which conforms with the textbook view of sporoderm stratification and chemistry (sporopollenin ektexine formed before the endexine). Only members of the subfamily Aroideae show a quite uncommon mode of exine formation pattern, with an endexine formed prior to the nonsporopollenin, polysaccharidic outer exine layer. The intine is formed simultaneously with this non-sporopollenin layer. From the differing timetable and especially from the different origin it is concluded that this outer exine layer is not homologous to the angiosperm ektexine. The fundamental question, why members of the Aroideae lack an elaborated sporopollenin ektexine, is discussed in terms of functionality of the nonsporopollenin outer exine layer. It seems that a major change in aroid evolution took place at the point when the family phylogenetically and ecologically shifted from bisexual (most subfamilies) to unisexual flowers (Aroideae only). The hypothesis is that ephemeral spathes and the absence of sporopollenin are the consequence of an adaptive syndrome for a short pollination time window in many members of the Aroideae, with short-lived pollen, an energetically not costly pollen wall, rapid germination of pollen tube, and brief receptivity of stigma.

Araceae↗

Caladium bicolor (Araceae) and Cyclocephala celata (Coleoptera, Dynastinae): a well-established pollination system in the Northern Atlantic rainforest of Pernambuco, Brazil.

Flowering, pollination ecology, and floral thermogenesis of Caladium bicolor were studied in the Atlantic Rainforest of Pernambuco, NE Brazil. Inflorescences of this species are adapted to the characteristic pollination syndrome performed by Cyclocephalini beetles. They bear nutritious rewards inside well-developed floral chambers and exhibit a thermogenic cycle which is synchronized to the activity period of visiting beetles. Heating intervals of the spadix were observed during consecutive evenings corresponding to the beginning of the female and male phases of anthesis. Highest temperatures were recorded during the longer-lasting female phase. An intense sweet odour was volatized on both evenings. Beetles of a single species, Cyclocephala celata, were attracted to odoriferous inflorescences of C. bicolor and are reported for the first time as Araceae visitors. All the inflorescences visited by C. celata developed into infructescences, whereas unvisited inflorescences showed no fruit development. Findings of previous studies in the Amazon basin of Surinam indicated that Cyclocephala rustica is a likely pollinator of C. bicolor. This leads to the assumption that locally abundant Cyclocephalini species are involved in the pollination of this species.

Animals↗

Biogeography of the Pistia clade (Araceae): based on chloroplast and mitochondrial DNA sequences and Bayesian divergence time inference.

Pistia stratiotes (water lettuce) and Lemna (duckweeds) are the only free-floating aquatic Araceae. The geographic origin and phylogenetic placement of these unrelated aroids present long-standing problems because of their highly modified reproductive structures and wide geographical distributions. We sampled chloroplast (trnL-trnF and rpl20-rps12 spacers, trnL intron) and mitochondrial sequences (nad1 b/c intron) for all genera implicated as close relatives of Pistia by morphological, restriction site, and sequencing data, and present a hypothesis about its geographic origin based on the consensus of trees obtained from the combined data, using Bayesian, maximum likelihood, parsimony, and distance analyses. Of the 14 genera closest to Pistia, only Alocasia, Arisaema, and Typhonium are species-rich, and the latter two were studied previously, facilitating the choice of representatives that span the roots of these genera. Results indicate that Pistia and the Seychelles endemic Protarum sechellarum are the basalmost branches in a grade comprising the tribes Colocasieae (Ariopsis, Steudnera, Remusatia, Alocasia, Colocasia), Arisaemateae (Arisaema, Pinellia), and Areae (Arum, Biarum, Dracunculus, Eminium, Helicodiceros, Theriophonum, Typhonium). Unexpectedly, all Areae genera are embedded in Typhonium, which throws new light on the geographic history of Areae. A Bayesian analysis of divergence times that explores the effects of multiple fossil and geological calibration points indicates that the Pistia lineage is 90 to 76 million years (my) old. The oldest fossils of the Pistia clade, though not Pistia itself, are 45-my-old leaves from Germany; the closest outgroup, Peltandreae (comprising a few species in Florida, the Mediterranean, and Madagascar), is known from 60-my-old leaves from Europe, Kazakhstan, North Dakota, and Tennessee. Based on the geographic ranges of close relatives, Pistia likely originated in the Tethys region, with Protarum then surviving on the Seychelles, which became isolated from Madagascar and India in the Late Cretaceous (85 my ago). Pistia and Protarum provide striking examples of ancient lineages that appear to have survived in unique or isolated habitats.

Araceae↗

Molecular characterization of five medicinally important species of Typhonium (Araceae) through random amplified polymorphic DNA (RAPD).

The interrelationship of five medicinally important species of Typhonium (Araceae) including T. venosum, which was previously placed under the genus Sauromatum, was inferred by analysis of random amplified polymorphic DNA (RAPD). DNA from pooled leaf samples was isolated and RAPD analysis was performed using 20 decamer oligonucleotide primers. Out of a total of 245 bands amplified, 12 were found to be monomorphic while 233 bands were polymorphic including 86 species-specific bands. The genetic similarities were analyzed from the dendrogram constructed by the pooled RAPD data using a similarity index. The dendrogram showed two distinct clades, one containing T. roxburghii, T. trilobatum and T. venosum and the other containing the remainder two species, i.e. T. diversifolium and T. flagelliforme. Both the clusters shared a common node approx. at 23.7% level of similarity. The maximum similarity of 31.2% was observed between T. venosum and T. trilobatum. In view of its close genetic similarity with other members of Typhonium, transfer of Sauromatum venosum to the genus Typhonium and merger of the two genera was supported.

Araceae↗

Cytophotometric estimation of in situ DNA content in several species of Araceae.

A wide variation in the in situ 4C DNA content, ranging from 15.02 pg to 54.09 pg was found in thirteen genera of the family Araceae. The obligate perennial species showed greater 4C DNA values compared with the facultative perennials and annuals. A remarkable heterogeneity in 4C nuclear DNA amounts was noted among obligate perennials. Intraspecific constancy in the amount of 4C DNA was recorded. The nuclear DNA content correlated positively with chromosome size, duration of the mitotic cycle, and annual to perennial growth forms. Despite an extensive variation in DNA content among the aroids investigated, each species was distinctly characterized by its specific nuclear DNA value, indicating its usefulness in taxonomic characterization and comparison of different aroids.

Araceae↗

Fatty acid composition of Dracunculus vulgaris Schott (Araceae) seed oil from Turkey.

Dracunculus vulgaris Schott is only one taxon of the genus Dracunculus (Araceae) in Turkey. The tubers and the fruits with the seeds of D. vulgaris have long been in use for the treatment of rheumatism and hemorrhoids, respectively. The fatty acid composition of D. vulgaris seeds have been analyzed as their methyl esters by GC and GC-mass spectrometry. C16:0, C16:1n-7, C18:1n-9, C18:1n-7 (cis -vaccenic acid), C18:2n-6 and 13-phenyl tridecanoic acids were found to be the main components in the seed oil.

Araceae↗

Cytotoxic activity of Typhonium flagelliforme (Araceae).

The plant Typhonium flagelliforme (Araceae), commonly known as the 'rodent tuber', is often included as an essential ingredient in various herbal remedies recommended for cancer therapies in Malaysia. Various extracts prepared from either the roots, tubers, stems or leaves were tested for cytotoxic activity on murine P388 leukaemia cells using the MTT assay method. Both the chloroform (IC50 = 6.0 microg/mL) and hexane (IC50 = 15.0 microg/mL) extract from the 'roots and tubers' exhibited weak cytotoxic activity. The hexane extract (IC50 = 65.0 microg/mL) from the 'stems and leaves' exhibited weaker cytotoxic activity than the chloroform extract (IC50 = 8.0 microg/mL). Although the juice extract from the 'roots and tubers' is frequently consumed for cancer treatment, it exhibited poor cytotoxic activity. Further analysis using an amino acid analyser revealed that the juice extract contained a high concentration of arginine (0.874%). A high tryptophan content (0.800%) was confirmed by NMR and HPLC analysis.

Amino Acids↗

Molecular phylogenetic relationships among Lemnaceae and Araceae using the chloroplast trnL-trnF intergenic spacer.

We test competing hypotheses of relationships among Aroids (Araceae) and duckweeds (Lemnaceae) using sequences of the trnL-trnF spacer region of the chloroplast genome. Included in the analysis were 22 aroid genera including Pistia and five genera of Lemnaceae including the recently segregated genus Landoltia. Aponogeton was used as an outgroup to root the tree. A data set of 522 aligned nucleotides yielded maximum parsimony and maximum likelihood trees similar to those previously derived from restriction site data. Pistia and the Lemnaceae are placed in two separate and well-supported clades, suggesting at least two independent origins of the floating aquatic growth form within the aroid clade. Within the Lemnaceae there is only partial support for the paradigm of sequential morphological reduction, given that Wolffia is sister to Wolffiella+Lemna. As in the results of the restriction site analysis, pantropical Pistia is placed with Colocasia and Typhonium of southeastern Asia, indicative of Old World affinities. Branch lengths leading to duckweed terminal taxa are much longer relative to other ingroup taxa (including Pistia), evidently as a result of higher rates of nucleotide substitutions and insertion/deletion events. Morphological reduction within the duckweeds roughly correlates with accelerated chloroplast genome evolution.

Chloroplasts↗

New lymphocyte stimulating monocot lectins from family Araceae.

Three monocot lectins from underground tubers of plants belonging to the family Araceae were investigated for their mitogenic potential towards human peripheral blood lymphocytes. All the three lectins turned out to be potent mitogens in the [3H]-thymidine uptake assay. Gonatanthus pumilus lectin was mitogenic at an optimum concentration of 25 micrograms/ml while Alocasia indica and Sauromatum guttatum lectins were most effective at a concentration of 50 micrograms/ml. [3H]-thymidine incorporation studies further revealed that the lectins were T-cell mitogens and did not induce any appreciable DNA synthesis in B-enriched lymphocytes. The proliferation kinetic studies detected maximum incorporation on day 3 and the mitogenic response was shown to be inhibited by asialofetuin in a concentration-dependent manner.

Adult↗

New lymphocyte stimulating monocot lectins from family Araceae. II.

Two lectins purified from the tubers of Arisaema consanguineum Schott (ACA) and A. curvatum Kunth (ACmA) belonging to the monocot family Araceae were mitogenic for human peripheral blood mononuclear cells (PBMC) in the [3H]-thymidine uptake assay. ACA and ACmA had an optimum stimulatory concentration of 10-25 micrograms/ml and 50-100 micrograms/ml, respectively, as observed in PBMC from five different individuals. The mitogenic response of PBMC was inhibitable in a dose-dependent manner by asialofetuin. The lectins were T-cell specific, and stimulation kinetic studies using ACA and ACmA showed that they induce maximum thymidine uptake in PBMC at day 4 and 3, respectively.

Humans↗

Size-dependent flowering behavior and heat production of a sequential hermaphrodite, Symplocarpus renifolius (Araceae).

We examined the flowering performance in a population of the protogynous perennial herb Symplocarpus renifolius (Araceae), with special consideration of plant size. Flowering of S. renifolius occurred in very early spring, soon after snow melt. The spadices generated heat throughout the pistillate (female) and bisexual phases, but heat production decreased quickly after the beginning of the staminate (male) phase. During the flowering season, the sex ratio within the population dramatically changed from a dominance of females to a dominance of males. The duration of the female phase was negatively correlated with the onset time of flowering, and the duration of the male phase was positively correlated with plant size. Larger plants began blooming earlier, produced more heat, made the transition from female to male phase more rapidly, and lasted longer as males than smaller ones. Such size-dependent flowering performance caused unidirectional pollen flow from large to small plants. The number of seeds produced per spadix was positively correlated with the duration of the female phase, although it was not correlated with plant size. However, the estimated number of seeds sired during the male phase was positively correlated with plant size. Early flowering, rapid gender change, and higher heat production of the spadices by larger plants were factors considered to promote the higher success of the male function without decreasing the success of the female function.

Journal Article↗