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Evolutionary dynamics of the chloroplast genome in Abutilon (Malvoideae, Malvaceae).

The genus Abutilon Mill. (Malvaceae) comprises approximately 178 species distributed across tropical and subtropical regions, many of which hold significant ornamental, economic, and medicinal value; yet its taxonomic classification remains challenging. In this study, six species were sequenced from herbarium specimens, and the chloroplast (cp.) genomes of ten additional species were assembled de novo from publicly available raw data. Three previously reported cp. genomes were also incorporated to characterise cp. genome structure, identify polymorphic loci, and perform phylogenetic analyses. The cp. genomes ranged from 159,458 to 160,454 bp and exhibited the typical quadripartite structure, with each genome containing 112 unique genes (78 protein-coding, 30 tRNA, and 4 rRNA) that showed conserved content and organisation. These genomes exhibited high similarity in GC content, inverted repeat boundaries, relative synonymous codon usage, amino acid frequencies, and substitution patterns. However, notable variation was observed in the total number of simple sequence repeats, ranging from 70 to 97 per genome. Selection analyses indicated predominant purifying selection, with evidence of episodic positive selection detected in rpoC2, rbcL, and ycf1. Two codons in rbcL were clade-specific and provided phylogenetic signal distinguishing Australian and Old World pantropical species. Nucleotide diversity analysis identified six highly polymorphic intergenic spacers (trnH-psbA, rps19-rpl2, psbT-pbf1, psaC-ndhD, trnR-atpA, and ndhJ-ndhK) that may be suitable for taxonomic studies. The phylogeny from maximum likelihood (ML) and Bayesian inference (BI) resolved two major clades: one comprising an exclusively Australian lineage occurring predominantly in arid and semi-arid environments, and the other a pantropical lineage spanning multiple continents. Abutilon grandifolium was recovered as sister to the remaining sampled Abutilon taxa in both ML and BI analyses, although no biogeographic origin inference can be drawn from this placement pending broader taxon sampling and integration of nuclear genomic data. These findings provide insights into the evolutionary dynamics of the cp. genome in Abutilon and offer a foundational genomic framework for refining Abutilon taxonomy.

Genome, Chloroplast

Localization of abutilon mosaic virus (AbMV) DNA within leaf tissue by in situ hybridization.

Abutilon mosaic virus (AbMV) is a whitefly-transmitted geminivirus with a a bipartite genome. Using in situ hybridization AbMV DNA was detected exclusively in the phloem of infected Abutilon sellovianum leaves and intracellularly predominant in the nuclei. No AbMV DNA was found in cells from palisade and spongy parenchyma, the tissues which show the predominant cytopathological effects. A hypothesis is discussed to account for the finding that, while AbMV accumulates in the phloem, symptoms are observed in other tissues. Shoot tips were analyzed by in situ hybridization to determine the earliest stage of leaf development in which AbMV is detectable. We found the first hybridization signals in the sixth to seventh leaf of the shoot tip, whereas we could not detect any viral DNA in younger leaves and meristems. These results are discussed with reference to the relation of AbMV multiplication to the cell cycle.

DNA, Viral

The nucleotide sequence of abutilon mosaic virus reveals prokaryotic as well as eukaryotic features.

The complete nucleotide sequence of abutilon mosaic virus (West Indian isolate, AbMVa) is presented. The resulting genomic structure resembles that of other geminiviruses which are transmitted by the whitefly Bemisia tabaci: AbMV possesses a bipartite circular genome with bidirectional orientation of the open reading frames (ORF). Both components have a common region of 180 bases with 99% homology while the rest of their sequence is distinct. Eukaryotic regulatory transcription elements precede most ORFs and polyadenylation signals are present at the end of most ORFs. However, two ORFs show features of prokaryotic genes. This chimaeric genome organisation is discussed with reference to the finding that AbMV DNA is present in plastids as well as in the nucleus of infected cells.

Amino Acid Sequence

Transcript mapping of Abutilon mosaic virus, a geminivirus.

The transcripts of the DNA of Abutilon mosaic virus, a geminivirus with a bipartite genome (DNA A and DNA B), were characterized by Northern blot hybridization, S1 nuclease assay, primer extension analysis, and sequencing of the 3' termini of cDNA clones. It was shown that transcription is bidirectional and that the transcripts are polyadenylated. Two overlapping transcripts of 1.6 and 0.7 kb were mapped to the complementary strand of DNA A and two of 1.3 and 1.2 kb to the complementary strand of DNA B. One transcript of 0.9 kb was mapped to the viral sense in DNA A and one of 1.0 kb to the viral sense in DNA B. The ends of complementary and viral mRNA overlapped in both genome parts in regions rich in polyadenylation signals.

Amino Acid Sequence

Abutilon mosaic geminivirus double-stranded DNA is packed into minichromosomes.

An intermediate form of Abutilon mosaic geminivirus, a complex of viral double-stranded DNA (dsDNA) and protein, was characterized by two different techniques. Cesium sulphate gradient centrifugation was used to show that the majority of this form comigrates with host chromatin. Micrococcus nuclease digestion experiments with isolated nuclei demonstrated that the viral dsDNA is organized in a manner comparable to that of host nucleosomes. Results from a previous electron microscopical work (Abouzid, A. M., Frischmuth, T., and H. Jeske, 1988, Mol. Gen. Genet. 212, 252-258) suggested to us that a part of the viral dsDNA must be free of nucleosomes. Whether this nucleosome-free space corresponds to the intergenic region which contains most of the promotor sequences and the putative origin of replication is discussed.

Centrifugation, Density Gradient

Determination of estrogenic/antiestrogenic potential of antifertility substances using rat uterine peroxidase assay.

The effect of three compounds (clomiphene citrate, centchroman, embelin) and plant-derived methanolic extracts (Abutilon indicum and Butea monosperma) was studied on uterotropic and uterine peroxidase activities in ovariectomized rats. It was observed that these two parameters were highly correlated in response to treatment with these test materials and also to estradiol. It was suggested that the uterine peroxidase assay could be utilized as a biochemical parameter in the screening of new antifertility agents for their estrogenic/antiestrogenic properties.

Animals

The nucleotide sequence of tomato mottle virus, a new geminivirus isolated from tomatoes in Florida.

A new geminivirus, tomato mottle virus (TMoV), affecting tomato production in Florida has been cloned and sequenced. Sequence analysis of the cloned replicative forms of TMoV revealed four potential coding regions for the A component [2601 nucleotides (nt)] and two for the B component (2541 nt). Comparisons of the nucleotide sequence of the TMoV genome with those of other whitefly-transmitted geminiviruses indicate that TMoV is a typical bipartite geminivirus of the New World and is closely related to but distinct from abutilon mosaic virus.

Base Sequence

Unraveling evolutionary relationships in the Sida generic alliance (Malvaceae, Malvoideae): a phylogenetic and cytotaxonomic overview.

Sida (Malvaceae), the largest Malveae-Abutilinae member, has poorly defined morphological limits which overlaps with 11 phylogenetically closely related genera that comprises the "Sida generic alliance". The 12 genera are distributed in the tropics especially in Brazil where one third of its species diversity is found. Evolutionary relationships within Sida generic alliance remain unresolved due to morphological convergence, limited taxon sampling, and lack of integrative approaches including cytogenetic data. We reconstructed the phylogeny of Sida and allied genera using a multilocus dataset (nuclear ITS and seven plastid loci) including 193 species classified in 19 genera and analyzed chromosome evolution using cytogenetic data (chromosome number) for 79 species of the 19 genera. The phylogeny recovered seven clades-Abutilon, Bakeridesia, Callianthe, Gaya, and three Sida clades (I-III)-and confirmed the polyphyly of Sida, the largest genera. We detected reticulate evolution, with incongruence between nuclear and plastid topologies. Chromosome number ranged from 2n = 12 to 60 and represented synapomorphies for most clades. Ancestral character reconstruction indicated that ascending dysploidy and polyploidy predominated in karyotype evolution of Sida and allied genera. Our results reveal taxonomic incongruence in current classifications probably related to reticulate evolution. A generic-level taxonomic revision is necessary and should rely on integrated phylogenetic and karyotypic evidence. This study provides a framework for phylogenetic systematics and emphasizes the role of Brazil as a hotspot for plant genomic research.

Phylogeny