Formally described species woefully underrepresent phylogenetic diversity in the common lichen photobiont genus Trebouxia (Trebouxiophyceae, Chlorophyta): An impetus for developing an integrated taxonomy.

Lichens provide valuable systems for studying symbiotic interactions. In lichens, these interactions are frequently described in terms of availability, selectivity and specificity of the mycobionts and photobionts towards one another. The lichen-forming, green algal genus Trebouxia Puymaly is among the most widespread photobiont, associating with a broad range of lichen-forming fungi. To date, 29 species have been described, but studies consistently indicate that the vast majority of species-level lineages still lack formal description, and new, previously unrecognized lineages are frequently reported. To reappraise the diversity and the evolutionary relationships of species-level lineages in Trebouxia, we assembled DNA sequence data from over 1600 specimens, compiled from a range of sequences from previously published studies, axenic algal cultures, and lichens collected from poorly sampled regions. From these samples, we selected representatives of the currently known genetic diversity in the lichenized Trebouxia and inferred a phylogeny from multi-locus sequence data (ITS, rbcL, cox2). We demonstrate that the current formally described species woefully underrepresent overall species-level diversity in this important lichen-forming algal genus. We anticipate that an integrative taxonomic approach, incorporating morphological and physiological data from axenic cultures with genetic data, will be required to establish a robust, comprehensive taxonomy for Trebouxia. The data presented here provide an important impetus and reference dataset for more reliably characterizing diversity in lichenized algae and in using lichens to investigate the evolution of symbioses and holobionts.

Evolution of complex symbiotic relationships in a morphologically derived family of lichen-forming fungi.

We studied the evolutionary history of the Parmeliaceae (Lecanoromycetes, Ascomycota), one of the largest families of lichen-forming fungi with complex and variable morphologies, also including several lichenicolous fungi. We assembled a six-locus data set including nuclear, mitochondrial and low-copy protein-coding genes from 293 operational taxonomic units (OTUs). The lichenicolous lifestyle originated independently three times in lichenized ancestors within Parmeliaceae, and a new generic name is introduced for one of these fungi. In all cases, the independent origins occurred c. 24 million yr ago. Further, we show that the Paleocene, Eocene and Oligocene were key periods when diversification of major lineages within Parmeliaceae occurred, with subsequent radiations occurring primarily during the Oligocene and Miocene. Our phylogenetic hypothesis supports the independent origin of lichenicolous fungi associated with climatic shifts at the Oligocene-Miocene boundary. Moreover, diversification bursts at different times may be crucial factors driving the diversification of Parmeliaceae. Additionally, our study provides novel insight into evolutionary relationships in this large and diverse family of lichen-forming ascomycetes.

The Lichen Genus Sticta (Lobariaceae, Peltigerales) in East African Montane Ecosystems.

The lichen flora of Africa is still poorly known. In many parts of the tropics, recent studies utilizing DNA methods have revealed extraordinary diversity among various groups of lichenized fungi, including the genus Sticta. In this study, East African Sticta species and their ecology are reviewed using the genetic barcoding marker nuITS and morphological characters. The studied regions represent montane areas in Kenya and Tanzania, including the Taita Hills and Mt. Kilimanjaro, which belong to the Eastern Afromontane biodiversity hotspot. Altogether 14 Sticta species are confirmed from the study region, including the previously reported S. fuliginosa, S. sublimbata, S. tomentosa, and S. umbilicariiformis. Sticta andina, S. ciliata, S. duplolimbata, S. fuliginoides, and S. marginalis are reported as new to Kenya and/or Tanzania. Sticta afromontana, S. aspratilis, S. cellulosa, S. cyanocaperata, and S. munda, are described as new to science. The abundance of new diversity detected and the number of taxa represented by only few specimens show that more comprehensive sampling of the region may be needed to reveal the true diversity of Sticta in East Africa. More generally, our results highlight the need for further taxonomic studies of lichenized fungi in the region.

An annotated plant checklist of the transboundary volcanic Mt Elgon, East Africa.

Mt Elgon is an ancient transboundary volcanic mountain found at the Kenya-Uganda boarder possessing high plant diversity. This study documents an updated checklist of the mountain's vascular plants obtained through random-walk field excursions and retrieval of herbarium specimen tracing back to 1900. We compiled 1709 species from 673 genera in 131 families. One new species of the family Cucurbitaceae was also reported. This checklist records respective habitat, habits, elevation ranges, voucher numbers and global distribution ranges of each species. Native and exotic species were also distinguished, where 8.4% of the total species in 49 families were exotic species. There were 103 endemic species, while 14 species were found to be both rare and endemic. IUCN conservation status revealed 2 Critically Endangered, 4 Endangered, 9 Vulnerable and 2 Near Threatened species. This study presents the first and most comprehensive plant inventory of Mt Elgon that will facilitate further ecological and phylogenetic studies.

A New Cryptic Lineage in Parmeliaceae (Ascomycota) with Pharmacological Properties.

We used molecular data to address species delimitation in a species complex of the parmelioid genus Canoparmelia and compare the pharmacological properties of the two clades identified. We used HPLC_DAD_MS chromatography to identify and quantify the secondary substances and used a concatenated data set of three ribosomal markers to infer phylogenetic relationships. Some historical herbarium specimens were also examined. We found two groups that showed distinct pharmacological properties. The phylogenetic study supported the separation of these two groups as distinct lineages, which are here accepted as distinct species: Canoparmelia caroliniana occurring in temperate to tropical ecosystems of a variety of worldwide localities, including America, Macaronesia, south-west Europe and potentially East Africa, whereas the Kenyan populations represent the second group, for which we propose the new species C. kakamegaensis Garrido-Huéscar, Divakar & Kirika. This study highlights the importance of recognizing cryptic species using molecular data, since it can result in detecting lineages with pharmacological properties previously overlooked.

Diversity of Leptogium (Collemataceae, Ascomycota) in East African Montane Ecosystems.

Tropical mountains and especially their forests are hot spots of biodiversity threatened by human population pressure and climate change. The diversity of lichens in tropical Africa is especially poorly known. Here we use the mtSSU and nuITS molecular markers together with morphology and ecology to assess Leptogium (Peltigerales, Ascomycota) diversity in the tropical mountains of Taita Hills and Mt. Kasigau in Kenya and Mt. Kilimanjaro in Tanzania. The sampled habitats cover a wide range of ecosystems from savanna to alpine heath vegetation and from relatively natural forests to agricultural environments and plantation forests. We demonstrate that Leptogium diversity in Africa is much higher than previously known and provide preliminary data on over 70 putative species, including nine established species previously known from the area and over 60 phylogenetically, morphologically, and/or ecologically defined Operational Taxonomic Units (OTUs). Many traditional species concepts are shown to represent morphotypes comprised of several taxa. Many of the species were only found from specific ecosystems and/or restricted habitats and are thus threatened by ongoing habitat fragmentation and degradation of the natural environment. Our results emphasize the importance of molecular markers in species inventories of highly diverse organism groups and geographical areas.

Complete Chloroplast Genomes of Acanthochlamys bracteata (China) and Xerophyta (Africa) (Velloziaceae): Comparative Genomics and Phylogenomic Placement.

Acanthochlamys P.C. Kao is a Chinese endemic monotypic genus, whereas Xerophyta Juss. is a genus endemic to Africa mainland, Arabian Peninsula and Madagascar with ca.70 species. In this recent study, the complete chloroplast genome of Acanthochlamys bracteata was sequenced and its genome structure compared with two African Xerophyta species (Xerophyta spekei and Xerophyta viscosa) present in the NCBI database. The genomes showed a quadripartite structure with their sizes ranging from 153,843 bp to 155,498 bp, having large single-copy (LSC) and small single-copy (SSC) regions divided by a pair of inverted repeats (IR regions). The total number of genes found in A. bracteata, X. spekei and X. viscosa cp genomes are 129, 130, and 132, respectively. About 50, 29, 28 palindromic, forward and reverse repeats and 90, 59, 53 simple sequence repeats (SSRs) were found in the A. bracteata, X. spekei, and X. viscosa cp genome, respectively. Nucleotide diversity analysis in all species was 0.03501, Ka/Ks ratio average score was calculated to be 0.26, and intergeneric K2P value within the Order Pandanales was averaged to be 0.0831. Genomic characterization was undertaken by comparing the genomes of the three species of Velloziaceae and it revealed that the coding regions were more conserved than the non-coding regions. However, key variations were noted mostly at the junctions of IRs/SSC regions. Phylogenetic analysis suggests that A. bracteata species has a closer genetic relationship to the genus Xerophyta. The present study reveals the complete chloroplast genome of A. bracteata and gives a genomic comparative analysis with the African species of Xerophyta. Thus, can be useful in developing DNA markers for use in the study of genetic variabilities and evolutionary studies in Velloziaceae.

An annotated checklist of the vascular plants of Aberdare Ranges Forest, a part of Eastern Afromontane Biodiversity Hotspot.

The Aberdare Ranges Forest, located in the Central highlands of Kenya, is an isolated volcanic mountain in the East African Rift Valley with unique flora. Despite its refugial importance to rare and endemic plant species, the diversity of plants in the Aberdare Ranges Forest remains poorly understood. The checklist presented here is a collation of data obtained from multiple floristic surveys and from herbarium specimen collections from the forest. A total of 1260 vascular plants taxa representing 136 families, 613 genera, 67 subspecies and 63 varieties are documented. The ferns comprised 84 species, lycophytes seven, gymnosperms six and angiosperms were 1163 taxa. This represents 17.9% of the Kenyan taxa, 1.7% of the African taxa and 0.3% of all the vascular plants known in the world. A total of 18 taxa were endemic and 14 taxa were found to be threatened globally. The life form, voucher specimen(s), habitat and distribution range of each taxon and a brief analysis of taxa diversity is presented in this checklist. This is the first comprehensive inventory of vascular plants in the entire Aberdare Ranges, providing a solid basis for more sustainable management and improved conservation of this montane forest. The checklist is also an important contribution to the world checklist of plants required by the Global Strategy for Plant Conservation.

Initial Complete Chloroplast Genomes of Alchemilla (Rosaceae): Comparative Analysis and Phylogenetic Relationships.

The genus Alchemilla L., known for its medicinal and ornamental value, is widely distributed in the Holarctic regions with a few species found in Asia and Africa. Delimitation of species within Alchemilla is difficult due to hybridization, autonomous apomixes, and polyploidy, necessitating efficient molecular-based characterization. Herein, we report the initial complete chloroplast (cp) genomes of Alchemilla. The cp genomes of two African (Afromilla) species Alchemilla pedata and Alchemilla argyrophylla were sequenced, and phylogenetic and comparative analyses were conducted in the family Rosaceae. The cp genomes mapped a typical circular quadripartite structure of lengths 152,438 and 152,427 base pairs (bp) in A. pedata and A. argyrophylla, respectively. Alchemilla cp genomes were composed of a pair of inverted repeat regions (IRa/IRb) of length 25,923 and 25,915 bp, separating the small single copy (SSC) region of 17,980 and 17,981 bp and a large single copy (LSC) region of 82,612 and 82,616 bp in A. pedata and A. argyrophylla, respectively. The cp genomes encoded 114 unique genes including 88 protein-coding genes, 37 transfer RNA (tRNA) genes, and 4 ribosomal RNA (rRNA) genes. Additionally, 88 and 95 simple sequence repeats (SSRs) and 37 and 40 tandem repeats were identified in A. pedata and A. argyrophylla, respectively. Significantly, the loss of group II intron in atpF gene in Alchemilla species was detected. Phylogenetic analysis based on 26 whole cp genome sequences and 78 protein-coding gene sequences of 27 Rosaceae species revealed a monophyletic clustering of Alchemilla nested within subfamily Rosoideae. Based on a protein-coding region, negative selective pressure (Ka/Ks < 1) was detected with an average Ka/Ks value of 0.1322 in A. argyrophylla and 0.1418 in A. pedata. The availability of complete cp genome in the genus Alchemilla will contribute to species delineation and further phylogenetic and evolutionary studies in the family Rosaceae.

The complete chloroplast genome sequence of Xerophyta spekei (Velloziaceae).

The complete chloroplast genome sequence of Xerophyta spekei Baker was reported in this study. The complete chloroplast genome showed a stereotypical quadripartite structure as observed in other angiosperms with a length of 155,235 bp and divided into four parts; a pair of IRs (27,109 bp) which is separated by a small single copy (SSC) region (17,388 bp) and a large single copy (LSC) region (83,629bp). The chloroplast genome had 132 genes, including 85 protein-coding genes, 38 tRNA genes, and 8 rRNA genes. Seven protein-coding genes were identified to have RNA editing.

An annotated checklist of vascular plants of Cherangani hills, Western Kenya.

Cherangani hills, located in Western Kenya, comprises of 12 forest blocks, maintaining great plant diversity. However, little attention to plant diversity studies has been paid to it in the past years. Here, we present a checklist of the vascular plants of this region obtained through intensive field investigations and matching of herbarium specimens. In total, 1296 species, including 17 endemic species are documented, belonging to 130 families and 608 genera. This flora represents 18.50%, 43.83% and 54.17% of the Kenyan species, genera and families, respectively. The habit, habitat and voucher specimens, as well as brief notes on the distribution of each taxon recorded are presented in this checklist. It is the first exhaustive inventory of the terrestrial vascular plants in Cherangani hills which is a significant regional centre for plant diversity.

Circumscription of the genus Lepra, a recently resurrected genus to accommodate the "Variolaria"-group of Pertusaria sensu lato (Pertusariales, Ascomycota).

Pertusarialean lichens include more than 300 species belonging to several independent phylogenetic lineages. Only some of these phylogenetic clades have been comprehensively sampled for molecular data, and formally described as genera. Here we present a taxonomic treatment of a group of pertusarialean lichens formerly known as "Pertusaria amara-group", "Monomurata-group", or "Variolaria-group", which includes widespread and well-known taxa such as P. amara, P. albescens, or P. ophthalmiza. We generated a 6-locus data set with 79 OTUs representing 75 species. The distinction of the Variolaria clade is supported and consequently, the resurrection of the genus Lepra is followed. Thirty-five new combinations into Lepra are proposed and the new species Lepra austropacifica is described from mangroves in the South Pacific. Lepra is circumscribed to include species with disciform ascomata, a weakly to non-amyloid hymenial gel, strongly amyloid asci without clear apical amyloid structures, containing 1 or 2, single-layered, thin-walled ascospores. Chlorinated xanthones are not present, but thamnolic and picrolichenic acids occur frequently, as well as orcinol depsides. Seventy-one species are accepted in the genus. Although the distinction of the genus from Pertusaria is strongly supported, the relationships of Lepra remain unresolved and the genus is tentatively placed in Pertusariales incertae sedis.