Multigene phylogeny and morphological descriptions of five species of Agaricus sect. Minores from subtropical climate zones of Pakistan.

The genus Agaricus includes more than 500 species mostly containing the edible and cultivated species worldwide. As part of the ongoing studies on the biodiversity of genus Agaricus in Pakistan, our objective was to focus on A. sect. Minores which is the largest section of the genus. In the first phylogenetic analyses based on the ITS region of the nuclear ribosomal DNA, our sample included specimens of 97 named species, 27 unnamed species, and 31 specimens (29 newly generated sequences in this study) from subtropical climate zones of Pakistan that likely belong to this section based on their morphology. The 31 specimens grouped into five distinct, well-supported clades corresponding to five species: A. glabriusculus already known from Pakistan and India, A. robustulus first recorded from Pakistan and briefly described here but already known from Bénin, Malaysia, China, and Thailand, and three possibly endemic new species described in detail A. badiosquamulosus sp. nov., A. dunensis sp. nov., and A. violaceopunctatus sp. nov. The sixth species currently known in Pakistan, including A. latiumbonatus also found in Thailand, were included in a multigene tree based on ITS, LSU, and Tef-1α sequence data. They all belong to a large pantropical paraphyletic group while most temperate species belong to a distinct clade, which includes about half of the species of the section. The current study aims to propose three novel species of genus Agaricus based on comprehensive morphological as well as molecular phylogenetic evidences from Pakistan.

Connecting nutritional facts with the traditional ranking of ethnobotanically used fodder grasses by local farmers in Central Punjab of Pakistan.

The local farmers of Central Punjab, Pakistan have been using indigenous grasses as vital components of ruminant diets, but little is reported about their nutritional potential. Hence this study investigated nutritive potential of a selection of ethnobotanically important fodder grasses. Multiple nutritional parameters (proximate components, fibre fractions), secondary metabolites (phenolics, tannins) and in vitro digestibility values were determined. Furthermore, the legitimacy of ethnobotanical knowledge of local inhabitants about these grasses was also verified. The results suggested that majority (77%) of these grasses can be regarded as good quality fodders because of their high protein (169 g/kg) and good digestibility (457 g/kg) with moderate fibre (≤ 602 g/kg), lignin (≤ 50 g/kg) and secondary metabolites (total phenols ≤ 87 g/kg, total tannins ≤ 78 g/kg, condensed tannins ≤ 61 g/kg). Pearson correlation between nutritional parameters indicated that in vitro digestibility values were positively correlated with crude proteins (IVDMD, r = + 0.83 and IVOMD, r = + 0.83 respectively) and negatively correlated with fibre (NDF, r = - 0.91), ADF, r = - 0.84 and ADL, r = - 0.82) contents. Moreover, a positive relationship was identified between ethnobotanical knowledge and laboratory findings for studied grasses. Spearman correlation test showed that ranking of grasses based on ethnobotanical preferences were highly correlated (r values) with the laboratory results for CP (0.85), NDF (- 0.76), ADF (- 0.72) and ADL (- 0.62). The resilient complementarities between ethnobotanical preferences and nutritive analysis authenticate farmer's traditional knowledge, which needed to be aligned with the corresponding scientific data. Farmers can use these findings for appropriate fodder selection and development of precise supplements for feeding ruminants within a sustainable and economically viable livestock industry for food security.

Light and scanning electron microscopic imaging of leaf transverse sections of indigenous fodder grasses of central Punjab, Pakistan.

The significance of grasses as fodder is enormous. However, it has been observed that different fodder grasses are identified locally with the similar vernacular names. Even same species recognized with different names. This happened due to their reduced and overlapped morphological characters. Hence comprehensive taxonomic keys were required to provide authentic identification of this grass flora. Current study established microscopic identification tools on the basis of leaf transverse anatomical features. Results demonstrated a great diversity in ribs and furrows arrangement among all studied grass members. Lolium temulentum, Phalaris minor, Poa annua, Arundo donax, Aristida adscensionis, Desmostachya bipinnata, Cenchrus biflorus and Digitaria ciliaris possessed ribs and furrows only at adaxial surfaces whereas all other species possessed this feature abaxially. Generally all the species contain three types of vascular bundles i.e. small, medium and large. Therefore, this qualitative feature is valuable only at family level, however, it found insignificant in taxa identification. Diversity was observed in bundle sheath type which could be taxonomically significant. Generally bundle sheath was found complete in most of species but in Bromus japonicus Thunb., Dactylis glomerata L., Acrachne racemosa (Heyne ex Roth) Ohwi, Chrysopogon aucheri (Boiss.) Stapf, Dichanthium annulatum (Forssk.) Stapf, Heteropogon contortus (L.) P Beauv. Ex. Roem & Schult. Eragrostis minor Host. and in Agrostis gigantea Roth. had incomplete type of sheath. Results reported great diversity in characters which could be taxonomically viable. RESEARCH HIGHLIGHTS: This research highlighted the significance of microscopic examination of leaf transverse anatomical features in field of taxonomy. Established anatomical tools would be of great importance for the identification of studied species.

An overview of Agaricus section Hondenses and Agaricus section Xanthodermatei with description of eight new species from Pakistan.

In a recent revision of the genus Agaricus, A. section Xanthodermatei was split into two sections A. sect. Hondenses and A. sect. Xanthodermatei. Our objectives were to investigate the species diversity of both sections in Pakistan and to give an overview of the major clades. Phylogenetic analyses based on the combined nucLSU, ITS and TEF1 dataset from 35 specimens of both sections revealed three major clades. Analyses based on ITS dataset and 106 specimens, including 33 from Pakistan, reveal eight new species and one new record species. These nine species are described in detail. It is noteworthy that intraspecific variability as well as interspecific variability between closely related species were very low in ITS sequences in many cases. In the case of the two new species A. xanthochromaticus and A. griseovariegatus, TEF1 sequence data were much more efficient than ITS to distinguish these species from each other. The other new species are A. atroumbonatus, A. fumidicolor, A. macropeplus, A. parviniveus, A. swaticus and A. bambusetorum. The latter is the only new species of A. sect. Hondenses in which it is morphologically atypical and also the unique (sub)tropical species. Agaricus gregariomyces is recorded for the first time in Pakistan. In addition, brief descriptions are provided not only for A. bisporiticus, A. endoxanthus and A. punjabensis, which are reported again in Pakistan, but also for A. californicus, which is reported for the first time in Spain and outside North America. In total 12 species of both sections were reported in Pakistan and half of them were from subtropical climatic areas, underlining the contribution of the climatic diversity to the high species richness in this country.