Micromorphological and phytochemical profiling of Delphinium suave Huth. from Hindukush range, Lower Dir Khyber Pakhtunkhwa Pakistan.

This is a very first attempt to study various parameters of a medicinal plant, Delphinium suave Huth. The plant is erect, geophytic, herbaceous, with tuberous root, trifid in a palmatipartite, strigose cuneate leaf and white spurred zygomorphic flower. The root was isodiametric phellem with single non-glandular trichomes. The stem revealed single-layered cuticle, multiseriate epidermis, cortex, pith ray and uniserate bowed non-glandular trichomes. The leaf was amphistomatic, showed tapering trichomes, prismatic crystals and ranunculaceous stomata with circumference 144.66-182.67 µm. Pollen grains in Light Microscopy (LM), were prolate, spheroidal trizonocolpate, isopolar, radiosymmetric, scabrate, elliptic and monads. Scanning Electron Microscope (SEM) pollen surface was scabrate, monad, size varied from 18.06 to 16.67 µm, colpus to inaperturate, tricolpate, ornamented, echinus, isopolar, isodiametric and circular. SEM roots showed sclerenchymatic tissues, stellate, glandular, non-glandular trichomes and crystals. The stem showed scalariform, pitted vessels, warty protuberances, unicellular, silicified, non-glandular trichomes. Leaves powder revealed, simple, unicellular, tapered headed, uniseriate, sessile, capitate, unbranched glandular, non-glandular, trichomes with crystals. Capitate, stellate, circular, unicellular, branchy trichomes were observed for the first time through SEM. Powder drug study of root, stem leaves through LM revealed different tissues. Preliminary phytochemical revealed alkaloids, anthocyanins, anthraquinones, coumarins, flavones, mucilages, saponins, steroids, terpenoids, volatile oils and proteins. GC/MS showed 36 compounds in roots, 33 in stem while 40 in leaves. Fluorescence analysis of roots, stem and leaves showed variations in color when treated with chemicals. This study will assist pharmacognostic exploration, authentication from adulterants/allied species for consistent quality, resulting in safe use, preservation and efficacy. RESEARCH HIGHLIGHTS: This was first attempt on pharmacognostic study on D. suave Huth. which could be used as a foundation for identifying and authenticating the specie from other allied species by these morphological, anatomical, GC/MS profiling, phytochemical analysis and fluorescence analysis.

Light and scanning electron microscopy-based foliar micro morphological tools for the identification of fodder grass taxa.

Fertile plain of Central Punjab Pakistan is rich with fodder grasses and from centuries the local inhabitants of this area have been using their regional grasses for ruminant feeding. However, they always faced difficulties in identification because of their overlapping vernacular names, more or less identical leaf shapes, indefinite variations in stem branching pattern, and reduced floral parts. Hence, the current study has provided a detailed and comprehensive micro-morphological analysis of 53 ethnobotanical fodder grass taxa. A variety of quantitative and qualitative leaf epidermal micromorphological traits was studied and results reported epidermal characters like stomatal index, silica bodies, prickles, microhairs, hook cells as most diagnostic in delimitation of species, and genera. As stomatal index was 79% in Poa annua while in its closely related species Poa infirmai was 85%. Similarly, Cenchrus ciliaris can be differentiated from Cenchrus pennisetiformis on the basis of silica body shape as butterfly shaped in former and dumbbell shaped in later one. Moreover, prickles were present in Chrysopogon aucheri while absent in Chrysopogon zizanioides. Hence, overall this study declared that diversity and variations in foliar micromorphological characters are valuable and supportive in the identification of grasses at the specific and generic level.

Significance of leaf morphoanatomical markers for the authentication of adultered drugs sold in herbal markets of district Lahore, Pakistan.

A number of herbal plants sold in herbal markets of Lahore are under adulteration threat which can pose harmful health effects to end-user. This adulteration problem of medicinal plants can be resolved by the implication of some valuable taxonomic parameter such as leaf epidermal anatomical characteristics. Hence, this research was aimed to provide viable anatomical markers in order to resolve this adulteration issue persisted in some common marketed medicinal plants of district Lahore, that is, Cinnamomum verum Presl., Cinnamomum tamala (Buuch.-Ham.) T.Nees&Eberm., Gymnema sylvestre (Retz.) R.Br.ex Sm., Sphaeranthus indicus Linn., Artemisia maritima Linn., Achillea millifolim L., Adhatoda vasica Nees, Butea monosperma (Lam.) Taub, and Morus nigra L. Overall multiple anatomical variations (epidermal cell shape, their length and width, type of stomata, length and width of guard cells along with presence or absence of trichomes) had been reported in the study that could be worthwhile for the correct identification of medicinal plants. Irregular shapes of epidermal cells were observed in Cinnamomum verum and Achillea millifolium while pentagonal and polygonal cells were found in their adultaerants, that is, Canella winterana and Adhatoda vasica, respectively. Types of stomata were also strikingly varied among genuine plant and its adulterant, for example, anisocytic stomata were observed in Artemisia maritima while in its adulterant (Artemisia absinthium) anomocytic stomata were found. Similarly, paracytic stomata were observed in Butea monosperma, whereas its adulterant plant (Averrhoa carambola) characteristically possessed anisocytic stomata. Hence, anatomical characteristics were proved to be a valuable taxonomic tool in resolving the adulteration issue of medicinal plants.

Microscopic and Phytochemical Comparison of the Three Leonurus Species L. cardiaca, L. japonicus, and L. sibiricus.

At least three Leonurus species, Leonurus cardiaca, Leonurus japonicus, and Leonurus sibiricus, are used in various traditional medicinal systems for different therapeutic purposes. The plant names "L. sibiricus" and "L. japonicus" are often used as synonyms, which causes confusion in literature and implies that several studies may be based on improperly identified plant material. To avoid further confusion, the current status of the identification of these three species is illustrated. Characteristics for their unequivocal identification are presented using stereo and light microscopy as well as HPTLC.The establishment of the species-specific anatomical, morphological, and phytochemical characters was based on reference specimens in comparison with wild collected or commercially obtained material. Morphologically, the species differed in shape and length ratio of the upper and lower lip of the corolla. Differentiating anatomical characters involve the presence, density, and dimension of trichomes on different organs. Detailed anatomical descriptions of the leaves and corollae of L. japonicus and L. sibiricus are given and compared to L. cardiaca. These three sets of characters facilitate fast and reliable identification. Complementary HPTLC fingerprints show type-specific patterns that allow the differentiation of L. japonicus from the other two species. For the distinction of L. cardiaca and L. sibiricus, flower morphology has to be considered.The presented data contribute to the quality control of the three Leonurus species and additionally meet a definite and timely need regarding the introduction of L. japonicus to the European Pharmacopoeia in addition to the already existing monograph of L. cardiaca.

Non-glandular trichomes of Solanum carolinense deter feeding by Manduca sexta caterpillars and cause damage to the gut peritrophic matrix.

Plant trichomes constitute a first line of defence against insect herbivores. The pre- and post-ingestive defensive functions of glandular trichomes are well documented and include direct toxicity, adhesion, antinutrition and defence gene induction. By contrast, the defensive functions of non-glandular trichomes are less well characterized, although these structures are thought to serve as physical barriers that impede herbivore feeding and movement. We experimentally varied the density of stellate non-glandular trichomes in several ways to explore their pre- and post-ingestive effects on herbivores. Larvae of Manduca sexta (Sphingidae) initiated feeding faster and gained more weight on Solanum carolinense (Solanaceae) leaves having lower trichome densities (or experimentally removed trichomes) than on leaves having higher trichome densities. Adding trichomes to artificial diet also deterred feeding and adversely affected caterpillar growth relative to controls. Scanning electron and light microscopy revealed that the ingestion of stellate trichomes by M. sexta caterpillars caused extensive damage to the peritrophic membrane, a gut lining that is essential to digestion and pathogen isolation. These findings suggest that, in addition to acting as a physical barrier to deter feeding, trichomes can inhibit caterpillar growth and development via post-ingestive effects.

Plant Responses to Brief Touching: A Mechanism for Early Neighbour Detection?

In natural habitats plants can be exposed to brief and light contact with neighbouring plants. This mechanical stimulus may represent a cue that induces responses to nearby plants. However, little is known about the effect of touching on plant growth and interaction with insect herbivores. To simulate contact between plants, a soft brush was used to apply light and brief mechanical stimuli to terminal leaves of potato Solanum tuberosum L. The number of non-glandular trichomes on the leaf surface was counted on images made by light microscope while glandular trichomes and pavement cells were counted on images made under scanning electronic microscope. Volatile compounds were identified and quantified using coupled gas chromatography-mass spectrometry (GC-MS). Treated plants changed their pattern of biomass distribution; they had lower stem mass fraction and higher branch and leaf mass fraction than untouched plants. Size, weight and number of tubers were not significantly affected. Touching did not cause trichome damage nor change their total number on touched terminal leaves. However, on primary leaves the number of glandular trichomes and pavement cells was significantly increased. Touching altered the volatile emission of treated plants; they released higher quantities of the sesquiterpenes (E)-ß-caryophyllene, germacrene D-4-ol and (E)-nerolidol, and lower quantities of the terpenes (E)-ocimene and linalool, indicating a systemic effect of the treatment. The odour of touched plants was significantly less preferred by the aphids Macrosiphum euphorbiae and Myzus persicae compared to odour of untouched plants. The results suggest that light contact may have a potential role in the detection of neighbouring plants and may affect plant-insect interactions.

Identification and QTL mapping of whitefly resistance components in Solanum galapagense.

Solanum galapagense is closely related to the cultivated tomato and can show a very good resistance towards whitefly. A segregating population resulting from a cross between the cultivated tomato and a whitefly resistant S. galapagense was created and used for mapping whitefly resistance and related traits, which made it possible to study the genetic basis of the resistance. Quantitative trait loci (QTL) for adult survival co-localized with type IV trichome characteristics (presence, density, gland longevity and gland size). A major QTL (Wf-1) was found for adult survival and trichome characters on Chromosome 2. This QTL explained 54.1 % of the variation in adult survival and 81.5 % of the occurrence of type IV trichomes. A minor QTL (Wf-2) for adult survival and trichome characters was identified on Chromosome 9. The major QTL was confirmed in F3 populations. Comprehensive metabolomics, based on GCMS profiling, revealed that 16 metabolites segregating in the F2 mapping population were associated with Wf-1 and/or Wf-2. Analysis of the 10 most resistant and susceptible F2 genotypes by LCMS showed that several acyl sugars were present in significantly higher concentration in the whitefly resistant genotypes, suggesting a role for these components in the resistance as well. Our results show that whitefly resistance in S. galapagense seems to inherit relatively simple compared to whitefly resistance from other sources and this offers great prospects for resistance breeding as well as elucidating the underlying molecular mechanism(s) of the resistance.