Structure and Histochemistry of Stomata and Epidermal Cells in Five Species of Polypodiaceae

The epidermal structure of the five species of ferns, Arthromeriswallichiana (Spr.) Ching., Drymoglossum piloselloides (Prest.),Drynaria quercifolia (L.) J. Smith, Lepisorus nudus (Hook.)Ching. and Pyrrosia nuda (Gies.) Ching., has been investigated.Fifteen types of stomatal structures have been identified ofwhich copolo-desmocytic and coperi-desmocytic are new types.Four more possible stomatal structures: ccpolo-peri-, codesmo-polo-,codesmo-peri- and duplodesmocytic, are suggested. Localizationof starch, insoluble polysaccharides, protein and lipids hasbeen examined histochemically in the guard cells, subsidiarycells and epidermal cells. In Drynaria starch plastids and plastidscontaining both starch and protein are present in guard cells.Starch plastids are present in the subsidiary cells of all speciesexcept in Arthromeris, whereas, they are present in epidermalcells of only Drymoglossum and Lepisorus. Granular or amorphousinsoluble polysaccharides (other than starch) are present inguard cells of all the species, in the subsidiary cells of Arthromeris,Drynaria and Pyrrosia, and in the epidermal cells of Pyrrosia.Except in Pyrrosia lipids are present in the guard cells. Subsidiarycells of Drynaria and the epidermal cells of Arthromeris andDrynaria show lipid bodies. The presence of plasmodesmata andectodesmata is demonstrated in the epidermal cells of Drymoglossum.     

Development of Vacuolar Volume in the Root Tips of Pea

Cell and vacuole areas were measured by light microscopy inlongitudinal and transverse sections cut at 0.4-mm intervalsalong the apical 7.2 mm of the primary root of pea. The vacuolararea (or volume) fraction — that is, vacuole area (orvolume) divided by cell area (or volume) — increased fromabout 15 % in cells 0.4 mm from the distal boundary of the apicalmeristem (the cap /root junction), to about 85% in cells situated6.8–7.2 mm from that boundary. At each distance, vacuoledevelopment tended to be greater in the cortex than in the stele.Vacuoles occupied about 22% of the tissue volume in the first1 mm length of root (measured from the cap/root junction), about31 % of the tissue volume in the first 2 mm, and about 45% whensummed over the apical 5-mm length of root. Phosphorus supplyor deprivation produced only minor and non-significant changesin vacuole development. The results have implications affectingprevious estimates of cytoplasmic and vacuolar phosphate concentrationsin pea root tips. Pisum sativum L., pea, root, vacuole, volume     

Effect of salinisation of soil on growth and macro- and micro-nutrient accumulation in seedlings of Acacia catechu (Mimosaceae)

The effects of salinisation of soil on Acacia catechu (Mimosaceae) were studied by means of emergence and growth of seedlings and pattern of mineral accumulation. A mixture of chlorides and sulphates of Na, K, Ca and Mg was added to the soil and salinity was maintained at 4.1, 6.3, 8.2,10.1 and 12.2 dSm⁻¹. A negative relationship between proportion of seed germination and salt concentration was obtained. Seedlings did not emerge when soil salinity exceeded 10.1 dSm⁻¹. Results suggested that this tree species is salt tolerant at the seed germination stage. Seedlings survived and grew up to soil salinity of 10.1 dSm⁻¹, which suggests that this species is salt tolerant at the seedling stage too. Elongation of stem and root was retarded by increasing salt stress. Among the tissues, young roots and stem were most tolerant to salt stress and were followed by old roots and leaves, successively. Leaf tissue exhibited maximum reduction in dry mass production in response to increasing salt stress. However, production of young roots and death of old roots were found to be continuous and plants apparently use this process as an avoidance mechanism to remove excess ions and delay onset of ion accumulation in this tissue. This phenomenon, designated “fine root turnover”, is of importance to the mechanisms of salt tolerance. Plants accumulated Na in roots and were able to regulate transfer of Na ions to leaves. Stem tissues were a barrier for translocation of Na from root to leaf. Moreover, K was affected in response to salinity; it rapidly decreased in root tissues with increased salinisation. Nitrogen content decreased in all tissues (leaf, stem and root) in response to low water treatment and salinisation of soil. Phosphorus content significantly decreased, while Ca increased in leaves as soil salinity increased. Changes in tissue and whole plant accumulation patterns of the other elements tested, as well as possible mechanisms for avoidance of Na toxicity in this tree species during salinisation, are discussed.     

Stomatal Features in the Leaf of Aganosma dichotoma (Roth) K. Schum

Paracytic and anisocytic types of mature stomata are found inthe leaf of Aganosma dichotoma. Stomata with one guard cell,stomata with degenerated guard cells, and contiguous stomataare common. Stomata with arrested pore development are alsofound in certain cases. A single guard cell without any porehas not been designated as a stoma with one guard cell in thepresent investigation. Ontogeny of contiguous stomata have beentraced. Subsidiary cells are, morphologically, just like theircontiguous guard cells. Subsidiary cells may retain their shapeand contents even when their contiguous stoma becomes mature,or may change their shape and lose their contents. They mayor may not divide. Subsidiary cells form a whorl of more thantwo subsidiary cells around a stoma by their divisions. Degenerationof guard cell(s)— their contents and nuclei—havebeen traced. In certain cases guard cells divide forming morethan two guard cells associated to a single pore. Cytoplasmicconnections are found between two guard cells of nearby stomata,and between a guard cell and an epidermal cell. Near the wound,the epidermal cells over the veins become meristermatic givingrise to new epidermal cells but no meristemoid.     

β-Carboline alkaloids induce structural plasticity and inhibition of SARS-CoV-2 nsp3 macrodomain more potently than remdesivir metabolite GS-441524: computational approach

The nsp3 macrodomain is implicated in the viral replication, pathogenesis and host immune responses through the removal of ADP-ribosylation sites during infections of coronaviruses including the SARS-CoV-2. It has ever been modulated by macromolecules including the ADP-ribose until Ni and co-workers recently reported its inhibition and plasticity enhancement unprecedentedly by remdesivir metabolite, GS-441524, creating an opportunity for investigating other biodiverse small molecules such as β-Carboline (βC) alkaloids. In this study, 1497 βC analogues from the HiT2LEAD chemical database were screened, using computational approaches of Glide XP docking, molecular dynamics simulation and pk-CSM ADMET predictions. Selectively, βC ligands, 129, 584, 1303 and 1323 demonstrated higher binding affinities to the receptor, indicated by XP docking scores of –10.72, –10.01, –9.63 and –9.48 kcal/mol respectively than remdesivir and GS-441524 with –4.68 and –9.41 kcal/mol respectively. Consistently, their binding free energies were –36.07, –23.77, –24.07 and –17.76 kcal/mol respectively, while remdesivir and GS-441524 showed –21.22 and –24.20 kcal/mol respectively. Interestingly, the selected βC ligands displayed better stability and flexibility for enhancing the plasticity of the receptor than GS-441524, especially 129 and 1303. Their predicted ADMET parameters favour druggability and low expressions for toxicity. Thus, they are recommended as promising adjuvant/standalone anti-SARS-CoV-2 candidates for further study.Key words: SARS-CoV-2, nsp3 macrodomain, ADP-ribose, β-carboline, bioinformatics, drug design     

Inadvertent gene silencing of argininosuccinate synthase (bcass1) in Botrytis cinerea by the pLOB1 vector system

For several years, researchers working on the plant pathogen Botrytis cinerea and a number of other related fungi have routinely used the pLOB1 vector system, based on hygromycin resistance, under the control of the Aspergillus nidulans oliC promoter and what was reported to be the β‐tubulin (tubA) terminator. Recently, it has been demonstrated that this vector contains a 446‐bp portion of the B. cinerea argininosuccinate synthase gene (bcass1) rather than the tubA terminator. As argininosuccinate synthase is essential for the production of l ‐arginine, inadvertent gene silencing of bcass1 may result in partial l ‐arginine auxotrophy and, indeed, may lead to altered phenotypes in planta. In this article, we report our findings relating to possible problems arising from this incorrect plasmid construction. As an absolute baseline, gene disruption of bcass1 was carried out and generated a strict auxotroph, unable to grow without exogenous arginine supplementation. The knockout displayed an alteration in host range in planta, showing a reduction in pathogenicity on strawberries, French bean leaves and tomatoes, but maintained wild‐type growth on grape, which is in accordance with the reported arginine availability in such tissues. Deliberate gene silencing of bcass1 mirrored these effects, with strongly silenced lines showing reduced virulence. The degree of silencing as seen by partial auxotrophy was correlated with an observed reduction in virulence. We also showed that inadvertent silencing of bcass1 is possible when using the pLOB1 vector or derivatives thereof. Partial arginine auxotrophy and concomitant reductions in virulence were triggered in approximately 6% of transformants obtained when expressing enhanced green fluorescent protein, luciferase, monomeric red fluorescent protein or β‐glucuronidase using the pLOB1‐based expression system, which inadvertently contains 446 bp of the bcass1 coding sequence. We recommend the testing of transformants obtained using this vector system for arginine auxotrophy in order to provide assurance that any observed effects on the development or virulence are a result of the desired genetic alteration rather than accidental bcass1 silencing.     

Complete mitochondrial genome of threatened mahseer <Emphasis Type="Italic">Tor tor</Emphasis> (Hamilton 1822) and its phylogenetic relationship within Cyprinidae family

The mahseers (Tor, Neolissochilus and Naziritor) are an important group of fishes endemic to Asia with the conservation status of most species evaluated as threatened. Conservation plans to revive these declining wild populations are hindered by unstable taxonomy. Molecular phylogeny studies with mitochondrial genome have been successfully used to reconstruct the phylogenetic tree and to resolve taxonomic ambiguity. In the present study, complete mitochondrial genome of Tor tor has been sequenced using ion torrent next-generation sequencing platform with coverage of more than 1000 ×. Comparative mitogenome analysis shows higher divergence value at ND1 gene than COI gene. Further, occurrence of a distinct genetic lineage of T. tor is revealed. The phylogenetic relationship among mahseer group has been defined as Neolissochilus hexagonolepis ((T. sinensis (T. putitora, T. tor), (T. khudree, T. tambroides)).     

Tracer studies with 32P on the distribution of functional roots of Arabica coffee in Kenya

Seven experiments were done over a period of 21 months at Ruiru, Kenya, using 4 mCi/tree of ³²P tracer to study the distribution of functional roots of mature Arabica coffee trees. Tracer was placed at sixteen equally spaced sites around individual trees at one of five depths (to 180 cm) and three distances (to 135 cm = mid-row). Thereafter, samples of three-leaf shoot tips were collected at 14-day intervals for up to seven occasions and the radioactivity assessed after drying and ashing. There was negligible activity at any time at 180 cm depth but at other depths, and at all three distances, the relative level of activity changed markedly with season. After prolonged drought relatively high root activity was found at mid-depth, near to the trunk; after the soil was re-wetted by rain most root activity occurred in the topsoil at the quarter-row distance; after the soil profile had been wet for some time there was a more general distribution of functional roots. Some departures from this general scheme are discussed as are practical implications and the need for further investigations.