Assessment of direct shoot organogenesis and genetic fidelity in Solanum viarum Dunal—a commercially important medicinal plant

Solanum viarum Dunal is an important medicinal plant with a high quantity of steroidal alkaloids used for the synthesis of contraceptives, corticosteroids, and sex hormones. It is also used by Indian tribal people for the treatment of leprosy, toothache, and diabetes. Therefore, to meet the existing needs for this plant, it is necessary to develop an efficient regeneration system useful for rapid and large-scale clonal propagation with ensured genetic fidelity. An efficient and improved regeneration protocol for prickly and prickleless genotypes of S. viarum has been developed using three explants, leaf, petiole, and internodes, under the influence of two plant growth regulators, thidiazuron (TDZ) and 6-benzyladenine (BA). Effects of genotype, explant type, and concentrations of TDZ and BA were studied. A higher percentage of shoot organogenesis (78.25% ± 2.53) and shoot number per explant (6.79 ± 1.04) were achieved in the leaf segments of prickly genotype cultured on modified Murashige and Skoog (MS) medium supplemented with TDZ (1.50 mg L⁻¹). Furthermore, basal leaf segments showed 100% regeneration from the prickly genotype. A significantly higher content of total phenolics was quantified in prickleless (3.66 μg mg⁻¹) than prickly genotypes (2.73 μg mg⁻¹). The monomorphic banding pattern of random amplified polymorphic DNA (RAPD) and simple sequence repeats (SSR) analysis confirmed the genetic fidelity of the regenerated plants. Additionally, flow cytometric analysis of regenerants showed no variation in the ploidy levels when compared to the mother (control) plants. These results clearly depicted the efficiency of developed protocol that can be utilized for generating genetically stable population of S. viarum.

     

Effects of NO3– and BAP on organogenesis in tissue-cultured shoot primordia induced from shoot apices of Utricularia praelonga St. Hil.

The effects of NO₃ ^– and BAP on organogenesis in shoot primordia of Utricularia praelonga subcultured in B5 liquid medium were studied. In B5 liquid basal medium supplemented with 24.73 mM KNO₃ and 2.0 mg/l BAP the subcultured shoot primordia continuously multiplied into numerous small, globular masses, while with dilution of the KNO₃ to 3 mM organogenesis was promoted. Pulse treatment of the shoot primordia with 3 mM KNO₃ in B5 liquid medium for 72 h and then transplantation to the B5 basal liquid-medium induced meristemoids in this tissue. When the shoot primordia regenerated meristemoids, they never reverted back into the proliferation cycle. The addition of BAP in the B5 liquid medium with 3 mM KNO₃ regulated the differentiation rate of the stems and leaves in the meristemoids induced in the masses of shoot primordia. The control produced 3 parts stems to 1 part leaves; medium with 0.02 mg/l BAP regenerated approximately 2 parts stems and 1 part leaves; that of 0.20 mg/l BAP 1 part stems and 2 parts leaves; and medium with 2.00 mg/l BAP regenerated leaves only. Received: 17 September 1997 / Revision received: 30 October 1997 / Accepted: 18 November 1997     

A filter paper-based liquid culture system for citrus shoot organogenesis—a mixture-amount plant growth regulator experiment

This study determined the effects of a static liquid culture system on shoot regeneration from citrus epicotyl explants. A mixture-amount experiment was used to determine the effects of zeatin riboside (ZR), 6-benzylaminopurine (BA), and indole-3-acetic acid (IAA) on two citrus types—citrange (Citrus sinensis ‘Washington’ L. Osbeck. × Poncirus trifoliata L. Raf var. Carrizo) and sweet orange (Citrus sinensis L. Osbeck var. ‘Ridge Pineapple’). A liquid culture system comprising a Petri dish, cellulose filter paper, and liquid culture medium was used. Shoot regeneration experiments were conducted over 6 wk that included 2 wk in the dark followed by 4 wk in the light. Three responses were measured: (1) number of explants forming buds and/or shoots, (2) number of explants with shoots >?2 mm, and (3) overall explant and shoot quality. The effects of paper disc number, liquid medium volume, and explant size on shoot regeneration were determined. High-quality shoots were produced from explants cultured in 5.25 to 12 mL medium volume and explant sizes ranging from 2 to 15 mm. The effects of the plant growth regulators were similar for the two citrus types and were as follows: (1) use of ZR or BA resulted in high-quality shoot production; (2) ZR and BA were not synergistic; (3) culture in 20 μM ZR resulted in the highest shoot production; (4) BA and IAA were strongly synergistic, with the greatest production with BA when IAA was included in the mixture; and (5) ZR and IAA were antagonistic, particularly with Ridge Pineapple.     

Induction of direct shoot organogenesis and in vitro flowering from shoot tip explants of cucumber (Cucumis sativus L. cv. ‘Green long’)

In vitro flowering is an alternative breeding tool for generating hybrid Cucumis spp. as it is able to overcome limitations caused by interspecific incompatibility. The present study describes an efficient method for induction of multiple shoots and in vitro flowering from shoot tip explants of cucumber (Cucumis sativus L.). Shoot tip explants were excised from 7-day-old seedlings and cultured on Murashige and Skoog (MS) medium fortified with different concentrations of 6-benzylaminopurine (BAP; 0.5–2.5 mg/L) alone or in combination with 0.5 mg/L kinetin (KIN). The highest frequency (93.1%) of multiple shoot formation with maximum number of shoots (15.2 shoots/explant) was achieved on MS medium supplemented with 1.0 mg/L BAP. For in vitro flowering, shoots were cultured on MS medium supplemented with 0.5 mg/L BAP and different concentrations of sucrose. Flowering occurred on about 95% of in vitro shoots cultured on MS medium fortified with 6% (w/v) sucrose and 0.5 mg/L BAP after 15 d. For rooting, shoots (>2 cm) were cultured on MS medium augmented with various concentrations of indole-3-butyric acid (IBA; 0.5–2.5 mg/L) alone or in combination with 0.5 mg/L KIN. Among the combinations tested, supplementation with IBA (1.5 mg/L) and KIN (0.5 mg/L) induced maximum rooting rates (95.4%) with 7.8 roots/shoot. Rooted plantlets were successfully transferred into plastic cups containing a mixture of soil and sand (1:1), established in the greenhouse, and subsequently acclimatized in the field. The in vitro flowering reported in this study may facilitate rapid hybridization in Cucumis species and offers a model system for studying the physiological mechanisms involved in flowering.     

Rapid in vitro differentiation of Sesbania bispinosa plants — a leguminous shrub

Multiple shoots differentiated from hypocotyl explants of Sesbania bispinosa (Jacq.) W.F. Wight, a leguminous woody shrub, when cultured on Gamborg's basal medium alone or in combination with 6-benzyl aminopurine (10^–7–10^–4 M). For cotyledonary explants 6-benzyl aminopurine (10^–6–10^–4 M) was necessary. The shoots rooted when cultured on Gamborg's basal medium containing indole-3-butyric acid (10^-5 M). Plantlets thus formed were transferred to soil where they have flowered and also set fruits.     

Development of shoots and roots in anther-derived callus of Azadirachta indica A. Juss.—a medicinal tree

Callus originated in microsporangial wall layers and connective tissues of anthers containing uninucleate microspores on Nitsch's or Murashige and Skoog's medium supplemented with growth regulators. A higher percentage of cultures (43) produced callus on Nitsch's medium containing 10 M indole-3-acetic acid + 1 M 6-benzyladenine. After 13–15 weeks, green nodular structures and prominent roots developed in 25% of the cultures on Murashige and Skoog's medium + 10 M -naphthaleneacetic acid + 1 M kinetin. Multiple shoots were induced in this anther-derived callus when subcultured on Murashige and Skoog's medium augmented with 4.44 M 6-benzyladenine + 0.53 M -naphthaleneacetic acid along with 18.75 M polyvinylpyrrolidone. The excised shoots formed roots after subculturing on Murashige and Skoog's medium + 4.90 M indole-3-butyric acid + 18.75 M polyvinylpyrrolidone, thus developing complete plantlets. Examination of callusing anthers also revealed two- to multi-celled pollen masses with intact exine.Abbreviations BA 6-benzyladenine - CW coconut water - 2,4-d 2,4-dichlorophenoxyacetic acid - GA₃ gibberellic acid - HCl hydrochloric acid - IAA indole-3-acetic acid - IBA indole-3-butyric acid - KMnO₄ potassium permanganate - MS Murashige & Skoog's medium - NAA -naphthaleneacetic acid - NB Nitsch's medium - PVP polyvinylpyrrolidone     

Enhanced phytostabilization of cadmium by a halophyte—Acanthus ilicifolius L.

Heavy metal pollution in mangrove wetlands has become a growing matter of concern as it serves as sink and source for toxic heavy metals including cadmium (Cd). The present study evaluates the phytostabilization potential of a halophyte, Acanthus ilicifolius L., toward Cd under hydroponic culture conditions. Accumulation, translocation, and effects of Cd on the antioxidant system of A. ilicifolius were studied. Results indicated that A. ilicifolius accumulated Cd mainly in roots (96.4%) as compared to stem (1.4%) and leaves (0.6%) and the accumulated Cd is retained in root rather than being translocated to shoots as indicated by TF < 0.26. Moreover, malondialdehyde (MDA) content increased upon Cd treatment, which is further detoxified by the enzymatic and nonenzymatic antioxidant mechanism. Antioxidants like proline, ascorbate, and amino acid recorded an increased accumulation in the Cd-treated plants followed by the upregulation of antioxidant enzymes like superoxide dismutase (SOD), catalase (CAT), guaiacol peroxidase (GPX), and ascorbate peroxidase (APX). Therefore, the rate of sugar accumulation was found to be decreased in plants treated with Cd as compared to the control plants. Thus, having relatively high BCF_root (69.3) and low TF_shoot (0.26) values, A. ilicifolius can be suggested as a potential candidate for phytostabilization of Cd in mangrove wetlands.     

Somatic embryogenesis and organogenesis in callus cultures of a tree legume — Albizia richardiana King

Hypocotyl explants of 1 and 10 mm lengths were excised from 12-day-old in vitro-grown seedlings of Albizia richardiana. The larger pieces, after 40 days of culture, developed shoots along with green calli on B₅ + BAP (10^–7–10^–5M), while the smaller segments produced only green calli on B₅+BAP (10^–7–10^–4M) medium. Some of the green calli turned morphogenic and started producing somatic embryos with the 2nd sub-culture and shoots from 7th sub-culture onwards. Calli retained the morphogenic potential even after repeated sub-culturing for over two years. The number of embryos in an embryogenic culture varied from 2 to 20 per callus mass of 5–6.5 cm³. Sucrose at the 2% level in MS medium was optimal for embryogenesis while 4% was optimal for shoot bud differentiation. Higher levels of sucrose (6–10%) caused browning of green calli and also inhibited differentiation into embryos and shoot buds. By selective sub-culturing of 0.1 cm³ pieces of embryogenic calli on MS+10^–5M BAP, 46% of the cultures produced somatic embryos. The latter germinated into plantlets on Knop's medium.Abbreviations BAP 6-benzylaminopurine - B₅ Gamborg et al., 1968 medium - IAA Indole-3-acetic acid - MS Murashige and Skoog's (1962) medium     

Leeches as a source of mammalian viral DNA and RNA—a study in medicinal leeches

Surveillance of wild vertebrates can be challenging in remote and inaccessible areas such as tropical rainforests. Blood-feeding parasites, such as leeches, can facilitate wild vertebrate monitoring by targeting residual DNA from the animals the leeches feed on. Successes in detecting host DNA from leeches suggest that host viruses may also be detectable. To systematically test this hypothesis, we performed a proof of concept study using quantitative PCR (qPCR) to detect DNA viruses (bovine herpesvirus [BHV], human adenovirus [HAdV]) and RNA viruses (influenza A [InfA] and measles morbillivirus [MeV]) from nucleic acids extracted from medicinal leeches fed with blood spiked with each virus. All viruses except BHV showed a gradual decline in concentration from day 1 to 50, and all except BHV were detectable in at least half of the samples even after 50 days. BHV exhibited a rapid decline at day 27 and was undetectable at day 50. Our findings in medicinal leeches indicate that leeches collected in the wild might be an untapped resource for detecting vertebrate viruses and could provide new opportunities to study wildlife viral diseases of rare species in challenging environments, where capturing and handling of animals is difficult.     

Plankton community cycling and recovery after drought — dynamics in a basin on a flood plain

Lake Merrimajeel, a shallow swamp, is part of the Murray-Darling, a flood plain river system with high plankton diversity. It fluctuates drastically in water level and often dries out. The dynamics and seasonality of zooplankton in the face of these fluctuations and the effects of a total drawdown and refilling were monitored between February 1977 and January 1980. During this period thirty species of rotifers, thirty four of microcrustacea and several other species were recorded. Despite erratic fluctuations in water level, temperature and turbidity a community cycle from dominance by rotifers in summer/autumn and crustacea in winter/spring was maintained. The drawdown disturbed this cycle but it was reestablished the following year. Diversity and density reached a maximum just before the drawdown, during hot, turbid conditions. During this drying phase the waters remained relatively fresh and well oxygenated. Phyllopods were not recorded. The recovery phase was characterised by a group of rare species. It is postulated that these rare, colonising or successional species are dependent on a brief flush of rich organic material produced on flooding.The generally depressive effect of increased turbidity does not hold in this lake and many limnetic species were abundant during periods of high turbidity. It is suggested that turbidity may be tolerated or even be advantageous to some species, depending on the nature of the suspended material.The ability of species to lead a fugitive existence by temporarily colonising basins after drawdowns and refilling may be an important factor in producing the high diversity of plankton in flood plain river systems such as the Murray-Darling.     

Silver nanoparticles as an effective stimulant in micropropagation of Panax vietnamensis—a valuable medicinal plant

Plant Cell, Tissue and Organ Culture (PCTOC) - Effects of silver nanoparticles (AgNPs) on somatic embryogenesis and plantlets with rhizome of Panax vietnamensis were presented in this study. The...     

On methods in hare Indian ethnology — a. rejoinder

Prelude to East African History, a collection of papers given at the First East African Vacation School in Pre‐European African History and Archaeology in December 1962, Merrick Posnansky, editor, Student's Library, Oxford University Press, London 1966. 186 pp., 9 s. 6 d.

Ethiopian Music. An Introduction. A Survey of Ecclesiastical and Secular Ethiopian Music and Instruments. Michael Powne. Oxford University Press, London 1966. xix+160 pp., 11 pl., 5 figs., 36 music ex. 1 map. Price 42 s. ‐ 2nd edition. London 1968. Price 45 s.

An Africans Religion. The Spirit of Nyajena. Case History of a Karanga People. Michael Gelfand. Juta &; Co., Limited. Cape Town, Wynberg, Johannesburg 1966. 135 pp. 1 Karte, Illustr. Price 37 s 6 d.

Mkhas Grub Rje's Fundamentals of the Buddhist Tantras. Ferdinand D. Lessing and Alex Wayman. Rgyud sde spyihi rnam par gzag pa rgyas par brjod. The Hague, Paris. 1968.

A Ceremonial Ox of India: The Mithan in Nature, Culture, and History. Frederick J. Simoons. University of Wisconsin Press Ltd., London 1968. 323 pp., pr. 104 s. 6 d.

Dor el Gussa und Gebel ben Ghnema. Zur nachpluvialen Besiedlungsgeschichte des Ostfezzan. Ergebnisse der Frobenius‐Expedition nach libyen 1963/64. Helmut Ziegert. Veröffentlichung des Frobenius‐Institutes Frankfurt. F. Steiner Verlag. Wiesbaden 1967. X+94 S. m. 19 Abb., 203 Taf., 3 Karten. Gebunden. DM 66: ‐.

Archäologische Untersuchungen am Mittleren Amazonas. Beiträge zur Vorgeschichte des südamerikanischen Tieflandes. Peter Paul Hilbert. Marburger Studien zur Völkerkunde I. Dietrich Reimer Verlag, Berlin 1968. 164 Abb., 11 Karten, 337 Seiten. Preis DM 60:‐.

Bibliography of Benin Art. Paula Ben‐Amos. Primitive Art Bibliographies. No. VI. The Museum of Primitive Art. New York 1968, 27 pp.

Anthropological Approaches to the Study of Religion. 1966. 176 pp. Price 30 s.

The Social Anthropology of Complex Societies. 1966. 156 pp. Price 30 s. A.S.A. Monographs 3 and 4. Michael Banton (general editor). Tavistock Publications. London.

Case Studies in Cultural Anthropology. George and Louise Spindler (general editors). Holt, Rinehart and Winston, Inc., New York, Chicago, San Francisco, Toronto, London.

Changing Japan. Edward Norbeck. 1965. 82 pp., 1 map, 1 pl.

The Dusun. A North Borneo Society. Thomas Rhys Williams. 1965. 100 pp., 1 pl., 1 map.

Understanding an African Kingdom: Bunyoro. John Beatty. New York … 1965. 61 pp., 1 map.

Bunyoro. An African Kingdom. John Beatty. New York … 1960. 86 pp., 1 pl., 1 map.     

Spontaneous somatic embryogenesis on in vitro root segment cultures of Rauvolfia micrantha Hook. F.—A rare medicinal plant

Summary Plant regeneration through direct somatic embryogenesis was achieved from root segments derived from in vitro shoots of Rauvolfia micrantha Hook. f. (Apocynaceae) grown for 6 wk in half-strength Murashige and Skoog (MS) medium with 3% sucrose, 100 mgl⁻¹ myo-inositol, and 0.5 mgl⁻¹ α-naphthaleneacetic acid (NAA). The effects of photoperiod and plant growth regulators (PGRs) in half-strength MS medium were studied for the rapid and maximum induction of somatic embryos. The characteristic globular or heart-shaped stages of somatic embryogenesis were not found and cotyledonary stage embryos occasionally appeared without the intervention of callus in total darkness and 16-h photoperiod. Root segments cultured in the medium containing 0.1 mgl⁻¹ NAA and 0.2 mgl⁻¹ 6-benzyladenine (BA) under 16-h photoperiod showed the maximum frequency (39%) of embryogenesis. The frequency of embryo formation was increased to 63% when they were cultured in medium with 0.1 mgl⁻¹ NAA and 0.2 mgl⁻¹ BA in the dark for 4wk, then grown under the 16-h photoperiod. Explants with developing embryos developed into plants after transfer to half-strength MS medium supplemented with 0.1 mgl⁻¹ BA and 0.05 mgl⁻¹ NAA. The well-developed plants were hardened and most plants (80%) survived and were phenotypically similar to the mother plants.     

Nectary structure of Labiatae in relation to their nectar secretion and characteristics in a Mediterranean shrub community — Does flowering time matter?

We studied the interrelation between nectary structure (13 parameters), nectar characteristics (yield, chemical composition), and flower size of 11 Labiatae species in a Mediterranean shrub community near Athens, Greece. We also explored whether the above attributes are affected by the Mediterranean summer drought constraints. Our findings show that among all nectary parameters studied, nectary size and stomatal opening are the most important in (positively) shaping nectar secretion, nectary size being the most meaningful. Nectary structure is correlated to quantity of the nectar secreted, not its quality. Wide flowers bear wide nectaries with large stomatal openings, whereas deep flowers are not related to any nectary size. Corolla size (both length and width) and nectary stomatal opening decrease with flowering time. This applies also to nectary size, nectar volume and sugar content of the perennials (9 species). All above cases of time dependence show that there is a constraint effect of Mediterranean climate on floral and nectary structure, reflected also as a decrease in nectar secretion. Nectary structure in Labiatae is largely shaped by both phylogenetic and climate constraints. On the other hand, although nectar is largely influenced by nectary structure, it is to a large extent ecologically biased, implying that, in addition to phylogeny, there are many other ecological parameters interfering in its secretion such as time within the season, life history, and light requirements.     

HvNax3—a locus controlling shoot sodium exclusion derived from wild barley (Hordeum vulgare ssp. spontaneum)

Previous work identified the wild barley (Hordeum vulgare ssp. spontaneum) accession CPI-71284-48 as being capable of limiting sodium (Na⁺) accumulation in the shoots under saline hydroponic growth conditions. Quantitative trait locus (QTL) analysis using a cross between CPI-71284-48 and a selection of the cultivated barley (H. vulgare ssp. vulgare) cultivar Barque (Barque-73, a moderate Na⁺ excluder) attributed the control of the Na⁺ exclusion trait from CPI-71284-48 to a single locus on the short arm of chromosome 7H, which was named HvNax3. The locus reduced shoot Na⁺ accumulation by 10–25% in plants grown in 150 mM NaCl. Markers generated using colinearity with rice and Brachypodium, together with the analysis of introgression lines and F₂ and F₃ families, enabled HvNax3 to be mapped to a 1.3-cM interval. Genes from the corresponding rice and Brachypodium intervals encode 16 different classes of proteins and include several plausible candidates for HvNax3. The potential of HvNax3 to provide a useful trait contributing to salinity tolerance in cultivated barley is discussed.     

Biochemical detection of a metallo-β-lactamase in carbapenem resistant strain ofStreptomyces sp. CN229 isolated from soil

Streptomyces sp. CN229 was isolated from Tunisia soil. This strain displayed antimicrobial activity against Gram positive and Gram negative bacteria. In addition it is resistant to most β-lactam antibiotics including imipenem and meropenem (MIC imipenem >70 μg/ml). Metallo-β-lactamase (MβL) production was confirmed by either imipenem MIC decrease in the presence of ethylene diamine tetraactic acid (EDTA) or the inhibition zone enhancement around EDTA-impregnated imipenem, or meropenem discs. Isolectric focusing analysis demonstrated the production of β-lactamase with pI of 5.8 that is inhibited by EDTA.Streptomyces sp. CN229 was screened for the imipenem resistance genes,bla _VIM andbla _IMP previously identified inPseudomonas aeruginosa. The presence of these genes was not confirmed by specific PCR analysis. We concluded that carbapenem resistance inStreptomyces sp. CN229 strain is mainly due to production of a novel carbapenemase. Our data show for the first time that MβL is produced byStreptomyces sp. MβL-mediated imipenem and meropenem resistance inStreptomyces is a cause for concern in the study of resistance evolution and antibiotic cluster biosynthetic genes.