Secondary wall deposition in tracheary elements of cucumber grown in vitro

It is a matter of controversy whether secondary wall deposition is dependent on lignification during the development of tracheary elements. To understand this, tracheary element differentiation was studied in the homogeneous calli obtained from the cotyledonary explants of Cucumis sativus subsequent to treatment with plant growth regulators, such as naphthalene acetic acid (NAA) and benzylamino purine (BAP), which are necessary for the induction of tracheary elements, along with metabolic blockers such as 2-aminoindan-2-phosphonic acid (AIP), 2,3,5-triiodobenzoic acid (TIBA) and nifedipine. Calli treated with AIP, a potential inhibitor of L-phenylalanine ammonia-lyase (PAL), have no PAL activity at any time during the culture period. There was a complete inhibition of lignification although secondary wall deposition was unaltered. Similar results were obtained using TIBA, an inhibitor of auxin transport, and nifedipine, a known calcium channel blocker. Thus the present study suggests that secondary wall deposition in the course of tracheary element differentiation need not to be dependent on lignification. This revised version was published online in July 2006 with corrections to the Cover Date.     

The secretory apparatus of Xerophyta viscosa (Velloziaceae): Epidermis anatomy and chemical composition of the secretory product

In this study, secretory activity on the adaxial surface of the leaves of the desiccation tolerant plant, Xerophyta viscosa Baker was investigated, using light, scanning and transmission electron microscopy. Glandular activity was associated with sunken cavities which appear to be modifications of infolded epidermal cells. The secretory cavity consisted of a globose lumen surrounded by two layers of cells. The cells of the outer layer were flattened with thickened walls, while those of the inner layer (epithelium), exhibited ultrastructural features of intense metabolic activity. Epithelial cells were larger with a large nucleus, numerous vacuoles, and a dense cytoplasm with abundant mitochondria, endoplasmic reticulum (ER), plastids and a few dictysomes. Lipophilic droplets were abundant in the cytoplasm, mitochondria, ER, plastids and in infolding sites of the plasmalemma outside the protoplast. ER appeared to be involved in the synthesis and transport of lipophilic substances. The mechanism of secretion in X. viscosa appeared to be granulocrine. The chemical composition of hexane and methanol extracts of the leaves, analysed by gas chromatography coupled with mass spectrometry (GC–MS), revealed the presence of diterpenes, phenolic compounds and fatty acids. Compounds in the hexane fraction included velloziolone, manoyl oxide, asperdiol and an unknown substance. Compounds in the methanol fraction included 5-(hydroxymethyl)-2-furancarboxaldehyde, 2,6-dimethoxyphenol, 4-(3-hydroxy-1-propenyl)-2-methoxyphenol and stearic acid.     

Molecular profiling of fungal assemblages in the healthy and infected roots of <Emphasis Type="Italic">Decalepis arayalpathra</Emphasis> (J. Joseph &amp; V. Chandras) Venter,an endemic and endangered ethnomedicinal plant from Western Ghats,India

Decalepis arayalpathra, an endangered, endemic ethnomedicinal plant from southern Western Ghats, India, is targeted for its aromatic and medicinal properties. This study aimed at to identify fungal endophyte populations associated with healthy and diseased roots of this perennial shrub. Healthy and rotted root samples of D. arayalpathra were collected, fungal endophytes assemblages were identified both by culture-dependent and culture-independent approaches, further sequenced and the retrieved sequences were analysed with the reference sequences in GenBank to know their phylogenetic relationships. Analysis of the ITS rDNA region generated 24 different Ascomycota and three Basidiomycota taxa. Trichoderma sp. was most abundant in healthy and diseased root samples, while Penicillium and Aspergillus were confined to healthy roots. Furthermore, Fusarium solani, Fusarium oxysporum and Mucor velutinosus were found to be the most frequent fungi identified from the rotted root samples, thus substantiated to be the cause for D. arayalpathra decline in the wild. Interestingly, the strains assigned to Fusarium sp. were isolated from diseased roots showing typical clearly visible symptoms, such as a severe brown discolouration on the taproot. Molecular profiling of all the pure fungal isolates, viz., Trichoderma, Penicillium, Aspergillus, Fusarium and Mucor, revealed high sequence similarities (≥ 98 %) to corresponding reference sequences. Sequencing of Trichoderma pure cultures isolated from healthy and diseased roots revealed sequence similarities to Trichoderma harzianum, T. hamatum, T. koningiopsis, T. asperellum, T. pubescens and Hypocrea sp. This confirms the morphological examinations, as Hypocrea is the teleomorph stage of Trichoderma sp. This study signifies the first work pertaining to the taxonomy of the fungal endophytic community of D. arayalpathra, and the results reported in this work may help to ascertain the cause of root rot disease often perceived in D. arayalpathra. Also, it could be useful to identify the promising endophytic communities against the root rot diseases occurring in D. arayalpathra.     

Micropropagation of <Emphasis Type="Italic">Harpagophytum procumbens</Emphasis>

An efficient protocol for micropropagation of Harpagophytum procumbens DC., an endangered African medicinal plant, was developed. Maximum shoot multiplication without callus was obtained from nodal explants cultured on Murashige and Skoog (MS) basal salts plus Gamborg’s (B₅) vitamins supplemented with 0.1 mg dm⁻³ indole-3-acetic acid and 5.0 mg dm⁻³ kinetin. The shoots were subsequently subcultured every 3 weeks on the same medium. Detached axillary shoots were transferred to MS basal salts plus B₅ vitamins supplemented with various concentrations of α-naphthalene-acetic acid or indole-3-butyric acid (IBA), ranging from 0.5 to 2.5 mg dm⁻³ and 100 % rooting and optimal subsequent acclimatization was achieved on 1.0 mg dm⁻³ IBA. After 4 weeks of culture, the rooted shoots (>5 cm) were planted in pots containing peat, vermiculite and bark (2:1:1), covered with plastic domes and maintained at 25 °C for 2 weeks before being transferred to a glasshouse. Plant survival was about 40 %.     

Quantitative trait loci (QTLs) for water use and crop production traits co-locate with major QTL for tolerance to water deficit in a fine-mapping population of pearl millet (<Emphasis Type="Italic">Pennisetum glaucum</Emphasis> L. R.Br.)

Key message

Four genetic regions associated with water use traits, measured at different levels of plant organization, and with agronomic traits were identified within a previously reported region for terminal water deficit adaptation on linkage group 2. Close linkages between these traits showed the value of phenotyping both for agronomic and secondary traits to better understand plant productive processes.

Abstract

Water saving traits are critical for water stress adaptation of pearl millet, whereas maximizing water use is key to the absence of stress. This research aimed at demonstrating the close relationship between traits measured at different levels of plant organization, some putatively involved in water stress adaptation, and those responsible for agronomic performance. A fine-mapping population of pearl millet, segregating for a previously identified quantitative trait locus (QTL) for adaptation to terminal drought stress on LG02, was phenotyped for traits at different levels of plant organization in different experimental environments (pot culture, high-throughput phenotyping platform, lysimeters, and field). The linkages among traits across the experimental systems were analysed using principal component analysis and QTL co-localization approach. Four regions within the LG02-QTL were found and revealed substantial co-mapping of water use and agronomic traits. These regions, identified across experimental systems, provided genetic evidence of the tight linkages between traits phenotyped at a lower level of plant organization and agronomic traits assessed in the field, therefore deepening our understanding of complex traits and then benefiting both geneticists and breeders. In short: (1) under no/mild stress conditions, increasing biomass and tiller production increased water use and eventually yield; (2) under severe stress conditions, water savings at vegetative stage, from lower plant vigour and fewer tillers in that population, led to more water available during grain filling, expression of stay-green phenotypes, and higher yield.

     

<Emphasis Type="Italic">In vitro</Emphasis> micropropagation of three rare,endangered, and endemic rhododendron species of Northeast India

In vitro propagation of three rare, endangered and endemic rhododendron species—Rhododendron dalhousiae var. rhabdotum, R. elliottii, and R. johnstoneanum—was attained. Nodal explants were used for multiple shoot induction studies. Three cytokinins (isopentenyladenine, benzyladenine, and kinetin) were evaluated in all three species. Isopentenyladenine performed better in all three species, especially at the concentration of 39.36 μM. Testing of combinations of growth regulators revealed that explants grown on Anderson medium supplemented with 39.36 μM isopentenyladenine and 4.90 μM indole butyric acid gave optimum results with 100% multiple shoot induction, 18 shoots per explant, and shoot length of 2.07 cm with R. johnstoneanum. Indole butyric acid was found to be the best auxin for root formation. Around 60% of the in vitro-raised plants of R. elliottii, R. johnstoneanum, and R. dalhousiae var. rhabdotum were able to establish ex vitro.