Enhanced exudation of malate in the rhizosphere due to AtALMT1 overexpression in blackgram (Vigna mungo L.) confers increased aluminium tolerance.

Worldwide, 50% of soil is acidic, which induces aluminium (Al) toxicity in plants, as the phyto-availability of Al3+ increases in acidic soil. Plants responds to Al3+ toxicity by exuding organic acids into the rhizosphere. The organic acid responsible for Al3+ stress response varies from species to species, which in the case of blackgram (Vigna mungo L.) is citrate. In blackgram, an Arabidopsis malate transporter, AtALMT1, was overexpressed with the motive of inducing enhanced exudation of malate. Transgenics were generated using cotyledon node explants through Agrobacterium tumefaciens-mediated transformation. The putative transgenics were initially screened by AtALMT1-specific genomic DNA PCR, followed by quantitative PCR. Two independent transgenic events were identified and functionally characterized in the T3 generation. The transgenic lines, Line 1 and 2, showed better root growth, relative water content and chlorophyll content under Al3+ stress. Both lines also accounted for less oxidative damage, due to reduced accumulation of ROS molecules. Photosynthetic efficiency, as measured in terms of Fv /Fm , NPQ and Y(II), increased when compared to the wild type (WT). Relative expression of genes (VmSTOP1, VmALS3, VmMATE) responsible for Al3+ stress response in blackgram showed that overexpression of a malate transporter did not have any effect on their expression. Malate exudation increased whereas citrate exudation did not show any divergence from the WT. A pot stress assay found that the transgenics showed better adaptation to acidic soil. This report demonstrates that the overexpression of a malate transporter in a non-malate exuding species improves adaptation to Al3+ toxicity in acidic soil without effecting its stress response mechanism.

Selection of salt tolerant plants of Nicotiana tabacum L. through in vitro and its biochemical characterization.

Sodium chloride tolerant organogenic callus lines of Nicotiana tabacum were developed in vitro on Murashige and Skoog [16] medium supplemented with BA, IAA and different concentration of NaCl. The maximum shoot bud regeneration was achieved from both tolerant and non-tolerant calluses on MS medium supplemented with 1.0 mg/l BA, 0.1 mg/l IAA with or without NaCl within 4 weeks of culture. Standard growth parameters such as fresh weight and dry weight of organogenic callus, growth tolerant index and enzyme activity (peroxidase and catalase) were used as indicators of salt tolerance. The growth tolerance index in the 4-week after the beginning of treatments yielded significant differences among the non-tolerant and tolerant organogenic callus lines. The regenerated shoots were rooted on half-strength MS basal salts supplemented with 2% sucrose but devoid of growth regulator. The regenerated plants from tolerant callus lines were capable of growing in vitro in presence of 175 mM NaCl. SDS-PAGE profile showed that the progenies derived from tolerant sources were tolerant to salt. This investigation may help in the selection and characterization of salt tolerance in plant improvement programme.

Tissue culture of ornamental pot plant: a critical review on present scenario and future prospects.

Recent modern techniques of propagation have been developed which could help growers to meet the demand of the horticultural industry in the next century. An overview on the in vitro propagation via thin cell layer, meristem culture, regeneration via organogenesis and somatic embryogenesis is presented. Available methods for the transfer of genes could significantly simplify the breeding procedures and overcome some of the agronomic and environmental problems, which other wise would not be achievable through conventional propagation methods. The development and remarkable achievements with biotechnology in ornamental pot plants made during the three decades have been reviewed. The usefulness of the pot plants in commercial industry as well as propagation techniques, screening for various useful characteristics and selection of somaclonal variation is also discussed.

Onset of in vitro rhizogenesis response and peroxidase activity in Zingiber officinale (Zingiberaceae)

The induction of rooting in microshoots of Zingiber officinale cvs. Suprava, Turia local, Suruchi and V3S18 was achieved on half-strength basal Murashige and Skoog's medium supplemented with 0.5-1.0 mg/l either indole-3-acetic acid (IAA) or indole-3-butyric acid (IBA) and 2 (w/v) sucrose within 7-9 days of culture. Rooting was inhibited when the microshoots were cultured under higher concentration of auxins. The microshoots cultured on medium supplemented with NAA induced large number of thin root hairs with friable calluses within 6-7 days. Peroxidase activity was determined during root induction (0-day to the 10th day at every 2 day interval) from microshoots derived in vitro. The activity was minimum in the inductive phase (primary) and at the maximum level during the root initiative phase. These finding may be useful in monitoring the rooting behaviour in microshoots derived from different subculture and peroxidase activity as a marker for root initiation.

In vitro micropropagation of Lawsonia inermis (Lythraceae)

A successful protocol was developed for mass propagation of Lawsonia inermis Linn., an important medicinal plant. Multiple shoots were induced in apical and axillary meristems derived from mature explants of L. inermis on Murashige and Skoog (1962) medium supplemented with 0.25 mg/l 6-benzylaminopurine (BA), 0.25 mg/l Kinetin (Kn), 0.5 mg/l ascorbic acid and 3 (w/v) sucrose. The rate of multiplication was higher when the cultures were incubated under continuous light rather than the 14 hr photoperiod. Rooting was readily achieved upon transferring the microshoots onto MS basal semi-solid medium supplemented with 0.25 mg/l indole-3-butyric acid (IBA) after ten days of culture. Micropropagated plantlets were acclimatized and successfully grown in soil.

Heavy metal and nutrient concentration in soil and plants growing on a metalliferous chromite minespoil.

Metal contamination in soil and plant samples from a chromite mine and its adjoining regions was determined. The metal concentration varied in stem, leaf and root of different tree species. In the case of shrubs, the highest concentration of iron (18.5 mg kg(-1) was detected in the stem of Combretum roxburghii. The concentration of aluminium varied from 1.8 - 5.3 mg kg(-1) dry weight, whereas the nickel content was found to be the highest in the stem of Calotropis gigantea. In the case of herbs, chromium concentration was highest (60.9 mg kg(-1) dry weight) in Evovulus alsenoides and the lowest (18.8 mg kg(-1) dry weight) in Andrographis paniculata. There was a significant correlation observed between chromium in soil with the root of tree species like Lagerstroemia parviflora, Madhuca longifolia, Anogeissus latifolia and Haldina cordyfolia. Nickel in soil was significantly correlated with the stem and leaf of all the tree species except Chlroxylon sweitenta. Iron in soil showed correlation with the stem and leaf of Chloroxylon sweitenia. Among the shrubs (Calotropis gigantea, Combretum roxburghii and Smilax zeylancia), chromium in soil showed a correlation with the root. Nickel in soil was positively correlated with the stem and leaf of Calotropis gigantea and Combretum roxburghii. Among the herbs, chromium in the whole plant of Evolvulus alsenoids, Solanum surattense and Phyllanthus fraternus showed significant positive correlation with soil; nickel in Solanum surattense showed significant positive correlation with soil. The positive correlation coefficient was observed between iron in the whole plant and soil on Phyllanthus virgatus, Phyllanthus fraternus and Andrographis paniculata. The above information would be useful for the establishment of a vegetation cover on the minewaste heaps.

In vitro micropropagation of Lawsonia inermis (Lythraceae).

A successful protocol was developed for mass propagation of Lawsonia inermis Linn., an important medicinal plant. Multiple shoots were induced in apical and axillary meristems derived from mature explants of L. inermis on Murashige and Skoog (1962) medium supplemented with 0.25 mg/l 6-benzylaminopurine (BA), 0.25 mg/l Kinetin (Kn), 0.5 mg/l ascorbic acid and 3% (w/v) sucrose. The rate of multiplication was higher when the cultures were incubated under continuous light rather than the 14 hr photoperiod. Rooting was readily achieved upon transferring the microshoots onto MS basal semi-solid medium supplemented with 0.25 mg/l indole-3-butyric acid (IBA) after ten days of culture. Micropropagated plantlets were acclimatized and successfully grown in soil.

Onset of in vitro rhizogenesis response and peroxidase activity in Zingiber officinale (Zingiberaceae).

The induction of rooting in microshoots of Zingiber officinale cvs. Suprava, Turia local, Suruchi and V3S18 was achieved on half-strength basal Murashige and Skoog's medium supplemented with 0.5-1.0 mg/l either indole-3-acetic acid (IAA) or indole-3-butyric acid (IBA) and 2% (w/v) sucrose within 7-9 days of culture. Rooting was inhibited when the microshoots were cultured under higher concentration of auxins. The microshoots cultured on medium supplemented with NAA induced large number of thin root hairs with friable calluses within 6-7 days. Peroxidase activity was determined during root induction (0-day to the 10th day at every 2 day interval) from microshoots derived in vitro. The activity was minimum in the inductive phase (primary) and at the maximum level during the root initiative phase. These finding may be useful in monitoring the rooting behaviour in microshoots derived from different subculture and peroxidase activity as a marker for root initiation.

Effects of chromium and nickel on germination and growth in tolerant and non-tolerant populations of Echinochloa colona (L.) Link.

The tolerance of populations of a grass, Echinochloa colona, growing abundantly on chromite minewaste dumps, was tested in two separate experiments. Seed-based experiments indicate that the populations growing naturally on uncontaminated sites, germinated better in nutrient solutions without metal than those collected from minewaste dumps. Metal tolerance indices were greater in the plant populations derived from metal contaminated sites and better growth of these plants was noted on mine spoil soil-mix in the ratio of 1:1; the percentage of seed germination and the rate of seedling growth, however, declined in a soil compost containing 25% mine spoil and 75% uncontaminated (control) soil. Populations of Echinochloa colona occurring naturally on chromite mine spoils, therefore, appear to have developed metal tolerance. It is maintained by a balanced and stable genetic system built up and adjusted by natural selection. Such material is very suitable to be used in restoration work designed to produce an effective vegetation cover to improve the derelict land and to reduce erosion. This finding might be useful in revegetation programmes on metalliferous minewastes.

In vitro manipulation and propagation of medicinal plants.

Well developed techniques are currently available to help growers meet the demand of the pharmaceutical industry in the next century. These protocols are designed to provide optimal levels of carbohydrates, organic compounds (vitamins), mineral nutrients, environmental factors (e.g. light, gaseous environment, temperature, and humidity) and growth regulators required to obtain high regeneration rates of many plant species in vitro and thereby facilitate commercially viable micropropagation. Well-defined cell culture methods have also been developed for the production of several important secondary products. An overview of the regeneration of medicinal plants by direct and indirect organogenesis and by somatic embryogenesis from various types of explants is presented, and the use of these techniques combined with other biotechnological approaches to improve medicinal plants through somaclonal variation and genetic transformation is reviewed.

Studies on cadmium toxicity in plants: a review.

This review emphasises cadmium toxicity on plants with regards to ecological, physiological and biochemical aspects. Cadmium toxicity in plants and problems concerning tolerance and ecological performance are discussed briefly. Efforts have been made to compare the relative sensitivity of various plant groups including micro-and macro-flora. This review may help in interdisciplinary studies to assess the ecological significance of metal stress.

In vitro somatic embryogenesis ofDalbergia sissoo Roxb. -a multipurpose timber-yielding tree.

Somatic embryogenesis was achieved in callus cultures dervied from 40-day-old semimature zygotic embryos ofDalbergia sissoo on semi-solid Murashige and Skoog (MS) salts and vitamins supplemented with 0.46-1.16 µM kinetin, 6.78-9.04 µM 2,4-dichlorophenoxy acetic acid (2,4-D) and 30 g/1 sucrose. Somatic embryos proliferated rapidly by secondary somatic embryogenesis after transfer to half-strength basal MS medium supplemented with 0.46-1.16 µM kinetin and 6.78-9.04 µM 2,4-D with 2% (w/v) sucrose. The light-green somatic embryos germinated on half-strength MS salts and vitamins supplemented with 0.5 mg/1 abscisic acid and 2% (w/v) sucrose. The developmental stages of somatic embryogenesis were studied by light and scanning electron microscopy.

In vitro somatic embryogenesis from callus culture of the timber yielding treeHardwickia binata Roxb.

Somatic embryogenesis was achieved in callus cultures derived from immature cotyledonary explants ofHardwickia binata Roxb., a multipurpose leguminous tree, on semisolid modified Murashige and Skoog's (mMS) medium containing 2900 mg/l potassium nitrate (KNO3) supplemented with 4.64 µM kinetin (Kn) and 5.37µM a-naphthaleneacetic acid (NAA). Somatic embryos proliferated rapidly after transfer to MS basal medium supplemented with 2052.6 µM L-glutamine and 0.084 µM gibberellic acid (GA3). Maturation of somatic embryos was achieved on half-strength MS basal medium supplemented with 1.23 µM IBA and 2% (w/v) sucrose. Histological studies confirmed different developmental stages of somatic embryogenesis inHardwickia binata.

Somatic embryogenesis and in vitro flowering of 3 species of bamboo.

Plant regeneration via somatic embryogenesis was achieved in callus cultures derived from nodal explants of in vitro grown seedlings and excised mature zygotic embryos of three bamboo species on Murashige and Skoog's (MS) basal medium supplemented with 0.5 mg/l kinetin (Kn), 2.0 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D), 10 mg/l adenine sulphate (Ads) and 3% (w/v) sucrose incubated in the light or in the dark. Somatic embryos germinated (95-98%) into normal plants and were transferred to soil with 95% success. In vitro flowering was induced on shoots developed from nodal explants taken from somatic embryo regenerated plants of Bambusa vulgaris, Dendrocalamus giganteus and Dendrocalamus strictus on half-strength MS basal medium supplemented with 0.25 mg/l indole-3-butyric acid (IBA), 0.5 mg/l Ads, 0.5 mg/l gibberellic acid (GA3) and 3% sucrose.

Micropropagation of Madhuca longifolia (Koenig) MacBride var. latifolia Roxb.

Bud break and multiple shoots were induced in apical and axillary meristems derived from 10-d old seedlings of Madhuca longifolia var. latifolia on Murashige and Skoog (MS) medium supplemented with 1.0 mg/l N(6)-benzyladenine (BA) singly or in combinatiobn with 1-naphthalene acetic acid (NAA), indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA). Excised shoots were rooted on half-strength MS with IBA (1.0 mg/l) after 18d of culture. Regenerated plantlets were acclimatized and successfully transferred to soil.