Comparative efficiency of phosphate-solubilizing bacteria under greenhouse conditions for promoting growth and aloin-A content of Aloe barbadensis

This study was conducted with Aloe barbadensis in order to investigate the efficacy of four phosphate-solubilizing bacteria (PSB), Pseudomonas synxantha 10223, Burkholderia gladioli 10242, Enterobacter hormaechei 10240 and Serratia marcescens 10241 to solubilize Mussorie rock phosphate (MRP) and to evaluate its effects on growth, soluble P content and P uptake compared with control, i.e. uninoculated plants. Pot experiments were conducted in a greenhouse, in soil supplemented with MRP. Each PSB treatment showed different effects on different plant growth parameters. The maximum increase in leaf length (23.7%), total number of leaves (33.33%) and dry rind weight (69.10%) was observed in plants treated with P. synxantha 10223 compared with control. Whereas, maximum increase in root length (23.43%), fresh leaves weight (79.03%), dry gel weight (113.08%) and total gel volume (112.10%), was observed in plants treated with S. marcescens 10241 compared with uninoculated plants. Maximum increase in aloin-A content [114.92% (per g dry gel weight) and 322.32% (per plant dry gel weight)] was observed in plants treated with P. synxantha 10223 compared with control plants. Root colonization by inoculated PSB as estimated by RAPD technique showed that all PSB were able to survive in the rhizosphere of Aloe plants.     

Improvement of growth and gymnemic acid production by altering the macro elements concentration and nitrogen source supply in cell suspension cultures of Gymnema sylvestre R. Br

Gymnema sylvestre is an important medicinal plant which bears bioactive compound namely gymnemic acids. The present work deals with optimization of cell suspension culture system of G. sylvestre for the production of biomass and gymnemic acid and we investigated effects of macro elements (NH₄NO₃, KNO₃, CaCl₂, MgSO₄ and KH₂PO₄ - 0.0, 0.5, 1.0, 1.5 and 2.0× strength) and nitrogen source [NH₄⁺/NO₃⁻ ratio of: 0.00/18.80, 7.19/18.80, 14.38/18.80, 21.57/18.80, 28.75/18.80, 14.38/0.00, 14.38/9.40, 14.38/18.80, 14.38/28.20 and 14.38/37.60 (mM)] of Murashige and Skoog medium on accumulation of biomass and gymnemic acid content. The highest accumulation of biomass (165.00 g l⁻¹ FW and 15.42 g l⁻¹ DW) was recorded in the medium with 0.5× concentration of NH₄NO₃ and the highest production of gymnemic acid content was recorded in the medium with 2.0× KH₂PO₄ (11.32 mg g⁻¹ DW). The NH₄⁺/NO₃⁻ ratio also influenced cell growth and gymnemic acid production; both parameters were greater when the NO₃⁻ concentration was higher than that of NH₄⁺. Maximum biomass growth (159.72 g l⁻¹ of FW and 14.95 g l⁻¹ of DW) was achieved at an NH₄⁺/NO₃⁻ ratio of 7.19/18.80, and gymnemic acid production was also greatest at the same concentration of NH₄⁺/NO₃⁻ ratio (11.35 mg g⁻¹ DW).     

Distribution of gymnemic acid in various organs of Gymnema sylvestre

The gymnemic acid contents in various organs of Gymnema sylvestre were investigated by High Performance Liquid Chromatography (HPLC) method. The results shows that the content of gymnemic acid in various organs, obviously different, was 54.29, 31.66, 28.82, 27.67, 25.39, 20.56 and 1.31 mg·g⁻¹ DW in shoot tips, flowers, nodes, leaves, internodes, roots and seeds, respectively. The highest gymnemic acid content (54.29 mg·g⁻¹ DW) was found in shoot tip, 1.96 fold higher than that in leaves (27.67 mg·g⁻¹ DW). Maximum quantity of gymnemic acid (35.39 mg·g⁻¹ DW) was observed in the young leaves, which was 1.52 times higher than that in old leaves (23.07 mg·g⁻¹ DW). The content of gymnemic acid in young, middle and old internodes was 26.47, 25.77 and 23.94 mg·g⁻¹ DW, respectively, all lower than that in leaves (27.67 mg·g⁻¹ DW), whereas the content of gymnemic acid in young, middle and old nodes was 27.96, 28.81 and 29.66 mg·g⁻¹ DW, respectively, all higher than that in leaves. The study provides the scientific evidences for the rational development and utilization of Gymnema sylvestre resources, since over exploitation of natural stands has caused depletion of these plants in nature. Foundation project: This work was supported by University Grants Commission, New Delhi (No. 33-88/2007 (SR))     

Expression of kit ligand and insulin‐like growth factor binding protein 3 during in vivo or in vitro development of ovarian follicles in sheep

Expression of Kit ligand (KL) and insulin‐like growth factor binding protein (IGFBP3) genes was studied at different in vivo and corresponding in vitro stages of development of the ovarian follicles in sheep. The expression of both KL and IGFBP3 was significantly higher in the primordial follicles relative to any other stage of development. Compared to the other stages, the KL expression in the cumulus cells from in vivo grown large antral follicles and that of IGFBP3 in COCs’ isolated from large antral follicles matured in vitro for 24 hr were significantly higher. In the oocytes from in vivo grown ovarian follicles, the expression of KL was the same at all the stages of development. Insulin‐like growth factor binding protein 3 expression was also the same in the oocytes at all the stages of the development except for a significantly lower expression in those from antral follicles. The expression of KL in the cumulus cells decreased significantly in the in vitro grown early antral follicles but did not change further as the development progressed. The expression of IGFBP3 in the cumulus cells from in vitro grown ovarian follicles appeared to increase as the development progressed although the increase was not significant between any two consecutive stages of development. In the oocytes in in vitro grown ovarian follicles, the expression levels of KL and IGFBP3 genes did not change with development. It is concluded that (i) KL and IGFBP3 genes follow specific patterns of expression during ovarian folliculogenesis and (ii) in vitro culture of preantral follicles compromises the development potential through alterations in the stage‐specific patterns of expression of these and other developmentally important genes.     

The extent of genetic diversity among Vanilla species: Comparative results for RAPD and ISSR

Vanilla is a large genus of about 110 species in the orchid family (Orchidaceae), including the species Vanilla planifolia from which commercial vanilla flavoring is derived. Since most species of vanilla are considered rare and endangered there is an urgent need to conserve them through genetic analysis and propagation/conservation studies on this crop.The present study investigated the genetic diversity among nine leafy- and leaf-less Vanilla species employing 30 decamer RAPD primers and 10 ISSR primers. The species under study were diverse and displayed a range of variability (0–66% and 0–81% for RAPD and ISSR, respectively). A total of 154 RAPD polymorphic markers (83.24%, h = 0.378) and 93 ISSR polymorphic markers (86.11%, h = 0.363) were used to generate a genetic similarity matrix followed by the cluster analysis. Specific groupings were revealed by each cluster analysis with slight variation between two different markers. Among the nine species studied, V. planifolia, Vanilla aphylla and Vanilla tahitensis revealed very low level of variation within their collections, thus indicating a narrow genetic base. The large genetic distance of Vanilla andamanica from other species suggests its different origin. A close genetic affinity was observed between the pairs V. planifolia, V. tahitensis and Vanilla albida, V. aphylla. These are the first comparative results for RAPD and ISSR reporting inter-relationship among nine cultivated, wild and hybrid Vanilla species.     

Characterization of Mannheimia (Pasteurella) haemolytica leukotoxin interaction with bovine alveolar macrophage beta2 integrins

Mannheimia (Pasteurella) haemolytica, the etiologic agent of bovine pneumonic mannheimiosis, produces an exotoxic leukotoxin. The leukotoxin (LktA) is a member of the RTX (repeats in toxin) family of bacterial cytolysins and is distinguished from other toxins by its unique target cell specificity to ruminant leukocytes occurring through binding to a specific receptor. We have demonstrated previously that the beta2 integrin LFA-1 is a receptor for LktA in bovine leukocytes and is involved in leukotoxin-induced biological effects. However the subunits within LFA-1 involved in binding to LktA, and post-binding signaling leading to cellular activation have not been well characterized. The purpose of our study was to pinpoint these precise subunits on bovine alveolar macrophages and to characterize their interaction with LktA. The results in this study indicate that although LktA can efficiently bind to the CD18 subunit of both LFA-1 and Mac-1, post-binding signaling events including elevation of intracellular calcium and CD18 tail phosphorylation are only observed through LFA-1. Furthermore, LktA also binds to the CD11a subunit of LFA-1. LktA binding to CD11a could be inhibited by a small molecule inhibitor of the I(inserted)-domain, the major ligand binding interface on CD11a. I-domain inhibition significantly blunts LktA-induced intracellular calcium elevation and tyrosine phosphorylation of the CD18 tail. Based on our results we suggest that LFA-1 serves as the functional leukotoxin receptor on bovine alveolar macrophages.     

Carotenoid Value Addition of Cereal Products by Monoculture and Mixed‐Culture Fermentation of Phaffia rhodozyma and Sporobolomyces roseus

Carotenoid value addition of corn whole stillage by red yeast fermentation has yielded astaxanthin‐ and β‐carotene‐enriched distillers dried grains with solubles (DDGS) for animal feed. In this study, commonly used animal feeds (rice bran, wheat bran, milo whole stillage, and soybean products) were subjected to carotenoid value addition. Phaffia rhodozyma and Sporobolomyces roseus monoculture and mixed‐culture submerged fermentation of these substrates supplemented with 5% glycerol were analyzed for astaxanthin, β‐carotene, and residual glycerol. Among all the substrates, full‐fat rice bran and full‐fat soy flour resulted in the highest astaxanthin (80 μg/g by P. rhodozyma) and β‐carotene yields (836 μg/g by S. roseus). P. rhodozyma produced the highest astaxanthin yield on each substrate, whereas depending on the substrate, either the mixed culture or S. roseus monoculture produced the highest β‐carotene yield. Soy hull was a poor substrate for carotenoid value addition. Both yeasts used glycerol as a carbon source for carotenoid production. This study shows that substrates influence the carotenoid yield. However, it is impossible to dissect the effect of specific nutrients on carotenoid production in complex biological substrates. Carotenoid value addition of these substrates provides as much as or more than the required daily dosage of carotenoids in animal feed.     

Rehabilitation of a lignite mine‐disturbed area in the Indian desert

Extensive lignite mining in the Indian (Thar) Desert commenced within the past decade. Accompanying extraction of this valuable resource there have been visible, important environmental impacts. The resultant land degradation has prompted concern from both public and regulatory bodies. This research assesses the success of rehabilitation plans implemented to revegetate a lignite mine‐disturbed area, near the village of Giral in western Rajasthan State. Rehabilitation success was achieved within the environmental constraints of this northwest Indian hot‐desert ecosystem using a combination of: (1) backfilling (abandoned pits) with minespoil and of covering the backfilled‐surfaces with fresh topsoil to a thickness of about 0·30 m; (2) use of micro‐catchment rainwater harvesting (MCWH) technique; (3) soil profile modification approaches; (4) plant establishment methodologies; and (5) the selection of appropriate germplasm material (trees, shrubs and grasses). Preliminary results indicate that the resulting vegetative cover will be capable of self‐perpetuation under natural conditions while at the same time meeting the land‐use requirements of the local people. The minespoil is alkaline in nature and has high electrical conductance. The average content of organic carbon, N, P and K is lower than in the regional topsoil. However, the concentration of Ca, Mg, Na and total S in the minespoil is much higher than in the topsoil. Further, the spoil material has no biological activity. Enhanced plant growth was achieved in MCWH plots, compared to control plots, where minespoil moisture storage was improved by 18–43 per cent. The rehabilitation protocol used at the site appears to have been successful because, in addition to the planted species, desirable native invasive species have become established. This study developed methods for the rehabilitation of lignite mine‐disturbed areas and has also resulted in an understanding of rehabilitation processes in arid regions with an emphasis on the long‐term monitoring of rehabilitation success. Copyright © 2004 John Wiley & Sons, Ltd.     

Changes in physico-chemical and biological properties of soil under conservation agriculture based pearl millet – mustard cropping system in rainfed semi-arid region

Poor soil health and low soil water content during crop growing period are major factor for low productivity of pearl millet – mustard rotation under rainfed semi-arid regions. The authors evaluated five different tillage and residue management practices for improving physico–chemical and biological properties of soil. Results showed that conservation agriculture (CA) practice (zero tillage (ZT) with 4 t ha^–1 residue retention) exhibited higher proportion of soil macro-aggregate. It also increased infiltration rate of about 15.2% over conventional tillage without residue but ZT increased soil penetration resistance in surface soil layer. In the residue applied plots, ~2–4% (w/w) higher soil water content was maintained throughout the season than the no-residue plots. CA practice had the highest soil organic carbon (4.96 g kg^–1) and microbial biomass carbon (188.3 μg g^–1 soil). Significant and positive correlation was also found between soil organic carbon with infiltration rate (r = 0.73**), mean weight diameter (r = 0.80**) and microbial biomass carbon (r = 0.86**). Thus, this study suggests that ZT with residue retention can be advocated in pearl millet – mustard rotations for improving, productivity, soil health and maintaining higher soil water content in rainfed semi-arid regions.