Antagonistic potential of Trichoderma longibrachiatum and T. hamatum resident on maize (Zea mays) plant against Fusarium verticillioides (Nirenberg) isolated from rotting maize stem

Antagonistic potential of Trichoderma longibrachiatum and T. hamatum against the pathogen Fusarium verticillioides was examined in the laboratory. This was done by pairing each Trichoderma species with the pathogen on 9 cm Petri plates of acidified potato dextrose agar (APDA). Three pairing methods were employed and gradings were assigned to different radial growth suppression of F. verticillioides by each Trichoderma species. Analysis was done using the GLM procedure of the SAS package. Both Trichoderma species significantly inhibited radial growth of F. verticillioides (P = 0.01, R ² = 0.99) irrespective of pairing method. ‘Inoculating antagonist before pathogen’ supported the best growth inhibition of F. verticillioides by both Trichoderma species. Both Trichoderma species differed significantly (P > 0.0029) in inhibiting radial growth of F. verticillioides. Growth inhibition differed significantly within (P > 0.0059) and among (P > 0.0001) pairing methods. T. longibrachiatum was significantly better than T. hamatum in inhibiting radial growth of F. verticillioides, even at P = 0.01. T. hamatum and T. longibrachiatum could thus be said to show promising antagonistic potential against F. verticillioides with the latter showing better prospects.     

A Simple One Pot Purification of Bacterial Amylase From Fermented Broth Based on Affinity Toward Starch-Functionalized Magnetic Nanoparticle

Surface-functionalized adsorbant particles in combination with magnetic separation techniques have received considerable attention in recent years. Selective manipulation on such magnetic nanoparticles permits separation with high affinity in the presence of other suspended solids. Amylase is used extensively in food and allied industries. Purification of amylase from bacterial sources is a matter of concern because most of the industrial need for amylase is met by microbial sources. Here we report a simple, cost-effective, one-pot purification technique for bacterial amylase directly from fermented broth of Bacillus megaterium utilizing starch-coated superparamagnetic iron oxide nanoparticles (SPION). SPION was prepared by co-precipitation method and then functionalized by starch coating. The synthesized nanoparticles were characterized by transmission electron microscopy (TEM), a superconducting quantum interference device (SQUID, zeta potential, and ultraviolet–visible (UV-vis) and Fourier-transform infrared (FTIR) spectroscopy. The starch-coated nanoparticles efficiently purified amylase from bacterial fermented broth with 93.22% recovery and 12.57-fold purification. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) revealed that the molecular mass of the purified amylase was 67 kD, and native gel showed the retention of amylase activity even after purification. Optimum pH and temperature of the purified amylase were 7 and 50°C, respectively, and it was stable over a range of 20°C to 50°C. Hence, an improved one-pot bacterial amylase purification method was developed using starch-coated SPION.     

Efficient micropropagation and chlorocholine chloride induced stevioside production of Stevia rebaudiana Bertoni

A promising method of micropropagation of Stevia rebaudiana Bertoni has been developed with an aim to increase the biomass, survivability of the plantlets and stevioside production, using chlorocholine chloride (CCC). Microshoots transferred to the MS medium containing different combinations CCC and IBA were found to be most effective in terms of growth pattern, hardening ability of the plantlets and stevioside content, compared to MS medium containing either IBA or CCC. Among other combinations tested, MS medium supplemented with 3 mg/l CCC and 3 mg/l IBA was found most effective in inducing significant changes like reduced shoot length, increased number of roots, higher leaf size, increased biomass and chlorophyll retaining capacity, higher survival percentage and most importantly the elevated stevioside content. Collectively, the major observations of this research indicate that application of CCC in micropropagation of S. rebaudiana Bertoni is a promising approach and has commercial prospects.     

The study of the electronic structures and properties of pure and transition metal-doped silicon nanoclusters: a density functional theory approach

This report presents the study of ab initio electronic structure and properties of pure and transition metal (TM = Ti, Zr and Hf)-doped silicon clusters, TM@Si(n), by using density functional theory with a polarised basis set (LanL2DZ) within the spin-polarised generalised gradient approximation for different values of n varying from 8 to 20. As the first step of the study, different optimised geometries of pure and doped clusters are calculated. These optimised clusters are then used to calculate different structural and physical parameters of the clusters, like binding energy, Highest Occupied Molecular Orbital – Lowest Unoccupied Molecular Orbital (HOMO–LUMO) gap, charge transfer, etc. In order to check the stability of the clusters, the second-order difference in the energy of the optimised structures is calculated. To study the optical behaviour of the clusters, infrared and Raman spectra are also calculated. Further calculations are also done on cation and anion clusters of both pure and doped nanoclusters to obtain their ionisation potential, electron affinity and chemical potential. An effort has been made to correlate the variation of different calculated parameters with the size of the clusters to explain the real existence and stabilities of different TM-doped clusters.     

Effects of culture conditions on multiple shoot induction from inflorescence and RAPD analysis of cloned plants in Limonium sinensis (Girard) Kuntze, var. Golden Diamond

The optimum physical and chemical microenvironment for micropropagation of Limonium sinensis (Girard) Kuntze, var. Golden Diamond was established from immature inflorescence segments as explant. The highest frequency (62 %) of axillary shoot induction was obtained on MS medium (Murashige and Skoog Physiol Plant 15:473–497, 1962) supplemented with 8.88 μM BA, 1.34 μM of NAA and two growth additives cysteine (142.33 μM), and glutamine (684.22 μM). In the subsequent culture maximum average number of shoots (11.13?±?0.34) were obtained from micro-shoot explant on MS medium supplemented with the same additives and 2.22 μM BA. During subcultures the problem of vitrification was mitigated through increasing agar concentration from 0.8 % to 1.0 % and providing better ventilation. The in vitro developed shoots were rooted on MS medium supplemented with 2.46 μM IBA and 0.88 μM BA. Rooted plants were acclimatized successfully in the greenhouse with 80 % survival rate. RAPD analysis using 15 random decamer primers generated monomorphic banding pattern in micropropagated plants and similar to those of mother plant revealing the genetic integrity of regenerants.     

In vitro and in vivo evaluation of L‐lactide/ε‐caprolactone copolymer scaffold to support myoblast growth and differentiation

Skeletal muscle regeneration involves the activation of satellite cells to myoblasts, followed by their proliferation and fusion to form multinucleated myotubes and myofibers. The potential of in vitro proliferated myoblasts to treat various diseases and tissue defects can be exploited using tissue‐engineering principles. With an aim to develop a biocompatible and biodegradable scaffold that supports myoblast growth and differentiation, we have developed a porous sponge with 70/30 L ‐lactide/ε‐caprolactone copolymer (PLC) using a phase inversion combined with particulate leaching method. Degradation studies indicated that the sponge retained its structural integrity for 5 months in vitro and had undergone complete biodegradation within 9 months in vivo. The sponge supported human myoblasts attachment and its proliferation. Myoblasts seeded on the PLC sponge differentiated and fused in vitro to form myotubes expressing myosin heavy chain. Histological and molecular analyses of the PLC scaffolds seeded with green fluorescent protein‐labeled human myoblasts and implanted ectopically under the skin in SCID mice demonstrated the presence of multinucleated myotubes expressing human muscle‐specific markers. Our results suggest that PLC sponges loaded with myoblasts can be used for skeletal muscle engineering or for inducing muscle repair. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 2013     

Soil organic carbon pools and productivity relationships for a 34 year old rice–wheat–jute agroecosystem under different fertilizer treatments

Soil organic carbon (SOC) pools are important in maintaining soil productivity and influencing the CO₂ loading into the atmosphere. An attempt is made here to investigate into the dynamics of pools of SOC viz., total organic carbon (C _tot), oxidisable organic carbon (C _oc) and its four different fractions such as very labile (C _{frac 1}), labile (C _{frac 2}), less labile (C _{frac 3}) and non-labile (C _{frac 4}), microbial biomass carbon (C _mic), mineralizable carbon (C _min), and particulate organic carbon (C _p) in relation to crop productivity using a 34 year old rice (Oryza sativa L)–wheat (Triticum aestivum L)–jute (Corchorus olitorius L) cropping system with different management strategies (no fertilization, only N, NP, NPK and NPK + FYM) in the hot humid, subtropics of India. A fallow treatment was also included to compare the impact of cultivation vis-à-vis no cultivation. Cultivation over the years caused a net decrease, while balanced fertilization with NPK maintained the SOC pools at par with the fallow. Only 22% of the C applied as FYM was stabilized into SOC, while the rest got lost. Of the analysed pools, C _{frac 1}, C _mic, C _p and C _min were influenced most by the treatments imposed. Most of the labile pools were significantly correlated with each other and with the yield and sustainable yield index (SYI) of the studied system. Of them, C _frac1, C _min, C _mic and C _p explained higher per cent variability in the SYI and yield of the crops. Results suggest that because of low cost and ease of estimation and also for upkeeping environmental conditions, C _frac1 may be used as a good indicator for assessment of soil as to its crop productivity. Responsible Editor: Hans Lambers.