Polymorphism of β-lacto globulin gene in Indian goats and its effect on milk yield

Polymorphism in the beta-LG gene in the Indian goat was investigated by the SDS-PAGE and PCR-RFLP method. SDS-PAGE was carried out in 1098 samples belonging to 8 different breeds of Indian goats. The electrophoretic pattern in the beta-LG locus showed the presence of AA and AB genotypes with frequencies of 0.81 and 0.19, 0.89 and 0.11, 0.50 and 0.50, 0.80 and 0.20, 0.84 and 0.16, 1.00 and 0.00, 0.98 and 0.02 and 0.950 and 0.050 in Jamunapari, Barbari, Marwari, Sirohi, Jakhrana, Beetal, Local UP and Local MP goats, respectively. A total of 358 individuals belonging to 13 different genetic groups were analyzed by the PCR-RFLP method. The amplified product was observed as 426 bp and the restriction digestion with SacII revealed three genotypes, namely S1S1, S1S2 and S2S2 at the beta-LG locus. The frequency of the S2S2 genotype ranged from 0.42 to 1.00 in the population. An analysis was carried out in Jamunapari and Barbari goats to observe the effect of the beta-LG genotype on 90-day milk yield. Least squares analysis of data showed that beta-LG AA animals had higher milk yield than the beta-LG AB genotype in both breeds (P < 0.01).     

Protease accessibility laddering: a proteomic tool for probing protein structure

Limited proteolysis is widely used in biochemical and crystallographic studies to determine domain organization, folding properties, and ligand binding activities of proteins. The method has limitations, however, due to the difficulties in obtaining sufficient amounts of correctly folded proteins and in interpreting the results of the proteolysis. A new limited proteolysis method, named protease accessibility laddering (PAL), avoids these complications. In PAL, tagged proteins are purified on magnetic beads in their natively folded state. While attached to the beads, proteins are probed with proteases. Proteolytic fragments are eluted and detected by immunoblotting with antibodies against the tag (e.g., Protein A, GFP, and 6xHis). PAL readily detects domain boundaries and flexible loops within proteins. A combination of PAL and comparative protein structure modeling allows characterization of previously unknown structures (e.g., Sec31, a component of the COPII coated vesicle). PAL's high throughput should greatly facilitate structural genomic and proteomic studies.     

The yeast nuclear pore complex and transport through it

Exchange of macromolecules between the nucleus and cytoplasm is a key regulatory event in the expression of a cell's genome. This exchange requires a dedicated transport system: (1) nuclear pore complexes (NPCs), embedded in the nuclear envelope and composed of proteins termed nucleoporins (or "Nups"), and (2) nuclear transport factors that recognize the cargoes to be transported and ferry them across the NPCs. This transport is regulated at multiple levels, and the NPC itself also plays a key regulatory role in gene expression by influencing nuclear architecture and acting as a point of control for various nuclear processes. Here we summarize how the yeast Saccharomyces has been used extensively as a model system to understand the fundamental and highly conserved features of this transport system, revealing the structure and function of the NPC; the NPC's role in the regulation of gene expression; and the interactions of transport factors with their cargoes, regulatory factors, and specific nucleoporins.     

Sodium nitroprusside enhances biomass and gymnemic acids production in cell suspension of Gymnema sylvestre (Retz.) R.Br. ex. Sm.

Plant Cell, Tissue and Organ Culture (PCTOC) - Gymnemic acids (a group of triterpenoid saponins) found in Gymnema sylvestre (Retz.) R.Br. ex Sm. works as the main hypoglycaemic active components....     

Copper-stress induced alterations in protein profile and antioxidant enzymes activities in the in vitro grown Withania somnifera L.

Withania somnifera L. seedlings were grown in half-strength MS (Murashige and Skoog) basal medium for 4 weeks and then transferred to full-strength MS liquid medium for 3 weeks. The sustainable plants were subcultured in the same medium but with different concentrations (0, 25, 50, 100 and 200 μM) of Cu for 7 and 14 days. The growth parameters (root length, shoot length, leaf length and total number of leaves per plant) showed a declining trend in the treated plants in a concentration dependant manner. Roots and leaves were analyzed for protein profiling and antioxidant enzymes [catalase (CAT, EC 1.11.1.6), superoxide dismutase (SOD, EC 1.15.1.1) and guaiacol peroxidase (GPX, EC 1.11.1.7)]. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of crude protein extracts showed the appearance of some new proteins due to Cu treatment. In plant samples grown with 25 and 50 μM of Cu, a rapid increase in antioxidant activities were noticed but at higher concentration (100 and 200 μM) the activities declined. Isoforms of CAT, SOD and GPX were separated using non-denaturing polyacrylamide gel electrophoresis and concentration specific new isoforms were noticed during the study. Isoforms of the antioxidant enzymes synthesized due to Cu stress may be used as biomarkers for other species grown under metal stress.     

The nuclear basket proteins Mlp1p and Mlp2p are part of a dynamic interactome including Esc1p and the proteasome

The basket of the nuclear pore complex (NPC) is generally depicted as a discrete structure of eight protein filaments that protrude into the nucleoplasm and converge in a ring distal to the NPC. We show that the yeast proteins Mlp1p and Mlp2p are necessary components of the nuclear basket and that they also embed the NPC within a dynamic protein network, whose extended interactome includes the spindle organizer, silencing factors, the proteasome, and key components of messenger ribonucleoproteins (mRNPs). Ultrastructural observations indicate that the basket reduces chromatin crowding around the central transporter of the NPC and might function as a docking site for mRNP during nuclear export. In addition, we show that the Mlps contribute to NPC positioning, nuclear stability, and nuclear envelope morphology. Our results suggest that the Mlps are multifunctional proteins linking the nuclear transport channel to multiple macromolecular complexes involved in the regulation of gene expression and chromatin maintenance.     

Early Jurassic gastropods from England

Abstract: Twenty‐five gastropod taxa are reported from the Early Jurassic (Hettangian to Toarcian) of England. Of these, 14 are identified to species level, and the remaining 11 are treated in open nomenclature. One genus (Lensataphrus) and six species are introduced as new. The new species are Lensataphrus tatei, Lensataphrus tenuis, Tricarilda toddi, Cylindrobullina dorsetensis, Cylindrobullina ventricosa and Consobrinella greeni. The following new combinations are introduced: Cassianopsis hebertana (d’Orbigny, 1852) for Neritopsis hebertana; Cryptaulax abscisum (Terquem and Piette, 1868) for Cerithium abscisum; and Cylindrobullina avena (Terquem, 1855) for Striactaeonina avena. Most of the genera and some of the species are also known from Central Europe (Germany and France). Typical vetigastropod genera that are present in England and Central Europe are Colpomphalus, Costataphrus, Ooliticia, Eucycloscala and Eucycloidea. The caenogastropod genera Levipleura and Cryptaulax are present in both regions, as are the heterobranchs Tricarilda and Cylindrobullina. The heterobranch genus Consobrinella is reported from England for the first time. Gastropods seem to follow the diversity trends of other marine invertebrates during the Early Jurassic. They diversify until the Late Pliensbachian but decrease sharply in number around the Pliensbachian–Toarcian boundary. This may reflect both regional anoxia and a global mass extinction event in the Early Toarcian.     

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 F_v/F_m, 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.

     

Plant Regeneration Through Somatic Embryogenesis in Leaf Derived Callus of Plumbago Rosea

Regeneration of Plumbago rosea L., a rare medicinal plant, via somatic embryogenesis in callus cultures derived from leaf explants was described. Optimum callus formation was achieved on semi-solid Murashige and Skoog (MS) medium supplemented with 0.25 mg dm^–3 kinetin and 2.0 mg dm^–3 1-naphthaleneacetic acid (NAA). Somatic embryogenesis was achieved upon transferring the 4-week-old callus to a medium containing 1.0 mg dm^–3 kinetic (Kn), 0.5 mg dm^–3 gibberellic acid (GA₃) and 0.1 mg dm^–3 NAA. Embryo maturation and germination was achieved on the half-strength MS basal salts supplemented with 0.01 – 0.25 mg dm^–3 Kn and 2 % (m/v) saccharose. An average of 50 – 60 plantlets were obtained from 150 mg of embryogenic callus within 4 week of subculture. Out of the 50 plantlets about 28 survived in the greenhouse.