In vitro differentiation in callus cultures of moth bean,Vigna aconitifolia (JACQ) marechal

Callus cultures of two cultivars of Vigna aconitifolia (IPCMO-926, RDM-120) were raised and their growth and differentiation studied. In IPCMO-926 callus cultures, numerous shoot buds differentiated on MS medium with BA (0.4–22.2 μM) alone or in combination with IAA (5.7 μM). In RDM-120 best differentiation of shoot buds was observed on a medium with K (23.2 μM) and IAA (5.7 μM). Kinetin alone, however, induced rhizogenesis in callus cultures. In suspension cultures of IPCMO-926 embryoids differentiated on MS medium with K (0.5 μM) and 2,4-D (0.4 and 0.9 μM).     

Effect of oilseed cakes on alpha-amylase production by Bacillus licheniformis CUMC305.

The effects of oilseed cakes on extracellular thermostable alpha-amylase production by Bacillus licheniformis CUMC305 was investigated. Each oilseed cake was made of groundnut, mustard, sesame, linseed, coconut copra, madhuca, or cotton. alpha-Amylase production was considerably improved in all instances and varied with the oilseed cake concentration in basal medium containing peptone and beef extract. Maximum increases were effected by a low concentration (0.5 to 1.0%) of groundnut or coconut, a high concentration (3%) of linseed or mustard, and an Rintermediate concentration (2%) of cotton, madhuca, or sesame. The oilseed cakes made of groundnut or mustard could completely replace the conventional peptone-beef extract medium as the fermentation base for the production of alpha-amylase by B. licheniformis. The addition of corn steep liquor to cotton, linseed, sesame, or madhuca cake in the medium improved alpha-amylase production.     

Interaction of the carboxamide of NADPH with Lactobacillus casei dihydrofolate reductase

Dihydrofolate reductase from Lactobacillus casei and its complexes with NADPH and methotrexate yield well-resolved Raman spectra. The 1685-cm^?1 Raman band assigned to the carboxamide of NADPH persists in the NADPH-enzyme binary complex but is absent from the NADPH-methotrexate-enzyme ternary complex. This is ascribed to stabilization of the polarized form of the carboxamide by H bonding to the NH and CO groups of Ala 6 and Ile 13 of the peptide backbone.     

Replication in Drosophila chromosomes

Prolongation of larval life in Drosophila melanogaster, by growing wild type larvae at lower temperature, or in animals carrying the X-linked mutation giant is known to result in a greater proportion of nuclei in salivary glands showing the highest level of polyteny. We have examined by autoradiography the patterns of ³H-thymidine incorporation during 10 min or 1 min pulses in salivary gland polytene chromosomes of older giant larvae and of wild type late third instar larvae of D. melanogaster grown since hatching either at 24 ° C or at 10 ° C. The various patterns of labelling and their relative frequencies are generally similar in glands from the warm-(24 ° C) or cold (10 ° C)-reared wild type larvae, except the interband (IB) labelling patterns which are very frequent in the later group but rare in the former. The IB type labelled nuclei in cold-reared wild type larvae show labelling ranging from only a few puffs/interbands labelled to nearly all puffs/interbands labelled. In warm-reared wild type larvae, very low labelled IB patterns are not seen. In older giant larvae, the ³H-thymidine labelling patterns are in most respects similar to those seen in cold-reared wild type larvae. In 1 min pulsed preparations from all larvae, the IB patterns are relatively more frequent than in corresponding 10 min pulsed preparations. No nuclei with the continuous (2C or 3C) type of labelling pattern, with all bands and interbands/puffs labelled, were seen in 1 min pulsed preparations from cold-reared wild type or in giant larvae, and only a few nuclei in 1 min pulsed preparations from warm-reared wild type larvae exhibited the 2C labelling pattern. Analysis of silver grain density on specific late replicating sites in late discontinuous (1D) type labelled nuclei suggests that the rate of DNA synthesis per chromosomal site is not different at the two developmental temperatures. It is suggested that correlated with the prolongation of larval life under cold-rearing conditions or in giant larvae, the polytene replication cycles are also prolonged. It is further suggested that the polytene S-period in these larvae is longer due to a considerable asynchrony in the initiation and termination of replication of different sites during a replication cycle.     

Identification and size variation of terminal fragments of sheep pox virus genome

Terminal fragments of sheep pox virus DNA identified by snap-back analysis showed terminal covalent cross-links. Southern blot hybridization using a terminal fragment probe confirmed the termini and terminal repeats (common sequences) of the sheep pox virus genome. Terminal fragment length variability was observed between virus isolates.     

Alkaloid production by plant cell cultures of Holarrhena antidysenterica: II. Effect of precursor feeding and cultivation in stirred tank bioreactor

Precursor feeding strategy for increasing the yield of conessine, a steroidal alkaloid of Holarrhena antidysenterica, was established in cell suspension culture. A total of 50 mg/L added cholesterol was converted into 43 mg/L of alkaloid, 90% of which constituted the conessine. By applying the precursor feeding policy to the cell suspension culture in modified Murashige and Skoog (MS) medium, a total of 143 mg/L of alkaloid was produced in 8 days. In this way the alkaloid content of the cells was increased more than six times compared to that obtained in the standard MS medium. The steps leading to biotransformation of cholesterol into alkaloids were unaffected by phosphate. The shake flask data were successfully transferred to a bench scale 6-L stirred tank bioreactor in which the specific biosynthetic rate of alkaloid production was 110 mg/100 g dry cell weight per day, about 160 times higher than that of whole plant.     

Nonradioactive in situ localization of poly(A)+ RNA during pollen development in anthers of tobacco (Nicotiana tabacum L.)

Summary A method is described for non-radioactive labeling of total mRNA [poly(A)+ RNA] in plastic-embedded plant tissue sections. Oligo-deoxythymidylic acid (oligo-dT) labeled with digoxigenin-conjugated dUTP was used for in situ hybridization to poly(A)+ RNA in sections of tobacco (Nicotiana tabacum) anthers. The digoxigenin was immuno-stained using antidigoxigenin IgG and gold-labeled protein-A, followed by silver enhancement of the gold label. Reproducibly similar positive staining patterns were obtained with digoxigenin-labeled oligo-dT and polyuridylic acid [poly(U)], but not with a similarly labeled sense probe, poly(A). In the developing anthers, from the onset of meiosis to the production of pollen grains, labeling patterns were compatible with a gradual depletion of nuclear and chromosome-associated sporophytic mRNA molecules during prophase of meiosis, followed by postmeiotic production of gametophytic mRNA in microspore nuclei and the vegetative nuclei of the pollen grains.Abbreviations BSA bovine serum albumin - DIG digoxigenin - IgG immunoglobulin-G - oligo-dT oligo-deoxythymidylic acid - PAS-ABB periodic acid Schiff-aniline blue black - PBS phosphate buffered saline - poly(A) polyadenylic acid - poly(U) polyuridylic acid - SSC standard saline citrate     

(ADP-ribosyl)ation pattern of chromosomal proteins during ageing.

(ADP-ribosyl)ation of chromosomal proteins was studied by incubating the nuclei of brain and liver of young and old rats with 14C-NAD+. In brain as well as in liver histone proteins show approximately 2-3 fold higher (ADP-ribosyl)ation than that of non-histone chromosomal (NHC) proteins of both the age groups. H1 seems to be the major target for (ADP-ribosyl)ation. Amongst nucleosomal histones H2B is the main acceptor of 14C-labelled ADP-ribose moieties. A sharp age related decline of (ADP-ribosyl)ation of chromosomal proteins was observed in both the tissues.     

Laminarinase from Penicillium funiculosum and its role in release of beta-glucosidase

An extracellular laminarinase (1----3)-beta-glucan glucohydrolase (EC 3.2.1.6) was purified from culture filtrates of Penicillium funiculosum. It was homogeneous on polyacrylamide gel electrophoresis in the presence and absence of sodium dodecyl sulfate. It had a Mr of 14,000 and isoelectric point of pH 4.2. The apparent Km value for lamimarinase was 8.3 mg/ml and Vmax was 8 mumol/min/mg. The distribution of beta-glucosidase activity in two different species of Penicillium showed that P. funiculosum had a higher ratio of extracellular to cell wall bound activity than Penicillium janthinellum. Treatment of mycelia of both species with NaCl, EDTA, Triton X-100, or proteolytic enzymes did not release the cell wall bound beta-glucosidase. Incubation of the mycelia with the laminarinase released 2-4 times more beta-glucosidase than the estimated cell bound activity in P. janthinellum and P. funiculosum.