Nanoscale organization of hedgehog is essential for long-range signaling

Hedgehog (Hh) plays crucial roles in tissue-patterning and activates signaling in Patched (Ptc)-expressing cells. Paracrine signaling requires release and transport over many cell diameters away by a process that requires interaction with heparan sulfate proteoglycans (HSPGs). Here, we examine the organization of functional, fluorescently tagged variants in living cells by using optical imaging, FRET microscopy, and mutational studies guided by bioinformatics prediction. We find that cell-surface Hh forms suboptical oligomers, further concentrated in visible clusters colocalized with HSPGs. Mutation of a conserved Lys in a predicted Hh-protomer interaction interface results in an autocrine signaling-competent Hh isoform--incapable of forming dense nanoscale oligomers, interacting with HSPGs, or paracrine signaling. Thus, Hh exhibits a hierarchical organization from the nanoscale to visible clusters with distinct functions.     

Soral crypsis: protective mimicry of a coccid on an Indian fern

Herbivory with crypsis is not well documented in ferns. The present record of cryptic coloration of coccid Saissetia filicum Boisduval (Homoptera: Coccidae) to the sori of a fern species Asplenium nidus L. (Aspleniaceae) is unique. Predatory beetles (Jauravia sp., Coleoptera: Coccinellidae) that feed on the coccids, are suggested to be selective pressure for the development of the present homopteran soral crypsis. A higher rate of effective predation is noticed in the vegetative leaves than the fertile leaves. Aggressive ants were found harvesting honeydew secretions from the coccids and defending the trophobionts as well as the host fern from their natural enemies. In addition, a possible three-way mutualistic relationship among the coccids, its host fern and the tending ant is suggested. Differential numbers of coccids on vegetative and fertile leaves is correlated with their phenol content and degree of predation by beetles. Such coloration mimicry by the coccids may enable them to obtain the necessary blend of sorus of the host fern needed to evade beetle detection and attack.     

Neem (Azadirachta indica) seed kernel powder retards urease and nitrification activities in different soils at contrasting moisture and temperature regimes

A laboratory experiment was conducted to examine the potentiality of a natural resource neem (Azadirachta indica) seed kernel powder (NSKP) to reduce the urease and nitrification activities in different soils (viz., normal, acid, and sodic) at contrasting moisture (1:1 soil to water and field capacity) and temperature regimes (10 degrees C and 37 degrees C). Results have revealed that application of NSKP with urea did not exhibit any urease inhibitory property in normal and sodic soils, but in acid soil it had maintained higher concentration of urea than the urea alone treated samples for two weeks after application. At 37 degrees C and under field capacity moisture level, urea hydrolysis was more rapid than at 10 degrees C and under waterlogged (1:1) conditions. The NSKP has showed variable effects (4-28%) to inhibit nitrification during 7-21 days after application, depending upon the soil types, temperature and moisture regimes. The nitrification activity was significantly low in acid soil followed by normal and sodic soils. The present study suggests that NSKP has the potential to retard the urease activity in acid soil, and nitrification in all the soils, and thus it may be used along with urea for the better use of applied -N.     

Molecular Cloning, Bacterial Overexpression and Characterization of L-myo- inositol 1- Phosphate Synthase from a Monocotyledonous Resurrection Plant, Xerophyta viscosa Baker

L-myo-inositol 1-phosphate synthase (EC 5.5.1.4; MIPS), an evolutionarily conserved enzyme-protein, catalyses the first and rate limiting step of inositol biosynthesis. Inositol and its derivatives play important roles in biological kingdom like growth regulation, membrane biogenesis, signal transduction and also acts as an osmolyte or osmoprotectant in abiotic stress tolerance. Here we report the cloning, sequencing and the characterization of the INO1 gene from Xerophyta viscosa (XINO1), a monocotyledonous resurrection plant. Nucleotide sequences of XINO1 show striking homology (70–99%) with a number of INO1 genes from plant sources particularly with the monocots. The gene is functionally identified through genetic complementation using a yeast inositol auxotrophic strain FY250. The gene is expressed in E. coli BL21, recombinant protein purified to homogeneity, biochemically characterized and compared with Oryza INO1 (RINO1) gene product. The XINO1 gene product is catalytically active in a broader range of lower temperature (between 10–40 °C) than the RINO1 gene- product. This is the first report of MIPS gene from any resurrection plant.     

Effect of plant growth regulators on regeneration of plantlets from bud cultures ofCymbopogon nardus L. and the detection of essential oils from thein vitro plantlets

Cymbopogon nardus L. could be propagated via tissue culture using axillary buds as explants. The aseptic bud explants obtained using double sterilization methods produced stunted abnormal multiple shoots when they were cultured on Murashige and Skoog medium (MS) supplemented with 1.0 mg L^-1 or 2.0 mg L^-1 benzyladenine (BA). Stunted shoots that cultured on MS + 1.0 mg L^-1 BA + 1.0 mg L^-1 N⁶-isopentenyl-adenine (2iP) could induce elongation of shoots from about 60% of the stunted shoots. Normal multiple shoots could be induced at the highest (19.7 shoots per bud) from the bud explants within six weeks when cultured on proliferation medium consisted of MS supplemented with 0.3 mg L^-1 BA and 0.1 mg L^-1 indole-3-butyric acid (IBA). The separated individual shoot produced roots when transferred to basic MS solid medium. The essential oils that were contained in the mature plants namely citronellal, geraniol and citronellol were also found in thein vitro C. nardus plantlets. Citronellal was the main essential oil component in the matured plants while geraniol was the main component in thein vitro plantlets.     

Evaluation of Ssp I polymerase chain reaction assay in the detection of Wuchereria bancrofti infection in field-collected Culex quinquefasciatus and its application in the transmission studies of lymphatic filariasis

Abstract: Ssp I polymerase chain reaction (PCR) assay, developed for Wuchereria bancrofti, was evaluated for its sensitivity in detecting infection in the vector, Culex quinquefasciatus, in the field. The evaluation of the assay was carried out using pools of vector mosquitoes collected from areas under filariasis survey and control trial projects, in comparison with the standard dissection and microscopy technique. In the filariasis survey area the infection rate as determined by the dissection and microscopy technique was 0.89% whereas it was 1.7% by PCR assay. In the Bacillus sphaericus trial area the infection rates as assessed by the conventional technique were 6.6 and 4.5% in the treated and check areas, respectively, whereas those obtained by the PCR assay were 4.7 to 2.2%. Although the infection rates determined by the PCR assay are slightly higher or lower than the rates obtained by the conventional technique, the difference was not statistically significant (P=0.451 for filariasis survey area, and P=0.203 and 0.161 for B. sphaericus trial area). When the pool size of Cx. quinquefasciatus was increased to 50 the sensitivity of the PCR was affected. The changes in infection rates as influenced by the antifilarial chemotherapy were similar when determined by PCR assay and the standard method. The major advantage of the PCR assay over the conventional technique is that thousands of mosquitoes can be processed within a short duration and this attribute has potential application in rapid assessment of disease prevalence and monitoring of the transmission dynamics.