Characterization and correlation of pathogenicity of Botryodiplodia theobromae isolates,the causal agent of black root rot of mulberry (Morus spp.)

Abstract

The study reports the characterization of 10 isolates of mulberry black root rot causing fungus, Botryodiplodia theobromae obtained from the infected gardens of Karnataka, Andhra Pradesh and Tamil Nadu. The analysis based on cultural, morphological, pathogenicity and molecular markers (RAPD and SSRs) revealed significant variations among the isolates. Based on the disease reaction on susceptible V-1 variety, isolates were grouped as pathogenic (60%), moderate pathogenic (20%) and non-pathogenic (20%). Among all isolates, RAPDs revealed higher marker polymorphism however, based on Shannon’s Information Index (I) SSRs were more informative (0.781) compared to the former (0.444). Stepwise Multiple Regression Analysis (MRA) indicated a total of 5, 5 and 3 molecular markers were found to correlate with disease symptoms. Screening of germplasm using multiple strains of virulent isolates will enhance possibilities of locating diverse resistant genes. Pyramiding of these genes will aid in development of mulberry variety with durable resistance and sustainable sericulture.     

Pseudomonas aeruginosa RRALC3 Enhances the Biomass,Nutrient and Carbon Contents of Pongamia pinnata Seedlings in Degraded Forest Soil

The study was aimed at assessing the effects of indigenous Plant Growth Promoting Bacterium (PGPB) on the legume Pongamia pinnata in the degraded soil of the Nanmangalam Reserve Forest (NRF) under nursery conditions. In total, 160 diazotrophs were isolated from three different nitrogen-free semi-solid media (LGI, Nfb, and JMV). Amongst these isolates, Pseudomonas aeruginosa RRALC3 exhibited the maximum ammonia production and hence was selected for further studies. RRALC3 was found to possess multiple plant growth promoting traits such as nitrogen accumulation (120.6ppm); it yielded a positive amplicon with nifH specific primers, tested positive for Indole Acetic Acid (IAA; 18.3μg/ml) and siderophore production, tested negative for HCN production and was observed to promote solubilization of phosphate, silicate and zinc in the plate assay. The 16S rDNA sequence of RRALC3 exhibited 99% sequence similarity to Pseudomonas aeruginosa JCM5962. Absence of virulence genes and non-hemolytic activity indicated that RRALC3 is unlikely to be a human pathogen. When the effects of RRALC3 on promotion of plant growth was tested in Pongamia pinnata, it was observed that in Pongamia seedlings treated with a combination of RRALC3 and chemical fertilizer, the dry matter increased by 30.75%. Nitrogen, phosphorus and potassium uptake increased by 34.1%, 27.08%, and 31.84%, respectively, when compared to control. Significant enhancement of total sugar, amino acids and organic acids content, by 23.4%, 29.39%, and 26.53% respectively, was seen in the root exudates of P. pinnata. The carbon content appreciated by 4-fold, when fertilized seedlings were treated with RRALC3. From the logistic equation, the rapid C accumulation time of Pongamia was computed as 43 days longer than the control when a combination of native PGPB and inorganic fertilizer was applied. The rapid accumulation time of N, P and K in Pongamia when treated with the same combination as above was 15, 40 and 33 days longer, respectively, as compared to the control.     

Impact of precursors and plant growth regulators on <Emphasis Type="Italic">in vitro</Emphasis> growth,bioactive lignans,and antioxidant content of <Emphasis Type="Italic">Phyllanthus</Emphasis> species

The production of specific secondary metabolites in vitro can be improved through medium supplementation with secondary metabolite precursors, plant growth regulators (PGRs), and abiotic and biotic elicitors. In the present study, node and internode explants of Phyllanthus amarus and P. urinaria collected from Karkala region, Udupi District, Karnataka, India, were inoculated aseptically onto Murashige and Skoog (MS) medium for callus induction. Uniform calluses were inoculated onto MS medium fortified with one of two precursor’s cinnamic acid (CA) or phenylalanine (PA), or with naphthalene acetic acid (NAA). After 30 d of treatment, calluses from treatment and control groups were harvested and quantitatively analyzed for three lignans (phyllanthin, hypophyllanthin and niranthin) and an antioxidant (ellagic acid). Increased amounts of the lignans and ellagic acid were obtained through supplementation with CA, PA, and NAA, and higher ellagic acid was present at higher amounts than the three lignans. These results demonstrated that the Phyllanthus species collected from Karkala region (designated “Accessions3”) show substantial response to CA, PA, and NAA treatment and represent a potential source of donor plants with higher amounts of lignans and antioxidants. These plants can be cultivated on a large scale both in vitro and in vivo for production of important bioactive compounds. Production of these compounds can be further enhanced through induction of somaclonal variant plants with higher amounts of bioactive molecule production and through production of transgenic plants overexpressing genes related to lignan- and phenolic-compound biosynthesis.     

Swapping the substrate specificities of the neuropeptidases neurolysin and thimet oligopeptidase

Thimet oligopeptidase (EC 3.4.24.15) and neurolysin (EC 3.4.24.16) are closely related zinc-dependent metallopeptidases that metabolize small bioactive peptides. They cleave many substrates at the same sites, but they recognize different positions on others, including neurotensin, a 13-residue peptide involved in modulation of dopaminergic circuits, pain perception, and thermoregulation. On the basis of crystal structures and previous mapping studies, four sites (Glu-469/Arg-470, Met-490/Arg-491, His-495/Asn-496, and Arg-498/Thr-499; thimet oligopeptidase residues listed first) in their substrate-binding channels appear positioned to account for differences in specificity. Thimet oligopeptidase mutated so that neurolysin residues are at all four positions cleaves neurotensin at the neurolysin site, and the reverse mutations in neurolysin switch hydrolysis to the thimet oligopeptidase site. Using a series of constructs mutated at just three of the sites, it was determined that mutations at only two (Glu-469/Arg-470 and Arg-498/Thr-499) are required to swap specificity, a result that was confirmed by testing the two-mutant constructs. If only either one of the two sites is mutated in thimet oligopeptidase, then the enzyme cleaves almost equally at the two hydrolysis positions. Crystal structures of both two-mutant constructs show that the mutations do not perturb local structure, but side chain conformations at the Arg-498/Thr-499 position differ from those of the mimicked enzyme. A model for differential recognition of neurotensin based on differences in surface charge distribution in the substrate binding sites is proposed. The model is supported by the finding that reducing the positive charge on the peptide results in cleavage at both hydrolysis sites.     

Oil cakes and their biotechnological applications--a review

Oil cakes have been in use for feed applications to poultry, fish and swine industry. Being rich in protein, some of these have also been considered ideal for food supplementation. However, with increasing emphasis on cost reduction of industrial processes and value addition to agro-industrial residues, oil cakes could be ideal source of proteinaceous nutrients and as support matrix for various biotechnological processes. Several oil cakes, in particular edible oil cakes offer potential benefits when utilized as substrate for bioprocesses. These have been utilized for fermentative production of enzymes, antibiotics, mushrooms, etc. Biotechnological applications of oil cakes also include their usages for vitamins and antioxidants production. This review discusses various applications of oil cakes in fermentation and biotechnological processes, their value addition by implementation in feed and energy source (for the production of biogas, bio-oil) as well.     

Development of anti-EGF receptor peptidomimetics (AERP) as tumor imaging agent

EGFR is over-expressed in several solid tumors including breast, prostate, pancreas, and lung cancers and is correlated to the metastasic potential of the tumor. Anti-EGFR receptor-binding peptidomimetics (AERP) were examined to assess the small molecule’s potential use as tumor-specific imaging agents. The aim of this work was to design and characterize the binding specificity of the radiolabeled peptidomimetics to EGFR over-expressing cell lysate and to A431 xenograft tumors. Our newly designed peptidomimetic, AERP, was conjugated to DTPA and labeled with ^99mTc. The in vivo tumor accumulation of [^99mTc] DTPA-AERP-2 was 1.6 ± 0.1 %ID/g and tumor to muscle ratio was 5.5. Our studies suggest that this novel peptidomimetic, AERP-2, warrants further development as an EGFR specific tumor-imaging agent.