Biocontrol potential of Pseudomonas azotoformans,Serratia marcescens and Trichoderma virens against Fusarium wilt of Dalbergia sissoo

Wilt disease caused by Fusarium solani is a serious constraint to Dalbergia sissoo (shisham) plantations in northern India. In this study, the antagonistic potential of 40 bacterial isolates recovered from rhizophere soil of healthy shisham trees, and a well‐characterized Trichoderma species (Trichoderma virens) were tested for their possibility as biocontrol agents for F. solani. Two promising isolates (S1 and S15) were identified which inhibited pathogen growth, caused chitin degradation, produced siderophores and solubilized phosphate in vitro. Isolate S15 scored highest for hydrogen cyanide (HCN) production while isolate S1 was a non‐HCN producer. These two isolates were identified as Serratia marcescens (S1) and Pseudomonas azotoformans (S15) following sequence analysis of 16S rDNA. In dual culture assays, T. virens caused 80% inhibition of mycelial growth of the test fungus. The three selected antagonists when tested in planta in the glasshouse completely suppressed production of wilt symptoms on 12‐month‐old shisham plants. Further work is needed to ascertain the potential of these isolates to be used as biocontrol agents to manage shisham wilt under field conditions.     

Fishmeal alternative from renewable CO2 for rainbow trout feed

The rapid global growth in fish farming and limited supply of fish meal (FM) has consequently reduced FM inclusion levels in compound feeds leading to a higher reliance on alternative protein sources. Sasya is a single cell protein (SCP) product that has a similar amino acid profile as FM. The aim of this study was to evaluate the effects of replacing FM with SCP on in vivo digestibility, growth, feed efficiency, whole‐body proximate/amino acid composition and gene expression levels of various hepatic enzymes in rainbow trout (Oncorhynchus mykiss). Three isonitrogenous (470 g/kg crude protein) and isolipidic (18 g/kg crude lipid) diets were formulated as follows: Diet 1: control (30% FM); Diet 2:24% FM + 6% SCP and Diet 3:18% FM + 12% SCP. Each diet was hand‐fed to triplicate tanks containing 30 rainbow trout fingerlings (4.99 ± 0.20 g) for 9 weeks. Apparent digestibility coefficients of SCP for dry matter, crude protein, lipid and energy were 60, 80, 93 and 74% respectively. Growth performance (final weight: 69–71 g), feed conversion ratios (0.91–0.94) as well as whole‐body protein and amino acid composition were unaffected by diets. However, Diet 3 significantly increased whole‐body crude fat and energy. Fish fed the SCP‐based diets had significantly higher expression for carnitine palmitoyltransferase‐1b (CPT1b), fatty acid delta 6 desaturase (FADS6) and fatty acid elongase 5 compared to the control. Overall, the quality of the SCP was similar as FM. Therefore, this product could enlarge the portfolio of alternative protein sources that can be used in fish diets and thus open a new market opportunity for use of a new feed resource in the feed industry.     

Genetic analysis of a CSR10 (indica) × Taraori Basmati F3 population segregating for salt tolerance using ISSR markers

Segregation for salinity tolerance and ISSR markers based molecular polymorphism were investigated in a F₃ plant population raised via single-seed descent method from a cross between salt-tolerant indica rice variety CSR10 and salt-susceptible premium traditional Basmati rice variety Taraori Basmati HBC19. A total of 130 F₃plants were evaluated individually for salinity tolerance on 1–9 scale on the basis of seedling growth parameters; the average score ranged between 1.7 to 8.3. Frequency distribution curve obtained using the salinity tolerance data of F₃ population and a chi-square analysis, showed a good fit to a normal distribution. Eleven plants each in the category of salt-tolerant and salt-susceptible were selected from the segregating F₃ population for ISSR marker analysis. A total of 149 bands (4–11 bands per primer) ranging from 200 to 3530 bp were scored for the two rice varieties and the selected CSR10 × HBC19 segregating F₃ plants using 26 ISSR primers. Of these, 89 were monomorphic and 60 were polymorphic. Of the 60 polymorphic bands,36 and 20 bands were specific to CSR10 andHBC19 respectively. The remaining four bands were amplified using UBC primers 810,848, 853 and 886 and present in only some of the CSR10 × HBC19 F₃ plants. Notably, ISSR primers with dinucleotide repeat motif and 5'-anchored end amplified more number of bands (7.0 bands/primer) compared to3'-anchored dinucleotide primers (5.4bands/primer), but 3'-anchored dinucleotide primers revealed higher level of polymorphism (2.6 polymorphic bands/primer) compared to 5'-anchoreddinucleotide primers (1.43 polymorphic bands/ primer). While distribution of majority of the polymorphic bands were more or less in the expected ratios in salt-tolerant and/or salt-sensitive F₃segregating plants, but some of the bands amplified using UBC ISSR primers 823, 825,826, 849, 853, 864, 866 and 884 showed highly skewed distribution. Such polymorphic bands stand greater chances of having a linkage with the genes/ QTLs for salinity tolerance and shall be the target for further studies. This revised version was published online in July 2006 with corrections to the Cover Date.     

Anthelmintic activity of Butea monosperma

The methanol extract of Butea monosperma seeds, tested in vitro, showed significant anthelmintic activity.     

Anaerobic Treatment of Brackishwater Aquaculture Sludge: An Alternative to Waste Stabilization Ponds

Environmental pressure, land utilization, and economic feasibility have resulted in the development of recirculating aquaculture systems (RAS). For many RAS, sludge is collected and washed from the system to waste stabilization ponds (WSPs). However, disposal of brackishwater aquaculture sludge into WSP is often prohibited because the high salinity can interfere with treatment. Moreover, there are problems associated with WSPs because of elevated salt content, such as the common practice of reusing treated water and land application of stabilized sludge. We tested and compared the treatment of brackishwater aquaculture sludge in an upflow anaerobic sludge blanket (UASB) reactor as an alternative to a WSP. In UASB, wastewater flows upward through a blanket of granular sludge and is treated by anaerobic micro‐organisms. Reduction in organic matter and 5‐d biochemical oxygen demand by 97 and 91%, respectively, was achieved in a UASB as compared to corresponding reductions of 22 and 41% in a WSP. During the UASB digestion process, methane is produced and recovered. Overall, a reduction in potential environmentally harmful factors such as salinization, land requirements, greenhouse gas emissions, as well as transportation costs are achieved, making the UASB reactor an attractive possible alternative for saline aquaculture sludge management.     

High frequency plantlet regeneration from nodal segment culture of female <Emphasis Type="Italic">Momordica dioica</Emphasis> (Roxb.)

An in vitro propagation method for female plants of Momordica dioica (Roxb.) has been established. The nodal segments were harvested and the cut ends of the explants were sealed with wax and then surface sterilized and cultured. Bud breaking occurred on Murashige and Skoog’s (MS) agar-gelled medium + 2.0 mg L⁻¹ 6-Benzylaminopurine (BAP) + 0.1 mg L⁻¹ Indole-3 acetic acid (IAA). The cultures were amplified by passages on MS medium supplemented with 1.0 mg L⁻¹ BAP + 0.1 mg L⁻¹ IAA. Further, shoot amplification (29.2 shoots per vessel) was achieved by subculturing of in vitro regenerated shoot clump on MS medium + 0.5 mg L⁻¹ BAP + 0.1 mg L⁻¹ IAA. The micropropagated shoots were subsequently transferred for root formation on half-strength MS medium + 2.0 mg L⁻¹ Indole-3 butyric acid (IBA) with 89% success rate. The in vitro-regenerated shoots were also rooted ex vitro with 34% success. These plantlets were hardened in the greenhouse and transferred to the field. The established protocol is suitable for true to type cloning of mature female plant of M. dioica.     

Concomitant production of lipase, protease and enterocin by Enterococcus faecium NCIM5363 and Enterococcus durans NCIM5427 isolated from fish processing waste

Enterococci are widely distributed in the environment ranging from foods to humans and are gaining industrial importance due to their technological traits. In the present study, enterococci (Enterococcus faecium NCIM5363 (EF-63) and Enterococcus durans NCIM5427 (ED-27)) which are native to fish processing waste with an ability to produce lipase, protease and enterocin concomitantly were characterised. Lipase assay was performed by titrimetry and protease activity and was estimated using haemoglobin and casein as substrates in the presence of buffers at acidic, basic and neutral pH. Furthermore, enterocin produced by the isolates was characterised. Enterocin was also checked for its stability at different pH, temperature and proteolytic enzymes. Lipase production was found to be 22 and 10 U/ml in the absence of tributryin and increased to 40 and 24 U/ml in its presence for EF-63 and ED-27, respectively, indicating that the lipase produced is substrate dependent. Protease production was confirmed by protease assay, and the protease produced showed more affinity towards the acidic substrate. Enterocin produced was stable at low pH (2 to 3) and high temperature (121°C, 15 min) and had a molecular weight of approximately 6 kDa. It exhibited antibacterial activity against both Gram-positive and Gram-negative food-borne pathogens. Proteinase K inactivated enterocin completely, whereas trypsin did not. Novelty of this work lies in the immense industrial importance these cultures hold as they are capable of producing lipase, protease and enterocin apart from being useful in recovering proteins and lipids from fish processing wastes.     

Comparative analysis of diversity based on morpho-agronomic traits and microsatellite markers in common bean

Morpho-agronomic traits and microsatellite markers were used to survey genetic diversity in 115 common bean genotypes that included 70 Indian landraces, 24 released varieties and 21 exotic accessions. Twelve morpho-agronomic traits, namely, days to 50% flowering, leaflet length, leaflet width, pod length, pod width, number of pods per plant, days to maturity, seed length, seed width, number of seeds per pod, 100 seed weight and seed yield per plant were studied. Field data of two consecutive years were subjected to multivariate analysis as proposed by Mahalanobis’s D²-statistics, Tochers method of clustering and combined analysis of variance. Seventeen microsatellite markers were also used to examine genetic diversity at molecular level that showed polymorphic information content (PIC) in the range of 0.00–0.684. Dendrograms based on Euclidean distances and UPGMA analysis showed the presence of majority of released varieties into single cluster, which pointed toward their low genetic base in comparison to indigenous landraces and exotic germplasm. Significant correlation existed between morphological genetic distance and microsatellite genetic distance tested by Mantel test (r = 0.876).     

Identification of PCR-based DNA Marker Linked to High Phytase Level of Wheat

The phytase is a key enzyme to hydrolyze phytic acid present in wheat grains and improves the bio-availability of micronutrients in monogastric animals. Phytase trait being contributed by specific regions of the genome requires identification of these regions, using suitable molecular markers. Hence, in the present investigation we attempted to develop a PCR-based marker that detects the phytase level in wheat. Six sets of PCR primers were designed on the basis of nucleotides sequence variation found in the sequence of both varieties. Out of six set of primers, one set amplified two different sized bands, i.e. 334 bp and 295 in two wheat cultivars C-306 (low phytase) and DBW 17 (high phytase), respectively. It exhibited a polymorphic banding pattern with length polymorphism and clearly separating low and high phytase genotypes. The primer set was also used for PCR of 46 synthetic hexaploids and 46 release varieties of wheat to validate the developed markers. Association among identified markers and phytase activity was found to be at 99.9% confidence level based on Fisher’s exact test (F-test). Therefore, this PCR primer set will be useful to select the wheat germplasm having high phytase levels and also in wheat breeding programs aimed at improving phytase levels in bread wheat cultivars.     

Dissolution of Dominant Soil Phosphorus Fractions in Phosphorus-Responsive Soils of Bihar,India: Effects of Mycorrhiza and Fertilizer Levels

We investigated the effects of Arbuscular Mycorrhiza (AM) fungi and various phosphorus (P) levels on the distribution and availability of P in dominant soils of Bihar, India. Potassium chloride (KCl)-P (labile P), sodium hydroxide (NaOH)-P (Fe-Al-bound P), hydrochloric acid (HCl)-P (Ca-bound P), and residual P (Res-P) fractions were analyzed in the soils under maize plant. Ca-bound P was the most abundant P fraction in the alkaline soils (65% of the total P) followed by neutral soil (35% of the total P), whereas it was less abundant (<4%) in the acidic soil type. Fe-Al-bound P was found to be highest for acidic soil (65% of the total P). Soils under the inoculation with Glomus mossae and control gave the highest and lowest values (15.63 mg kg^?1 and 10.74 mg kg^?1 respectively) for the labile fraction which was similar to the organically bound residual fractions of P (200.17 mg kg^?1 and 193.66 mg kg^?1 respectively. Inoculation of the soils with AM fungi leads to the redistribution of P fractions in different soils which consequently helps in improvement of available P in soil conducive for plant uptake.