Salt-tolerance in Brassica juncea L. I. In vitro selection,agronomic evaluation and genetic stability

Summary In vitro selection of salt tolerant plants of Brassica juncea L. (Indian mustard) cv. Prakash has been accomplished by screening highly morphogenic cotyledon explant cultures on high NaCl media. Out of a total of 2,620 cotyledons cultured on high salt medium, 3 survived, showed sustained growth and regenerated shoots. They were multiplied by axillary bud culture on NaCl free medium. The salt-selected shoots retained salt tolerance following 3 month of growth and multiplication on control medium. While two of these somaclones flowered and set seeds, third one grew slowly, had abnormal leaf morphology and was sterile. The seed of the two fertile plants were sown in the field to raise R₁ segregating generation. Data were recorded for field, other agronomic components and oil content. The somaclonal lines, both selected salt-tolerant and non-selected, showed tremendous amount of variation for all the characters studied. One of the two tolerant somaclones invariably showed reduced height, longer reproductive phase and higher 1000 seed weight. Based on the agronomic performance of R₁ plants of these somaclones, some plants were selected and their progeny were evaluated for agronomic performance under standard field conditions and salt-tolerance in the greenhouse using sand pot culture method. Most of the lines bred true for their specific characteristics. In the greenhouse, selected salt-tolerant somaclones (SR-2 and SR-3) performed better for plant growth, yield and other agronomic traits at higher salt treatments, indicating thereby that salt-tolerance trait selected in vitro was expressed in the whole plants and is genetically stable and transmitted onto the progeny. The two tolerant lines, however, differed in their salt-tolerance during vegetative and reproductive phases as indicated by their salt-tolerance and stress susceptibility indices. The mechanism of salt-tolerance is not clear and needs to be further investigated.     

Molecular mapping of Aegilops speltoides derived leaf rust resistance gene Lr28 in wheat

In a segregating homozygous F₂ population of bread wheat involving a leaf rust resistance gene Lr28 derived from Aegilops speltoides, six randomly amplified polymorphic DNA (RAPD) markers, three each in coupling and repulsion phase were identified as linked to Lr28, mapped to a region spanning 32 cM including the locus. The F₂ and F₃ populations were studied in the phytotron challenged with the most virulent pathotype 77-5 of leaf rust. A coupling phase linked RAPD marker S464₇₂₁ and a repulsion phase linked RAPD marker S326₅₅₀ flanked the gene Lr28 by a distance of 2.4± 0.016 cM on either side. The flanking markers genetically worked as co-dominant markers when analyzed together after separate amplification in the F₂ population by distinguishing the homozygotes from the heterozygotes and increased the efficiency of marker assisted selection by reducing the false positives and negatives. One of the three RAPD markers, S421₆₄₀ was converted to locus specific SCAR marker SCS421₆₄₀ which was further truncated by designing primers internal from both ends of the original RAPD amplicon to eliminate a non-specific amplification of nearly same size. The truncated polymorphic sequence characterized amplified region marker (TPSCAR) SCS421₅₇₀ was 70 bp smaller, but resulted in a single band polymorphism specific to Lr28 resistance. The TPSCAR marker was validated for its specificity to the gene Lr28 in nine different genetic backgrounds and on 43 of the 50 Lr genes of both native and alien origin, suggesting the utility of the SCAR markers in pyramiding leaf rust resistance genes in wheat.     

Burrow characteristics of the co-existing sibling species Mus booduga and Mus terricolor and the genetic basis of adaptation to hypoxic/hypercapnic stress

Background  

The co-existing, sibling species Mus booduga and Mus terricolor show a difference in site-preference for burrows. The former build them in flat portion of the fields while the latter make burrows in earthen mounds raised for holding water in cultivated fields. In northern India which experiences great variation in climatic condition between summer and winter, M. booduga burrows have an average depth of 41 cm, as against 30 cm in southern India with less climatic fluctuation.     

Micropropagation of yellow mangosteen: a valuable endemic tree of India

We developed a method for in vitro regeneration of Garcinia xanthochymus(yellow mangosteen) from matured seed segments. Multiple shoots were induced on woody plant(WP) medium supplemented with cytokinins.An average of 11 shoots per explant were regenerated from mature seed segments on WP medium containing 20 μM 6-benzylaminopurine. Histological analysis revealed that hypodermal cells of seed segments were initially involved in active division, which later developed into meristemoids, subsequently leading to the formation of shoot buds.Shoot elongation was achieved by repeated subculturing of seed explants in shoot regeneration medium. Rooting of shoots was achieved on WP medium supplemented with indole-3-butyric acid or a-naphthalene acetic acid. Plantlets were transplanted to pots containing soil: compost(1:1) and survival rate was 90%.     

A preliminary study to evaluate the immune responses induced by immunization of dogs with inactivated Ehrlichia canis organisms

Ehrlichia canis is an intracellular pathogen that causes canine monocytic ehrlichiosis. Although the role of antibody responses cannot be discounted, control of this intracellular pathogen is expected to be by cell mediated immune responses. The immune responses in dogs immunized with inactivated E. canis organisms in combination with Quil A were evaluated. Immunization provoked strong humoral and cellular immune responses, which were demonstrable by Western blotting and lymphocyte proliferation assays. By Western blotting antibodies to several immunodominant E. canis proteins were detected in serum from immunized dogs and antibody titres increased after each immunization. The complement of immunogenic proteins recognized by the antisera were similar to those recognized in serum from infected dogs. Upon challenge with live E. canis, rapid anamnestic humoral responses were detected in the serum of immunized dogs and primary antibody responses were detected in the serum from control dogs. Following immunization, a lymphocyte proliferative response (cellular immunity) was detected in peripheral blood mononuclear cells (PBMNs) of immunized dogs upon stimulation with E. canis antigens. These responses were absent from non-immunized control dogs until after infection with live E. canis, when antigen specific-lymphocyte proliferation responses were also detected in the PBMNs of the control dogs. It can be thus concluded that immunization against canine monocytic ehrlichiosis may be feasible. However, the immunization regimen needs to be optimized and a detailed investigation needs to be done to determine if this regimen can prevent development of acute and chronic disease.     

Enzymatic Peeling of Potato: A Novel Processing Technology

Development of an innovative biotechnological method for potato peeling, closer to the ‘ideal’ peeling conditions of the product, was the main objective of the present research. Studies on enzymatic hydrolysis of potato peel were conducted. The efficacy of enzymatic peel hydrolysis was expressed in terms of reducing sugar content of the enzyme solution in which potato peel was incubated. Enzyme screening revealed that an enzyme solution containing a cellulase-xylanase mixture and amylase in a ratio of 1:1 showed good peel hydrolyzing efficiency of peeled peels. To further maximise the peel hydrolysis, condition parameters were optimised using response surface methodology (RSM). At the optimised conditions of 60 °C, pH 6 and 4 h, the reducing sugar yield in the solution was maximum. Characterization of the potato peel using microscopic techniques (scanning electron microscopy (SEM), atomic force microscopy (AFM)) further illustrated degradation of cell wall and reduction in the surface roughness of the peel after enzymatic treatment, which could enhance peel loosening from the intact potato. To further ascertain the efficiency of the process, studies were conducted with the selected enzyme on intact potato tubers under optimised conditions. Easy removal of peel was observed in enzyme-treated potato tubers, which showed 0.52% peel loss by abrasive peeling. The present process employing enzymes could be applied for peeling of intact potato as an alternative to conventional peeling process.     

Genetic diversity and relationship between Bradyrhizobium strains isolated from blackgram and cowpea

The genetic diversity of bradyrhizobial strains associated with blackgram and cowpea grown in two different agricultural soils (non-saline and saline) along the coastline of Tamil Nadu has been analysed. Phenotypically indistinguishable isolates were analysed for DNA polymorphism using random amplification of polymorphic DNA (RAPD) and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of 16S rDNA and nifD. Although these bacteria belong to a group with a broad host range, RAPD analysis showed a considerable level of genetic diversity among the strains isolated from different host plants. Soil pH and salinity seem to have an effect on the selection of natural populations as revealed by PCR-RFLP of 16S rDNA. A combination of PCR-RFLP genotyping with nodulation studies indicates that monocropping of blackgram and the salinity of the soil have made ineffective rhizobia the dominant genotype, thereby creating an ecological burden on their other compatible hosts. A group of strains and a type strain sharing three different 16S PCR-RFLP types were shown to have the same set of symbiotic genes as inferred from the PCR-RFLP pattern of nifD. Another group of cowpea rhizobia that were found to be effective nitrogen fixers and sharing distinct 16S profiles were found to have a different set of symbiotic genes.     

Water productivity and nutrient status of rice soil in response to cultivation techniques and nitrogen fertilization

Three methods of rice cultivation were compared in a field experiment at New Delhi, India during 2012 for their water use and changes in nutrient availability of soil. The experiment was laid out in a split plot design with conventional transplanting (CT), system of rice intensification (SRI), and aerobic rice (AR) cultivation technologies. Five doses of nitrogen included 100 % (120 kg N ha^?1), 125, and 150 % recommended dose of N(RDN) through urea, 75 % of RDN through urea (90 kg N ha^?1) + 25 % of RDN (30 kg ha^?1) through farm yard manure (FYM), and 100 % of RDN through FYM. Results revealed that status of available N in soil under rice at 45 and 90 days after sowing (DAS) was significantly higher in CT and SRI compared to AR method. Application of the highest dose of nitrogen through urea resulted in the highest availability of N (188.9, 174.2, and 135.2 kg ha^?1 for 45 and 90 DAS and at harvest stage, respectively). The soil under AR recorded significantly low availability of phosphorus and iron. However, availability of K in soil was not affected significantly under adopted production techniques and nitrogen management. The recorded irrigation water productivity was maximum in AR cultivation (9.16 kg ha mm^?1) followed by SRI (7.02 kg ha mm^?1) with irrigation water saving of 54 and 36 %, respectively compared to CT.     

Genetic tagging of Azotobacter chroococcum for colonization on wheat (Triticum aestivum) and cotton (Gossypium sp.) roots

Abstract

Azotobacter chroococcum strains E12, HT57 were genetically tagged with lac Z, gfp to study the colonization behaviour on wheat (Triticum aestivum) and cotton (Gossypium sp.) in soil under controlled conditions. 10³ – 10⁴ cfu g^?1 soil of HT57 lac Z were found to colonize roots of both cotton and wheat crops whereas 1.7 × 10⁴ – 7.2 × 10⁴ cfu g^?1 soil of E12 gfp was colonizing wheat roots and 1.6 × 10⁴ – 9.3 × 10⁴ cfu g^?1 soil of E12 gfp colonized cotton roots respectively. Tagged strains colonized mostly on root tips compared to basal roots in both the crops.     

Nitric oxide accumulation and actin distribution during auxin-induced adventitious root development in sunflower

Auxin-mediated adventitious root (AR) formation from the basal ends of hypocotyl segments in sunflower (Helianthus annuus L., cv. Morden) is associated with gradual subcellular and anatomical changes. Pre-mitotic events (induction phase) are evident within 1 d (24 h) in the interfascicular parenchyma. Evidence for nitric oxide (NO) accumulation in the interfascicular cells 2 d after indole-3-acetic acid (IAA) treatment, highlights the involvement of NO specifically during root initiation, following AR induction phase. Thus, adventitious root induction and initiation can be distinguished as NO-independent and NO-dependent phases. Treatment of hypocotyl explants with 10 μM 1-napthylphthlamic acid (NPA), an inhibitor of polar auxin transport in plant cells, leads to complete abolition of NO-associated fluorescence, detected upon incubation of hypocotyl sections with 4,5-diaminofluorescein diacetate (DAF-2DA). This indicates a signaling link between auxin transport and NO accumulation in the target cells. Latrunculin B (Lat B), an actin depolymerizing agent, leads to substantial inhibition of IAA-induced AR response and NO accumulation in the responding cells. Aminoguanidine [an inhibitor of inducible form of nitric oxide synthase (iNOS)] brings about a reduction in the tissue NO content, indicating a significant activity of putative NOS leading to endogenous NO accumulation. Confocal laser scanning microscope (CLSM) imaging has revealed bundling of actin with NPA and Lat B treatments. A probable interaction is thus evident between actin and NO during auxin-mediated AR formation in the initiation and extension phase. Present work, thus, provides novel observations on an interaction among endogenous NO, IAA and actin at specific stages of AR formation.