RAPD and ISSR fingerprinting in cultivated chickpea (<Emphasis Type="Italic">Cicer arietinum</Emphasis> L.) and its wild progenitor <Emphasis Type="Italic">Cicer reticulatum</Emphasis> Ladizinsky

Detection of genetic relationships between 19 chickpea cultivars and five accessions of its wild progenitor Cicer reticulatum Ladizinsky were investigated by using RAPD and ISSR markers. On an average, six bands per primer were observed in RAPD analysis and 11 bands per primer in ISSR analysis. In RAPD, the wild accessions shared 77.8% polymorphic bands with chickpea cultivars, whereas they shared 79.6% polymorphic bands in ISSR analysis. In RAPD analysis 51.7% and 50.5% polymorphic bands were observed among wild accessions and chickpea cultivars, respectively. Similarly, 65.63% and 56.25% polymorphic bands were found in ISSR analysis. The dendrogram developed by pooling the data of RAPD and ISSR analysis revealed that the wild accessions and the ICCV lines showed similar pattern with the dendrogram of RAPD analysis. The ISSR analysis clearly indicated that even with six polymorphic primers, reliable estimation of genetic diversity could be obtained, while nearly 30 primers are required for RAPD. Moreover, RAPD can cause genotyping errors due to competition in the amplification of all RAPD fragments. The markers generated by ISSR and RAPD assays can provide practical information for the management of genetic resources. For the selection of good parental material in breeding programs the genetic data produced through ISSR can be used to correlate with the relationship measures based on pedigree data and morphological traits to minimize the individual inaccuracies in chickpea.     

Long-term manuring and fertilization influence soil inorganic phosphorus transformation vis-a-vis rice yield in a rice–wheat cropping system

A study on the rice–wheat cropping system was conducted at Bidhan Chandra Krishi Viswavidyalaya, West Bengal, India, to assess the effects of long-term manuring and fertilization on transformation of the inorganic phosphorus (P) fraction in soil after 22 years of the crop cycle. Soil samples were collected after Kharif from seven treated plots having different types of organic amendments like farm yard manure, paddy straw and green manuring with 50% substitution of nitrogen levels in rice crop only. The result showed that the yield trend of rice was maintained due to the buildup of P from various organic inputs. Although cultivation for 22 years without adding any fertilizer caused a significant decrease in almost all the forms of P viz. avail-P, saloid P, iron phosphorus fraction (Fe–P), aluminum phosphorus fraction (Al–P), calcium phosphorus fraction (Ca–P) and total P in control. Partial substitution of inorganic fertilizer N (50%) with organics, however, caused a significant increase in almost all the P fractions in soil over the control. The relative abundance of all the fractions of inorganic P irrespective of treatments was as follows: Fe–P > reductant soluble P fraction > occluded P > Al–P > Ca–P > saloid P. Saloid and Fe–P were the dominating fractions responsible for 92% variation of available P and total P levels, respectively.     

Fijian adolescents’ understanding and evaluation of climate change: Implications for enabling effective future adaptation

Pacific Island countries are particularly vulnerable to future manifestations of climate change due to high coastline‐to‐land‐area ratios, and high dependence of inhabitants on natural ecosystems. While everyone in the Pacific Islands should participate in climate change adaptation activities, it is the young people, given they are the generation likely to not only bear the burden of climate change, but to lead and live effective climate change adaptation activities and strategies specific to their region, the involvement of youth is critical. Pacific Island youths are often marginalised within traditional decision‐making hierarchies, therefore they are typically excluded from participating in meaningful discussions at community and government levels. Discussions were held with 30 adolescents aged 14–18 years in Fiji to explore knowledge and experiences regarding climate change. Participants revealed their dismay at their inability to talk to family – who they consider are not doing enough – about what they consider as appropriate responses to climate change, recommending the help of an authoritative outsider who could speak to their community leaders and family. Discussions also revealed that Fijian youth could not distinguish between changes in the climate and normal weather events, attesting to the importance of climate‐change education and awareness‐raising efforts within the Pacific Islands more generally.     

Characterization and molecular mapping of EMS-induced brittle culm mutants of diploid wheat (<Emphasis Type="Italic">Triticum monococcum</Emphasis> L.)

Diploid wheat (Triticum monococcum L, A^mA^m) is an ideal material for induced mutations which can be easily characterized and transferred to polyploid wheats. The EMS-induced brittle culm mutants, brc1, brc2, and brc3 used in the present investigation, were isolated from T. monococcum. All the brittle mutants had brittle roots, leaves, leaf sheaths, culms, and spikes, and were also susceptible to lodging. The mutants had 47–57% reduced α-cellulose in the secondary cell walls than that of T. monococcum indicating that all of them had defective synthesis of cellulose. All the mutants were monogenic recessive. Bulk segregation analysis of the mutants, using A^m genome anchored SSR markers in their F ₂ populations with T. boeoticum, located the mutants, brc1, brc2, and brc3 on chromosome 6A, 3A, and 1A of T. monococcum, respectively. Molecular analysis of the putatively linked markers showed that brc1 mapped on chromosome 6AS between Xbarc37 and Xbarc113 markers, brc2 on chromosome 3AL between Xcfd62 and Xcfa2170 markers whereas brc3 mapped on chromosome 1AL between Xgwm135 and Xwmc470 markers. Isolation and mapping of three different brittle culm mutants in wheat for the first time shows that there might be many more genes in wheat which affect synthesis and deposition of cellulose.     

Identification of biomarker for determining genotypic potential of nitrogen-use-efficiency and optimization of the nitrogen inputs in crop plants

Worldwide, the nitrogen use efficiency (NUE) for crop plants is of great concern. The burgeoning world population needs crop genotypes that respond to higher nitrogen and show a direct relationship to yield with use of nitrogen inputs, i.e. high nitrogen-responsive genotypes. However, for fulfilling the high global demand of organic produce, it requires the low nitrogen responsive genotypes with greater NUE and grain yields. The lack of knowledge about precise regulatory mechanisms to explain NUE in crop plants hampers the goal of agricultural productivity. Understanding the molecular basis of NUE will enable to provide handle for crop improvement through biotechnological means. With the advent of modern genomics and proteomics approaches such as subtractive hybridization, differential display, and microarray techniques are revolutionizing to identify the candidate genes which play a pivotal role in the regulation of NUE. Beside it, quantitative real-time polymerase chain reaction technology is also being used to establish marker-trait association for NUE. The identification of potential candidate genes/proteins in the regulation of NUE will serve as biomarker(s) for screening genotypes for their nitrogen responsiveness for optimization of nitrogen input in agriculture. This paper describes the molecular basis of NUE with respect to nitrogen metabolism and its intimate relationship with carbon metabolism, use of molecular-physiological-genetics approaches for understanding the role of various genes/proteins, and their validation to use as biomarker(s) for determining genotypic potential for NUE. Since NUE in plants is a complex trait which not only involves the primary process of nitrogen uptake and assimilatory pathways but also a series of events, including metabolite partitioning, secondary remobilization, C-N interactions, as well as molecular signalling pathways and regulatory control outside the metabolic cascades. Therefore, identification of novel nitrogen responsive genes and their cis- and trans-acting gene elements is essential. Thus, fishing out a single gene, biomarker or a master regulator controlling complex trait of NUE could serve as an appropriate strategy for nitrogen management in agriculture.     

Comparisons among cultivars of wheat,hulless and hulled oats: Dry matter,N and P accumulation and partitioning as affected by N supply

A better understanding of the impact of fertilizer nitrogen (N) on biomass and N accumulation, and their partitioning into different plant components is needed to optimize crop yield and quality. A field experiment with spring wheat (Triticum aestivum), hulless (Avena nuda), and hulled (Avena sativa) oats was conducted for 3 years in Ottawa, ON, Canada, to determine the crop responses to N addition (0, 75, and 150 kg N ha^–1). Biomass, N, and phosphorus (P) accumulation and partitioning into different plant components were examined during the growth season. Lodging score was determined for all crops when it occurred and again at harvest. During the growth season, both hulless and hulled oats and the wheat cultivar showed almost similar patterns of N and P accumulation with maximum contents at late grain filling or at harvest. Plant N concentration was up to 60 g kg^–1 during the seedling stage, decreased gradually with advancing growth stages, and was lowest at harvest. Nitrogen treatments significantly increased plant N and P contents. At heading stage, N treatments enhanced dry matter (24%–45%), N (35%–135%), and P (27%–45%) contents in plant components (i.e., culm, leaf, and head), but also enhanced crop lodging, especially in oats. Both hulled and hulless oats had higher total plant N (5%–35%), N : P ratio, and dry‐matter content in leaf (6%–43%) and head (0%–129%) along with higher P (up to 27%) in culm than the wheat cultivar. The wheat cultivar accumulated greater dry matter and higher N content in kernels than both hulled and hulless oats at harvest. Both hulled and hulless oat cultivars exhibited similar lodging susceptibility to N addition (75 or 150 kg N ha^–1), produced lower dry weight and lower kernel N, and hence lower grain yield than the wheat cultivar. The larger vegetative dry‐matter accumulation at heading coupled with higher P content in culms under high‐N‐supply conditions may be related to severe lodging in oat cultivars.     

Protection of mice against <Emphasis Type="Italic">Brucella abortus 544</Emphasis> challenge by vaccination with recombinant OMP28 adjuvanted with CpG oligonucleotides

Brucella abortus, a gram negative, facultative intracellular pathogen causes brucellosis in many animal species and humans. Although live, attenuated vaccines are available against this infection, they suffer from certain limitations. Therefore, the development of an effective subunit vaccine against brucellosis is an area of intense research. The outer membrane proteins (OMPs) of Brucella species have been extensively studied for its immunogenicity and protective ability. We have investigated the potential of CpG ODN to enhance the immunogenicity and protective efficacy of recombinant 28 kDa outer membrane protein (rOMP28) of Brucella melitensis. The study demonstrated vigorous immunoglobulin G (IgG) response of OMP28. The administration of rOMP28 with CpG caused increased cell mediated immune response in terms of induced IgG2a, T-cell proliferation and up-regulation of type I cytokine expression. In contrast, the free antigen suppressed the interferon gamma (type I cytokine) production on in-vitro stimulation of spleenocytes. The result indicates the role of OMP28 in the down regulation of IFN-γ production. Moreover, the B. abortus S-19 vaccinated mice showed highest production of IL-4 and IFN-γ. The protective ability of the antigen was evaluated by systemic bacterial clearance after challenging the mouse with B. abortus 544 pathogen. The level of protection was significant in rOMP28+CpG treated mice but was lower than the required level. The results of the present study indicate that rOMP28 could be an immunogen capable of inducing both humoral and cellular immune response. The humoral response was biased towards Th1 type when it was co-administered with CpG.     

Cryopreservation of silver barb Puntius gonionotus (Bleeker) spermatozoa: effect of extender composition,cryoprotective agents and freezing rate on their postthawing fertilization ability

The effects of extender composition, cryoprotectant concentration and freezing and thawing on the fertilization efficiency of cryopreserved spermatozoa of Puntius gonionotus were evaluated. Computer‐aided motility analysis of semen was conducted to check the suitability of spermatozoa for cryopreservation after mixing with different extenders and cryoprotective agents (CPAs). Extender‐4 with an osmolality 260 mOsmol kg⁻¹and pH 7.6 was used for the cryopreservation study. Among the CPAs, dimethyl sulphoxide (DMSO) was least toxic and more than 60% fertilization was achieved when used at 1.4 M at 0 °C for 10 and 30 min, whereas the toxicity of all CPAs to spermatozoa was evident when tested at 30 °C. Semen frozen at −16 °C min⁻¹ with 1.4 M DMSO showed 70% fertilization, which was significantly higher (P<0.05) than other freezing rates. Samples thawed at 35 °C water showed a fertilization rate comparable with that of fresh semen. Computer‐assisted semen analysis of fresh and frozen semen after thawing showed variations in different types of motility in spermatozoa and in their class. There was no significant difference in motility before or after cryopreservation; however, significant differences could be observed in the average path velocity (VAP), straight line velocity (VSL) and curve linear velocity (VCL). Semen of silver barb could be cryopreserved with extender‐4 by addition of 1.4 M DMSO to a final cryopreservation medium (MED 2) cooled at a rate of −16 °C min⁻¹, stored in liquid nitrogen (−196 °C) and utilized after thawing at 35±2 °C.     

Feed consumption, nutrient utilization and serum metabolite profile of captive blackbucks (Antelope cervicapra) fed diets varying in crude protein content

A feeding trial was conducted to determine the optimum level of crude protein (CP) in the diet of captive blackbuck (Antelope cervicapra) in which feed consumption and nutrient utilization are maximal. Fifteen blackbucks (BW 25-34 kg) were distributed into three groups of five each in an experiment of 75-days duration including a digestion trial of 5-day collection period. All the animals were offered 200 g of concentrates and fresh maize fodder ad libitum. The overall CP content of the three respective diets was 6.9%, 10.4% and 12.7%. Blood samples were collected on the last day of the experiment. Intake and digestibility of CP increased (p < 0.01) with the increased level of CP in the diet. Feed consumption and nutrient intake were not significantly different among the groups. However, digestibilities of most of the nutrients were higher in the 10.4% CP diet than in the 6.9% CP diet. The endogenous loss of nitrogen was similar among the groups. Based on the endogenous losses, minimum N requirement was calculated to be 776 mg/kg BW(0.75) /day, and to meet this requirement, diet must contain at least 8.27% CP. Serum urea nitrogen concentration increased (p < 0.01) with increased level of dietary CP. Serum level of glutamate oxaloacetate transaminase and glutamate pyruvate transaminase was higher (p < 0.05) in the group fed 6.9% CP diet. Animals in the group fed low protein diet also lost body mass during the experimental period. It was concluded that a diet containing 10.4% CP was optimum for maximizing nutrient utilization without any adverse effect on voluntary feed consumption and serum metabolite profile of blackbucks.