Dissecting Arabidopsis thaliana DICER function in small RNA processing, gene silencing and DNA methylation patterning.

Small RNAs have several important biological functions. MicroRNAs (miRNAs) and trans-acting small interfering RNAs (tasiRNAs) regulate mRNA stability and translation, and siRNAs cause post-transcriptional gene silencing of transposons, viruses and transgenes and are important in both the establishment and maintenance of cytosine DNA methylation. Here, we study the role of the four Arabidopsis thaliana DICER-LIKE genes (DCL1-DCL4) in these processes. Sequencing of small RNAs from a dcl2 dcl3 dcl4 triple mutant showed markedly reduced tasiRNA and siRNA production and indicated that DCL1, in addition to its role as the major enzyme for processing miRNAs, has a previously unknown role in the production of small RNAs from endogenous inverted repeats. DCL2, DCL3 and DCL4 showed functional redundancy in siRNA and tasiRNA production and in the establishment and maintenance of DNA methylation. Our studies also suggest that asymmetric DNA methylation can be maintained by pathways that do not require siRNAs.

MicroRNAs and other small RNAs enriched in the Arabidopsis RNA-dependent RNA polymerase-2 mutant.

The Arabidopsis genome contains a highly complex and abundant population of small RNAs, and many of the endogenous siRNAs are dependent on RNA-Dependent RNA Polymerase 2 (RDR2) for their biogenesis. By analyzing an rdr2 loss-of-function mutant using two different parallel sequencing technologies, MPSS and 454, we characterized the complement of miRNAs expressed in Arabidopsis inflorescence to considerable depth. Nearly all known miRNAs were enriched in this mutant and we identified 13 new miRNAs, all of which were relatively low abundance and constitute new families. Trans-acting siRNAs (ta-siRNAs) were even more highly enriched. Computational and gel blot analyses suggested that the minimal number of miRNAs in Arabidopsis is approximately 155. The size profile of small RNAs in rdr2 reflected enrichment of 21-nt miRNAs and other classes of siRNAs like ta-siRNAs, and a significant reduction in 24-nt heterochromatic siRNAs. Other classes of small RNAs were found to be RDR2-independent, particularly those derived from long inverted repeats and a subset of tandem repeats. The small RNA populations in other Arabidopsis small RNA biogenesis mutants were also examined; a dcl2/3/4 triple mutant showed a similar pattern to rdr2, whereas dcl1-7 and rdr6 showed reductions in miRNAs and ta-siRNAs consistent with their activities in the biogenesis of these types of small RNAs. Deep sequencing of mutants provides a genetic approach for the dissection and characterization of diverse small RNA populations and the identification of low abundance miRNAs.

Role of RNA polymerase IV in plant small RNA metabolism.

In addition to the three RNA polymerases (RNAP I-III) shared by all eukaryotic organisms, plant genomes encode a fourth RNAP (RNAP IV) that appears to be specialized in the production of siRNAs. Available data support a model in which dsRNAs are generated by RNAP IV and RNA-dependent RNAP 2 (RDR2) and processed by DICER (DCL) enzymes into 21- to 24-nt siRNAs, which are associated with different ARGONAUTE (AGO) proteins for transcriptional or posttranscriptional gene silencing. However, it is not yet clear what fraction of genomic siRNA production is RNAP IV-dependent, and to what extent these siRNAs are preferentially processed by certain DCL(s) or associated with specific AGOs for distinct downstream functions. To address these questions on a genome-wide scale, we sequenced approximately 335,000 siRNAs from wild-type and RNAP IV mutant Arabidopsis plants by using 454 technology. The results show that RNAP IV is required for the production of >90% of all siRNAs, which are faithfully produced from a discrete set of genomic loci. Comparisons of these siRNAs with those accumulated in rdr2 and dcl2 dcl3 dcl4 and those associated with AGO1 and AGO4 provide important information regarding the processing, channeling, and functions of plant siRNAs. We also describe a class of RNAP IV-independent siRNAs produced from endogenous single-stranded hairpin RNA precursors.

Impact of small indigenous species of fish (SIS) on livelihood of local fishing community in two upazilas of Mymensingh

The fisher folk used to catch small indigenous species of fish (SIS) from rivers, canals, wetlands and floodplains at little or no cost for their livelihood. Surplus fish was sold at the local market to generate some little capital for the households. The livelihood and consumption of SIS in fishing community of two upazilas viz. Trisal and Ishwarganj under Mymensingh district were studied for 3 months in 2004. Most of the fisher folk of the study areas belong to resource-poor section of the society living below the poverty level. Majority of them had no cultivable land. As professional fishers they face many problems during lean fishing period from January to April due to little or non-availability of fish. Majority of the fisher households consumed SIS three to four days a week. The fisher households of Trisal upazila consumed more small fish than those of Ishwargonj upazila. More than 50% respondents consumed <20 g SIS/day and 20% consumed >40 g SIS/day in Trisal upazila. On the other hand, in Ishwargonj upzila, most of the fisher households (66%) were found to consume <20 g SIS/day. SIS was mostly available from July-December in rivers, wetlands (beels), and canals, and income from fishing was reported to be good. The dominant SIS was Puntius spp., Mystus spp., Anabas testudineus, catfishes, mola, and small prawns. Non-indigenous species like tilapia was also dominant in Trisal upzila where aquaculture practices were well established.

Marketing of small indigenous species of fish (SIS) in three selected fish markets of Mymensingh

A socio-economic survey was conducted round the year in three fish markets at Mymensingh, Bangladesh. The selected markets were categorized as rural market (Sutiakhali market), a peri-urban market (Kamal Ranjeet market, BAU) and an urban market (Notun Bazar market, Mymensingh town). It was learnt from the survey that the availability of Small Indigenous Fish Species (SIS) declined to a great extent over the last few years and at presently many of such fish species are either threatened or at the edge of extinction. The supply of SIS was highest in KR market (37% of total) and more or less similar in Notun Bazar and Sutiakhali fish market (25 and 27% respectively). The total supply of SIS fluctuated from 25% to 35% throughout the year in these markets. About 48 SIS were found in the sampled markets over the survey period. The highest number of species (45) was found in KR market followed by Notun Bazar (42) and Sutiakhali (37) fish markets. During the survey, three critically endangered species namely, schilbid catfish, garua catfish and rita were found in these markets. Beside these, other 11 and 10 species were listed to be endangered and vulnerable respectively. The biodiversity of 21 SIS found in three markets were no threat at all. Three species (guntea loach, Indian glass barb and flying barb) were 'data deficient' as reported by the IUCN Red Book (IUCN-Bangladesh 2000). From the supply point of view small prawn, spotted snakehead, stinging catfish, pool barb, striped dwarf catfish, Gangetic mystus, walking catfish and tank goby were the prominent fish. The least available species found in this survey were lesser spiny eel, barred spiny eel, Gangetic ailia, freshwater garfish, zig-zag eel, flying barb, Ganges river sprat, freshwater river shad and dwarf gourami. The weight of SIS available in Notun bazar was highest and nearly double than other two markets. There was no significant difference recorded in the supply of SIS in Sutiakhali and KR markets. The average monthly SIS supply was 185, 192 and 467 kg in KR, Sutiakhali and Notun Bazar, respectively; therefore, the cumulative average supply was 844 kg per month in three markets. The price of SIS ranged widely from taka 50-450/kg depending on species, location of market, time of purchase and the condition of fish. In general small prawn, ticto barb, dwarf gourami, Gangetic leaffish, and Annandale loach were sold at a lower price (ranged taka 50-100/kg) and these species could be considered at the bottom of the market-price list. Other SIS like walking catfish, climbing parch, butter catfish, cotio and schilbid catfish valued as highest price (ranged taka 150-450/kg). There was no specific marketing chain for SIS in Mymensingh region. The components of marketing channels and their expansion varied with seasons and locations. The general pattern, however, was as this - after buying fish from fish farmer/fishermen, middlemen (locally known as Foria) used to buy fish to wholesale market and sell to the wholesalers. The retailers used to buy fish from wholesaler through auction to the highest bidders. The retailers then send the fish to particular market where the fish reached the consumers. The livelihood strategy of SIS retailers in three fish markets showed that socio-economic constraints such as low income, poor educational background, low economic status and lack of capital are the main constrains [sic]. Most of the retailers proposed that government should control the fish price throughout the year, so that the producers can get reasonable and stable price. Construction of cold storage and preservation facilities at market sites, improvement of road and communication, improvement of physical market facilities and reduction of market chain is essential. Credit facilities, improvement of their standard of living, health and sanitary condition, housing condition, children education and access to drinking water facilities were identified as additional aspects to improve socio-economic condition of SIS retailers.

Probing small molecule binding to amyloid fibrils.

Much effort has focussed in recent years on probing the interactions of small molecules with amyloid fibrils and other protein aggregates. Understanding and control of such interactions are important for the development of diagnostic and therapeutic strategies in situations where protein aggregation is associated with disease. In this perspective article we give an overview over the toolbox of biophysical methods for the study of such amyloid-small molecule interactions. We discuss in detail two recently developed techniques within this framework: linear dichroism, a promising extension of the more traditional spectroscopic techniques, and biosensing methods, where surface-bound amyloid fibrils are exposed to solutions of small molecules. Both techniques rely on the measurement of physical properties that are very directly linked to the binding of small molecules to amyloid aggregates and therefore provide an attractive route to probe these important interactions.

A review of experimental trawling from small and medium sized mechanized vessels off Kanikada

The most suitable otter trawl for small boats was found to be a 10.9 to 15m 2-seam trawl with 100cm x 50cm x 35kg horizontally curved otter boards together with long single sweep line. For operation from medium sized trawls, 18.26m 2-seam 18.3m 4-seam and 29.26m long wing trawl were found suitable. An 18.3m 4-seam trawl was netting a considerable quantity of off-bottom fishes. Shrimps predominated in the catches of the 29.26m trawl. Productive grounds for Cynagris species, Psenus species and Decapterus species within 50 to 100m depth ranges off Kakinada were available for profitable exploitation.