Identification and validation of cold responsive microRNAs of <Emphasis Type="Italic">Hevea brasiliensis</Emphasis> using high throughput sequencing

Cold stress is one of the major abiotic factors that influence the productivity and geographical distribution of many agriculturally important crops like Hevea brasiliensis. Cultivation of H. brasiliensis in India is being extended to northeastern regions, where low temperature during winter adversely affects its survival, growth, and productivity. Developing cold-tolerant genotypes is a primary requisite to maximize the productivity under such challenging environmental conditions. However, lack of methods for early evaluation of cold tolerance in the newly developed clones and the extensive time required for assessing their tolerance in the field are major constraints for clonal selection. The present study was initiated with an objective to identify and characterize cold stress responsive miRNAs from H. brasiliensis that show stronger association with cold tolerance. Next generation sequencing using Illumina HiSeq method revealed the expression of 21 and 29 conserved miRNA (from clone RRIM 600) families in cold-stressed and control samples, respectively. Forty-two novel miRNAs were identified from this study. Upon differential expression analysis, eight conserved miRNAs were found commonly expressed in both the samples. When expression analyses were performed subsequently with six selected miRNAs in two Hevea clones (viz. RRII 105 and RRIM 600), miR169 showed a strong association with cold tolerance. miRNAs such as miR482 and miR159 also exhibited association with cold tolerance. This study suggests the possibility of employing these miRNAs as markers for cold tolerance after validation in more number of genotypes with varying levels of cold tolerance.     

Dysgenesis of the tubal system in an infertile mare karyotype 63,X:64,XY:65,XXY

A Thoroughbred mare, barren during three years at stud, unresponsive to therapy, and not palpably abnormal anatomically was found to have hypoplasia of the uterine tubes after excision at laparotomy. The cause was an unbalanced genotype. The mosaic karyotype, determined on a blood sample, was indispensible in diagnosis and the case extends the potential for cytogenetic prognostic procedures.     

OSTEOCHONDROSIS IN THE LATERAL FEMORAL CONDYLES OF A HORSE

Osteochondrosis of the lateral femoral condyles was diagnosed radiographically in an 8-month-old, female Arabian horse, which had been presented with a hindlimb lameness. The diagnosis was confirmed by gross and microscopic pathology. The location of the lesions was considered unusual for osteochondrosis in the horse.     

Recombinant Bone Morphogenetic Proteins: Novel Substances for Enhancing Bone Healing

Bone morphogenetic proteins (BMPs) are differentiative factors whose principal function is to induce transformation of undifferentiated mesenchymal cells into chondroblasts and osteoblasts in a dose-dependent manner. Bone morphogenetic proteins have been isolated postnatally in mammals from bone matrix, periosteal cells, mesenchymal cells of marrow stroma, tooth anlagen, and cells of osteosarcoma and chondrosarcoma. Distribution in additional embryonic tissues implies a broader organogenic function. Bone morphogenetic proteins are the only differentiative factors able to singularly induce de novo bone formation in vitro and in vivo. Recombinant DNA technology allows their production in large and highly purified quantities. The BMPs' osteoinductive ability has been shown with a variety of carriers including collagens and polymers at heterotopic and orthotopic sites in a wide range of species. They are presently being readied for clinical use as alternatives to bone grafts. Other potential applications include use as pulp capping agents, promoters of implant osteointegration and soft tissue reunion with bone, treatments for nonadaptive bone disease, and implants for use with mitotically expanded skeletal stem cell populations. Errors in the genetic coding of BMPs may manifest as clinical disease entities.     

Importance of sepals in fruit and seed development in linseed (Linum usitatissimum L.)

Summary The green sepals in linseed (Linum usitatissimum L.) persist till almost the time of fruit ripening. Since the plant retains very few leaves and the fruits are better exposed to receive sunlight, the green sepals may be important in fruit and seed development. Influence of the removal of 0, 1, 3 and 5 sepals on the day of anthesis, 7, 15 and 25 days after anthesis on fruit and seed development was examined. Excision of all the fine sepals on the day of anthesis stopped fruit and seed development in the varieties M 10 and NP (RR) 45 but in Mukta the reduction was about 55%. Removal of three sepals also decreased fruit growth in Mukta and NP (RR) 45. The effect of removal of sepals was more pronounced on the number of seeds per fruit than on seed weight. Sepals fixed ¹⁴CO₂ and translocated ¹⁴C to seeds. However, the ¹⁴CO₂ fixation was maximum at early stages of fruit development, whereas a higher percentage of the fixed ¹⁴C was translocated at later stages of growth. It is suggested that the sepal size in linseed could be a useful selection criterion.     

Molecular tagging of Asiatic soybean rust resistance in exotic genotype EC 241780 reveals complementation of two genes

Asian soybean rust (ASR) caused by Phakopsora pachyrhizi severely reduces seed yield in soybean. Molecular tagging of ASR resistance can help in the process of resistance breeding. In this study, an F₂ population of cross (susceptible cultivar ‘NRC 7’ × resistant exotic genotype EC 241780) was used for bulked segregant analysis (BSA) with 25 SSR (simple sequence repeat) primers linked with six Rpp genes. Among them, five polymorphic SSR markers, viz., Sct 187, SSR 1859, Satt 191 (Rpp1b like loci) and Satt 215, Sat_361 (Rpp2 loci) distinguished the ASR resistant and susceptible bulks and individuals. In combined marker analysis, the markers Satt 191 (Rpp1b like loci) and Satt 215 (Rpp2 loci) were linked with ASR severity score and were also confirmed in individual 110 F₂ segregants. Hence, these markers could be utilized in the marker assisted rust resistance breeding of Rpp1b like and Rpp2 genes. In silico candidate gene analysis for hypersensitive response revealed that Satt 191 linked region was rich in genes encoding apoptotic ATPase having leucine‐rich repeat (LRR) domain.     

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.     

Follow-up of mercury levels in fish,human hair and urine in the Madeira and Tapajós basins,Amazon, Brazil

Informal economy of gold mining has contaminated some important river basins in Amazon. Follow-up studies on critical compartments showed some areas with high Hg levels in fish as well as in human hair samples. Average Hg in piscivorous fish in the Madeira river itself was 846 ppb (N=284) with a maximum of 3921 ppb. Mercury in fish from non polluted areas in this basin shows high variability, even for single species. A seasonal variation in Hg content was observed, with higher values at the end of the dry season. In the upper Tapajós basin comparable values were found for fish but with a definite decreasing trend downstream. Average value for piscivorous fish in the whole Tapajós basin is 482 ppb (N=122) with a maximum value of 3770 ppb. Hair Hg was higher in fishing villages in the Tapajós (average: 17 ppm; with N=432 and maximum value of 176 ppm) than in the Madeira (average: 9 ppm; N=169; maximum 71 ppm), and data from some areas of the Tapajós suggest a decrease with time. Mercury was much higher in urine of goldshop workers in Santarém (low Tapajós) than in Alta Floresta (high Tapajbs) and show a decreasing trend in both cases, probably related to the significant decline in gold mining activities during the study period (1986–1994).