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.     

Integration of pre- and postharvest treatments for the control of black spot caused by Alternaria alternata in stored persimmon fruit

In Israel, black spot caused by Alternaria alternata is the main postharvest factor that impairs the quality and reduces the storability of persimmon fruit (Diospyros kaki cv. Triumph). The fungus infects the fruit in the orchard and remains quiescent until harvest. After harvest, the pathogen slowly colonizes the fruit during storage at 0 °C, which elicits black spot symptom development 2–3 months after storage entry. A commercial postharvest dip treatment in chlorine at 500 mg L^?1, released from sodium troclosene tablets, effectively controlled black spot in fruit stored for up to 2 months. However, decay incidence increased as the length of storage was extended beyond 2.5 months. The long incubation period that precedes black spot symptom development after harvest enabled the development of a series of integrative approaches for application at the pre- and postharvest stages, in combination with the commercial chlorine dip treatment, to improve the control of black spot disease. Preharvest treatments included treatment with the cytokinin-like N₁-(2-chloro-4-pyridyl)-N₃-phenylurea (CPPU) 30 d after fruit set, or a single spray with the curative fungicide polyoxin B 14 d before harvest, and when one of these was applied in combination with the postharvest chlorine dip treatment, the black spot infected area was reduced by 3 and 60%, respectively, compared with the chlorine dip alone. At the postharvest stage, fogging during storage, or post-storage on-line spraying with sodium troclosene, when applied in combination with the postharvest chlorine dip, improved the percentage of marketable fruit by 2 or 10%, respectively, compared with the chlorine dip alone. The results indicate that postharvest pathogens that show a slow colonization pattern might enable the integration of pre- and postharvest disease control methods to improve quality and reduce postharvest disease development.     

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.     

采前与采后处理结合控制柿子贮藏过程中链格孢菌(Alternaria alternata)引起的黑斑病的研究

由链格孢菌引起的黑斑病是导致以色列柿子(Diospyros kaki cv.Triumph)贮藏过程中果实品质下降、贮期缩短的主要因素。其病原真菌在采收前的果园中即开始侵染果实,但一直处于休眠状态。采收后,在0℃贮藏环境中病原菌开始繁殖,当贮藏2～3个月后,柿果表面出现黑斑症状。商业上采用二氯异氰尿酸钠片剂溶解后生成500 mg.L-1的氯水浸果,可在2个月的贮期内有效控制黑斑病的发生。然而,当贮期超过2个半月后,柿子果实的腐烂率则不断增加。在柿子采后出现黑斑症状之前病原真菌较长的潜伏期内,采前与采后阶段可采取一系列的综合技术措施,并与商业氯水浸果处理相结合来提高对贮藏柿果黑斑病的防控效果。采前处理包括柿子坐果30 d后,以1-(2-氯-4-吡啶)-3-苯基脲(CPPU)处理或采收前14 d以治疗性杀菌剂多抗霉素(polyoxin B)进行喷果处理。试验结果表明,与采后单独使用氯水浸果处理相比,将CPPU或polyoxin B处理与采后商业处理相结合,可使柿果黑斑浸染面积分别减小3%和60%;采收后贮藏期间二氯异氰尿酸钠喷雾处理或采后立即用二氯异氰尿酸钠喷果处理与采后商业氯水处理相结合比单独使用氯水处理可分别提高柿子好果率2%和10%。研究结果表明,对于具有较长潜伏侵染方式的采后病原菌来说,可应用采前和采后处理相结合的方法来提高果实的贮藏品质及减轻采后病害的发生。     

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).     

Clinical cytogenetics in equine stud practice: Consideration of the present situation and future application

The situation in Thoroughbred breeding in relation to cytogenetics is considered and it is indicated where screening of horses will be useful in practice. The techniques involved are explained and karyotypes are illustrated. The present state of knowledge in horse cytogenetics is reviewed. The types of abnormality are described in relation to phenotype. Future promise is discussed in relation to the human field, concluding that with the refined methods now available a wider rangeof meaningful aberrations will be uncovered.