Analysis of the transcript levels of VvAdh1, VvAdh2 and VvGrip4, three genes highly expressed during Vitis vinifera L. berry development

Well defined promoter sequences are required for the targeted expression of genes in transgenic grapevines. This paper describes a detailed study of the expression of three genes with potentially useful promoters. We have used real-time RT-PCR to evaluate the transcript levels of VvAdh1, VvAdh2 and VvGrip4 genes in various tissues (root, bud, tendril, inflorescence, fruit and embryogenic callus) of Vitis vinifera L. at different stages of development. Transcript levels of the three genes were highest in berry tissues but each had a distinct pattern of expression. VvAdh1 showed higher transcript levels during the early stages of berry development, while levels of VvAdh2 and VvGrip4 were higher during ripening. However, none of these genes was expressed in a strictly fruit-specific manner. In particular, significant levels of VvAdh1 and VvGrip4 transcripts were observed during late tendril development and in the inflorescence, respectively. Transcript levels of all three genes were similar in both flesh and skin, indicating no tissue-specificity within the berry. Promoter sequences of the VvAdh1, VvAdh2 and VvGrip4 genes could be very useful to drive ectopic gene expression in berries of transgenic plants

Correlates of Social, Demographic and Behavioral Factors affecting Adolescent Sexuality in a Traditional Society in India: Perspectives and Challenges

Background: Adolescent risk behavior is a major concern.Objectives: The study assessed the effect of school background, personal, social and other deviant behavior on school going adolescent sexual behavior. Material and Methods: The sample covered 3069 randomly selected adolescents (students) from 9th to 12th standard in various governments, private and missionary run schools from two districts namely; Aizawl being state capital and Champhai being distant under developed district.Summary: School background, leisure and entertainment practices, influence of taking alcohol, tobacco, drug, peer influence were found to be the major risk factor for indulgence in unsafe sex practices among adolescents. About 10% accepted involvement in premarital sex and majority of them (70%) of them had premarital sex between age group 15-19 years. The schools lacked in organizing awareness program and counseling activities on consequences of adolescent sex. The ARSH Program needs to synergize with school health program for desired results. Conclusions: Adolescent sexual risk behavior is a part of overall deviant behavior and can be managed in holistic manner

A large Indian family with rearrangement of chromosome 4p16 and 3p26.3 and divergent clinical presentations

BACKGROUND: The deletion of the chromosome 4p16.3 Wolf-Hirschhorn syndrome critical region (WHSCR-2) typically results in a characteristic facial appearance, varying intellectual disability, stereotypies and prenatal onset of growth retardation, while gains of the same chromosomal region result in a more variable degree of intellectual deficit and dysmorphism. Similarly the phenotype of individuals with terminal deletions of distal chromosome 3p (3p deletion syndrome) varies from mild to severe intellectual deficit, micro- and trigonocephaly, and a distinct facial appearance.METHODS AND RESULTS: We investigated a large Indian five-generation pedigree with ten affected family members in which chromosomal microarray and fluorescence in situ hybridization analyses disclosed a complex rearrangement involving chromosomal subregions 4p16.1 and 3p26.3 resulting in a 4p16.1 deletion and 3p26.3 microduplication in three individuals, and a 4p16.1 duplication and 3p26.3 microdeletion in seven individuals. A typical clinical presentation of WHS was observed in all three cases with 4p16.1 deletion and 3p26.3 microduplication. Individuals with a 4p16.1 duplication and 3p26.3 microdeletion demonstrated a range of clinical features including typical 3p microdeletion or 4p partial trisomy syndrome to more severe neurodevelopmental delay with distinct dysmorphic features.CONCLUSION: We present the largest pedigree with complex t(4p;3p) chromosomal rearrangements and diverse clinical outcomes including Wolf Hirschorn-, 3p deletion-, and 4p duplication syndrome amongst affected individuals.<br/

Hypomorphic PCNA mutation underlies a novel human DNA repair disorder

A number of human disorders, including Cockayne syndrome, UV-sensitive syndrome, xeroderma pigmentosum and trichothiodystrophy, result from the mutation of genes encoding molecules important for nucleotide excision repair. Here, we describe a novel syndrome in which the cardinal clinical features include postnatal growth retardation, hearing loss, premature aging, telangiectasia, neurological signs and photosensitivity, resulting from a homozygous missense (p.Ser228Ile) sequence alteration of the proliferating cell nuclear antigen (PCNA). PCNA is a highly conserved sliding clamp protein essential for DNA replication and repair. Due to this fundamental role, mutations in PCNA that profoundly impair protein function would be incompatible with life. Interestingly, while the p.Ser228Ile alteration appears to have no effect on protein levels or DNA replication, patient cells exhibit significant abnormalities in response to UV irradiation displaying substantial reductions in both UV survival and RNA synthesis recovery. The p.Ser228Ile change also profoundly alters PCNA’s interaction with Flap endonuclease 1 and DNA Ligase 1, DNA metabolism enzymes. Taken together our findings detail the first mutation of PCNA in humans, associated with a unique neurodegenerative disease displaying clinical and molecular features common to other DNA repair disorders, which we show to be attributable to a hypomorphic amino acid alteration

Next Generation Mapping of Enological Traits in an F2 Interspecific Grapevine Hybrid Family

In winegrapes (Vitis spp.), fruit quality traits such as berry color, total soluble solids content (SS), malic acid content (MA), and yeast assimilable nitrogen (YAN) affect fermentation or wine quality, and are important traits in selecting new hybrid winegrape cultivars. Given the high genetic diversity and heterozygosity of Vitis species and their tendency to exhibit inbreeding depression, linkage map construction and quantitative trait locus (QTL) mapping has relied on F1 families with the use of simple sequence repeat (SSR) and other markers. This study presents the construction of a genetic map by single nucleotide polymorphisms identified through genotyping-by-sequencing (GBS) technology in an F2 mapping family of 424 progeny derived from a cross between the wild species V. riparia Michx. and the interspecific hybrid winegrape cultivar, ‘Seyval’. The resulting map has 1449 markers spanning 2424 cM in genetic length across 19 linkage groups, covering 95% of the genome with an average distance between markers of 1.67 cM. Compared to an SSR map previously developed for this F2 family, these results represent an improved map covering a greater portion of the genome with higher marker density. The accuracy of the map was validated using the well-studied trait berry color. QTL affecting YAN, MA and SS related traits were detected. A joint MA and SS QTL spans a region with candidate genes involved in the malate metabolism pathway. We present an analytical pipeline for calling intercross GBS markers and a high-density linkage map for a large F2 family of the highly heterozygous Vitis genus. This study serves as a model for further genetic investigations of the molecular basis of additional unique characters of North American hybrid wine cultivars and to enhance the breeding process by marker-assisted selection. The GBS protocols for identifying intercross markers developed in this study can be adapted for other heterozygous species

Identification and Characterization of a Protease Encoding Metagenome Clone from Dairy Effluent

583-586Metagenomic DNA was isolated from dairy effluent collected from a milk processing unit. The Hpa I digests of metagenome were cloned in pEZ BAC vector. The transformed DH5 colonies were selected by blue white screening on chloramphenicol (25μg/ml) containing plates. Functional screening of the library on skim milk agar showed a clone having protease activity. This clone was confirmed to have a metagenome insert of size approx. 9kb. The crude cell extract of this clone at 24th h of growth showed protease activity of 166.99U/ml (111.32U/mg protein). The protease was found to be alkaline and had maximum activity at 400C and pH 9.0

Analysis of the transcript levels of VvAdh1, VvAdh2 and VvGrip4, three genes highly expressed during Vitis vinifera L. berry development

International audienceWell defined promoter sequences are required for the targeted expression of genes in transgenic grapevines. This paper describes a detailed study of the expression of three genes with potentially useful promoters. We have used real-time RT-PCR to evaluate the transcript levels of VvAdh1, VvAdh2 and VvGrip4 genes in various tissues (root, bud, tendril, inflorescence, fruit and embryogenic callus) of Vitis vinifera L. at different stages of development. Transcript levels of the three genes were highest in berry tissues but each had a distinct pattern of expression. VvAdh1 showed higher transcript levels during the early stages of berry development, while levels of VvAdh2 and VvGrip4 were higher during ripening. However, none of these genes was expressed in a strictly fruit-specific manner. In particular, significant levels of VvAdh1 and VvGrip4 transcripts were observed during late tendril development and in the inflorescence, respectively. Transcript levels of all three genes were similar in both flesh and skin, indicating no tissue-specificity within the berry. Promoter sequences of the VvAdh1, VvAdh2 and VvGrip4 genes could be very useful to drive ectopic gene expression in berries of transgenic plants