RESUMO
Suppression subtractive hybridization complementary DNA libraries identified differentially expressed genes in liver tissue of winter flounder collected from the highly impacted Raritan-Hudson estuary versus those from less industrialized estuaries farther south in New Jersey. Distinct transcript profiles emerged in the fish from these different habitats. A total of 251 clones from the forward (upregulated with anthropogenic impact) and reverse (downregulated with anthropogenic impact) subtracted libraries were sequenced. In the upregulated library immune response transcripts, including complement C-3, C-7, factor H, factor Bf/C2, differentially regulated trout protein 1, and the antimicrobial hepcidin, indicated the pollution-impacted fish were under a high viral or bacterial load. Transcripts for cytochrome P450 1A, P450 3A, and glutathione S-transferase, important components of phase I and II metabolism of xenobiotics, were found in the upregulated-with-pollution library. Vitellogenins I and II and egg envelope protein (zp) appeared to be downregulated. A homologue of the tumor suppressor p33(ING1) (down) and hepatocyte growth factor-like protein (up) may indicate liver damage or hepatocellular carcinoma or hepatoma. These expression patterns, confirmed by quantitative polymerase chain reaction, indicate that transcript analysis is a useful method for assessing the health of local habitats and the organisms therein.
Assuntos
Meio Ambiente , Linguado/genética , Regulação da Expressão Gênica , Fígado/metabolismo , Poluição Química da Água , Animais , Sequência de Bases , Primers do DNA , Linguado/metabolismo , Biblioteca Gênica , Marcadores Genéticos/genética , Dados de Sequência Molecular , New Jersey , Hibridização de Ácido Nucleico/métodos , Reação em Cadeia da Polimerase/métodos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNARESUMO
Tissue cultures of the halophytic saltmarsh grass Sporobolus virginicus were initiated from unemerged immature inflorescence tissue. Typical graminaceous embryogenic and nonembryogenic callus and cell types were noted. Embryogenic callus was compact golden yellow. Histological evidence indicated that proliferation of the ovary tissue of the immature pistil was the source for embryogenic callus. Plants regenerated after first reducing and then eliminating auxin from the culture medium. Regeneration was observed both through the concerted development of bipolar meristems from somatic embryos and by the formation of multiple shoot meristems that were either connected through callus tissue to root meristems or which later adventitiously rooted. The main mode of regeneration appeared to be somatic embryogenesis with additional multiple shoot formation probably due to precocious germination of somatic embryos. Plants recovered from culture were acclimated to soil, grown up in a greenhouse, and planted in field plots with saline irrigation to ensure stability of salt tolerance.
