In situ loop-mediated isothermal amplification （in situ LAMP） combines in situ hybridization and loop-mediated isothermal amplification （LAMP） techniques for chromosomal localization of DNA sequences. In situ LAMP is a method that is generally more specific and sensitive than conventional techniques such as fluorescence in situ hybridization （FISH）, primed in situ labeling （PRINS）, and cycling primed in situ labeling （C-PRINS）. Here, we describe the development and application of in situ LAMP to identify the chromosomal localization of DNA sequences. To benchmark this technique, we successfully applied this technique to localize the major ribosomal RNA gene on the chromosomes of the Zhikong scallop （ Chlarnys farreri）.
Toll-like receptor(TLR) signaling pathway plays a pivotal role in the innate immune system. Studies on TLR signaling pathway genes in Zhikong scallop(Chlamys farreri) have mainly focused on sequence analysis and expression profiling, no research has been carried out on their localization. The chromosomal position of TLR signaling pathway genes can be valuable for assemblying scallop genome and analysizing gene regulatory networks. In the present study, five key TLR signaling pathway genes(Cf TLR, Cf Myd88, Cf TRAF6, Cf NFκB, and Cf IκB) containing bacterial artificial chromosomes(BACs) were isolated and physically mapped through fluorescence in situ hybridization on five non-homologous chromosome pairs, showing a similar distribution to another five model species. The isolation and mapping of these key immune genes of C. farreri will aid to the research on innate immunity, assignment of interested genes to chromosomes, and integration of physical, linkage and cytogenetic maps of this species.
Patinopecten yessoensis is a commercial valuable species. This study deals with the effect of nutrient effects of the broodstock(mainly ovaries) on the larvae. Concentrations of total carbohydrate, total protein and total lipid in the gonads of P. yessoensis from three Hatcheries(Hatchery 1, Hatchery 2, and Hatchery 3) were determined before and after spawning. The relationship between the nutrient concentration in ovaries before spawning(BC) and that of larvae(LC) was assessed as well as the change in nutrient levels in ovaries after spawning(DC). Results indicate that the BC of total carbohydrate(7.66%) and total lipid(14.48%) in ovaries were significantly higher than in testes(5.20%, 5.20% respectively), whereas the BC of total protein in the ovaries was lower(61.76%) than in the testes(81.67%). The different gonadal composition suggests the different nutrient demands between male and female broodstocks in breeding season. P atinopecten yessoensis gonads contained a higher proportion of lipids, in comparison to other bivalves, which might be a response to the low ambient water temperatures. Further analysis of fatty acids showed that the concentrations of n-3PUFA, EPA and DHA in larvae(LC) were positively correlated with BC and DC, indicating the significant nutrient influence of broodstocks on the larvae. As these fatty acids are important in metabolism, and have been demonstrated to be influential to the viability of the larvae, larval growth and the settlement, spat growth, and juvenile survival in many bivalves, they could possibly be used as indexes to evaluate, and predict condition of broodstocks and larvae.
