The complicated life cycle ofAurelia spp., comprising benthic asexually-reproducing polyps and sexually-reproducing medusae, makes it hard for researchers to identify and track them, especially for early stage individ...The complicated life cycle ofAurelia spp., comprising benthic asexually-reproducing polyps and sexually-reproducing medusae, makes it hard for researchers to identify and track them, especially for early stage individuals, such as planulae. To solve this problem, we developed a real-time PCR assay (SYBR Green I) to identify planulae in both cultured and natural seawater samples. Species-specific primers targeting Aurelia sp.1 mitochondrial 16S rDNA (mr 16S rDNA) regions were designed. Using a calibration curve constructed with plasmids containing the Aurelia sp. 1 mt 16S rDNA fragment and a standard curve for planulae, the absolute number of mt 16S rDNA copies per planula was determined and from that the total number ofplanulae per sample was estimated. For the field samples, a 100-fold dilution of the sample DNA combined with a final concentration of 0.2 μg/μL BSA in the PCR reaction mixture was used to remove real- time PCR inhibitors. Samples collected in Jiaozhou Bay from July to September 2012 were subsequently analyzed using this assay. Peak Aurelia sp.1 planula abundance occurred in July 2012 at stations near Hongdao Island and Qingdao offshore; abundances were very low in August and September. The real-time PCR assay (SYBR Green I) developed here negates the need for traditional microscopic identification, which is laborious and time-consuming, and can detect and quantify jellyfish planulae in field plankton samples rapidly and specifically.展开更多
Weng,an phosphates of the Precambrian Dou-shantuo Formation, Guizhou (southwestern China) preserve a large number of exquisite biological structures, which are mostly micro-spherical and represent seaweeds, acritarchs...Weng,an phosphates of the Precambrian Dou-shantuo Formation, Guizhou (southwestern China) preserve a large number of exquisite biological structures, which are mostly micro-spherical and represent seaweeds, acritarchs and developing eggs related to various groups of metazoans. Here is a report of a variety of developing eggs and larvae, which are most probably of Cnidarian affinity. The eggs ex-amined in the study are composed of early cleavage embryos and two-layered gastrulae. The early cleavage embryos are radial and total cleavage with equal-size blastomeres. The gastrulae mostly bear a large archenteron, which is filled with yolk-degrading organic matter. Ovoid to fusiform planula-like larvae identified in thin sections under light microscope are mostly mouthless and their gastrovascularcavity is filled with possible yolk-degrading organic matter. They are likely representatives of non-feeding larva. The uncommon planula-like structures are hollow, with each having a mouth-like structure on its narrow end. We inter-pret them as feeding larva. Study of these embryos with pos-sible Cnidarian affinities shed new insight on the origin ofmetazoans.展开更多
基金Supported by the National Basic Research Program of China(973 Program)(No.2011CB403602)the National Natural Science Foundation of China(No.41076085)the National Special Research Fund for Non-Profit Marine Sector(No.201205031)
文摘The complicated life cycle ofAurelia spp., comprising benthic asexually-reproducing polyps and sexually-reproducing medusae, makes it hard for researchers to identify and track them, especially for early stage individuals, such as planulae. To solve this problem, we developed a real-time PCR assay (SYBR Green I) to identify planulae in both cultured and natural seawater samples. Species-specific primers targeting Aurelia sp.1 mitochondrial 16S rDNA (mr 16S rDNA) regions were designed. Using a calibration curve constructed with plasmids containing the Aurelia sp. 1 mt 16S rDNA fragment and a standard curve for planulae, the absolute number of mt 16S rDNA copies per planula was determined and from that the total number ofplanulae per sample was estimated. For the field samples, a 100-fold dilution of the sample DNA combined with a final concentration of 0.2 μg/μL BSA in the PCR reaction mixture was used to remove real- time PCR inhibitors. Samples collected in Jiaozhou Bay from July to September 2012 were subsequently analyzed using this assay. Peak Aurelia sp.1 planula abundance occurred in July 2012 at stations near Hongdao Island and Qingdao offshore; abundances were very low in August and September. The real-time PCR assay (SYBR Green I) developed here negates the need for traditional microscopic identification, which is laborious and time-consuming, and can detect and quantify jellyfish planulae in field plankton samples rapidly and specifically.
基金This work was supported jointly by the National Natural Science Foundation of China (Grant No. 40132010, 40302004);Chinese Acad-emy of Sciences (Grant No. KZCX3 SW-14l) ; Ministry of Science and Technology of China (Grant No. G2000077700).
文摘Weng,an phosphates of the Precambrian Dou-shantuo Formation, Guizhou (southwestern China) preserve a large number of exquisite biological structures, which are mostly micro-spherical and represent seaweeds, acritarchs and developing eggs related to various groups of metazoans. Here is a report of a variety of developing eggs and larvae, which are most probably of Cnidarian affinity. The eggs ex-amined in the study are composed of early cleavage embryos and two-layered gastrulae. The early cleavage embryos are radial and total cleavage with equal-size blastomeres. The gastrulae mostly bear a large archenteron, which is filled with yolk-degrading organic matter. Ovoid to fusiform planula-like larvae identified in thin sections under light microscope are mostly mouthless and their gastrovascularcavity is filled with possible yolk-degrading organic matter. They are likely representatives of non-feeding larva. The uncommon planula-like structures are hollow, with each having a mouth-like structure on its narrow end. We inter-pret them as feeding larva. Study of these embryos with pos-sible Cnidarian affinities shed new insight on the origin ofmetazoans.