用FIASCO(Fast Isolation by AFLP of Sequences Containing Repeats)技术开展了合浦珠母贝Pinctada fucata基因组微卫星标记的分离与筛选研究。合浦珠母贝基因组DNA经限制性内切酶MseI酶切后与接头连接,用生物素标记的(CA)15探针与其杂...用FIASCO(Fast Isolation by AFLP of Sequences Containing Repeats)技术开展了合浦珠母贝Pinctada fucata基因组微卫星标记的分离与筛选研究。合浦珠母贝基因组DNA经限制性内切酶MseI酶切后与接头连接,用生物素标记的(CA)15探针与其杂交,然后用磁珠富集、洗脱获得单链目的片段,经PCR扩增后形成双链,最后进行克隆转化,构建微卫星富集文库。挑选克隆用探针引物(CA)15和载体引物进行第2次筛选,获得阳性克隆357个,测序结果表明,297个克隆(83.2%)含有微卫星序列,包括479个微卫星DNA结构域。其中完美型微卫星有370个(77.3%),非完美型95个(19.8%),复合型14个(2.9%)。合成引物49对,有31对(63%)扩增出目的产物,其中9对在种群中(n=32)具有扩增多态性,其多态信息含量PIC值在0.375―0.809之间,平均为0.536;等位基因数在2―9个之间,平均为4.889个;观测杂合度介于0.200―0.600之间,平均为0.415;期望杂合度的变化范围为0.454―0.844,平均为0.598。表明FIASCO技术适合于合浦珠母贝微卫星标记的分离与筛选。展开更多
Oceanic uptake of anthropogenic carbon dioxide results in a decrease in seawater pH, a process known as "ocean acidification". The pearl oyster Pinctada fucata, the noble scallop Chlamys nobilis, and the gre...Oceanic uptake of anthropogenic carbon dioxide results in a decrease in seawater pH, a process known as "ocean acidification". The pearl oyster Pinctada fucata, the noble scallop Chlamys nobilis, and the green-lipped mussel Perna viridis are species of economic and ecological importance along the southern coast of China. We evaluated the effects of seawater acidification on clearance, respiration, and excretion rates in these three species. The animals were reared in seawater at pH 8.1 (control), 7.7, or 7.4. The clearance rate was highest at pH 7.7 for P. fucata and at pH 8.1 for C. nobilis and P. viridis. The pH had little effecton the respiration rate of P. fucata and P. viridis. In contrast, the respiration rate was significantly lower atpH 7.4 in C. nobilis. The excretion rate was significantly lower at pH 7.4 than pH 8.1 for all species. Theresults indicate that the reduction in seawater pH likely affected the metabolic process (food intake, oxygenconsumption, and ammonia excretion) of these bivalves. Different species respond differently to seawateracidification. Further studies are needed to demonstrate the exact mechanisms for this effect and evaluateadaptability of these bivalves to future acidified oceans.展开更多
In this study, we examined the effect of elevated temperature on the expression patterns of genes, i.e., nacrein, irr, n16, n19, and hsp70 in the pearl oyster Pinctada fucata. The experiment was carried out at 4 tempe...In this study, we examined the effect of elevated temperature on the expression patterns of genes, i.e., nacrein, irr, n16, n19, and hsp70 in the pearl oyster Pinctada fucata. The experiment was carried out at 4 temperatures, i.e., 20℃(ambient, control), 24, 28℃, and 32℃. The expression levels of target genes in P. fucata were assayed at 0, 6, 24, 48, and 96 h via real-time polymerase chain reaction. Results showed that the expression levels of nacrein and irr had no significant variations among different time points below 28℃, but significantly increased over time at 32℃. The expression levels of n16 and n19 did not change markedly at 20℃. The former increased significantly at 6 h and 24 h while the latter substantially decreased during 6–96 h at 24, 28 and 32℃. Among different temperatures, the level of n16 was significantly lower at 20℃ than at other temperatures during 6–96 h, and the level of n19 significantly varied among different temperatures at 48 h and 96 h. The expression level of hsp70 was significantly higher at 32℃ than at 20, 24 and 28℃ at 24 h. These results demonstrated that elevated temperature impacted the physiological processes of P. fucata and potentially influenced its adaptability to thermal stress.展开更多
文摘用FIASCO(Fast Isolation by AFLP of Sequences Containing Repeats)技术开展了合浦珠母贝Pinctada fucata基因组微卫星标记的分离与筛选研究。合浦珠母贝基因组DNA经限制性内切酶MseI酶切后与接头连接,用生物素标记的(CA)15探针与其杂交,然后用磁珠富集、洗脱获得单链目的片段,经PCR扩增后形成双链,最后进行克隆转化,构建微卫星富集文库。挑选克隆用探针引物(CA)15和载体引物进行第2次筛选,获得阳性克隆357个,测序结果表明,297个克隆(83.2%)含有微卫星序列,包括479个微卫星DNA结构域。其中完美型微卫星有370个(77.3%),非完美型95个(19.8%),复合型14个(2.9%)。合成引物49对,有31对(63%)扩增出目的产物,其中9对在种群中(n=32)具有扩增多态性,其多态信息含量PIC值在0.375―0.809之间,平均为0.536;等位基因数在2―9个之间,平均为4.889个;观测杂合度介于0.200―0.600之间,平均为0.415;期望杂合度的变化范围为0.454―0.844,平均为0.598。表明FIASCO技术适合于合浦珠母贝微卫星标记的分离与筛选。
基金Supported by National Natural Science Foundation of China (No.41006090)the Knowledge Innovation Program of Chinese Academy of Sciences (No. KZCX2-YW-Q07-03)the National High Technology Research and Development Program of China (863 Program) (No.2006AA10A409)
文摘Oceanic uptake of anthropogenic carbon dioxide results in a decrease in seawater pH, a process known as "ocean acidification". The pearl oyster Pinctada fucata, the noble scallop Chlamys nobilis, and the green-lipped mussel Perna viridis are species of economic and ecological importance along the southern coast of China. We evaluated the effects of seawater acidification on clearance, respiration, and excretion rates in these three species. The animals were reared in seawater at pH 8.1 (control), 7.7, or 7.4. The clearance rate was highest at pH 7.7 for P. fucata and at pH 8.1 for C. nobilis and P. viridis. The pH had little effecton the respiration rate of P. fucata and P. viridis. In contrast, the respiration rate was significantly lower atpH 7.4 in C. nobilis. The excretion rate was significantly lower at pH 7.4 than pH 8.1 for all species. Theresults indicate that the reduction in seawater pH likely affected the metabolic process (food intake, oxygenconsumption, and ammonia excretion) of these bivalves. Different species respond differently to seawateracidification. Further studies are needed to demonstrate the exact mechanisms for this effect and evaluateadaptability of these bivalves to future acidified oceans.
基金supported by the National Natural Science Foundation of China (41006090)Joint Program of NSFC-Guangdong (U0831001)the Funds of Knowledge Innovation Program of Chinese Academy of Sciences (ZCX2-EW-Q21)
文摘In this study, we examined the effect of elevated temperature on the expression patterns of genes, i.e., nacrein, irr, n16, n19, and hsp70 in the pearl oyster Pinctada fucata. The experiment was carried out at 4 temperatures, i.e., 20℃(ambient, control), 24, 28℃, and 32℃. The expression levels of target genes in P. fucata were assayed at 0, 6, 24, 48, and 96 h via real-time polymerase chain reaction. Results showed that the expression levels of nacrein and irr had no significant variations among different time points below 28℃, but significantly increased over time at 32℃. The expression levels of n16 and n19 did not change markedly at 20℃. The former increased significantly at 6 h and 24 h while the latter substantially decreased during 6–96 h at 24, 28 and 32℃. Among different temperatures, the level of n16 was significantly lower at 20℃ than at other temperatures during 6–96 h, and the level of n19 significantly varied among different temperatures at 48 h and 96 h. The expression level of hsp70 was significantly higher at 32℃ than at 20, 24 and 28℃ at 24 h. These results demonstrated that elevated temperature impacted the physiological processes of P. fucata and potentially influenced its adaptability to thermal stress.
