摘要
为获取高活力的外套膜细胞,研究通过常压室温等离子体(Atmospheric and room temperature plasma,ARTP)诱变和流式细胞术等技术分析了不同诱变气量组(10、12和15 SLM组)和处理时间对三角帆蚌(Hyriopsis cumingii)体外培养的外套膜细胞的细胞活性及生物矿化相关功能的影响。结果表明:ARTP诱变360—900s能显著升高各组三角帆蚌外套膜细胞活力,且在900s时达到最大值(P<0.05);渗透压稳定剂的添加,显著提高了诱变过程中12和15 SLM组在360—900s作用时间下的细胞活力(P<0.05),其中12 SLM组外套膜细胞增殖指数显著上升至最大值(P<0.05);诱变后细胞体外培养24h时结果显示,12 SLM组720s的外套膜细胞活力显著达到最高(P<0.05);15 SLM组(诱变时间为720s,下同)SOD活力随着诱变气量的增大呈显著下降趋势,且在15 SLM组显著降至最低水平(P<0.05),相反,微核率在15 SLM组达到最大值;生物矿化分析表明,外套膜细胞Ca^(2+)的浓度、生物矿化相关的关键酶(碳酸酐酶、碱性磷酸酶)和钙调蛋白基因(Calmodulin,CAM)基因均在12 SLM组达到最大值(P<0.05),而EFCB1(EF-hand calcium-binding domain-containing protein 1)基因结果显示在10 SLM组达到最大值(P<0.05),12 SLM次之;以上分析表明,氦气诱变在气量为12 SLM,处理720s时与渗透压稳定剂连用对外套膜细胞活性及其他生物学活性影响最为显著,暗示氦气诱变可有效作用于外套膜细胞的离体培养,为三角帆蚌建立细胞系提供生物学基础与新思路。
The mantle of pearl oyster is one of the indispensable tissues for pearl cultivation.In order to obtain highactivity mantle cells,the effects of different gas volume groups[10,12 and 15 SLM(standard liter per minute;SLM)]and treatment time on cell activity and biomineralizated functions of mantle cells of Hyriopsis cumingii were analyzed using ARTP mutagenesis and flow cytometry.The results showed that the activity of mantle cells of Hyriopsis cumingii increased significantly in 360—900s by ARTP,and reached the maximum value at 900s(P<0.05),which was consistent with the cell activity results of osmotic stabilizer groups.Mantle cells activity in 12 and 15 SLM groups increased significantly at 360—900s(P<0.05),and the proliferation index(PI)of mantle cells reached the maximum value in 12 SLM group(P<0.05);in addition,cells were cultured in vitro for 24h after mutagenesis,the viability of mantle cells in 12 SLM group at 720s was the highest(P<0.05),and the SOD activity(mutagenesis time was 720s,the same as below)showed a significant decreasing trend with the increase of mutagenesis gas volume,and it decreased significantly to the lowest level in the 15 SLM group(P<0.05).On the contrary,the micronucleus rate reached the maximum value;biomineralization analysis showed that the concentration of Ca^(2+),the key enzymes related to biomineralization(carbonic anhydrase,alkaline phosphatase)and calmodulin(CAM)genes of mantle cells reached the maximum in the 12 SLM group(P<0.05),but expression levels of EFCB1(EF-hand calcium-binding domain-containing protein 1)gene reached the maximum in the 10 SLM group(P<0.05),followed by 12 SLM group.The above findings indicated that ARTP mutation had the most significant effect on the cell activity and other biological activity of mantle cells when the gas volume is 12 SLM for 720s and the osmotic pressure stabilizer is used together with ARTP,indicating that ARTP mutagenesis can be effective in vitro culture of mantle cells to provide a biological basis and a new idea for the establ
作者
陆阿利
曹玉香
李文娟
施志仪
夏煌慧
杨婧漪
LU A-Li;CAO Yu-Xiang;LI Wen-Juan;SHI Zhi-Yi;XIA Huang-Hui;YANG Jing-Yi(Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources,Ministry of Education,Shanghai Ocean University,Shanghai 201306,China;Key Laboratory of Freshwater Aquatic Genetic Resources,Ministry of Agriculture and Rural Affairs,Shanghai Ocean University,Shanghai 201306,China;National Demonstration Center for Experimental Fisheries Science Education,Shanghai Ocean University,Shanghai 201306,China)
出处
《水生生物学报》
CAS
CSCD
北大核心
2021年第2期341-351,共11页
Acta Hydrobiologica Sinica
基金
国家自然科学基金(31201991)
国家重点研发计划(2018YFD0901406)资助。
关键词
三角帆蚌
外套膜
ARTP
诱变
生长活性
生物矿化
Hyriopsis cumingii
mantle
ARTP
Mutagenesis
Growth activity
Biomineralization
