不同培养条件下纳米碳溶胶对烟草细胞生长的影响  

Effects of nano-carbon sol on tobacco cell growth under different culture conditions

作　　者：赵振杰[1] 梁太波[1] 王宝林 陈千思[1] 胡利伟[1] 张艳玲[1] 孟祥宇 尹启生[1] ZHAO Zhenjie;LIANG Taibo;WANG Baolin;CHEN Qiansi;HU Liwei;ZHANG Yanling;MENG Xiangyu;YIN Qisheng(Zhengzhou Tobacco Research Institute of CNTC, Zhengzhou 450001, China;Technology Center, China Tobacco Henan Industrial Co., Ltd., Zhengzhou 450000, China)

机构地区：[1]中国烟草总公司郑州烟草研究院 [2]河南中烟工业有限责任公司技术中心

出　　处：《烟草科技》2018年第5期1-7,共7页Tobacco Science & Technology

基　　金：中国烟草总公司郑州烟草研究院科技项目"纳米碳对烟草促生作用的机理研究"(112013CZ0580);中国烟草总公司郑州烟草研究院院长基金项目"基于非损伤微测技术的烟草根系钾离子流速研究"(112017CA0090)

摘　　要：为进一步明确纳米碳溶胶的促生效果及其生理机制,在固体和液体两种培养条件下,以烟草BY-2细胞为试验材料,对纳米碳溶胶处理后细胞生物量、营养元素积累量、水通道蛋白基因和细胞周期蛋白基因的表达量以及悬浮细胞生长特性等指标进行了测定。结果表明,固体培养基中添加纳米碳溶胶能有效促进烟草愈伤组织生长,增加N、P、K养分积累量;其中50mg/L浓度的纳米碳溶胶处理,愈伤组织N、P、K养分积累量增加幅度最大,分别为41.3%、27.6%和47.0%。同时,纳米碳溶胶能显著提高培养前期水通道蛋白基因NtPIP1和细胞周期蛋白基因CycB的表达量,这是其促进愈伤组织生长的基础。在悬浮培养体系中,纳米碳溶胶处理对培养前期和中期悬浮细胞密度、密实体积以及死亡率无显著影响,表现出良好的生物相容性;在培养后期,10mg/L浓度的纳米碳溶胶处理对细胞生长产生一定抑制作用,表现为细胞密度下降和细胞活力降低。可见,纳米碳溶胶对烟草细胞生长的作用效果受培养条件和纳米碳溶胶浓度的共同影响。In order to investigate any promoting effect and associated physiology by nano-carbon sol, culture experiments were carried out under solid and liquid culture conditions. The effects of nano-carbon sol application on the expression of related genes including water channel protein gene and cell division gene,biomass, nutrient accumulation and suspension cell growth characteristics of tobacco BY-2 cells were studied.The results showed that the addition of nano-carbon sol enhanced the growth of tobacco callus and increased the accumulation of N, P and K nutrients. In all the treatments, 50mg/L concentration of nano-carbon sol had the largest increase rates of nutrient accumulation in callus, namely 41.3%, 27.6% and 47.0%, respectively. At the same time, nano-carbon sol significantly increased the expression levels of aquaporin gene NtPIP1 and cyclin gene Cyc B at early culture stage, which played an important role for promoting callus growth. In suspension culture experiments, nano-carbon sol presented a good biocompatibility at the early and middle culture stages without significantly affecting the density, packed cell volume and mortality of the suspension cells. At the late culture stage, nano-carbon sol had an inhibitory effect at the concentration of 10mg/L, which decreased the cell density and cell viability, which is likely to be associated with the aging of cultured cells. In conclusion, the effects of nano-carbon sol on the growth of tobacco cells were influenced by the culture conditions and application concentration.

关 键 词：烟草 纳米碳溶胶 愈伤组织 悬浮细胞 生物量 

分 类 号：S572.01[农业科学—烟草工业]