碘乙酰胺和紫外线对烟草原生质体的钝化作用  

Passivation effect of iodacetamide and ultraviolet light on tobacco protoplasts

作　　者：聂琼[1] 朱俊[1] 徐如宏[1] 刘仁祥[1] 

机构地区：[1]贵州大学农学院/贵州省烟草品质研究重点实验室,贵州贵阳550025

出　　处：《西北农林科技大学学报（自然科学版）》2013年第4期101-107,共7页Journal of Northwest A&F University(Natural Science Edition)

基　　金：贵州省烟草公司项目(200912);贵州省自然科学基金项目([2009]2097)

摘　　要：【目的】探索烟草原生质体钝化的临界处理剂量,为不对称体细胞杂交技术改良和创新烟草种质奠定基础。【方法】以不同碘乙酰胺(IOA)浓度(0.1,0.25,0.5,0.75,1.0mmol/L)和3种处理方法(不静置直接700r/min离心5min,静置5min后700r/min离心5min,静置3min后700r/min离心2min),以及紫外线(UV)照射(2根15W紫外灯,距灯管25cm处照射0.5,1,4,5,7,10,15min)处理烟草原生质体,并进行融合培养,研究烟草原生质体钝化的临界处理条件。【结果】烟草原生质体对IOA十分敏感,以0.1mmol/L IOA静置3min后700r/min离心2min可使烟草原生质体钝化停止分裂,临界状态原生质体比例最高为92.9%;UV连续照射15min的烟草叶肉原生质体失活,但大多数原生质体处于临界状态,比例为91.4%;2种临界处理后的原生质体融合后形成杂种细胞,能进行正常的细胞分裂。【结论】0.1mmol/L IOA静置3min后700r/min离心2min或2根15W紫外灯连续照射25cm处的烟草原生质体15min,是烟草原生质体钝化的临界处理,2种临界处理后的原生质体融合体由于生理代谢上得到互补形成杂种细胞,恢复了分裂能力。[Objective] This study aimed to explore the critical dose of tobacco protoplast passivation for improvement and innovation of tobacco germplasm by asymmetric somatic hybridization. [Method] To- bacco protoplasts were treated with iodacetamide （IOA） of different concentrations （0.1,0.25,0.5,0.75, and 1. 0 mmol/L）,three centrifuging methods （centrifuging for 5 min at 700 r/rain without standing, standing for 5 min and then centrifuging for 5 min at 700 r/min,and standing for 3 min and then centrifuging for 2 min at 700 r/min）, and irradiation of ultraviolet light （UV） （exposure by 0. 5,1,4,5,7,10, and 15 min in two ultraviolet lamp of 15 W）to investigate the critical dose for passivation of tobacco protoplast. [Result] Tobacco protoplast was very sensitive to IOA. The treatment with 0. 1 mmol/L IOA before standing for 3 min and centrifuging for 2 min at 700 r/min had the highest percentage of the protoplast under the critical condition （92.9 % ）, which became the best to tobacco protoplast passivation. Continuous irradiation 15 min was the critical treatment to tobacco rriesophyll protoplasts,and the protoplast ratio under the critical condition was 91.4%. The fused protoplast can divide and form cell colonies. [Conclusion] The treatment with 0.1 mmol/L IOA,standing for 3 min and then centrifuging for 2 min at 700 r/min and continuous irradiation for 15 min was the critical dose of tobacco protoplast passivation, two kinds of critical dose of two tobacco protoplasts after being treated with critical dose restored ability of division due to cornplementation after fusing and forming hybrid.

关 键 词：烟草 原生质体 钝化作用 碘乙酰胺 紫外线 

分 类 号：S572.035.2[农业科学—烟草工业]