| 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（0.1,0.25,0.5,0.75,1.0 mmol/L)和3种处理方法（不静置直接700 r/min 离心5 min，静置5 min后700 r/min离心5 min，静置3 min后700 r/min离心2 min）,以及紫外线（UV）照射（2根15 W紫外灯,距灯管25 cm处照射0.5,1,4,5,7,10，15 min）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0.1 mmol/L IOA静置3 min后700 r/min离心2 min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15 min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】0.1 mmol/L IOA静置3 min后700 r/min离心2 min或2根15 W紫外灯连续照射25 cm处的烟草原生质体15 min，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。 |
| 关键词: 烟草 原生质体 钝化作用 碘乙酰胺 紫外线 |
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| 基金项目:贵州省烟草公司项目（200912）；贵州省自然科学基金项目（［2009］2097） |
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| Passivation effect of iodacetamide and ultraviolet light on tobacco protoplasts |
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| Abstract: |
| 【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】Tobacco 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/min 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 mesophyll 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 complementation after fusing and forming hybrid. |
| Key words: tobacco protoplast passivation iodacetamide ultraviolet light |
