| GFP标记拟南芥突变体 sad2微管及F2代幼苗的检测与分析 |
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| 引用本文：陈炳佑，武仁文，刘广志，侍福梅.GFP标记拟南芥突变体 sad2微管及F2代幼苗的检测与分析[J].西北农业学报,2015,24(10):131~136 |
| DOI:10.7606/j.issn.1004-1389.2015.10.019 |
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| 基金项目:国家自然科学基金（31240035）；山东省自然科学基金（ZR2010CQ002）。 |
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| 中文摘要:为观察拟南芥突变体 sad2（sensitive to ABA and drought）的微管列阵，以拟南芥突变体 sad2-1和 sad2-2为母本，转GFP（green fluorescence protein）-α-tubulin野生型拟南芥为父本杂交，并对F2代幼苗进行叶表型分析、卡那霉素抗性筛选和荧光镜检。表型分析显示，拟南芥 sad2-2突变体与转GFP-α-tubulin的杂交F2代幼苗叶片出现有毛和无毛2种性状，二者分离比为2.81∶1。卡那霉素抗性筛选显示，部分F2代幼苗在卡那霉素培养基上出现白化死亡，大部分可正常生长。荧光镜检显示，卡那霉素阳性苗的子叶出现GFP绿色荧光。此外，共聚焦显微镜观察显示，拟南芥突变体子叶细胞微管列阵清晰可见，且 sad2-1和 sad2-2两种突变体的微管比野生型更加致密，但 sad2-1和 sad2-2两突变体间无明显差异。说明：采用杂交法将GFP-α-tubulin引入突变体来分析微管是一种简便可靠的方法，且 sad2基因影响细胞微管列阵，可用于 sad2基因与微管功能的进一步研究。 |
| 中文关键词:绿色荧光蛋白 微管 杂交 sad2-1 sad2-2 拟南芥 |
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| Identification and Analysis of F2 Seedlings Microtubule with GFP-tagged sad2 Mutants in Arabidopsis |
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| Abstract:In order to observe the microtubule arrays of sensitive to ABA and drought ( sad2) mutants in Arabidopsis, both sad2-1 and sad2-2 mutant lines were hybridized as female parent with the pollens from GFP-α-tubulin tagged wild type as male donor. Then F2 seedlings were tested with phenotypic analysis, antibiotic resistant screening and fluorescence detection. The results of phenotype analysis indicated that seedlings of hybrid F2 generation between mutant sad2-2 and GFP-α-tubulin appear two leaf phenotypes with or without trichome and the separation ratio of hairy：hairless were 2.81∶1. The kanamycin resistance screening showed that seedlings grown on the medium containing kanamycin were part of dead contrast to most part of surviving. The green fluorescence of GFP was founded in kanamycin resistant seedlings under the fluorescence microscope. In addition, clear microtubule arrays in leaf epidermal cells were observed via the laser scanning confocal microscopy, and cortical arrays in both sad2-1 and sad2-2 mutants are denser and more ordered than that of wild type. While there were no significant difference between the two mutant lines. Therefore, it is convenient and liable for observing microtubule array of mutants introduced with GFP tagged tubulin via hybridization technique. And sad2 were turned out to involve in cortical array regulation in Arabidopsis, which would be good candidates for further analysis the potential links between microtubule and mutant gene. |
| keywords:Green fluorescent protein Microtubule Hybridization sad2-1 sad2-2 Arabidopsis |
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