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首页> 外文学位 >A tissue culture system facilitates examination of gene expression during breakage of vegetative dormancy in the potato tuber shoot apical meristem.
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A tissue culture system facilitates examination of gene expression during breakage of vegetative dormancy in the potato tuber shoot apical meristem.

机译：组织培养系统有助于检查马铃薯块茎茎尖分生组织营养休眠期间的基因表达。

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While the shoot apical meristem's role during dormancy emergence in seeds is relatively well-understood, molecular factors governing vegetative endodormancy and sprouting in tuberous plants have not been well studied. A microtuber culture system was developed in order to study dormancy emergence in the shoot apical meristem of the potato (Solanum tuberosum). Microtubers were induced from subcultured shoot internodes, grown for 9 weeks, then harvested. The microtubers were hardened at 20° for 2 weeks then stored in the dark at 4°C. Subsequently, at 4-week intervals, microtubers were transferred to 20°C in the dark for 7 days. Meristem length was measured prior to RNA extraction. Two genes, SHOOTMERISTEMLESS (STM) and WUSCHEL (WUS), are known to regulate the development of the shoot apical meristem in Arabidopsis thaliana. Primers for quantitative PCR (qPCR) were designed to amplifty putative potato STM and WUS homologs using expressed sequence tag (EST) sequences obtained during a basic local alignment search tool (BLAST) search of known petunia (Petunia hybrida) and tomato ( S. Lycopersicum) STM and WUS sequences. Primer pairs for reference genes were obtained from the literature. Using qPCR we measured the expression level of STM normalized to two potato meristem reference genes, L2 and ef1-alpha. L2 expression increased 1.5 to 6 fold during the post harvest period while that of ef1-alpha expression was between 1 and 1.4 fold the levels found in dormant meristems immediately after harvest. By contrast, STM expression increased from 0.17 to 0.75 fold of week 2 values 7 weeks after harvest from 17.4 to 42.5 fold 11 weeks after harvest and increased further thereafter. The meristem length showed a statistically significant increase from 0.23 (+/- 0.01) mm to 0.33 (+/- 0.07) mm (mean +/- Standard Error) length between 7 and 11 weeks after harvest. Taken together, this data suggests dormancy break of shoot apical meristems in this microtuber culture system occurs between 7 and 11 weeks after harvest. This system will be used in the study of the molecular events occurring during dormancy break in the potato shoot apical meristem.
机译：尽管种子在休眠中的芽顶分生组织的作用是相对容易理解的，但对块茎植物中营养内在的气味和发芽的分子因素尚未进行充分的研究。为了研究马铃薯（Solanum tuberosum）的茎尖分生组织中休眠的出现，开发了一种微型块茎培养系统。从传代的茎节间诱导微块茎，生长9周，然后收获。将微块茎在20°硬化2周，然后在4°C的黑暗环境中保存。随后，每隔4周将微块茎在黑暗中转移至20°C，持续7天。在提取RNA之前测量分生组织长度。已知两个基因SHOOTMERISTEMLESS（STM）和WUSCHEL（WUS）可以调节拟南芥中茎尖分生组织的发育。使用已知的矮牵牛（Petunia hybrida）和番茄（S. Lycopersicum ）STM和WUS序列。从文献中获得了参考基因的引物对。使用qPCR，我们测量了标准化为两个马铃薯分生组织参考基因L2和ef1-alpha的STM的表达水平。在收获后的期间，L2表达增加了1.5至6倍，而ef1-alpha表达的水平是收获后立即在休眠分生组织中发现的水平的1至1.4倍。相比之下，STM表达从收获后第7周的第2周值的0.17增至0.75倍，从收获后第11周的17.4增至42.5倍，此后进一步增加。在收获后的7至11周之间，分生组织的长度显示出统计学上的显着增加，从0.23（+/- 0.01）mm增长到0.33（+/- 0.07）mm（平均+/-标准误差）。两者合计，该数据表明在该微型块茎培养系统中茎尖分生组织的休眠破裂发生在收获后的7至11周之间。该系统将用于研究马铃薯茎尖分生组织休眠中断期间发生的分子事件。

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