摘要
【目的】基于转录组水平分析芜菁雌蕊退化突变(turnip pistil aborted,tpa)发生的可能原因,为揭示芸薹属作物雌蕊发育的分子遗传调控机制提供研究依据。【方法】利用tpa及其野生型植株的开放花制备的mRNA反转录成cDNA与拟南芥ATH1芯片进行杂交,筛选出在tpa及其野生型植株中表达有差异的基因,并利用RT-PCR技术对芯片筛选出的基因进行验证。【结果】获得了152个在野生型(W1)和完全退化株(M3)转录组中差异表达的基因,61个在W1和部分退化株(I2)转录组中差异表达的基因,以及24个在I2和M3转录组中差异表达的基因,上述差异基因分别属于细胞壁合成与调控、防卫与胁迫反应、转录调控、蛋白质代谢以及普通新陈代谢等9个不同的功能类别。通过对41个基因的RT-PCR验证,获得了At2g42840、At1g57750、At5g20630、At2g03090、At3g08030、At5g08000、At2g28790、At5g63310和At2g24270等9个在tpa及野生型植株中具显著不同的时空表达特性的基因。【结论】拟南芥cDNA芯片可用于分析tpa及其野生型基因的转录差异,筛选获得的9个显著差异基因不但在雌蕊发育等生殖发育过程起重要作用,还参与了植物营养生长的生理生化过程。
【Objective】The possible reasons of the pistil abortion in turnip have been studied at the transcriptional level and it will be helpful to provide some useful clues to illuminate the molecular genetics mechanism of the gynoecium development in Brassica crops.【Method】The Arabidopsis ATH1 genome array was employed to analyze the transcriptome differences between the tpa mutants and their wild type flowers.Subsequently,the RT-PCR was used to confirm the genes with transcriptional difference.【Result】Data analysis results indicated that there were differences in 152 genes between the wild type（W1） and the pistil aborted completely（M3）,likewise,61 genes with transcriptional difference between the W1and the pistil aborted partially（I2）,and 24 genes with transcriptional difference between the I2 and M3.All of the previous genes were classified into the following in functional categoryies： cell wall biosynthesis and regulation,defense and stressing reaction,transcriptional regulation,protein metabolism and normal metabolism,respectively.Out of the 41 genes,9 genes with significant temporal and spatial transcription difference（At2g42840,At1g57750,At5g20630,At2g03090,At3g08030,At5g08000,At2g28790,At5g63310,At2g24270） were obtained through the RT-PCR screening and confirmation.【Conclusion】The Arabidopsis ATH1 genome array can be successfully utilized to analyze the transcriptome differences between the tpa mutants and their wild type.And the 9 genes with significant temporal and spatial transcription difference,are not only important to the reproductive process including gynoecium development,but also involve in the vegetative processes.
出处
《中国农业科学》
CAS
CSCD
北大核心
2011年第5期972-981,共10页
Scientia Agricultura Sinica
基金
国家自然科学基金项目(30771377)
浙江省科技计划项目(2009C32029)
浙江省自然科学基金项目(Y3090294)
教育部留学回国人员科研启动基金项目(2009-1001)
浙江省人事厅留学回国基金项目(J20090028)
关键词
芜菁
拟南芥cDNA芯片
雌蕊发育
基因表达
转录
Brassica campestris L.ssp.rapifera Sinsk
syn.B.rapa L.ssp.Rapifera
Arabidopsis ATH1 genome array
gynoecium development
gene expression
transcription
