Transcriptome differences between two vitamin C-deficient mutants and wild type

The Arabidopsis mutant vtc2 is vitamin C-deficient and high light sensitive (Mueller-Moule et al., 2002. Plant Physiol 128: 970-977; Mueller-Moule et al., 2003. Plant Physiol 133: 748-760). Even though the VTC2 gene has been cloned (Jander et al., 2002. Plant Physiol 129: 440-450), its function is still unclear. The goal of this experiment is to learn more about the function of the VTC2 gene by determining differences in the transcriptomes of wild type and the vtc2-1 mutant. In order to differentiate between VTC2 gene-specific differences and vitamin C-induced differences, a second vitamin C-deficient mutant vtc1-1 (Conklin et al., 1996. PNAS 93: 9970-9974) is included in the experiment. The plants will be grown on agar plates in a growth chamber with a 16 hour light (about 40 umol photons m(-2)s(-1))/8 hour dark regime at 20C for 14 days to stage 1.04. Whole plants will be collected and total RNA will be isolated using the Qiagen Rneasy plant mini kit (Qiagen, Hilden, Germany). 9 samples were used in this experiment.

拟南芥（Arabidopsis）突变体vtc2表现为维生素C缺乏且对高光敏感（Mueller-Moule等，2002，《植物生理学》128: 970-977；Mueller-Moule等，2003，《植物生理学》133: 748-760）。尽管VTC2基因已被克隆（Jander等，2002，《植物生理学》129: 440-450），但其功能仍未明确。本实验旨在通过分析野生型与vtc2-1突变体的转录组（transcriptome）差异，深入解析VTC2基因的功能。为区分VTC2基因特异性差异与维生素C诱导的差异，实验纳入了另一株维生素C缺乏突变体vtc1-1（Conklin等，1996，《美国国家科学院院刊》93: 9970-9974）。实验植株将接种于琼脂平板，置于光照培养箱中培养，采用16小时光照（光强约40 μmol·m⁻²·s⁻¹）/8小时黑暗的周期，培养温度为20℃，持续培养14天至生长阶段1.04。收获整株植株后，使用Qiagen RNeasy植物小提试剂盒（德国希尔德市Qiagen公司）提取总RNA。本实验共使用9个样本。