Global gene expression of Arabidopsis lines with altered ANAC102 expression under normal and low oxygen conditions. Arabidopsis thaliana

Global gene expression was compared between Arabidopsis lines with altered expression of ANAC102 (over-expressed and knocked-out) and wild-type. ANAC102 is a putative NAC domain transcription factor. Gene expression was compared between an ANAC102 over-expressing line and parental ecotype C24 under ambient atmosphere to determine which genes ANAC102 is capable of regulating. Gene expression was also compared between three week old plants of an ANAC102 knock-out line and parental ecotype Col-0 under 0.1% Oxygen and ambient atmosphere conditions to determine which genes may require ANAC102 for appropriate expression under these conditions. Gene expression was also compared between imbibed seeds of an ANAC102 knock-out line and parental ecotype Col-0 following a 0.1% Oxygen treatment. Keywords: Genetic modification Overall design: Three related experiments were performed. In the first two, three week old Arabidopsis seedlings (4-6 leaves) were used. In the first experiment one line of ANAC102 over-expressing plants were compared to ecotype C24 (the parental line used in the creation of the ANAC102 over-expressing line). Two biological replicates were used for each line and for each biological replicate, five Arabidopsis seedlings grown on the same petri dish were bulked. RNA was extracted from whole plants. In the second experiment, one line of plants bearing a T-DNA insertion in the second exon of the ANAC102 gene was compared to ecotype Col-0 (the parental line used for the T-DNA mutagenesis) both without treatment and following treatment with 0.1% Oxygen for four hours. Two biological replicates were used for each line and for each biological replicate, five Arabidopsis seedlings grown on the same petri dish were bulked. RNA was extracted from root tissue only. In the third experiment imbibed seeds of one Arabiopsis line bearing a T-DNA insertion in the second exon of the ANAC102 gene was compared to seeds of ecotype Col-0 (the parental line used for the T-DNA mutagenesis) following treatment with 0.1% Oxygen for six days. Three biological replicates were used for each line and for each biological replicate, ~5mg (pre-imbibition) of seed were bulked. RNA was extracted from whole seed.

本数据集对ANAC102表达量改变（过表达与敲除）的拟南芥（Arabidopsis）品系与野生型之间的全局基因表达（global gene expression）谱进行了比较。ANAC102是一种推定的NAC结构域转录因子（NAC domain transcription factor）。

为明确ANAC102可调控的靶基因，本研究在大气环境下，比较了ANAC102过表达品系与其构建所用的亲本生态型C24的基因表达差异。为确定在0.1%氧气与大气环境条件下哪些基因的正常表达依赖ANAC102，本研究比较了三周龄ANAC102敲除品系与其亲本生态型Col-0的基因表达差异。此外，经0.1%氧气处理6天后，本研究还比较了ANAC102敲除品系与亲本生态型Col-0的吸胀种子（imbibed seeds）的基因表达差异。

关键词：遗传修饰

整体实验设计：共开展3组相关实验，前两组实验均采用三周龄、具4~6片真叶的拟南芥幼苗。
第一组实验：将ANAC102过表达植株品系与其构建时所用的亲本生态型C24进行对照比较。每个品系设置2个生物学重复（biological replicate），每个生物学重复选取同一培养皿中生长的5株拟南芥幼苗混合取样，提取全植株总RNA。
第二组实验：将ANAC102基因第2外显子区域带有T-DNA插入（T-DNA insertion）的敲除植株品系，与其构建时所用的亲本生态型Col-0，分别在未处理及0.1%氧气处理4小时的条件下进行基因表达比较。每个品系设置2个生物学重复，每个生物学重复选取同一培养皿中生长的5株拟南芥幼苗混合取样，仅提取根部组织总RNA。
第三组实验：经0.1%氧气处理6天后，将ANAC102基因第2外显子区域带有T-DNA插入的敲除植株的吸胀种子，与其构建时所用的亲本生态型Col-0的种子进行对照比较。每个品系设置3个生物学重复，每个生物学重复选取约5mg（吸胀前）的种子混合取样，提取全种子总RNA。