USDA-ARS-SRRC, 1100 Robert E. Lee Blvd., New Orleans, LA, 70124, USA.
Theor Appl Genet. 2010 May;120(7):1347-66. doi: 10.1007/s00122-010-1260-6. Epub 2010 Jan 20.
Gene expression profiles of developing cotton (Gossypium hirsutum L.) fibers from two near-isogenic lines (NILs) that differ in fiber-bundle strength, short-fiber content, and in fewer than two genetic loci were compared using an oligonucleotide microarray. Fiber gene expression was compared at five time points spanning fiber elongation and secondary cell wall (SCW) biosynthesis. Fiber samples were collected from field plots in a randomized, complete block design, with three spatially distinct biological replications for each NIL at each time point. Microarray hybridizations were performed in a loop experimental design that allowed comparisons of fiber gene expression profiles as a function of time between the two NILs. Overall, developmental expression patterns revealed by the microarray experiment agreed with previously reported cotton fiber gene expression patterns for specific genes. Additionally, genes expressed coordinately with the onset of SCW biosynthesis in cotton fiber correlated with gene expression patterns of other SCW-producing plant tissues. Functional classification and enrichment analysis of differentially expressed genes between the two NILs revealed that genes associated with SCW biosynthesis were significantly up-regulated in fibers of the high-fiber quality line at the transition stage of cotton fiber development. For independent corroboration of the microarray results, 15 genes were selected for quantitative reverse transcription PCR analysis of fiber gene expression. These analyses, conducted over multiple field years, confirmed the temporal difference in fiber gene expression between the two NILs. We hypothesize that the loci conferring temporal differences in fiber gene expression between the NILs are important regulatory sequences that offer the potential for more targeted manipulation of cotton fiber quality.
使用寡核苷酸微阵列比较了两个近等基因系（NIL）的发育棉纤维（Gossypium hirsutum L.）基因表达谱，这两个 NIL 在纤维束强度、短纤维含量以及少于两个遗传基因座方面存在差异。在纤维伸长和次生细胞壁（SCW）生物合成的五个时间点上比较了纤维基因表达。从随机完全区组设计的田间小区中收集纤维样本，每个 NIL 在每个时间点有三个空间上不同的生物学重复。微阵列杂交采用循环实验设计进行，允许比较两个 NIL 之间的纤维基因表达谱随时间的变化。总体而言，微阵列实验揭示的发育表达模式与先前报道的特定基因的棉花纤维基因表达模式一致。此外，与棉纤维次生细胞壁生物合成起始相关的基因与其他次生细胞壁产生植物组织的基因表达模式协调表达。两个 NIL 之间差异表达基因的功能分类和富集分析表明，与次生细胞壁生物合成相关的基因在高纤维品质系的纤维中在棉纤维发育的过渡阶段显著上调。为了独立证实微阵列结果，选择了 15 个基因进行纤维基因表达的定量反转录 PCR 分析。这些分析在多个田间年度进行，证实了两个 NIL 之间纤维基因表达的时间差异。我们假设，赋予 NIL 之间纤维基因表达时间差异的基因座是重要的调节序列，为更有针对性地操纵棉花纤维品质提供了潜力。
