Near-isogenic cotton germplasm lines that differ in fiber-bundle strength have temporal differences in fiber gene expression patterns as revealed by comparative high-throughput profiling
Near-isogenic cotton germplasm lines that differ in fiber-bundle strength have temporal differences in fiber gene expression patterns as revealed by comparative high-throughput profiling
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DOI：
10.1007/s00122-010-1260-6
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发表时间：
2010-05-01
影响因子：
5.4
通讯作者：
Triplett, Barbara A.
中科院分区：
文献类型：
--
作者：
Hinchliffe, Doug J.;Meredith, William R.;Triplett, Barbara A.
棉花（Gossypium hirsutum L.）使用寡核苷酸微阵列比较来自两个近等基因系（NIL）的纤维，所述近等基因系在纤维束强度、短纤维含量和少于两个遗传基因座上不同。纤维基因表达进行了比较，在五个时间点跨越纤维伸长和次生细胞壁（SCW）的生物合成。纤维样品从田间小区以随机化、完全区组设计收集，每个NIL在每个时间点具有三个空间上不同的生物重复。微阵列杂交进行循环实验设计，允许比较纤维基因表达谱作为两个近等离子体之间的时间的函数。总体而言，发育表达模式揭示的微阵列实验同意与先前报道的特定基因的棉花纤维基因表达模式。此外，与棉花纤维中SCW生物合成的开始协调表达的基因与其他产生SCW的植物组织的基因表达模式相关。对两个近等基因系差异表达基因的功能分类和富集分析表明，在棉纤维发育的过渡期，与SCW生物合成相关的基因在高纤维品质系纤维中显著上调。为了独立确证微阵列结果，选择15个基因用于定量逆转录PCR分析纤维基因表达。这些分析，进行了多个领域的年，证实了两个近等离子体之间的纤维基因表达的时间差异。我们推测，赋予近等基因系之间的纤维基因表达的时间差异的位点是重要的调控序列，提供了更有针对性的操纵棉花纤维品质的潜力。
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.