Gene expression patterns to define stages of post-harvest senescence in Alstroemeria petals
Gene expression patterns to define stages of post-harvest senescence in Alstroemeria petals
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DOI：
10.1111/j.1467-7652.2004.00059.x
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发表时间：
2004-03-01
影响因子：
13.8
通讯作者：
Buchanan-Wollaston, V
中科院分区：
文献类型：
--
作者：
Breeze, E;Wagstaff, C;Buchanan-Wollaston, V
许多物种的花瓣衰老受乙烯调节，但有些花，如单子叶植物Alstroemeria，var。丽贝卡对乙烯不敏感利用几种不同的技术研究了在花采后衰老过程中基因表达的变化。抑制性消减杂交（SSH）被用来获得丰富的基因表达在花瓣衰老的选定阶段的cDNA文库。对获得的EST克隆进行测序，得到超过1000个序列，代表大约500个不同的基因。对这些基因的潜在功能的分析提供了花瓣发育过程的快照。细胞壁相关基因和参与代谢的基因在早期阶段都以较高的比例存在。编码金属结合蛋白（主要是金属硫蛋白样）的基因是衰老增强文库的主要组成部分。这限制了在后期鉴定出的显示差异表达的基因的多样性。使用微阵列杂交分析所有基因表达的变化，并鉴定显示上调或下调的基因。使用北方杂交证实了选择的基因的表达模式。北方杂交证实了金属硫蛋白在花开放后的上调，然而，这是没有检测到的微阵列分析，表明使用组合的方法来研究基因表达模式的重要性。相当多的基因被上调而不是下调。这可能反映了在Alstroemeria花瓣衰老过程中需要表达一组全新的基因，这些基因与降解和动员有关。表达谱的潜在用途，以提高花卉质量的育种计划或作为一种诊断工具进行了讨论。
Petal senescence in many species is regulated by ethylene but some flowers, such as those on the monocotyleclonous plant Alstroemeria, var. Rebecca are ethylene insensitive. Changes in gene expression during the post-harvest senescence of Alstroemeria flowers were investigated using several different techniques. Suppressive subtractive hybridization (SSH) was used to obtain cDNA libraries enriched for genes expressed at selected stages of petal senescence. Sequencing of the EST clones obtained resulted in over 1000 sequences that represent approximately 500 different genes. Analysis of the potential functions of these genes provides a snapshot of the processes that are taking place during petal development. Both cell wall related genes and genes involved in metabolism were present at a higher proportion in the earlier stages. Genes encoding metal binding proteins (mostly metallothionein-like) were the major component of senescence enhanced libraries. This limited the diversity of genes identified showing differential expression at the later stages. Changes in the expression of all genes were analysed using microarray hybridization, and genes showing either up or down-regulation were identified. The expression pattern of a selection of genes was confirmed using Northern hybridization. Northern hybridization confirmed the up-regulation of metallothioneins after floral opening, however, this was not detected by the microarray analysis, indicating the importance of using a combination of methods to investigate gene expression patterns. Considerably more genes were upregulated than down-regulated. This may reflect the need during Alstroemeria petal senescence for the expression of a whole new set of genes involved with degradation and mobilization. The potential uses of expression profiling to improve floral quality in breeding programmes or as a diagnostic tool are discussed.