Hoeberichts Frank A, van Doorn Wouter G, Vorst Oscar, Hall Robert D, van Wordragen Monique F
Agrotechnology and Food Science, Wageningen University and Research Centre, PO Box 17, 6700 AA Wageningen, The Netherlands.
J Exp Bot. 2007;58(11):2873-85. doi: 10.1093/jxb/erm076. Epub 2007 Jul 13.
cDNA microarrays were used to characterize senescence-associated gene expression in petals of cut carnation (Dianthus caryophyllus) flowers, sampled from anthesis to the first senescence symptoms. The population of PCR fragments spotted on these microarrays was enriched for flower-specific and senescence-specific genes, using subtractive hybridization. About 90% of the transcripts showed a large increase in quantity, approximately 25% transiently, and about 65% throughout the 7 d experiment. Treatment with silver thiosulphate (STS), which blocks the ethylene receptor and prevented the normal senescence symptoms, prevented the up-regulation of almost all of these genes. Sucrose treatment also considerably delayed visible senescence. Its effect on gene expression was very similar to that of STS, suggesting that soluble sugars act as a repressor of ethylene signal transduction. Two fragments that encoded a carnation EIN3-like (EIL) protein were isolated, some of which are key transcription factors that control ethylene response genes. One of these (Dc-EIL3) was up-regulated during senescence. Its up-regulation was delayed by STS and prevented by sucrose. Sucrose, therefore, seems to repress ethylene signalling, in part, by preventing up-regulation of Dc-EIL3. Some other transcription factors displayed an early increase in transcript abundance: a MYB-like DNA binding protein, a MYC protein, a MADS-box factor, and a zinc finger protein. Genes suggesting a role in senescence of hormones other than ethylene encoded an Aux/IAA protein, which regulate transcription of auxin-induced genes, and a cytokinin oxidase/dehydrogenase, which degrades cytokinin. Taken together, the results suggest a master switch during senescence, controlling the co-ordinated up-regulation of numerous ethylene response genes. Dc-EIL3 might be (part of) this master switch.
利用cDNA微阵列技术,对从开花期到出现最初衰老症状的切花康乃馨(香石竹)花瓣中衰老相关基因的表达进行了表征。通过消减杂交,富集了这些微阵列上所点样的PCR片段中的花特异性和衰老特异性基因。约90%的转录本数量大幅增加,其中约25%为短暂增加,约65%在整个7天的实验过程中持续增加。用硫代硫酸银(STS)处理可阻断乙烯受体并防止出现正常的衰老症状,该处理几乎阻止了所有这些基因的上调。蔗糖处理也显著延迟了可见衰老。其对基因表达的影响与STS非常相似,表明可溶性糖作为乙烯信号转导的抑制因子发挥作用。分离出了两个编码康乃馨EIN3样(EIL)蛋白的片段,其中一些是控制乙烯反应基因的关键转录因子。其中一个(Dc-EIL3)在衰老过程中上调。其上调被STS延迟,并被蔗糖阻止。因此,蔗糖似乎部分通过阻止Dc-EIL3的上调来抑制乙烯信号传导。其他一些转录因子的转录本丰度早期增加:一种MYB样DNA结合蛋白、一种MYC蛋白、一种MADS盒因子和一种锌指蛋白。暗示除乙烯外其他激素在衰老中起作用的基因编码一种Aux/IAA蛋白(调节生长素诱导基因的转录)和一种细胞分裂素氧化酶/脱氢酶(降解细胞分裂素)。综合来看,结果表明在衰老过程中存在一个主开关,控制着众多乙烯反应基因的协同上调。Dc-EIL3可能是这个主开关的(一部分)。