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Flower proteome_ changes in protein spectrum during the advanced stages of rose petal development

Flower proteome_ changes in protein spectrum during the advanced stages of rose petal development

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ORIGINAL ARTICLE Mery Dafny-Yelin Æ Inna Guterman Æ Naama Menda Mariana Ovadis Æ Moshe Shalit Æ Eran Pichersky Dani Zamir Æ Efraim Lewinsohn Æ Zach Adam David Weiss Æ Alexander Vainstein Flower proteome: changes in protein spectrum during the advanced stages of rose petal development Received: 30 November 2004 / Accepted: 3 February 2005 / Published online: 10 May 2005 Ó Springer-Verlag 2005 Abstract Flowering is a unique and highly programmed process, but hardly anything is known about the devel- opmentally regulated proteome changes in petals. Here, we employed proteomic technologies to study petal development in rose ( Rosa hybrida ). Using two-dimen- sional polyacrylamide gel electrophoresis, we generated stage-specific (closed bud, mature flower and flower at anthesis) petal protein maps with ca. 1,000 unique pro- tein spots. Expression analyses of all resolved protein spots revealed that almost 30% of them were stage- specific, with ca. 90 protein spots for each stage. Most of the proteins exhibited differential expression during pe- tal development, whereas only ca. 6% were constitu- tively expressed. Eighty-two of the resolved proteins were identified by mass spectrometry and annotated. Classification of the annotated proteins into functional groups revealed energy, cell rescue, unknown function (including novel sequences) and metabolism to be the largest classes, together comprising ca. 90% of all identified proteins. Interestingly, a large number of stress-related proteins were identified in developing petals. Analyses of the expression patterns of annotated proteins and their corresponding RNAs confirmed the importance of proteome characterization. Keywords Flower development Æ Petals Æ Proteome Æ Rose Abbreviations 2D-PAGE: Two-dimensional polyacryl- amide gel electrophoresis Æ EST: Expression sequence tag Æ IEF: Isoelectric focusing Æ ESI-MS/MS: Electrospray ionization mass spectrometry Æ MALDI-TOF: Matrix-assisted laser-desorption time-of-flight Æ CID: Collision-induced dissociation Introduction The flower’s basic architecture is rather simple, despite its almost unlimited variability in showy traits, such as color, fragrance, shape, etc. The constant development of processes generating and fine-tuning these traits has strongly contributed to the evolutionary success of flowering plants. Nevertheless, in many plant systems, petals are the main source of this enormous variability. Flower development is a uniquely programmed and tightly regulated process (Rolland-Lagan et al. 2003 ). Two discrete growth phases are distinguishable in petal development: one involves cell division, the other cell expansion (Martin and Gerats 1993 ; Yu et al. 2004 ). Very little is known about the genetic machinery involved in corolla-shape determination in general and phase tran- sition in particular: in Gerbera , GEG has been proposed to be involved in the former (Kotilainen et al. 1999 ) and in Arabidopsis , NAP has been shown to function in the transition from active cell division to cell expansion Electronic Supplementary Material Supplementary
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