Harrison E P, McQueen-Mason S J, Manning K
Horticulture Research International, Wellesbourne, Warwick CV35 9EF, UK.
J Exp Bot. 2001 Jul;52(360):1437-46. doi: 10.1093/jexbot/52.360.1437.
Expansins are proteins which have been demonstrated to induce cell wall extension in vitro. The identification and characterization of six expansin cDNAs from strawberry fruit, termed FaExp3 to FaExp7, as well as the previously identified FaExp2 is reported here. Analysis of expansin mRNAs during fruit development and in leaves, roots and stolons revealed a unique pattern of expression for each cDNA. FaExp3 mRNA was present at much lower levels than the other expansin mRNAs and was expressed in small green fruit and in ripe fruit. FaExp4 mRNA was present throughout fruit development, but was more strongly expressed during ripening. FaExp5 was the only clone to show fruit specific expression which was up-regulated at the onset of ripening. FaExp6 and FaExp7 mRNAs were present at low levels in the fruit with highest expression in stolon tissue. During fruit development FaExp6 had the highest expression at the white, turning and orange stages whereas expression of FaExp7 was highest in white fruit. The expression profiles of FaExp2 and FaExp5 in developing fruit were similar except that FaExp2 was induced at an earlier stage. Analysis of expansin protein by Western blotting using an antibody raised against CsExp1 from cucumber hypocotyls identified two bands of 29 and 31 kDa from developing fruit. Protein extracts from developing fruit were assayed for extension activity. Considerable rates of extension were observed with extracts from ripening fruit, but no extension was observed with protein from unripe green fruit. These results demonstrate the presence of at least six expansin genes in strawberry fruit and that during ripening the fruit acquires the ability to cause extension in vitro, characteristic of expansin action.
扩张蛋白是一类已被证明能在体外诱导细胞壁伸展的蛋白质。本文报道了从草莓果实中鉴定和表征的6个扩张蛋白cDNA,命名为FaExp3至FaExp7,以及之前鉴定的FaExp2。对果实发育过程中以及叶片、根和匍匐茎中扩张蛋白mRNA的分析揭示了每个cDNA独特的表达模式。FaExp3 mRNA的水平远低于其他扩张蛋白mRNA,在小绿果和成熟果中表达。FaExp4 mRNA在果实发育过程中均有存在,但在成熟过程中表达更强。FaExp5是唯一显示果实特异性表达的克隆,在成熟开始时上调。FaExp6和FaExp7 mRNA在果实中的水平较低,在匍匐茎组织中表达最高。在果实发育过程中,FaExp6在白色、转色和橙色阶段表达最高,而FaExp7在白色果实中表达最高。FaExp2和FaExp5在发育中的果实中的表达谱相似,只是FaExp2在更早阶段被诱导。使用针对黄瓜下胚轴的CsExp1产生的抗体通过蛋白质印迹分析扩张蛋白,从发育中的果实中鉴定出两条29 kDa和31 kDa的条带。对发育中的果实的蛋白质提取物进行伸展活性测定。观察到成熟果实提取物有相当高的伸展率,但未成熟绿色果实的蛋白质未观察到伸展。这些结果表明草莓果实中至少存在6个扩张蛋白基因,并且在成熟过程中果实获得了在体外引起伸展的能力,这是扩张蛋白作用的特征。