del Campillo E, Bennett A B
Department of Plant Biology, University of Maryland, College Park 20742, USA.
Plant Physiol. 1996 Jul;111(3):813-20. doi: 10.1104/pp.111.3.813.
Six cellulase genes were isolated from total RNA of the ethylene-treated tomato (Lycopersicon esculentum Mill.) flower abscission zone by reverse-transcription polymerase chain reaction using degenerate primers to conserved amino acid sequences from known plant cellulases. Four of the gene fragments are homologous to fruit pericarp cellulases. The other two are novel cellulase genes, referred to as Cel5 and Cel6. Breakstrength and cellulase gene expression were then analyzed in naturally abscising flowers and flower explants. In both naturally abscising flowers and flower explants induced to abscise in air or ethylene, both new cellulase mRNAs were correlated with flower shedding. Whereas the Cel5 mRNA increased in later stages of abscission, the Cel6 mRNA was present in nonabscising flowers and then decreased in the final stage of abscission. A third cellulase, Cel1, increased during the final stage of abscission in flower explants and yet did not increase during shedding in planta, although it was detectable at low levels in all abscission stages. Cel1 and Cel5 mRNA decreased 99% when indole-3-acetic acid was added during ethylene treatment, consistent with low levels of abscission (3%). In contrast, Cel6 mRNA increased slightly when indole-3-acetic acid was added. These results suggest that abscission is a multistep process involving both activated and repressed cellulase genes and that the relative importance of each cellulase in the process depends on the physiological conditions under which abscission takes place.
通过逆转录聚合酶链反应,利用针对已知植物纤维素酶保守氨基酸序列的简并引物,从乙烯处理过的番茄(Lycopersicon esculentum Mill.)花脱落区的总RNA中分离出六个纤维素酶基因。其中四个基因片段与果实果皮纤维素酶同源。另外两个是新的纤维素酶基因,分别称为Cel5和Cel6。然后对自然脱落的花朵和花外植体中的抗断强度和纤维素酶基因表达进行了分析。在自然脱落的花朵以及在空气中或乙烯诱导下脱落的花外植体中,两种新的纤维素酶mRNA都与花朵脱落相关。虽然Cel5 mRNA在脱落后期增加,但Cel6 mRNA在未脱落的花朵中存在,然后在脱落的最后阶段减少。第三种纤维素酶Cel1在花外植体脱落的最后阶段增加,但在植株体内脱落过程中并未增加,尽管在所有脱落阶段都能检测到低水平的Cel1。在乙烯处理期间添加吲哚-3-乙酸时,Cel1和Cel5 mRNA下降了99%,这与低水平的脱落率(3%)一致。相反,添加吲哚-3-乙酸时,Cel6 mRNA略有增加。这些结果表明,脱落是一个多步骤过程,涉及激活和抑制的纤维素酶基因,并且每个纤维素酶在该过程中的相对重要性取决于脱落发生时的生理条件。