Xu F X, Chye M L
Department of Botany, University of Hong Kong, China.
Plant J. 1999 Feb;17(3):321-7. doi: 10.1046/j.1365-313x.1999.00370.x.
Here we show that the expression of a cysteine proteinase coincides with several developmental events associated with programmed cell death (PCD) in Solanum melongena (brinjal), i.e. during leaf senescence, fruit senescence, xylogenesis, nucellar cell degeneration and anther senescence. We have isolated a cDNA encoding brinjal cysteine proteinase (SmCP) that shares high (90-92%) amino acid identity to cysteine proteinases of tobacco (CYP-8) and tomato (LCYP-2) that have not been previously reported to be senescence-associated. In contrast, SmCP shows lower (39-41%) amino acid identity to other senescence-related cysteine proteinases and, unlike most of them, it is not preferentially expressed in certain organs or cell types. Northern analysis of leaves, fruits and flowers at different stages of development showed that SmCP expression increased significantly at senescence in leaf and fruit, but was highly expressed throughout flower development. In situ hybridization studies on flower sections using an antisense RNA probe localized the SmCP mRNA to the xylem, the epidermis and the endothecium of the anther and the nucellar cells, suggesting its involvement in PCD during xylogenesis, anther senescence and ovule development, respectively. Its expression during nucellar cell degeneration suggests that protein reserves of the nucellus are released to the developing embryo. Polarity in its pattern of expression in the nucellus of the developing seed (40DAP) further implies a directional flow of these nutrients.
我们在此表明,在茄子中,一种半胱氨酸蛋白酶的表达与几个与程序性细胞死亡(PCD)相关的发育事件相吻合,即在叶片衰老、果实衰老、木质部形成、珠心细胞退化和花药衰老期间。我们分离出了一个编码茄子半胱氨酸蛋白酶(SmCP)的cDNA,它与烟草(CYP-8)和番茄(LCYP-2)的半胱氨酸蛋白酶具有较高的氨基酸同一性(90-92%),而此前尚未报道这些蛋白酶与衰老相关。相比之下,SmCP与其他衰老相关的半胱氨酸蛋白酶的氨基酸同一性较低(39-41%),并且与它们中的大多数不同,它并非在某些器官或细胞类型中优先表达。对不同发育阶段的叶片、果实和花朵进行的Northern分析表明,SmCP的表达在叶片和果实衰老时显著增加,但在整个花朵发育过程中都高度表达。使用反义RNA探针在花切片上进行的原位杂交研究将SmCP mRNA定位到木质部、花药的表皮和内皮以及珠心细胞,表明它分别参与了木质部形成、花药衰老和胚珠发育过程中的PCD。它在珠心细胞退化过程中的表达表明珠心的蛋白质储备被释放到发育中的胚中。其在发育种子(授粉后40天)珠心中的表达模式的极性进一步暗示了这些营养物质的定向流动。