Staiger D, Kappeler S, Müller M, Apel K
Swiss Federal Institute of Technology, Institute for Plant Sciences, Zürich.
Planta. 1994;192(2):221-31.
By differential screening of a copy DNA (cDNA) library from flowering Sinapis alba L. apices against cDNAs from vegetative apices, two cDNA clones were isolated representing transcripts that are expressed transiently at an early stage of tapetum development. The Sa tap35 cDNA encodes a polypeptide with a predicted molecular weight of 12.7 kDa and an isoelectric point of 10.4. The sa tap44 cDNA codes for a putative 12.4-kDa polypeptide with an isoelectric point of 7.5. The deduced amino-acid sequences display 76% sequence identity and contain an N-terminal stretch of hydrophobic amino acids which has characteristics of secretory signal sequences. In-vitro transcription of the cDNAs and translation of the resulting RNAs in the presence of canine pancreatic microsomes demonstrates that the two proteins are translocated into the microsomes and that the putative preproteins are proteolytically processed to the mature forms. By immunoelectron microscopy the Sa TAP35 and Sa TAP44 proteins were detected at the developing peritapetal membrane between the tapetal cytoplasm and the adjacent middle layer of the anther wall. Furthermore, labelling was observed within the locule in association with globules resembling pro-Ubisch bodies which appeared at the tetrad stage. During the early vacuolate stage of microspore development the young exine was strongly labelled. The exine and the peritapetal membrane both are composed of sporopollenin, and the pro-Ubisch bodies are thought to contain sporopollenin precursors. Thus, Sa TAP35 and Sa TAP44 might be involved in sporopollenin formation and/or deposition.
通过用开花期白芥(Sinapis alba L.)顶端的互补DNA(cDNA)文库与营养顶端的cDNA进行差异筛选,分离出两个cDNA克隆,它们代表在绒毡层发育早期瞬时表达的转录本。Sa tap35 cDNA编码一种预测分子量为12.7 kDa、等电点为10.4的多肽。sa tap44 cDNA编码一种推定的12.4 kDa多肽,等电点为7.5。推导的氨基酸序列显示76%的序列同一性,并且含有一段具有分泌信号序列特征的N端疏水氨基酸序列。cDNA的体外转录以及所得RNA在犬胰腺微粒体存在下的翻译表明,这两种蛋白质都被转运到微粒体中,并且推定的前体蛋白被蛋白水解加工成成熟形式。通过免疫电子显微镜在花药壁绒毡层细胞质和相邻中层之间发育中的围绒毡膜上检测到了Sa TAP35和Sa TAP44蛋白。此外,在药室内观察到标记与类似于原乌氏体的小球有关,这些小球出现在四分体阶段。在小孢子发育的早期液泡化阶段,年轻的花粉外壁被强烈标记。花粉外壁和围绒毡膜都由孢粉素组成,并且原乌氏体被认为含有孢粉素前体。因此,Sa TAP35和Sa TAP44可能参与孢粉素的形成和/或沉积。