Sasaki Shinya, Baba Kei'ichi, Nishida Tomoaki, Tsutsumi Yuji, Kondo Ryuichiro
Department of Forest and Forest Products Sciences, Kyushu University, 6-10-1 Hakozaki, Higashiku, Fukuoka, 812-8581, Japan.
Plant Mol Biol. 2006 Dec;62(6):797-807. doi: 10.1007/s11103-006-9057-3. Epub 2006 Sep 27.
Previously we reported that purified Cell Wall Peroxidase-Cationic (CWPO-C) from poplar callus (Populus alba L.) oxidizes sinapyl alcohol and polymeric substrate unlike other plant peroxidases and proposed that this isoenzyme is a conceivable lignification specific peroxidase. In this study, we cloned full-length cDNA of CWPO-C and investigated the transcription of CWPO-C gene in various organs and the localization of CWPO-C protein in the differentiating xylem of poplar stem.Real-time PCR analyses indicated that CWPO-C gene is constitutively expressed in the developing xylem, leaf, and shoot but not affected by many stress treatments. Immunohistochemical analysis showed that CWPO-C locates in the middle lamellae, cell corners, and secondary cell walls of the fiber cells during the lignification. The intensity of the CWPO-C labeling increased gradually from the cell wall thickening stage to mature stage of fiber cells, which is very consistent with the increase of lignin content in the developing xylem. These results strongly support that CWPO-C is responsible for the lignification of the secondary xylem. Interestingly, immuno-labeling of CWPO-C was also observed inside of the ray parenchyma cells instead no signals were detected within the developing fiber cells. This suggests that CWPO-C is biosynthesized in the parenchyma cells and provided to the middle lamellae, the cell corners, and the cell walls to achieve lignin polymerization.
此前我们报道,从杨树愈伤组织(银白杨)中纯化得到的细胞壁过氧化物酶-阳离子型(CWPO-C)与其他植物过氧化物酶不同,它能氧化芥子醇和聚合底物,并提出这种同工酶可能是一种木质化特异性过氧化物酶。在本研究中,我们克隆了CWPO-C的全长cDNA,并研究了CWPO-C基因在杨树各器官中的转录情况以及CWPO-C蛋白在杨树茎分化木质部中的定位。实时PCR分析表明,CWPO-C基因在发育中的木质部、叶片和嫩枝中组成性表达,但不受多种胁迫处理的影响。免疫组织化学分析表明,在木质化过程中,CWPO-C定位于纤维细胞的中层、细胞角和次生细胞壁。从纤维细胞壁增厚阶段到成熟阶段,CWPO-C标记的强度逐渐增加,这与发育中的木质部中木质素含量的增加非常一致。这些结果有力地支持了CWPO-C负责次生木质部的木质化。有趣的是,在射线薄壁细胞内部也观察到了CWPO-C的免疫标记,而在发育中的纤维细胞内未检测到信号。这表明CWPO-C在薄壁细胞中生物合成,并被提供到中层、细胞角和细胞壁以实现木质素聚合。