Ohmiya Yasunori, Nakai Tomonori, Park Yong Woo, Aoyama Takashi, Oka Atsuhiro, Sakai Fukumi, Hayashi Takahisa
Wood Research Institute, Kyoto University, Uji, Kyoto 611-0011, Japan.
Plant J. 2003 Mar;33(6):1087-97. doi: 10.1046/j.1365-313x.2003.01695.x.
Poplar calli transcribed two cellulase (endo-1,4-beta-glucanase) genes, PopCel1 and PopCel2, whose mRNAs were differentially located in the growing leaves of poplar during cell wall synthesis. Histochemical and RT-PCR analyses of promoter-GUS fusion gene activities in transgenic poplar demonstrated that PopCel1 promoter-derived GUS activity was localized in the petiole and leaf veins, whereas PopCel2 was confined to mesophyll cells and disappeared from the tip during the development of leaves. Autoradiography of the leaf showed that the radioactivity of [14C]sucrose incorporated into cellulose corresponded to the combination of the sucrose-induced tissue-specific patterns of PopCel1 and PopCel2. Interestingly, 2,6-dichlorobenzonitrile (DCB) not only inhibited the incorporation of the radioactivity into cellulose, but also repressed the induction of both cellulase genes. Suppression of cellulases by expression of PopCel1 antisense cDNA or co-suppression of PopCel1 mRNA by overexpression of PopCel1 sense cDNA reduced leaf growth. Therefore, we came to the conclusion that PopCel1 and PopCel2 probably function to promote leaf growth in poplar by the endohydrolysis of 1,4-beta-glucan.
杨树愈伤组织转录了两个纤维素酶(内切-1,4-β-葡聚糖酶)基因PopCel1和PopCel2,其mRNA在杨树细胞壁合成过程中于生长叶片中的定位存在差异。对转基因杨树中启动子-GUS融合基因活性的组织化学和RT-PCR分析表明,PopCel1启动子驱动的GUS活性定位于叶柄和叶脉,而PopCel2局限于叶肉细胞,并在叶片发育过程中从叶尖消失。叶片的放射自显影显示,掺入纤维素中的[14C]蔗糖的放射性与蔗糖诱导的PopCel1和PopCel2的组织特异性模式的组合相对应。有趣的是,2,6-二氯苯腈(DCB)不仅抑制放射性掺入纤维素,还抑制两个纤维素酶基因的诱导。通过PopCel1反义cDNA的表达或通过PopCel1正义cDNA的过表达共抑制PopCel1 mRNA来抑制纤维素酶,会降低叶片生长。因此,我们得出结论,PopCel1和PopCel2可能通过1,4-β-葡聚糖的内切水解作用来促进杨树叶片的生长。