Zeba Naheed, Isbat Mohammad, Kwon Nak-Jung, Lee Mi Ok, Kim Seong Ryong, Hong Choo Bong
School of Biological Sciences and Institute of Molecular Biology and Genetics, Seoul National University, Seoul, 151-742, South Korea.
Planta. 2009 Mar;229(4):861-71. doi: 10.1007/s00425-008-0884-0. Epub 2009 Jan 6.
Capsicum annuum RING Zinc Finger Protein 1 (CaRZFP1) gene is a novel C3HC4-type RING zinc finger protein gene which was previously isolated from a cDNA library for hot pepper plants treated of heat-shock. The CaRZFP1 was inducible to diverse environmental stresses in hot pepper plants. We introduced the CaRZFP1 into the Wisconsin 38 cultivar of tobacco (Nicotiana tabacum) by Agrobacterium mediated transformation under the control of the CaMV 35S promoter. Expression of the transgene in the transformed tobacco plants was demonstrated by RNA blot analyses. There appeared no adverse effect of over-expression of the transgene on overall growth and development of transformants. The genetic analysis of tested T(1) lines showed that the transgene segregated in a Mendelian fashion. Transgenic tobacco lines that expressed the CaRZFP1 gene were compared with several different empty vector lines and they exhibited enhanced growth; they have larger primary root, more lateral root, larger hypocotyls and bigger leaf size, resulting in heavier fresh weight. Enhanced growth of transgenic lines accompanied with longer vegetative growth that resulted in bigger plants with higher number of leaves. Microarray analysis revealed the up-regulation of some growth related genes in the transgenic plants which were verified by specific oligomer RNA blot analyses. These results indicate that CaRZFP1 activates and up-regulates some growth related proteins and thereby effectively promoting plant growth.
辣椒环指锌指蛋白1(CaRZFP1)基因是一种新型的C3HC4型环指锌指蛋白基因,先前从经热激处理的辣椒植株的cDNA文库中分离得到。CaRZFP1在辣椒植株中可被多种环境胁迫诱导。我们通过农杆菌介导的转化,在CaMV 35S启动子的控制下,将CaRZFP1导入烟草(Nicotiana tabacum)的Wisconsin 38品种中。通过RNA印迹分析证实了转基因在转化烟草植株中的表达。转基因的过表达对转化体的整体生长和发育未出现不利影响。对测试的T(1)代株系的遗传分析表明,转基因以孟德尔方式分离。将表达CaRZFP1基因的转基因烟草株系与几个不同的空载体株系进行比较,它们表现出增强的生长;它们有更大的主根、更多的侧根、更大的下胚轴和更大的叶片尺寸,从而导致鲜重更重。转基因株系生长增强伴随着营养生长延长,从而产生叶片数量更多的更大植株。微阵列分析揭示了转基因植物中一些与生长相关基因的上调,这通过特异性寡聚体RNA印迹分析得到验证。这些结果表明,CaRZFP1激活并上调一些与生长相关的蛋白质,从而有效地促进植物生长。