Characteristic and expression analysis of a metallothionein gene, OsMT2b, down-regulated by cytokinin suggests functions in root development and seed embryo germination of rice.
Metallothioneins (MTs) are low molecular mass and cysteine-rich metal-binding proteins known to be mainly involved in maintaining metal homeostasis and stress responses. But, their functions in higher plant development are scarcely studied. Here, we characterized rice (Oryza sativa) METALLOTHIONEIN2b (OsMT2b) molecularly and found that its expression was down-regulated by cytokinins. OsMT2b was preferentially expressed in rice immature panicles, scutellum of germinating embryos, and primordium of lateral roots. In contrast with wild-type plants, OsMT2b-RNA interference (RNAi) transgenic plants had serious handicap in plant growth and root formation, whereas OsMT2b-overexpressing transformants were dwarfed and presented more adventitious roots and big lateral roots. The increased cytokinin levels in RNAi plants and decreased cytokinin levels in overexpressing plants were confirmed by high-performance liquid chromatography quantitative analysis in the roots of wild-type and transgenic plants. In RNAi plants, localization of isopentenyladenosine, a kind of endogenous cytokinin, in roots and germinating embryos expanded to the whole tissues, whereas in overexpressing plants, the isopentenyladenosine signals were very faint in the vascular tissues of roots and scutellum cells of germinating embryos. In vitro culture of embryos could largely resume the reduced germination frequency in RNAi plants but had no obvious change in overexpressing plants. Taken together, these results indicate a possible feedback regulation mechanism of OsMT2b to the level of endogenous cytokinins that is involved in root development and seed embryo germination of rice.
Yuan
Jing
J
College of Life Sciences, Wuhan University, Wuhan 430072, China.
Chen
Dan
D
Ren
Yujun
Y
Zhang
Xuelian
X
Zhao
Jie
J
D000595:Amino Acid Sequence; D001483:Base Sequence; D003583:Cytokinins / Q000502:physiology; D017931:DNA Primers; D015536:Down-Regulation; D018506:Gene Expression Regulation, Plant; D018525:Germination; D008668:Metallothionein / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D012275:Oryza / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D018517:Plant Roots / Q000254:growth & development*; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012639:Seeds / Q000254:growth & development*; D017386:Sequence Homology, Amino Acid
D003583:Cytokinins; D017931:DNA Primers; D012333:RNA, Messenger; D008668:Metallothionein