Molecular and Physiological Characterisation of an Insertion Mutant in the ARR21 Putative Response Regulator Gene from Arabidopsis thaliana

Abstract

In our search for insertion mutants of Arabidopsis response regulator (ARR) genes, we identified a candidate for an ARR21 dSpm transposon insertion line in the SLAT collection by searching the SINS sequence database. Molecular characterisation of this line revealed that the transposon is integrated as a single copy 1727 bases downstream of the ATG signal, within the third intron of the ARR21 gene. The transposon insertion segregated in a Mendelian fashion from heterozygous plants that were allowed to self-pollinate. RT-PCR-based expression analysis showed that ARR21 transcript predominantly accumulates in siliques. In contrast, the full-length ARR21 transcript was not detectable in the ARR21 transposon insertion line, indicating that it harbours an insertion of dSpm transposon in the ARR21 gene. The ARR21 insertion mutant (arr21-1) was subjected to several physiological tests for a possible insertion-linked phenotype. However, our results revealed that the insertion in the ARR21 gene did not cause any alterations in viability and fertility, flowering time, sensitivity to ethylene, cytokinin or red light. We discuss these results in the light of recent findings about the function of the other members of the response regulator gene family of Arabidopsis.

Key words

SLAT - insertion lines - two-component system - response regulator - signal transduction - Arabidopsis thaliana

References

• 1 Brandstatter I. B., Kieber J. J.. Two genes with similarity to bacterial response regulators are rapidly and specifically induced by cytokinin in Arabidopsis. .  The Plant Cell. (1998);  10 1009-1019
• 2 Chang C., Kwok S. F., Bleecker A. B., Meyerowitz E. M.. Arabidopsis ethylene response gene ETR1: similarity of product to two-component regulators.  Science. (1993);  262 539-544
• 3 Chang C., Stadler R.. Ethylene hormone receptor action in Arabidopsis. .  Bioassays. (2001);  23 619-627
• 4 D'Agostino I. B., Deruère J., Kieber J. J.. Characterization of the response of the Arabidopsis response regulator gene family to cytokinin.  Plant Physiology. (2000);  124 1706-1717
• 5 Hosoda K., Imamura A., Katoh E., Hatta T., Tachiki M., Yamada H., Mizuno T., Yamazaki T.. Molecular structure of the GARP family of the plant myb-related DNA binding motifs of the Arabidopsis response regulators.  The Plant Cell. (2002);  14 2015-2029
• 6 Hutchinson C. E., Kieber J. J.. Cytokinin signaling in Arabidopsis. .  The Plant Cell. Supplement. (2002);  2002 S47-S59
• 7 Hwang I., Chen H. C., Sheen J.. Two-component signal transduction pathways in Arabidopsis. .  Plant Physiology. (2002);  129 500-515
• 8 Hwang I., Sheen J.. Two-component circuity in Arabidopsis signal transduction.  Nature. (2001);  413 383-389
• 9 Imamura A., Hanaki N., Nakamura A., Suzuki T., Taniguchi M., Kiba T., Ueguchi C., Sugiyama T., Mizuno T.. Compilation and characterization of Arabidopsis thaliana response regulators implicated in His-Asp phosphorelay signal transduction.  Plant and Cell Physiology. (1999);  40 733-742
• 10 Inoue T., Higuchi M., Hashimoto Y., Seki M., Kobayashi M., Kato T., Tabata S., Shinozaki K., Kakimoto T.. Identification of CRE1 as a cytokinin receptor from Arabidopsis. .  Nature. (2001);  409 1060-1063
• 11 Kiba T., Yamada H., Mizuno T.. Characterization of the ARR15 and ARR16 response regulators with special reference to the cytokinin signaling pathway mediated by the AHK4 histidine kinase in roots of Arabidopsis thaliana. .  Plant and Cell Physiology. (2002);  43 1059-1066
• 12 Kubo M., Kakimoto T.. The cytokinin-hypersensitive genes of Arabidopsis negatively regulate the cytokinin-signaling pathway for cell division and chloroplast development.  Plant Journal. (2000);  23 385-394
• 13 Lohrmann J., Bucholz G., Keitel C., Sweere U., Kircher S., Bäurle I., Kudla J., Schäfer E., Harter K.. Differential expression and nuclear localization of response regulator-like proteins from Arabidopsis thaliana. .  Plant Biology. (1999);  1 495-505
• 14 Lohrmann J., Harter K.. Plant two-component signaling systems and the role of response regulators.  Plant Physiology. (2002);  128 363-369
• 15 Lohrmann J., Sweere U., Zabaleta E., Bäurle I., Keitel C., Kozma-Bognar L., Brennicke A., Schäfer E., Kudla J., Harter K.. The response regulator ARR2: A pollen-specific transcription factor involved in the expression of nuclear genes for components of mitochondrial complex I in Arabidopsis. .  Molecular Genetics and Genomics. (2001);  265 2-13
• 16 Parkinson J. S., Kofoid E. C.. Communication modules in bacterial signaling proteins.  Annual Review in Genetics. (1992);  26 71-112
• 17 Riechmann J. L., Heard J., Martin G., Reber L., Jiang L., Keddie J., Adam L., Pineda O., Ratcliffe J., Samaha R. R., Creelman R., Pilgrim M., Broun P., Zhang J. Z., Gandehari D., Sherman B. K., Yu G.. Arabidopsis transcription factors: genome-wide comparative analysis among eukaryotes.  Science. (2000);  290 2105-2110
• 18 Sakai H., Ayoma T., Oka A.. Arabidopsis ARR1 and ARR2 response regulators operate as transcriptional activators.  Plant Journal. (2000);  24 703-711
• 19 Sakai H., Honma T., Aoyama T., Sato S., Kato T., Tabata S., Oka A.. ARR1, a transcription factor for genes immediately responsive to cytokinins.  Science. (2001);  294 1519-1521
• 20 Schaller G. E., Mathews D., Gribskov M., Walker J. C.. Two-component signaling elements and histidyl-to-aspartyl phosphorelays. Somerville, C. and Meyerowitz, E., eds. The Arabidopsis Book. Washington; American Society of Plant Biologists (2002)
• 21 Suzuki T., Miwa K., Ishikawa K., Yamada H., Aiba H., Mizuno T.. The Arabidopsis sensor His-kinase, AHK4, can respond to cytokinins.  Plant and Cell Physiology. (2001);  42 107-113
• 22 Suzuki T., Ishikawa K., Mizuno T.. An Arabidopsis histidine-containing phosphotransfer (Hpt) factor implicated in phosphorelay signal transduction: Overexpression of AHP2 in plants results in hypersensitiveness to cytokinin.  Plant and Cell Physiology. (2002);  43 123-129
• 23 Sweere U., Eichenberg K., Lohrmann J., Mira-Rodado V., Bäurle I., Kudla J., Nagy F., Schäfer E., Harter K.. Interaction of the response regulator ARR4 with phytochrome B in modulating red light signaling.  Science. (2001);  294 1108-1111
• 24 Taniguchi M., Kiba T., Sakakibara H., Ueguchi C., Mizuno T., Sugiyama T.. Expression of Arabidopsis response regulator homologs is induced by cytokinins and nitrate.  FEBS Letters. (1998);  429 259-262
• 25 Tissier A. F., Marillonnet S., Klimyuk V., Patel K., Torres M. A., Murphy G., Jones J. D. G.. Multiple independent defective suppressor-mutator transposon insertions in Arabidopsis: a tool for functional genomics.  The Plant Cell. (1999);  11 1841-1852
• 26 Ueguchi C., Sato S., Kato T., Tabata S.. The AHK4 gene involved in the cytokinin-signaling pathway as a direct receptor molecule in Arabidopsis thaliana. .  Plant and Cell Physiology. (2001);  42 751-755
• 27 Urao T., Yakubov B., Satoh R., Yamaguchi-Shinozaki K., Seki M., Hirayama T., Shinozaki K.. A transmembrane hybrid-type histidine kinase in Arabidopsis functions as an osmosensor.  The Plant Cell. (1999);  11 1743-1754
• 28 Urao T., Yamaguchi-Shinozaki K., Shinozaki K.. Two-component systems in plant signal transduction.  Trends in Plant Science. (2000);  5 67-74
• 29 Yamada H., Suzuki T., Terada K., Takei K., Ishikawa K., Miwa K., Yamashino T., Mizuno T.. The Arabidopsis AHK4 histidine kinase is a cytokinin-binding receptor that transduces cytokinin signals across the membrane.  Plant and Cell Physiology. (2001);  42 1017-1023

Section Editor: W. Frommer