FOX-superroots of Lotus corniculatus, overexpressing Arabidopsis full-length cDNA, show stable variations in morphological traits

• Published in: Journal of plant physiology Vol. 168; no. 2; pp. 181 - 187
• Main Authors: Himuro, Yasuyo, Tanaka, Hidenori, Hashiguchi, Masatsugu, Ichikawa, Takanari, Nakazawa, Miki, Seki, Motoaki, Fujita, Miki, Shinozaki, Kazuo, Matsui, Minami, Akashi, Ryo, Hoffmann, Franz
• Format: Journal Article
• Language: English
• Published: Munich Elsevier GmbH 15.01.2011; Elsevier
• Subjects: Agrobacterium; Arabidopsis; Arabidopsis - genetics; Biological and medical sciences; DNA, Complementary - genetics; FOX-hunting system; Full-length cDNA; Fundamental and applied biological sciences. Psychology; Loteae - genetics; Loteae - growth & development; Lotus corniculatus; Plant physiology and development; Plant Roots - genetics; Plant Roots - growth & development; Plants, Genetically Modified - genetics; Plants, Genetically Modified - growth & development; Polymerase Chain Reaction; Rhizobium - genetics; Superroots; Transgenic plants; RT; RARGE; Superroots; FOX-hunting system; FOX hunting system; Lotus corniculatus; hpt; Transgenic plants; BAP; fl-cDNA; FOX-SR lines; Full-length cDNA; FSL; SR; Gene overexpression; Variations; System; Arabidopsis thaliana; Leguminosae; Complementary DNA; Plant physiology; Cruciferae; Length; Dicotyledones; Angiospermae; Morphological character; Spermatophyta; Transgenic plant; Fodder crop; Experimental plant
• ISSN: 0176-1617; 1618-1328
• DOI: 10.1016/j.jplph.2010.10.003

Using the full-length cDNA overexpressor (FOX) gene-hunting system, we have generated 130 Arabidopsis FOX-superroot lines in bird's-foot trefoil ( Lotus corniculatus) for the systematic functional analysis of genes expressed in roots and for the selection of induced mutants with interesting root growth characteristics. We used the Arabidopsis-FOX Agrobacterium library (constructed by ligating pBIG2113SF) for the Agrobacterium-mediated transformation of superroots (SR) and the subsequent selection of gain-of-function mutants with ectopically expressed Arabidopsis genes. The original superroot culture of L. corniculatus is a unique host system displaying fast root growth in vitro, allowing continuous root cloning, direct somatic embryogenesis and mass regeneration of plants under entirely hormone-free culture conditions. Several of the Arabidopsis FOX-superroot lines show interesting deviations from normal growth and morphology of roots from SR-plants, such as differences in pigmentation, growth rate, length or diameter. Some of these mutations are of potential agricultural interest. Genomic PCR analysis revealed that 100 (76.9%) out of the 130 transgenic lines showed the amplification of single fragments. Sequence analysis of the PCR fragments from these 100 lines identified full-length cDNA in 74 of them. Forty-three out of 74 full-length cDNA carried known genes. The Arabidopsis FOX-superroot lines of L. corniculatus, produced in this study, expand the FOX hunting system and provide a new tool for the genetic analysis and control of root growth in a leguminous forage plant.