Publications (3)7.22 Total impact
Article: Large-scale screening of Arabidopsis enhancer-trap lines for seed germination-associated genes.[show abstract] [hide abstract]
ABSTRACT: Enhancer trap is a powerful approach for identifying tissue- and stage-specific gene expression in plants and animals. For Arabidopsis research, beta-glucuronidase (GUS) enhancer-trap lines have been created and successfully used to identify tissue-specific gene expression in many plant organs. However, limited applications of these lines for seed germination research have been reported. This is probably due to the impermeability of the testa to the GUS substrate. By focusing on the stages between testa and endosperm rupture, we were able to circumvent the testa barrier to the GUS substrate and observe diverse tissue-specific gene expression during germination sensu stricto. One hundred and twenty-one positive subpools of 10 lines out of 1130 were isolated. Approximately 4500 plants from these subpools were grown in a greenhouse and one to seven individual plants exhibiting GUS expression in seeds were isolated for each subpool. This library of the Arabidopsis seed enhancer-trap lines is an efficient tool for identifying seed germination-associated genes. The individual lines from this library will be provided to the international seed biology research community. International collaboration to identify the trapped genes using genome-walking PCR and to characterize the gene functions using knockout plants will significantly enhance our understanding of the molecular mechanisms of seed germination.The Plant Journal 04/2005; 41(6):936-44. · 6.16 Impact Factor
Article: Endo-β-mannanase activity is associated with the completion of embryogenesis in imbibed carrot (Daucus carota L.) seeds[show abstract] [hide abstract]
ABSTRACT: Development of the rudimentary embryo in mature carrot (Daucus carota) seed during imbibition was characterized. The small embryo in the carrot seed, located in the micropylar region, elongated into the lateral part during imbibition and attained about two-thirds the length of the seed before radicle protrusion. Developing embryos excised from imbibed seeds were only capable of germinating in both water and Murashige–Skoog (MS) medium when they reached maximum size. The corrosion cavity into which the embryo grew enlarged concomitantly with endosperm degradation. The expression of endo-β-mannanase (EC 220.127.116.11), which is assumed to be involved in endosperm degradation, was characterized. A cDNA encoding an endo-β-mannanase was obtained by reverse transcription polymerase chain reaction (RT-PCR) using total RNA extracted from 24-h-imbibed carrot seeds. The full-length cDNA (DcMAN1) exhibited 64% deduced amino acid sequence identity with tomato (Lycopersicon esculentum) seed germination-associated mannanase (LeMAN2). DcMAN1 mRNA and endo-β-mannanase activity were first detected in the micropylar-half seed and then in the lateral-half seed. The timing of the appearance of DcMAN1 mRNA and endo-β-mannanase activity in the lateral-half seed corresponded with that of embryo development into this region. These results suggest that the expression of DcMAN1 and endo-β-mannanase activity in imbibed carrot seeds is associated with the enlargement of the corrosion cavity, which accompanies embryogenesis.Seed Science Research 08/2003; 13(03):219 - 227. · 1.06 Impact Factor
Article: Regulation of seed germination and sand establishment - - importance of repression of developmental programs.