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Arabidopsis EMBRYOMAKER encoding an AP2 domain transcription factor plays a key role in developmental change from vegetative to embryonic phase

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Abstract

Although several types of plant cells retain the competence to enter into embryonic development without fertilization, the molecular mechanism(s) underlying ectopic embryogenesis is largely unknown. To gain insight into this mechanism, in a previous study we identified 136 ESTs specifically expressed in microspore embryogenesis of Brassica napus. Here, we describe the characterization of the Arabidopsis EMBRYOMAKER (EMK) gene, which is homologous to one of the identified Brassica ESTs (BnGemb-58) and encodes an AP2 domain transcription factor. The AtEMK was expressed in developing and mature embryos, but its rapid disappearance occurred during germination. After germination, the expression of AtEMK was found in the root apical meristem and the distal parts of cotyledons. Although a mutant lacking AtEMK exhibited no distinctive defects in the embryo, ectopic expression of AtEMK induced embryo-like structures from cotyledons. The embryo-like structures contained high concentration of lipids, expressed several embryo-specific genes, and could convert into independent plants, indicating that the structures are somatic embryos. In vitro culture, AtEMK enhanced the efficiency of somatic embryogenesis. Furthermore, ectopic expression of AtEMK caused the formation of trichomes on cotyledons, dedifferentiated several tissues into calli, and retarded root development, demonstrating that AtEMK is harmful for the normal development of plants after germination. From these results, we conclude that the AtEMK is a key player to maintain embryonic identity, and the rapid disappearance of AtEMK expression during germination is essential for the developmental transition between the embryonic and vegetative phases in plants.

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Acknowledgments

We thank Prof. Yasuo Niwa for providing the pTH2 vector. This work was supported by a Grant-in-Aid for Scientific Research (KAKENHI) from the Ministry of Education, Science, Culture, Sports and Technology, Japan.

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  1. Ryo Tsuwamoto

    Present address: Faculty of Agriculture and Life Science, Hirosaki University, Hirosaki, 036-8561, Japan

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  1. Laboratory of Plant Breeding, Faculty of Agriculture, Iwate University, 3-18-8 Ueda, Morioka, 020-8550, Japan

    Ryo Tsuwamoto, Shuji Yokoi & Yoshihito Takahata

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  1. Ryo Tsuwamoto
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Correspondence to Yoshihito Takahata.

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11103_2010_9634_MOESM2_ESM.tif

Fig. S1 emk bbm double mutant analysis. a Insertional location of T-DNA. The first intron of AtEMK and last exon of BBM are interrupted by inserted T-DNA in their corresponding mutant. b Confirmation of transcriptional knockout. In emk, bbm and emk bbm mutant, transcripts for AtEMK and BBM were detected by RT-PCR using cDNA from mature seeds. As a result, knockout of corresponding gene(s) in each plant line was attested. c Developing embryos in each line. Developing embryos at 16-cell, globular, triangular, heart, torpedo and cotyledonary stage were cleared with Hoyer’s solution and observed using differential interference contrast microscopy. d Root tip of each line. Root of seedlings grown under sterile condition at 14 days after stratification was stained by Lugol’s solution, cleared with Hoyer’s solution and observed. No distinct defect was found. Bars = 50 μm. (TIFF 1,146 kb)

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Tsuwamoto, R., Yokoi, S. & Takahata, Y. Arabidopsis EMBRYOMAKER encoding an AP2 domain transcription factor plays a key role in developmental change from vegetative to embryonic phase. Plant Mol Biol 73, 481–492 (2010). https://doi.org/10.1007/s11103-010-9634-3

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