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Expression analysis of two rice pollen-specific promoters using homologous and heterologous systems

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Abstract

Tissue-specific promoters are a very useful tool for manipulating gene expression in a target tissue or organ; however, their range of applications in other plant species has not been determined, to date. In this study, we identified two late pollen-specific rice promoters (ProOsLPS10 and ProOsLPS11) via meta-anatomical expression analysis. We then investigated the expression of both promoters in transgenic rice (a homologous system) and Arabidopsis (a heterologous system) using ProOsLPS10 or ProOsLPS11::GFP-GUS constructs. As predicted by microarray data, both promoters triggered strong GUS expression during the late stages of pollen development in rice, with no GUS signals detected in the examined microspores and sporophytic tissues. Interestingly, these promoters exhibited different GUS expression patterns in Arabidopsis. While in Arabidopsis, the OsLPS10 promoter conferred GUS expression at the uni- and bi-cellular pollen stages, as well as at the shoot apical region during the seedling stage, the OsLPS11 promoter was not active in the pollen at any stage, or in the examined sporophytic tissues. Furthermore, by performing a complementation analysis using a sidecar pollen (scp) mutant that displays developmental defects at the microspore stage, we found evidence that OsLPS10 promoter, which can be applied in Arabidopsis, is useful for directing gene expression in the early stages of pollen development. Our results indicate that the OsLPS10 and OsLPS11 promoters can drive the expression of target genes during the late stages of pollen development in rice, but not in Arabidopsis. Our results also emphasize the necessity of confirming the applicability of an established promoter to heterologous systems.

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Acknowledgments

This research was supported by the Next-Generation BioGreen21 Program, of the Rural Development Administration in the Republic of Korea (The National Center for GM Crops, Grant No. PJ01128401; Plant Molecular Breeding Center No. PJ008137), and the Basic Science Research Program through the National Research Foundation of Korea (NRF), funded by the Ministry of Education, Science and Technology (NRF-2013R1A1A2006040 to KHJ, and NRF-2014R1A1A2005312 to SAO).

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  1. Hyun-Kyung Bae

    Present address: Research and Development Division, Gimcheon Agricultural Technology Center, Gimchen, 740-924, Republic of Korea

Authors and Affiliations

  1. School of Applied Biosciences, Kyungpook National University, Daegu, 702–701, Republic of Korea

    Tien Dung Nguyen, Moe Moe Oo, Hyun-Kyung Bae, Sung Aeong Oh, Jong Tae Song & Soon Ki Park

  2. Department of Molecular Biology, Sejong University, Seoul, 143-747, Republic of Korea

    Moon-Soo Soh

  3. Department of Biology, Kyungpook National University, Daegu, 702-701, Republic of Korea

    Jeong Hoe Kim

  4. Graduate School of Biotechnology and Crop Biotech Institute, Kyung Hee University, Yongin, 446-701, Republic of Korea

    Sunok Moon & Ki Hong Jung

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T. D. Nguyen and M. M. Oo contributed equally to this work.

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11816_2015_365_MOESM3_ESM.jpg

Supplemental Figure 3. Complementation analysis of scp-2 homozygotes (a-j). Pollen phenotypes in scp-2hm. a- normal tricellular pollen; b- uninucleate pollen; c, d- binucleate pollen; e- larger tricellular pollen; f-i- four-nucleate pollen with (g-i) or without (f) the extracellular compartment marked with an arrowhead; j- aborted pollen. Pollen images were all taken at the same magnification. (k-m) Confirmation of T-DNA insertion in the ProOsLPS10-SCP:dHA lines. (n) 1,316 bp SCP-dHA fragment was amplified from the DNA of fourteen transgenic lines (lane 1-14) using gene-specific primers (SCP forward and reverse). (o) scp-2 allele was detected using the LBb1 and LOBRP primer set. (p) SCP wild-type gene was checked in scp-2 hm background and transgenic lines using the LBb1 and 47870-R primers. scp- 2 hm and Col were used as the control. (JPEG 278 kb)

Supplemental Figure 4. Sequence aligment of OsLPS10 and OsLPS11. (JPEG 594 kb)

Supplementary material 5 (DOCX 14 kb)

Supplementary material 6 (DOCX 14 kb)

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Nguyen, T.D., Oo, M.M., Moon, S. et al. Expression analysis of two rice pollen-specific promoters using homologous and heterologous systems. Plant Biotechnol Rep 9, 297–306 (2015). https://doi.org/10.1007/s11816-015-0365-4

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