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Silencing of meiosis-critical genes for engineering male sterility in plants

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Abstract

The potential for pollen-mediated transgene flow into wild or closely related species has provoked unease in terms of transgenic modification of agricultural plant species. One approach to remedy this situation in species whose seeds and fruits are not of particular value is to engineer male sterility into the transgenic lines. In this study, three meiosis-critical genes, namely AHP2, AtRAD51C and SWITCH1 (SWI), were chosen as silencing targets to test the feasibility of incorporating sterility into plants using an RNAi-based approach. Our results indicated that the silencing of each of these genes via hairpin RNA (termed AHPi, RAD51Ci and SWIi lines) in Arabidopsis thaliana yielded a proportion of transgenic plants exhibiting a similar ‘partially sterile’ phenotype in which less than 50% of pollen was viable. In addition, a ‘sterile’ phenotype was also evident in a minority of RAD51Ci and SWIi, but not AHPi, lines in which plants yielded no seeds and either produced inviable pollen (RAD51Ci lines) or displayed a complete absence of pollen (SWIi lines). This suggests that AtRAD51C and SWI may function at distinct stages of meiosis. Further analyses of SWIi lines demonstrated that the ‘sterile’ phenotype was associated with a substantial reduction in the level of targeted gene transcript in floral tissues and resulted from sterility of the male, but not female gametes. This work demonstrates that generating male sterility through the silencing of key genes involved in the regulation of meiosis is feasible, and its advantages and potential applications for transgene containment are discussed.

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Abbreviations

AHP2:

Histidine-containing phosphotransmitter 2

AHPi:

AHP2 silencing construct/transgenic line

DT-A:

Diptheria toxin A

RAD51Ci:

AtRAD51C silencing construct/transgenic line

SWI:

SWITCH1

SWIi:

SWI silencing construct/transgenic line

SWIi-F:

SWI silencing line bearing a fertile phenotype

SWIi-S:

SWI silencing line bearing a ‘sterile’ phenotype

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Acknowledgments

We would like to thank Mr. Dennis Bennett for his excellent technical assistance. This study was funded by a USDA Cooperative State Research, Education, and Extension Service Biotechnology Risk Assessment Research grant (2007-03054).

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Author notes

  1. Xiping Wang

    Present address: College of Horticulture, Northwest A&F University, Yangling, Shaanxi, 712100, People’s Republic of China

  2. Stacy D. Singer

    Present address: Department of Plant Pathology and Plant-Microbe Biology, New York State Agricultural Experiment Station, Cornell University, Geneva, NY, 14456, USA

Authors and Affiliations

  1. USDA-ARS Appalachian Fruit Research Station, 2217 Wiltshire Road, Kearneysville, WV, 25430, USA

    Xiping Wang, Stacy D. Singer & Zongrang Liu

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  1. Xiping Wang
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  2. Stacy D. Singer
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Correspondence to Zongrang Liu.

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Communicated by R. Schmidt.

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Wang, X., Singer, S.D. & Liu, Z. Silencing of meiosis-critical genes for engineering male sterility in plants. Plant Cell Rep 31, 747–756 (2012). https://doi.org/10.1007/s00299-011-1193-9

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  • DOI: https://doi.org/10.1007/s00299-011-1193-9

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