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Published on November 26, 2008; 10.1104/pp.108.128728

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Received August 29, 2008
Accepted November 15, 2008

GS52 ecto-apyrase plays a critical role during soybean nodulation

Manjula Govindarajulu , Sung-Yong Kim , Marc Libault , R. Howard Berg , Kiwamu Tanaka , Gary Stacey , and Christopher G. Taylor *

Donald Danforth Plant Science Center, 975 North Warson Road, St. Louis, Missouri 63132; Division of Plant Sciences, National Center for Soybean Biotechnology, C.S. Bond Life Sciences Center, University of Missouri-Columbia, Columbia, Missouri 65211; Division of Biochemistry and Department of Molecular Microbiology and Immunology, C.S. Bond Life Sciences Center, University of Missouri-Columbia, Columbia, Missouri 65211

Apyrases are nonenergy-coupled NTPases that hydrolyze nucleoside tri- (NTP) and di-phosphates (NDP) to nucleoside mono-phosphates (NMP) and orthophosphate(s). GS52, a soybean ecto-apyrase, was previously shown to be induced very early in response to inoculation with the symbiotic bacterium Bradyrhizobium japonicum. Over-expression of the GS52 ecto-apyrase in Lotus japonicus increased the level of rhizobial infection and enhanced nodulation. These data suggest a critical role for the GS52 ecto-apyrase during nodulation. To further investigate the role of GS52 during nodulation, we used RNA interference (RNAi) to silence GS52 expression in soybean roots using Agrobacterium rhizogenes-mediated root transformation. Transcript levels of GS52 were significantly reduced in GS52 silenced roots and exhibited reduced numbers of mature nodules. Development of the nodule primordium and subsequent nodule maturation were significantly suppressed in GS52 silenced roots. Transmission electron micrographs of GS52 silenced root nodules showed early senescence and infected cortical cells were devoid of symbiosome containing bacteroids. Application of exogenous adenosine di-phosphate (ADP) to silenced GS52 roots restored nodule development. Restored nodules contained bacteroids, thus indicating that extracellular ADP is important during nodulation. These results clearly suggest that GS52 ecto-apyrase catalytic activity is critical for the early rhizobium infection process, initiation of nodule primordium development and subsequent nodule organogenesis in soybean.




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