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Infection and Colonization of Rice Seedlings by the Plant Growth-Promoting Bacterium Herbaspirillum seropedicae Z67

¹International Rice Research Institute, DAPO Box 7777, Metro Manila, Philippines; ²Institute of Biology, University of the Philippines, Diliman, Quezon City 1101, Philippines; ³Genes to Products, Quality, Health and Nutrition, Scottish Crops Research Institute, Invergowrie, Dundee DD2 5DA, U.K.; ⁴Setor de Citologia Vegetal, Lab. Biologia Celular e Tecidual, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense, Campos dos Goytacazes, RJ 28015-620, Brazil

A β-glucoronidase (GUS)-marked strain of Herbaspirillum seropedicae Z67 was inoculated onto rice seedling cvs. IR42 and IR72. Internal populations peaked at over 10⁶ log CFU per gram of fresh weight by 5 to 7 days after inoculation (DAI) but declined to 10³ to 10⁴ log CFU per gram of fresh weight by 28 DAI. GUS staining was most intense on coleoptiles, lateral roots, and at the junctions of some of the main and lateral roots. Bacteria entered the roots via cracks at the points of lateral root emergence, with cv. IR72 appearing to be more aggressively infected than cv. IR42. H. seropedicae subsequently colonized the root intercellular spaces, aerenchyma, and cortical cells, with a few penetrating the stele to enter the vascular tissue. Xylem vessels in leaves and stems were extensively colonized at 2 DAI but, in later harvests (7 and 13 DAI), a host defense reaction was often observed. Dense colonies of H. seropedicae with some bacteria expressing nitrogenase Fe-protein were seen within leaf and stem epidermal cells, intercellular spaces, and substomatal cavities up until 28 DAI. Epiphytic bacteria were also seen. Both varieties showed nitrogenase activity but only with added C, and the dry weights of the inoculated plants were significantly increased. Only cv. IR42 showed a significant (approximately 30%) increase in N content above that of the uninoculated controls, and it also incorporated a significant amount of ¹⁵N₂.

Additional keywords: nitrogen fixation , immunogold labeling , electron microscopy , endophytic diazotrophs.