Gluconacetobacter diazotrophicus:: a natural endophytic diazotroph of Nile Delta sugarcane capable of establishing an endophytic association with wheat

Gluconacetobacter- like diazotrophs were encountered as dense populations inside the root and stem tissues of sugarcane cultivated in ancient agricultural fields of the Nile Delta. Counts of >10(5) cells g(-1) were recorded in root and stem samples. The leaves contained a smaller population (<10(3) g(-1)). The typical dark-orange colonies which developed on LGIP agar plates were purified. Identification was performed with the API microtube systems: API 20E for Enterobacteriaceae and API 20NE for non-Enterobacteriaceae. API profiles of the local isolates were closely related to those of the type culture Gluconacetobacter diazotrophicus (ATTC 49037). The isolates successfully reduced C2H2 and produced appreciable amounts of ethylene in the presence of cane juice. This suggested that the local isolates are closely related to the type strain G. diazotrophicus. Wheat seedlings were inoculated with a number of isolates under gnotobiotic conditions. Both optical and scanning electron microscopy showed that endophytic Gluconacetobacter spp. were present in all the samples tested. They were observed in apparently intact and enlarged epidermal root cells, and also in stem tissues, indicating that the bacterium was able to migrate upward into the shoot tissues. Although Gluconacetobacter inoculation did not stimulate the growth of the cereal plant, the results obtained are particularly interesting because this bacterial species was capable of colonizing the internal tissues of wheat, not considered a natural host until now.