A systematic analysis of the role of GGDEF-EAL domain proteins in virulence and motility in Xanthomonas oryzae pv. oryzicola

The second messenger c-di-GMP is implicated in regulation of various aspects of the lifestyles and virulence of Gram-negative bacteria. Cyclic di-GMP is formed by diguanylate cyclases with a GGDEF domain and degraded by phosphodiesterases with either an EAL or HD-GYP domain. Proteins with tandem GGDEF-EAL domains occur in many bacteria, where they may be involved in c-di-GMP turnover or act as enzymatically-inactive c-di-GMP effectors. Here, we report a systematic study of the regulatory action of the eleven GGDEF-EAL proteins in Xanthomonas oryzae pv. oryzicola, an important rice pathogen causing bacterial leaf streak. Mutational analysis revealed that XOC_2335 and XOC_2393 positively regulate bacterial swimming motility, while XOC_2102, XOC_2393 and XOC_4190 negatively control sliding motility. The ΔXOC_2335/XOC_2393 mutant that had a higher intracellular c-di-GMP level than the wild type and the ΔXOC_4190 mutant exhibited reduced virulence to rice after pressure inoculation. In vitro purified XOC_4190 and XOC_2102 have little or no diguanylate cyclase or phosphodiesterase activity, which is consistent with unaltered c-di-GMP concentration in ΔXOC_4190. Nevertheless, both proteins can bind to c-di-GMP with high affinity, indicating a potential role as c-di-GMP effectors. Overall our findings advance understanding of c-di-GMP signaling and its links to virulence in an important rice pathogen.

Analysis of the failure of cracked biscuits

Cracks or checks in biscuits weaken the material and cause the product to break at low load levels that are perceived as injurious to product quality. In this work, the structural response of circular digestive biscuits, with diameter 72 mm and thickness 7.2 mm, simply supported around the circumference and loaded by a central concentrated force was investigated by experiment and theory. Tests were conducted to quantify the distribution in breakage strength for structurally sound biscuits, biscuits with natural checks and biscuits with a single known part-through crack. For sound biscuits the breakage force is Normally distributed with a mean of 12.5 N and standard deviation of 1.2 N. For biscuits with checks, the corresponding statistics are 9.6 N ± 2.62 N respectively. The presence of a crack weakens the biscuit and strength, as measured by breakage force falls almost linearly with crack length and crack depth. The orientation of the crack, whether radial or tangential, and its location (i.e. position of the crack mid-point on the biscuit surface) are also important. Deep, radial, cracks located close to the biscuit centre can reduce the strength by up to 50%. Two separate failure criteria were examined for sound and cracked biscuits respectively. The results from these tests were in good accord with theory. For a biscuit without defects, breakage occurred when maximum biscuit stress reached or exceeded the failure stress of 420 kPa. For a biscuit with cracks, breakage occurred as above or alternatively when its critical stress intensity factor of 18 kPam0.5 was reached.