Instituto de Ciencia y Tecnología Dr. César Milstein, Fundación Pablo Cassará, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Saladillo 2468 (C1440FFX), Ciudad de Buenos Aires, Argentina.
Centro de Investigaciones y Transferencia (CIT Villa María), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET)-Instituto de Ciencias Básicas y Aplicadas, Universidad Nacional de Villa María. Av. Arturo Jauretche 1555, (5900), Villa María, Córdoba, Argentina.
Glycobiology. 2019 Mar 1;29(3):269-278. doi: 10.1093/glycob/cwy114.
Xanthan is a virulence factor produced by Xanthomonas spp. We previously demonstrated that this exopolysaccharide is not only essential for pathogenicity by contributing with bacterial survival but also its pyruvate substituents interfere with some plant defense responses. Deepening our studies about xanthan properties and structure, the aim of this work was to analyze the characteristics of xanthan produced by Xanthomonas in different culture media. We analyzed the xanthan produced by Xanthomonas citri subsp. citri (Xcc) in leaf extracts from grapefruit (a susceptible host of this bacterium) and compared it with the xanthan produced in a synthetic culture medium. We found that the xanthan produced in the grapefruit extract (Xan-GLE) presented shorter and more disordered molecules than xanthan produced in the synthetic medium (Xan-PYM). Besides, Xan-GLE resulted less viscous than Xan-PYM. The disordered molecular conformation of Xan-GLE could be attributed to its higher pyruvilation degree and lower acetylation degree compared with those detected in Xan-PYM. Meanwhile, the difference in the viscosity of both xanthans could be due to their molecules length. Finally, we cultured Xcc in the presence of the Xan-GLE or Xan-PYM and observed the formation of biofilm-like structures in both cases. We found significant differences in biofilm architecture between the two conditions, being the biofilm produced in presence of Xan-GLE similar to that formed in canker lesions developed in lemon plant leaves. Together, these results show how xanthan structure and properties changed when Xcc grew in a natural substrate and can contribute to better understand the biological role of xanthan.
黄原胶是黄单胞菌属产生的一种毒力因子。我们之前证明,这种胞外多糖不仅通过促进细菌存活对致病性至关重要,而且其丙酮酸取代基还会干扰一些植物防御反应。为了深入研究黄原胶的特性和结构,本工作旨在分析黄单胞菌在不同培养基中产生的黄原胶的特性。我们分析了柑橘溃疡病菌(柑橘溃疡病的易感宿主)在葡萄柚叶片提取物中产生的黄原胶,并将其与在合成培养基中产生的黄原胶进行了比较。结果发现,在葡萄柚提取物中产生的黄原胶(Xan-GLE)的分子比在合成培养基中产生的黄原胶(Xan-PYM)更短且更无序。此外,Xan-GLE 的粘性比 Xan-PYM 低。与 Xan-PYM 相比,Xan-GLE 的分子构象无序可能归因于其较高的丙酮酸化程度和较低的乙酰化程度。同时,两种黄原胶的粘度差异可能与其分子长度有关。最后,我们在存在 Xan-GLE 或 Xan-PYM 的情况下培养 Xcc,并观察到两种情况下都形成了类似生物膜的结构。我们发现两种条件下生物膜结构存在显著差异,在存在 Xan-GLE 的情况下形成的生物膜类似于在柠檬叶片溃疡病中形成的生物膜。综上所述,这些结果表明当 Xcc 在天然基质中生长时,黄原胶的结构和性质如何发生变化,并有助于更好地理解黄原胶的生物学作用。