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Slr2019,脂多糖A转运蛋白同源物,对集胞藻PCC6803的耐酸性至关重要。

Slr2019, lipid A transporter homolog, is essential for acidic tolerance in Synechocystis sp. PCC6803.

作者信息

Matsuhashi Ayumi, Tahara Hiroko, Ito Yutaro, Uchiyama Junji, Ogawa Satoru, Ohta Hisataka

机构信息

Graduate School of Mathematics and Science Education, Tokyo University of Science, Shinjuku-ku, Tokyo, 162-8601, Japan,

出版信息

Photosynth Res. 2015 Aug;125(1-2):267-77. doi: 10.1007/s11120-015-0129-6. Epub 2015 Mar 31.

Abstract

Living organisms must defend themselves against various environmental stresses. Extracellular polysaccharide-producing cells exhibit enhanced tolerance toward adverse environmental stress. In Synechocystis sp. PCC6803 (Synechocystis), lipopolysaccharide (LPS) may play a role in this protection. To examine the relationship between stress tolerance of Synechocystis and LPS, we focused on Slr2019 because Slr2019 is homologous to MsbA in Escherichia coli, which is related to LPS synthesis. First, to obtain a defective mutant of LPS, we constructed the slr2019 insertion mutant (slr2019) strain. Sodium deoxycholate-polyacrylamide gel electrophoresis indicated that slr2019 strain did not synthesize normal LPS. Second, to clarify the participation of LPS in acid tolerance, wild type (WT) and slr2019 strain were grown under acid stress; slr2019 strain growth was significantly weaker than WT growth. Third, to examine influences on stress tolerance, slr2019 strain was grown under various stresses. Under salinity and temperature stress, slr2019 strain grew significantly slower than WT. To confirm cell morphology, cell shape and envelope of slr2019 strain were observed by transmission electron microscopy; slr2019 cells contained more electron-transparent bodies than WT cells. Finally, to confirm whether electron-transparent bodies are poly-3-hydroxybutyrate (PHB), slr2019 strain was stained with Nile Blue A, a PHB detector, and observed by fluorescence microscopy. The PHB granule content ratio of WT and slr2019 strain grown at BG-11 pH 8.0 was each 7.18 and 8.41 %. At pH 6.0, the PHB granule content ratio of WT and slr2019 strain was 2.99 and 2.60 %. However, the PHB granule content ratio of WT and slr2019 strain grown at BG-11N-reduced was 10.82 and 0.56 %. Because slr2019 strain significantly decreased PHB under BG-11N-reduced compared with WT, LPS synthesis may be related to PHB under particular conditions. These results indicated that Slr2019 is necessary for Synechocystis survival in various stresses.

摘要

生物必须保护自己免受各种环境压力的影响。产生胞外多糖的细胞对不利环境压力表现出更强的耐受性。在集胞藻属PCC6803(集胞藻)中,脂多糖(LPS)可能在这种保护作用中发挥作用。为了研究集胞藻的应激耐受性与LPS之间的关系,我们重点研究了Slr2019,因为Slr2019与大肠杆菌中的MsbA同源,而MsbA与LPS合成有关。首先,为了获得LPS缺陷突变体,我们构建了slr2019插入突变体(slr2019)菌株。脱氧胆酸钠-聚丙烯酰胺凝胶电泳表明,slr2019菌株不能合成正常的LPS。其次,为了阐明LPS在耐酸性中的作用,将野生型(WT)和slr2019菌株在酸性胁迫下培养;slr2019菌株的生长明显弱于WT菌株。第三,为了研究对胁迫耐受性的影响,将slr2019菌株在各种胁迫下培养。在盐度和温度胁迫下,slr2019菌株的生长明显比WT菌株慢。为了确认细胞形态,通过透射电子显微镜观察了slr2019菌株的细胞形状和包膜;slr2019细胞比WT细胞含有更多的电子透明体。最后,为了确认电子透明体是否是聚-3-羟基丁酸酯(PHB),用PHB检测器尼罗蓝A对slr2019菌株进行染色,并通过荧光显微镜观察。在BG-11 pH 8.0条件下生长的WT和slr2019菌株中PHB颗粒含量比分别为7.18%和8.41%。在pH 6.0时,WT和slr2019菌株的PHB颗粒含量比分别为2.99%和2.60%。然而,在BG-11N还原条件下生长的WT和slr2019菌株中PHB颗粒含量比分别为10.82%和0.56%。因为与WT相比,slr2019菌株在BG-11N还原条件下PHB显著减少,所以在特定条件下LPS合成可能与PHB有关。这些结果表明,Slr2019是集胞藻在各种胁迫下生存所必需的。

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