Guo Dao-Jun, Li Dong-Ping, Singh Rajesh Kumar, Singh Pratiksha, Sharma Anjney, Verma Krishan K, Qin Ying, Khan Qaisar, Lu Zhen, Malviya Mukesh K, Song Xiu-Peng, Xing Yong-Xiu, Li Yang-Rui
College of Agriculture, Guangxi University, Nanning, China.
Key Laboratory of Sugarcane Biotechnology and Genetic Improvement, Ministry of Agriculture and Rural Affairs, Sugarcane Research Center, Chinese Academy of Agricultural Sciences, Guangxi Key Laboratory of Sugarcane Genetic Improvement, Sugarcane Research Institute, Guangxi Academy of Agricultural Sciences, Nanning, China.
Front Plant Sci. 2021 Nov 23;12:727741. doi: 10.3389/fpls.2021.727741. eCollection 2021.
Plant endophytic bacteria have many vital roles in plant growth promotion (PGP), such as nitrogen (N) fixation and resistance to biotic and abiotic stresses. In this study, the seedlings of sugarcane varieties B8 (requires a low concentration of nitrogen for growth) and GT11 (requires a high concentration of nitrogen for growth) were inoculated with endophytic diazotroph ED5, which exhibits multiple PGP traits, isolated from sugarcane roots. The results showed that the inoculation with ED5 promoted the growth of plant significantly in both sugarcane varieties. N detection at 60 days post-inoculation proved that the inoculation with strain ED5 increased the total nitrogen concentration in the leaf and root than control in both sugarcane varieties, which was higher in B8. Biochemical parameters and phytohormones in leaf were analyzed at 30 and 60 days after the inoculation. The results showed that the inoculation with ED5 improved the activities of superoxide dismutase (SOD), catalase (CAT), NADH-glutamate dehydrogenase (NADH-GDH), glutamine synthetase (GS), and endo-β-1,4-glucanase, and the contents of proline and indole acetic acid (IAA) in leaf, and it was generally more significant in B8 than in GT11. Tandem Mass Tags (TMT) labeling and liquid chromatography-tandem mass spectrometry (LC-MS/MS) were used to perform comparative proteomic analysis in the sugarcane leaves at 30 days after inoculation with strain ED5. A total of 27,508 proteins were detected, and 378 differentially expressed proteins (DEPs) were found in the treated sugarcane variety B8 (BE) as compared to control (BC), of which 244 were upregulated and 134 were downregulated. In contrast, a total of 177 DEPs were identified in the treated sugarcane variety GT11 (GE) as compared to control (GC), of which 103 were upregulated and 74 were downregulated. The DEPs were associated with nitrogen metabolism, photosynthesis, starch, sucrose metabolism, response to oxidative stress, hydrolase activity, oxidative phosphorylation, glutathione metabolism, phenylpropanoid metabolic process, and response to stresses in Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) database. To the best of our knowledge, this is the first proteomic approach to investigate the molecular basis of the interaction between N-fixing endophytic strain ED5 and sugarcane.
植物内生细菌在促进植物生长（PGP）方面具有许多重要作用，例如固氮以及抵抗生物和非生物胁迫。在本研究中，对甘蔗品种B8（生长需要低浓度氮）和GT11（生长需要高浓度氮）的幼苗接种了从甘蔗根部分离出的具有多种PGP特性的内生固氮菌ED5。结果表明，接种ED5显著促进了两个甘蔗品种的植株生长。接种后60天的氮检测证明，接种菌株ED5后，两个甘蔗品种叶片和根部的总氮浓度均高于对照，其中B8中的总氮浓度更高。在接种后30天和60天分析了叶片中的生化参数和植物激素。结果表明，接种ED5提高了超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、NADH-谷氨酸脱氢酶（NADH-GDH）、谷氨酰胺合成酶（GS）和内切β-1,4-葡聚糖酶的活性，以及叶片中脯氨酸和吲哚乙酸（IAA）的含量，并且在B8中通常比在GT11中更显著。采用串联质谱标签（TMT）标记和液相色谱-串联质谱（LC-MS/MS）对接种菌株ED5后30天的甘蔗叶片进行了比较蛋白质组学分析。共检测到27508种蛋白质，与对照（BC）相比，在处理后的甘蔗品种B8（BE）中发现了378种差异表达蛋白（DEP），其中244种上调，134种下调。相比之下，与对照（GC）相比，在处理后的甘蔗品种GT11（GE）中鉴定出177种DEP，其中103种上调，74种下调。这些DEP在基因本体论（GO）和京都基因与基因组百科全书（KEGG）数据库中与氮代谢、光合作用、淀粉、蔗糖代谢、氧化应激反应、水解酶活性、氧化磷酸化作用、谷胱甘肽代谢、苯丙烷代谢过程以及胁迫反应相关。据我们所知，这是首次采用蛋白质组学方法研究固氮内生菌株ED5与甘蔗相互作用的分子基础。
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