Welcome to Chinese Journal of Tropical Crops,

Chinese Journal of Tropical Crops ›› 2021, Vol. 42 ›› Issue (10): 2819-2827.DOI: 10.3969/j.issn.1000-2561.2021.10.010

• Omics & Biotechnology • Previous Articles     Next Articles

Cloning of Phosphorus-soluble Gene GDH and pqqE from Sugarcane Endogenous Nitrogen-fixing Bacteria Klebsiella variicola DX120E

CHEN Jiongyu1, QIN Ying1, XIE Xianqiu1, HUANG Yuyan1, DONG Dengfeng1, XING Yongxiu1,*(), LI Yangrui1,2,*()   

  1. 1. College of Agriculture, Guangxi University, Nanning, Guangxi 530004, China
    2. Guangxi Key Laboratory of Sugarcane Genetic Improvement / Key Laboratory of Sugarcane Biotechnology and Genetic Improvement (Guangxi), Ministry of Agriculture and Rural Affairs / Sugarcane Research Center, Chinese Academy of Agricultural Sciences / Sugarcane Research Institute, Guangxi Academy of Agricultural Sciences, Nanning, Guangxi 530007, China
  • Received:2020-12-28 Revised:2021-03-08 Online:2021-10-25 Published:2021-11-25
  • Contact: XING Yongxiu,LI Yangrui

Abstract:

Glucose dehydrogenase (GDH) and pyrroloquinoline quinone (PQQE) play an important role in the dissolution of inorganic phosphorus by phosphorus-soluble microorganisms. The phosphorus-soluble mechanism of the nitrogen-fixing bacteria Klebsiella variicola DX120E in sugarcane was investigated. The ORF of the phosphorous-soluble gene GDH and pqqE were cloned from the bacterium and analysised by bioinformatics. At the same time, the utilization ability of the bacteria to different phosphorus sources was studied. The ORF of GDH and pqqE was 2373 bp and 1143 bp, the coding amino acids were 790 and 380, respectively. Bioinformatics analysis showed that GDH was a stable protein, but pqqE was an unstable one. GDH was a membrane receptor protein related to cell signaling, with PQQ_membr_ DH, PQQ_mGDH functional domain and PQQ_DH_like superfamily protein structure. pqqE encoded proteins called intracellular proteins, structure containing PQQ_syn_pqqE functional domain and Radical SAM superfamily proteins. The utilization of different phosphorus sources and the qRT-PCR analysis of GDH and pqqE in different phosphorus sources showed that under different conditions, the ability of the bacteria to dissolve phosphorus was FePO4>AlPO4> Ca3(PO4)2 and the ability to dissolve FePO4 was significantly different from that of Ca3(PO4)2 and AlPO4(P<0.05). Under the condition of several phosphorus sources, the relative expression of GDH and pqqE increased, and the expression change trend of the genes was the same. The expression of GDH and pqqE in phosphorus source was significantly different from that in Ca3(PO4)2(P<0.05). This study would lay a foundation for further study on the interaction and phosphorus dissolution mechanism between endophytic nitrogen-fixing bacteria and sugarcane.

Key words: sugarcane, Klebsiella variicola DX120E, GDH, pqqE, phosphorus-soluble characteristics

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