Abstract

YxaL is conserved within the Bacillus subtilis species complex associated with plants and soil. The mature YxaL protein contains a repeated beta-propeller domain, but the subcellular location and function of YxaL has not been determined. The gene encoding the mature YxaL protein was PCR amplified from genomic DNA of B. velezensis strain GH1-13 and used for recombinant protein production. A rabbit polyclonal antibody against the purified YxaL was generated and used for western blotting to determine the constitutive expression and secretion of YxaL. During normal culture growth of strain GH1-13, levels of the constitutively secreted YxaL were slowly rising to 100 μg L-1, and degraded with a half-life of 1.6 h in the culture medium. When the effects of YxaL on plant seed germination and seedling growth were examined, it was shown that seed treatment of Arabidopsis thaliana and rice (Oryza sativa L.) with purified YxaL at the optimal concentration of 1 mg L-1 was effective at improving the root growth of plants. Seedlings from the treated Arabidopsis seeds markedly increased transcription of a 1-aminocyclopropane-1-carboxylate synthetase marker gene (ACS11) but reduced expression of auxin- and abscisic acid-responsive marker genes (IAA1, GH3.3, and ABF4), especially when provided with exogenous auxin. Horticulture experiments showed that pepper (Capsicum annuum) seeds treated with 1 mg L-1 YxaL in a soaking solution increased shoot growth and improved tolerance to drought stress. We hypothesize that YxaL secreted from plant growth-promoting Bacillus cells has a significant impact on plant roots, with the potential to improve plant growth and stress tolerance.

Highlights

  • The protein YxaL has been observed to interact with the DNA helicase PcrA in Bacillus subtilis [1]

  • Evolutionary relationships of YxaL with beta-propeller domains in bacteria A database search for YxaL homologs in bacteria showed that the amino acid sequence of YxaL obtained from B. velezensis strain GH1-13 is highly conserved in Bacillus species

  • The YxaL homologs are diverged into two types, tentatively named YxaL1 and YxaL2, which are present in distinct operational taxonomic groups of B. amyloliquefaciens-siamensis-velezensis and B. halotolerans-nakamurai-tequilensis (Fig 1 and S1 Fig)

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Summary

Introduction

The protein YxaL has been observed to interact with the DNA helicase PcrA in Bacillus subtilis [1]. Based on the sequence homology and structural similarity, it is postulated that beta-propeller homologs with different structure architectures of the blades may have originated from one ancestral blade, most likely that of a PQQ motif beta-propeller [4]. This betapropeller domain is ubiquitous in diverse proteins with a similar beta-propeller fold observed in methanol dehydrogenase, which uses PQQ as cofactor [5]. YxaL homologs in Bacillus species have no conserved cysteine residue, which suggest that their functions differ from that of PQQ-containing enzymes

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