Genome of Bacillus sp. strain QHF158 provides insights into its parasporal inclusions encoded by the S-layer gene
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ENVIRONMENTAL MICROBIOLOGY - RESEARCH PAPER
Genome of Bacillus sp. strain QHF158 provides insights into its parasporal inclusions encoded by the S-layer gene Yinghong Jin 1 & Junhui Wang 1 & Qian Liu 1 & Haifang Qiu 1 & Yangyang Wang 1 & Jiayi Zhou 1 & Jie Sun 1 & Peng Li 1 & Ruiping Wang 1 & Wenfei Zhang 1 Received: 31 July 2019 / Accepted: 1 December 2019 # Sociedade Brasileira de Microbiologia 2019
Abstract Bacillus sp. strain QHF158, a Gram-positive, spore-forming and parasporal crystal–secreting bacterium, was isolated from soil of Limushan National Forest Park in China. Here we present the significant feature of parasporal inclusions of this organism, together with the draft genome sequence and annotation. Phylogenetic analysis suggested that strain QHF158 is possibly a novel species, most closely related to Bacillus mycoides. Genome annotation results revealed that strain QHF158 did not contain any typical Cry or Cyt toxin coding gene. Furthermore, the mass spectrometry analyses demonstrated that the parasporal crystalline inclusions were encoded by the orf_05273 gene, with 95% similarity to the S-layer protein (SLP) EA1 of B. mycoides, which indicated that the parasporal crystal from Bacillus sp. strain QHF158 was mainly formed by SLP, instead of the typical Cry or Cyt toxin proteins. Keywords Genome . Parasporal crystal . S-layer protein . Mass spectrometry
Introduction Bacillus thuringiensis (Bt), as a microbial insecticide or biopesticide, has been applied widely and successfully around the world to reduce the amount of chemical pesticides [1, 2]. Within the genus Bacillus, Bacillus thuringiensis is very similar to other species such as Bacillus cereus, Bacillus anthracis, and Bacillus wiedmannii [3, 4]. Bt is generally known as the parasporal crystal producer, which synthesizes and secretes Cry or Cyt proteins during the sporulation phase. The common method of screen Bt isolates is to get the observation of parasporal crystals by microscope. However, not all the parasporal crystalline inclusions are consisted of Cry or Cyt toxins; some are consisted of surface layer proteins (S-
layer protein, abbreviated as SLP) [5, 6]. During the vegetative growth phase, Bt also synthesizes insecticidal proteins such as vegetative insecticidal proteins (Vip) and secreted insecticidal protein (Sip) with insecticidal activity against lepidopteran and coleopteran pests, which have no primary sequence homology to the Cry and Cyt toxins [7–9]. In this research, a strain QHF158 was screened and isolated with observation of parasporal crystals by scanning electron microscope (SEM). To further gain the insights into parasporal crystal inclusions of strain QHF158, the draft genome was sequenced using the Illumina HiSeq PE1500 platform. Interestingly, genome annotation and mass spectrometry (MS) analysis indicated that the parasporal crystals secreted by strain QHF158 were confirmed as SLP, instead of Cry or Cyt toxins.
Responsible Editor: Rodrigo Galhardo. Electronic supplementary material The online version of this artic
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