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Characterization of a Plant Growth-Promoting Endohyphal Bacillus subtilis in Fusarium acuminatum from Spiranthes sinensis Cover

Characterization of a Plant Growth-Promoting Endohyphal Bacillus subtilis in Fusarium acuminatum from Spiranthes sinensis

Polish Journal of Microbiology
Volume 72 (2023): Issue 1 (March 2023)
By: LAN FANG,  XIAO ZHENG,  ZHENGXIANG SUN,  YANYAN LI,  JIANXIN DENG and  YI ZHOU  
Open Access
|Mar 2023

Figures & Tables

Fig. 1.

The endohyphal bacteria in Fusarium acuminatum YZU 172038. (A, B) Fluorescent SYTO-9 nucleic acid stained hyphae under fluorescent microscopy (FN – fungal nucleus, BD – bacterial DNA), scale bars = 5 μm; (C, D) bacteria (green signals pointed by white arrows) in hypha detected by the FISH method, scale bars = 5 μm; (E) PCR amplification of the 16S RNA gene (M – 2000 bp maker, 1 – YZU 172038); (F) colony morphology of EHB YZASR384 on LA medium; (G) scanning electron microscopy of EHB YZASR384; (H) PCR amplification of endoglucanase gene (1 – EHB YZSR384, 2 – YZU 172038).
The endohyphal bacteria in Fusarium acuminatum YZU 172038. (A, B) Fluorescent SYTO-9 nucleic acid stained hyphae under fluorescent microscopy (FN – fungal nucleus, BD – bacterial DNA), scale bars = 5 μm; (C, D) bacteria (green signals pointed by white arrows) in hypha detected by the FISH method, scale bars = 5 μm; (E) PCR amplification of the 16S RNA gene (M – 2000 bp maker, 1 – YZU 172038); (F) colony morphology of EHB YZASR384 on LA medium; (G) scanning electron microscopy of EHB YZASR384; (H) PCR amplification of endoglucanase gene (1 – EHB YZSR384, 2 – YZU 172038).

Fig. 2.

Phylogenetic tree of endohyphal strain YZSR384 using the neighbor-joining method based on the 16S rRNA gene sequences. Bootstrap values (%) presented at the branches were calculated from 1,000 replications.
Phylogenetic tree of endohyphal strain YZSR384 using the neighbor-joining method based on the 16S rRNA gene sequences. Bootstrap values (%) presented at the branches were calculated from 1,000 replications.

Fig. 3.

Plant growth promoting (PGP) assays for rice seeds using the culture broth of endophytic Fusarium acuminatum YZU 172038 (A, B) and its EHB Bacillus subtilis YZSR384 (C, D). The controls were PDB broth (A, B) and demineralized LB liquid (C, D).
Plant growth promoting (PGP) assays for rice seeds using the culture broth of endophytic Fusarium acuminatum YZU 172038 (A, B) and its EHB Bacillus subtilis YZSR384 (C, D). The controls were PDB broth (A, B) and demineralized LB liquid (C, D).

Fig. 4.

Detection of the IAA production in EHB YZSR384 culture broth. (A) Salkowski test of the negative control (left) and the EHB YZSR384 broth (right). (B) The IAA concentration in different incubation times.
Detection of the IAA production in EHB YZSR384 culture broth. (A) Salkowski test of the negative control (left) and the EHB YZSR384 broth (right). (B) The IAA concentration in different incubation times.

Genes associated with plant growth-promoting (PGP) traits for EHB Bacillus subtilis YZSR384_

PGP activityGeneFunctionChromosome location
IAA productiontrpSTryptophanyl-tRNA synthetase1218699–1217707–
trpATryptophan synthase subunit alpha2371911–2371108–
trpBTryptophan synthase subunit beta2373106–2371904–
trpFPhosphoribosylanthranilate isomerase2373734–2373087–
trpCIndole-3-glycerol phosphate synthase2374491–2373739–
trpDAnthranilate phosphoribosyltransferase2375500–2374484–
trpEAnthranilate synthase2377019–2375472–
Phosphate solubilizationphoDAlkaline phosphatase283609–285360+
phoBAlkaline phosphatase621083–619695–
phoAAlkaline phosphatase1018051–1016675–
phoHPhosphate starvation-inducible protein2615049–2614090–
PhoPAlkaline phosphatase synthesis transcriptional regulatory protein2978117–2977395–
phoRTwo-component sensor histidine kinase3236753–3238354+
pstBPhosphate ABC transporter ATP-binding protein2577585–2576803–2578405–2577596–
pstAPhosphate ABC transporter permease2579310–2578426–
pstCPhosphate ABC transporter permease2580239–2579310–
pstSPhosphate ABC transporter substrate-binding protein2581210–2580308–
Siderophore productionentBIsochorismatase552212–552754+3288208–3287270–
entCIsochorismate synthase3153313–3151898–3153500–3153375–3291891–3291106–
entE2,3-dihydroxybenzoate-AMP ligase3289855–3288236–
entA2,3-dihydro-2,3-dihydroxybenzoate dehydrogenase3291080–3289884–

Genome properties of EHB Bacillus subtilis YZSR384_

FeatureBacillus subtilis YZSR384
Genome size (bp)4,215,636
GC content (%)43.51
Total number of genes4444
Number of CDSs4216
tRNA genes87
rRNA genes30
tmRNA genes1
Genomic islands4
Genes allocated to Uniprot4,157
Genes allocated to Refseq4,180
Genes allocated to KEGG2,330
Genes allocated to Pfam3,761
Genes allocated to GO3,359
Genes allocated to COG2,816
DOI: https://doi.org/10.33073/pjm-2023-007 | Journal eISSN: 2544-4646 | Journal ISSN: 1733-1331
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Language: English
Page range: 29 - 37
Submitted on: Nov 10, 2022
Accepted on: Feb 3, 2023
Published on: Mar 24, 2023
Published by: Polish Society of Microbiologists
In partnership with: Paradigm Publishing Services
Publication frequency: 4 issues per year
Keywords:
endohyphal bacterium,
plant growth-promoting ability
Related subjects:
Life sciences,
Microbiology and virology

© 2023 LAN FANG, XIAO ZHENG, ZHENGXIANG SUN, YANYAN LI, JIANXIN DENG, YI ZHOU, published by Polish Society of Microbiologists
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

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