Five novel halophilic archaeal strains, named BND22
T
, ZY10
T
, ZY41
T
, ZY58
T
, and ZY43
T
, were isolated from the coastal saline sediment of the intertidal zone located in Qingdao and the ...natural sea salt produced from Huanghua marine solar saltern, PRChina. These five strains demonstrated the typical morphology and growth characteristics of haloarchaea. The comparison of 16S rRNA gene revealed that strain BDN22
T
was associated with
Salinirubellus salinus
ZS-35-S2
T
(95.2% similarity), strain ZY10
T
was related to
Halosegnis rubeus
F17-44
T
(95.1% similarity), both strains ZY41
T
and ZY58
T
were closely related to
Haloglomus irregulare
F16-60
T
(98.1% and 98.2% similarities, respectively), and strain ZY43
T
was close to
Natronomonas salina
YPL13
T
(98.0% similarity). In addition to the 16S rRNA gene, the
rpoB′
gene is undoubtedly another important molecular marker for the identification of halophilic archaea, and concatenated-conserved-protein phylogeny was widely used in archaeal classification in recent years. Phylogenetic and phylogenomic analyses based on
rpoB′
genes and 122 concatenated archaeal protein genes showed that these haloarchaea construct three different clades and gathered together with the current members of
Haloarculaceae
. Strains BND22
T
and ZY10
T
formed two distinct clades separated from
Salinirubellus salinus
ZS-35-S2
T
and
Halosegnis
members, strains ZY41
T
and ZY58
T
clustered with
Haloglomus irregulare
F16-60
T
, and strain ZY43
T
gathered with the current members of
Natronomonas.
The ANI, DDH, and AAI values of these five strains against phylogenetic neighbours were no more than 91%, 45%, and 92%, respectively, far below the cut-off values for species delineation, supporting their placements in new taxa. Based on the phenotypic, chemotaxonomic, phylogenetic, and phylogenomic properties, these five strains represent five novel taxa of the family
Haloarculaceae
,
Halorarius litoreus
gen. nov., sp. nov. (type strain BND22
T
= CGMCC 1.18780
T
= JCM 34966
T
),
Halorarius halobius
sp. nov. (type strain ZY10
T
= CGMCC 1.17475
T
= JCM 34319
T
),
Haloglomus halophilum
sp. nov. (type strain ZY41
T
= CGMCC 1.17030
T
= JCM 34161
T
),
Haloglomus salinum
sp. nov. (type strain ZY58
T
= CGMCC 1.17216
T
= JCM 34163
T
), and
Natronomonas marina
sp. nov. (type strain ZY43
T
= CGMCC 1.17202
T
= JCM 34162
T
). This is the first report of description of a novel haloarchaeon isolated from a marine intertidal zone.
Allostimulated CD8
+
T cells (aCD8
+
T cells), as the main mediators of acute liver rejection (ARJ), are hyposensitive to apoptosis due to the inactivation of death receptor FAS-mediated pathways and ...fail to allow tolerance induction, eventually leading to acute graft rejection. Although tacrolimus (FK506), the most commonly used immunosuppressant (IS) in the clinic, allows tolerance induction, its use is limited because its target immune cells are unknown and it is associated with increased incidences of malignancy, infection, and nephrotoxicity, which substantially impact long-term liver transplantation (LTx) outcomes. The dark agouti (DA)-to-Lewis rat LTx model is a well-known ARJ model and was hence chosen for the present study. We show that both hepatocyte growth factor (HGF) (cHGF, containing the main form of promoting HGF production) and recombinant HGF (h-rHGF) exert immunoregulatory effects mainly on allogeneic aCD8
+
T cell suppression through FAS-mediated apoptotic pathways by inhibiting cMet to FAS antagonism and Fas trimerization, leading to acute tolerance induction. We also showed that such inhibition can be abrogated by treatment with neutralizing antibodies against cMet (HGF-only receptor). In contrast, we did not observe these effects in rats treated with FK506. However, we observed that the effect of anti-rejection by FK506 was mainly on allostimulated CD4
+
T cell (aCD4
+
T cell) suppression and regulatory T cell (Treg) promotion, in contrast to the mechanism of HGF. In addition, the protective mechanism of HGF in FK506-mediated nephrotoxicity was addressed. Therefore, HGF as a tolerance inducer, whether used in combination with FK506 or as monotherapy, may have good clinical value. Additional roles of these T-cell subpopulations in other biological systems and studies in these fields will also be meaningful.
Halophilic archaeal strain YGHS32T was isolated from the Yinggehai marine solar saltern near Shanya city of Hainan Province, China. Cells of the strain were pleomorphic and lysed in distilled water, ...stained Gram-negative and formed red-pigmented colonies. Strain YGHS32T was able to grow at 20-50 °C (optimum 37 °C), in the presence of 0.9-4.8 M NaCl (optimum 2.1 M NaCl), with 0.005-1.0 M MgCl2 (optimum 0.3 M MgCl2) and at pH 6.0-8.5 (optimum pH 7.5). The minimal NaCl concentration to prevent cell lysis was 5% (w/v). The major polar lipids of the strain were phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate and four major glycolipids chromatographically identical to sulfated mannosyl glucosyl diether, mannosyl glucosyl diether, glucosyl mannosyl glucosyl diether and a diglycosyl diether. Strain YGHS32T had two dissimilar 16S rRNA genes and both of them were phylogenetically related to those of Halomicroarcula pellucida JCM 17820T (92.9-96.3% sequence similarity). The rpoB' gene sequence similarity between strain YGHS32T and Halomicroarcula pellucida JCM 17820T was 91.3%. The DNA G+C content of strain YGHS32T was 64.0 mol%. The DNA-DNA hybridization value between strain YGHS32T and Halomicroarcula pellucida JCM 17820T was 45%. It was concluded that strain YGHS32T (=CGMCC 1.12129T=JCM 18640T) represents a novel species of the genus Halomicroarcula, for which the name Halomicroarcula limicola sp. nov. is proposed. An emended description of the genus Halomicroarcula is also presented.
Four halophilic archaeal strains, designated TNN18T, TBN12, TNN28T and TBN19, were isolated from brines sampled from two artificial marine solar salterns in eastern China. Strains TNN18T and TNN28T ...were isolated from the Tainan marine solar saltern, whereas strains TBN12 and TBN19 were from the Taibei marine solar saltern. Colonies of the four strains were red-pigmented and their cells were pleomorphic, motile, Gram-reaction-negative rods. Strains TNN18T and TBN12 were able to grow at 25–50 °C (optimum 37 °C), in 10–30 % (w/v) NaCl (optimum 15 %), with 0–1.0 M MgCl2 (optimum 0.05 M) and at pH 5.5–9.0 (optimum pH 7.0–7.5), while strains TNN28T and TBN19 were able to grow at 20–50 °C (optimum 37 °C), in 15-30 % (w/v) NaCl (optimum 18–20 %), in 0.005–1.0 M MgCl2 (optimum 0.01–0.3 M) and at pH 6.0–9.0 (optimum pH 7.0–7.5). Cells of these strains lyse in distilled water; minimal NaCl concentrations to prevent cell-lysis are 10 % (w/v) for strains TNN18T and TBN12 and 12 % (w/v) for strains TNN28T and TBN19. The major polar lipids of strains TNN18T and TBN12 were phosphatidylglycerol (PG), phosphatidylglycerol phosphate methyl ester (PGP-Me), phosphatidylglycerol sulfate (PGS) and one major glycolipid (GL1), which was chromatographically identical to sulfated mannosyl glucosyl diether (S-DGD-1). Minor amounts of other lipids (GL0, GL2, GL3 and GL4) were also detectable. The polar lipid profiles of strains TNN28T and TBN19 contained PG, PGP-Me, GL1, which was chromatographically identical to S-DGD-1, and three to four minor unidentified glycolipids (GL2–GL5). Phylogenetic analyses revealed that strains TNN18T and TBN12 formed a distinct clade with strains of the closest related species, Haloquadratum walsbyi (91.5–91.8 % 16S rRNA gene sequence similarity) and strains TNN28T and TBN19 formed a distinct clade with strains of the species Halosimplex carlsbadense (89.9–93.3 % similarity) and two members of the genus Halorhabdus (92.5–93.3 % similarity). The DNA G+C contents of strains TNN18T, TBN12, TNN28T and TBN19 were 61.5, 62.4, 61.9 and 61.5 mol%, respectively. DNA–DNA hybridization values between strains TNN18T and TBN12, and strains TNN28T and TBN19 were 82.9 % and 88.2 %, respectively. The phenotypic, chemotaxonomic and phylogenetic properties suggest that the four strains represent two novel species of two new genera within the family Halobacteriaceae, for which the names Halobellus clavatus gen. nov., sp. nov. (type strain TNN18T = CGMCC 1.10118T = JCM 16424T) and Halorientalis regularis gen. nov., sp. nov. (type strain TNN28T = CGMCC 1.10123T = JCM 16425T) are proposed.
Based on ITS sequences, the molecular identification of Cordyceps cicadae and Tolypocladium dujiaolongae was carried out, and high-performance liquid chromatography(HPLC) fingerprint combined with ...chemical pattern recognition method was established to differentiate C. cicadae from its adulterant T. dujiaolongae. The genomic DNA from 10 batches of C. cicadae and five batches of T. dujiaolongae was extracted, and ITS sequences were amplified by PCR and sequenced. The stable differential sites of these two species were compared and the phylogenetic tree was constructed via MEGA 7.0. HPLC was used to establish the fingerprints of C. cicadae and T. dujiaolongae, and similarity evaluation, cluster analysis(CA), principal component analysis(PCA), and partial least squares discriminant analysis(PLS-DA) were applied to investigate the chemical pattern recognition. The result showed that the sources of these two species were different, and there were 115 stable differential sites in ITS sequences of C. cicadae and T. d
Three halophilic archaeal strains, XD46
T
, YJ-63-S1 and ZS-1-H, were isolated from three Chinese marine solar salterns. All were observed to have pleomorphic cells that lysed in distilled water, ...stained Gram-negative and formed red-pigmented colonies. They were found to grow optimally at 37 °C, at pH 7.0 and in the presence of 2.6 M NaCl and 0.05 M Mg
2+
. The major polar lipids were identified as those typical for members of the
Halobacteriaceae
but also included major glycolipids chromatographically identical to sulfated mannosyl glucosyl diether (S-DGD-1), mannosyl glucosyl diether (DGD-1) and two unidentified ones. The 16S rRNA gene sequences of the three strains were 99.8–100 % identical, showing most similarity to sequences of members of the family
Halobacteriaceae
, and clustering together as a distinct clade in phylogenetic tree reconstructions. The
rpoB′
gene similarities between the three strains were 98.7–100 % and lower to the sequences of other halobacteria. Their DNA G+C contents were determined to be 65.1–65.5 mol%. The phenotypic, chemotaxonomic and phylogenetic properties suggest that strains XD46
T
(=CGMCC 1.12237
T
= JCM 18649
T
), YJ-63-S1 (=CGMCC 1.12574) and ZS-1-H (=CGMCC 1.12544) represent a novel species in a new genus within the family
Halobacteriaceae
, for which the name
Salinarubrum litoreum
gen. nov., sp. nov. is proposed.
A halophilic archaeal strain, SKJ47
T
, was isolated from a commercial preparation of the brown alga
Laminaria
produced at Dalian, Liaoning Province, China. Cells of the strain were observed to be ...short rods, stain Gram-negative, and to form red-pigmented colonies on solid media. Strain SKJ47
T
was found to be able to grow at 20–50 °C (optimum 37 °C), at 0.9–4.8 M NaCl (optimum 2.6–3.1 M), at pH 6.0–9.5 (optimum pH 7.0). The cells lysed in distilled water and the minimal NaCl concentration to prevent cell-lysis was found to be 5 % (w/v). The major polar lipids of the strain were identified as phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate and two glycolipids chromatographically identical to those of
Halopenitus persicus
IBRC 10041
T
. The 16S rRNA gene and
rpoB′
gene of strain SKJ47
T
were found to be phylogenetically related to the corresponding genes of
Halopenitus malekzadehii
IBRC-M 10418
T
(96.3 and 91.9 % nucleotide identity, respectively) and
Hpt. persicus
IBRC 10041
T
(96.2 and 93.8 %). The DNA G+C content of strain SKJ47
T
was determined to be 65.0 mol%. The phenotypic, chemotaxonomic and phylogenetic properties suggested that strain SKJ47
T
(=CGMCC 1.12229
T
= JCM 18641
T
) represents a new species of the genus
Halopenitus
, for which the name
Halopenitus salinus
sp. nov. is proposed.
Two moderately halophilic strains, designated SL013A34A2ᵀand SL013A24A, were isolated from oil-contaminated saline soil from Shengli Oilfield, eastern China. Cells were found to be Gram-staining ...negative, aerobic, rod-shaped with a single polar flagellum. The isolates were found to grow at 10–40 °C (optimum 35 °C), pH 6.0–9.0 (optimum pH 8.0), and NaCl concentrations of 0.5–18.0 % (w/v) (optimum 3.0–6.0 NaCl). The 16S rRNA gene sequence analysis indicated that the isolates belong to the genus Marinobacter. Strain SL013A34A2ᵀshares the highest 16S rRNA gene sequence similarities with strain SL013A24A (99.3 %), followed by M. hydrocarbonoclasticus CGMCC 1.7683ᵀ(97.8 %), M. vinifirmus CGMCC 1.7265ᵀ(97.8 %), and M. excellens KMM 3809ᵀ(97.4 %), respectively, but low similarities (93.8–96.4 %) with type strains of the other numbers of genus Marinobacter. DNA–DNA relatedness values of strain SL013A34A2ᵀwith strains SL013A24A, M. hydrocarbonoclasticus CGMCC 1.7683ᵀ, M. vinifirmus CGMCC 1.7265ᵀand M. excellens KMM 3809ᵀwere 88.7, 29.2, 33.4 and 29.4 %, respectively. The major fatty acids of strain SL013A34A2ᵀwere identified as C₁₈:₁ω9c, C₁₆:₀, C₁₂:₀3-OH, C₁₂:₀, C₁₆:₁ω9c and 10-methyl C₁₈:₀. The major respiratory quinone of strain SL013A34A2ᵀwas found to be ubiquinone-9, and its predominant polar lipids were identified as diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol and unidentified glycolipid. The genomic DNA G + C content was found to be 56.1 mol %. Based on the phenotypic, genetic and chemotaxonomic characteristics, these two isolates are representatives of a novel species of the genus Marinobacter, for which the name Marinobacter shengliensis sp. nov. is proposed. The type strain is SL013A34A2ᵀ(=LMG 27740ᵀ = CGMCC 1.12758ᵀ).
The halophilic archaeal strain, YC82 ᵀ, was isolated from Yuncheng salt lake in Shanxi, PR China. Cells from strain YC82 ᵀ were Gram-stain negative, pleomorphic rods, which lysed in distilled water ...and formed light-red colonies on solid media. Strain YC82 ᵀ grew at 25–50 °C (optimum 37 °C), in 1.4–4.8 M NaCl (optimum 2.0 M), in 0–1.0 M MgCl ₂ (optimum 0.05 M) and at pH 6.0–9.5 (optimum pH 7.5). The major polar lipids of strain YC82 ᵀ were phosphatidic acid, phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate and three glycolipids, which were chromatographically identical to those of Haloarchaeobius iranensis IBRC-M 10013 ᵀ and Haloarchaeobius litoreus CGMCC 1.10390 ᵀ. 16S rRNA gene analysis revealed that strain YC82 ᵀ had two dissimilar 16S rRNA genes and that it was phylogenetically related to Hab. iranensis IBRC-M 10013 ᵀ (94.3–99.0 % nucleotide identity) and Hab. litoreus CGMCC 1.10390 ᵀ (94.1–98.8 % nucleotide identity). The rpoB′ gene similarities between strain YC82 ᵀ and Hab. iranensis IBRC-M 10013 ᵀ and Hab. litoreus CGMCC 1.10390 ᵀ were 96.5 % and 95.7 %, respectively. The DNA G+C content of strain YC82 ᵀ was 63.7 mol%. Strain YC82 ᵀ showed low DNA–DNA relatedness with Hab. iranensis IBRC-M 10013 ᵀ and Hab. litoreus CGMCC 1.10390 ᵀ. The phenotypic, chemotaxonomic and phylogenetic properties of strain YC82 ᵀ (= CGMCC 1.12232 ᵀ = JCM 18644 ᵀ) suggest that it represents a novel species of the genus Haloarchaeobius , for which the name Haloarchaeobius salinus sp. nov. is proposed. An emended description of the genus Haloarchaeobius is also presented.
Two extremely halophilic archaeal strains GX1
T
and GX60 were isolated from the Gangxi marine solar saltern, China. Cells from the two strains were observed to be rod-shaped and stained ...Gram-negative, with red-pigmented colonies. Strains GX1
T
and GX60 were found to be able to grow at 25–50 °C (optimum 37 °C), at 1.4–4.8 M NaCl (optimum 2.6 M), at pH 5.5–9.5 (optimum pH 7.0) and neither strain required Mg
2+
for growth. The cells lysed in distilled water and the minimal NaCl concentration to prevent cell-lysis was found to be 8 % (w/v). The major polar lipids of the two strains were identified as phosphatidic acid, phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate and three glycolipids chromatographically identical to those of
Haloarchaeobius iranensis
IBRC-M 10013
T
. 16S rRNA gene analysis revealed that each strain had two dissimilar 16S rRNA genes and both strains were phylogenetically related to
Hab. iranensis
IBRC-M 10013
T
(94.9–98.9 % nucleotide identity). The
rpoB′
gene similarity between strains GX1
T
and GX60, and between these strains and
Hab. iranensis
IBRC-M 10013
T
were found to be 99.6, 96.0 and 95.8 %, respectively. The DNA G + C content of strain GX1
T
and GX60 were determined to be 67.7 and 67.8 mol %, respectively. The DNA–DNA hybridization value of strains GX1
T
and GX60 was 86 % and the two strains showed low DNA–DNA relatedness with
Hab. iranensis
IBRC-M 10013
T
(38 and 32 %). It was concluded that strain GX1
T
(= CGMCC 1.10390
T
= JCM 17114
T
) and strain GX60 (= CGMCC 1.10389 = JCM 17120) represent a new species of
Haloarchaeobius
, for which the name
Haloarchaeobius litoreus
sp. nov. is proposed.
PDF
