Yadav Anu, Maertens Laurens, Meese Tim, Van Nieuwerburgh Filip, Mysara Mohamed, Leys Natalie, Cuypers Ann, Janssen Paul Jaak
Interdisciplinary Biosciences, Microbiology Unit, Belgian Nuclear Research Centre (SCKCEN), 2400 Mol, Belgium.
Environmental Biology, Centre for Environmental Sciences, Hasselt University, 3590 Diepenbeek, Belgium.
Microorganisms. 2021 Jul 30;9(8):1626. doi: 10.3390/microorganisms9081626.
Two morphotypes of the cyanobacterial (formerly sp.) strain PCC 8005, denoted as P2 (straight trichomes) and P6 (helical trichomes), were subjected to chronic gamma radiation from spent nuclear fuel (SNF) rods at a dose rate of ca. 80 Gy·h for one mass doubling period (approximately 3 days) under continuous light with photoautotrophic metabolism fully active. Samples were taken for post-irradiation growth recovery and RNA-Seq transcriptional analysis at time intervals of 15, 40, and 71.5 h corresponding to cumulative doses of ca. 1450, 3200, and 5700 Gy, respectively. Both morphotypes, which were previously reported by us to display different antioxidant capacities and differ at the genomic level in 168 SNPs, 48 indels and 4 large insertions, recovered equally well from 1450 and 3200 Gy. However, while the P2 straight type recovered from 5700 Gy by regaining normal growth within 6 days, the P6 helical type took about 13 days to recover from this dose, indicating differences in their radiation tolerance and response. To investigate these differences, P2 and P6 cells exposed to the intermediate dose of gamma radiation (3200 Gy) were analyzed for differential gene expression by RNA-Seq analysis. Prior to batch normalization, a total of 1553 genes (887 and 666 of P2 and P6, respectively, with 352 genes in common) were selected based on a two-fold change in expression and a false discovery rate FDR smaller or equal to 0.05. About 85% of these 1553 genes encoded products of yet unknown function. Of the 229 remaining genes, 171 had a defined function while 58 genes were transcribed into non-coding RNA including 21 tRNAs (all downregulated). Batch normalization resulted in 660 differentially expressed genes with 98 having a function and 32 encoding RNA. From PCC 8005-P2 and PCC 8005-P6 expression patterns, it emerges that although the cellular routes used by the two substrains to cope with ionizing radiation do overlap to a large extent, both strains displayed a distinct preference of priorities.
对蓝藻(原 种)菌株PCC 8005的两种形态型进行了研究,分别记为P2(直形藻丝)和P6(螺旋形藻丝)。在光自养代谢完全活跃的连续光照条件下,以约80 Gy·h的剂量率,用乏核燃料(SNF)棒对其进行慢性γ辐射,持续一个质量倍增期(约3天)。在对应于约1450、3200和5700 Gy累积剂量的15、40和71.5小时时间间隔采集样本,用于辐照后生长恢复和RNA-Seq转录分析。我们之前报道过这两种形态型表现出不同的抗氧化能力,并且在168个单核苷酸多态性(SNP)、48个插入缺失(indel)和4个大插入方面存在基因组水平差异。P2和P6从1450 Gy和3200 Gy的辐射中恢复情况相同。然而,P2直形类型在6天内恢复正常生长,从5700 Gy的辐射中恢复,而P6螺旋形类型从该剂量恢复则需要约13天,这表明它们在辐射耐受性和反应方面存在差异。为了研究这些差异,通过RNA-Seq分析对暴露于中等剂量γ辐射(3200 Gy)的P2和P6细胞进行了差异基因表达分析。在批次标准化之前,基于表达量两倍变化和错误发现率FDR小于或等于0.05,共选择了1553个基因(P2中有887个,P6中有666个,其中352个基因是共有的)。这1553个基因中约85%编码功能未知的产物。在其余229个基因中,171个具有确定的功能,58个基因转录为非编码RNA,包括21个转运RNA(均下调)。批次标准化后得到660个差异表达基因,其中98个具有功能,32个编码RNA。从PCC 8005 - P2和PCC 8005 - P6的表达模式可以看出,尽管两个亚菌株应对电离辐射所采用的细胞途径在很大程度上重叠,但两个菌株都表现出明显的优先顺序偏好。