Effects of light and hydrogen peroxide on gene expression of newly identified antioxidant enzymes in the harmful algal bloom species Chattonella marina

Figures & data

Table 1. Primers and probes used in this experiment

Name		Sequence 5’-3’
Cu/Zn SOD	Primer (F)	GGACCGATTGGTGAAGCTCATAGG
	Primer (R)	GAGGATGCAATCCCAACAACTCCTC
	Probe	56-FAM/GTGGACATG/ZEN/AACTTTCGCTGCT/3IABkFQ
GPX	Primer (F)	GGTGAGTGGCTTCACGTACAATC
	Primer (R)	CCGAAGCAACGTTAACTACAAGG
	Probe	56-FAM/TGGAAACAT/ZEN/GAAAGGCCTTCTCGCAC/3IABkFQ
CAT	Primer (F)	GCACATTTTGATCGTGAGCGTATCC
	Primer (R)	TTCACTGCAAACCCACGAGGATC
	Probe	56-FAM/GGGTACTTT/ZEN/GAGGTCACAACTCTC/3IABkFQ
APX	Primer (F)	GCACTGACATGCCACAAGAGAAATG
	Primer (R)	TCAAAGACCAATGGCTCTTGAGTCC
	Probe	56-FAM/ACATCTTTG/ZEN/GCCGCATGGAAT/3IABkFQ
TRX	Primer (F)	TCTCCGATCTCCGTGTTGATTTTGC
	Primer (R)	GGTTTCCTGATTGCCTTTTGCGC
	Probe	56-FAM/ACCAAAGGT/ZEN/GTACAGGAATCCTC/3IABkFQ
18S	Primer (F)	CATCAACTCCCAACTGCTTC
	Primer (R)	CTCCGTTACCCGTTAAAGCC
	Probe	56-FAM/GGTTCATTC/ZEN/AAGTTTCTGCCC/3IABkFQ

Table 2. Results of homology search using blastx

							Predicted protein localization
Selected unigene	Number of residue	Target protein	Accession no.	Reference species	Homology (%)	E-value	HECTAR	ASAFind	Plant-mPLoc
c3928	288	Cu/Zn-SOD	ABB88583	Ulva fasciata	55.7	2e-34	^aOther	^bNegative	Chloroplast
			EEH52862	Micromonas pusilla	44.9	1e-30			Cytoplasm
			AAD10208	Arabidopsis thaliana	41.2	9e-38
c14615	314	GPX	EFJ51396	Volvox carteri	50.0	1e-43	Chloroplast	Plastid	Mitochondria
			EDP05620	Chlamydomonas reinhardtii	47.6	3e-41
			AFX60859	Pyropia haitanensis	46.8	2e-35
c24484	539	CAT	AAB70006	C. reinhardtii	44.7	3e-129	Other	Negative	Mitochondria
			EFJ48656	V. carteri	44.6	3e-129			Peroxisome
			EFN55455	Chlorella vulgaris	42.1	2e-123
c5141	364	APX	AJA71275	P. haitanensis	61.9	1e-104	Other	Negative	Peroxisome
			EED87598	Thalassiosira pseudonana	46.7	2e-70
			CAA11265	C. reinhardtii	45.4	2e-67
c13659	302	TRX	WP061430753	Microcystis aeruginosa	41.0	1e-12	Chloroplast	Plastid	Chloroplast
			YP007947754	P. haitanensis	38.4	1e-24
			EDP10052	C. reinhardtii	37.1	1e-21

^aLocalization other than chloroplast and mitochondria. ^bNon-detection of signal peptide.

Fig. 1. Changes in expression level of (a) Cu/Zn-SOD, (b) GPX, (c) CAT, (d) APX, (e) TRX and (f) PRX in Chattonella marina cells grown under three different light intensities for 3 days. Values shown are mean ± SD (N = 4). **, p < 0.01 (two-way ANOVA and Tukey-Kramer and Scheffe’s F test comparing different conditions on each day).

Fig. 2. Changes in Chattonella marina cell density over 2 days under different oxidative stress conditions after dark acclimation (N = 4).

Fig. 3. Changes of gene expression in Chattonella marina antioxidant enzymes over 2 days under different oxidative stress conditions after dark acclimation. Values shown are mean ± SD (N = 4). **, p < 0.01 (two-way ANOVA, Tukey-Kramer and Scheffe’s F test comparing different conditions on each day).