Cloning and characterization of nitrate reductase gene in kelp Saccharina japonica (Laminariales, Phaeophyta)

doi:10.1186/s12870-023-04064-7

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	Cloning and characterization of nitrate reductase gene in kelp Saccharina japonica (Laminariales, Phaeophyta)
	Wang, Zhenghua 1,2,3,4; Wu, Chunhui1,2,3 ; Jiang, Peng 1,2,3
	2023-02-06
发表期刊	BMC PLANT BIOLOGY
ISSN	1471-2229
卷号	23 期号:1 页码:14
通讯作者	Jiang, Peng(jiangpeng@qdio.ac.cn)
摘要	BackgroundBrown macroalgae dominate temperate coastal ecosystems, and their productivity is typically limited by nitrate availability. As an economically important kelp, Saccharina japonica is the most productive farmed seaweed and needs to be supplemented with sufficient nitrate throughout the cultivation process. However, molecular characterization of genes involved in nitrogen assimilation has not been conducted in brown macroalgae.ResultsHere, we described the identification of the nitrate reductase (NR) gene from S. japonica (SjNR). Using two different cloning methods for SjNR, i.e. rapid amplification of cDNA ends (RACE) and cDNA cloning alone, a single fragment was obtained respectively. According to results of sequence analysis between these two fragments, the tentative coding sequence in two clones, SjNR-L and SjNR-S, were suggested to represent two transcripts of the single copy SjNR, and the ATG of SjNR-S was located inside the third exon of SjNR-L. In the 5 ' upstream sequence of each transcript, promoter core elements, response elements, especially multiple N response elements which occurred in microalgal NR, were all predicted. Further sequence analysis revealed that both transcripts encoded all five domains conserved in eukaryotic plant NRs. RT-qPCR results showed that the transcription level of SjNR in juvenile sporophytes could be significantly induced by nitrate and inhibited by ammonium, which was in line with plant NRs. The recombinant SjNR-L and SjNR-S were all proved to have NR activity, suggesting that the single-copy gene SjNR might be regulated on transcription level based on alternative promoters and multiple transcriptional start sites. Moreover, both NADH and NADPH were found to be able to act as electron donors for SjNR alone, which is the first confirmation that brown algal NR has a NAD(P)H-bispecific form.ConclusionThese results will provide a scientific basis for understanding the N demand of kelp in various stages of cultivation and evaluating the environmental remediation potential of kelp in eutrophic sea areas.
关键词	Function verification Gene cloning Kelp Nitrate reductase Saccharina japonica
DOI	10.1186/s12870-023-04064-7
收录类别	SCI
语种	英语
WOS研究方向	Plant Sciences
WOS类目	Plant Sciences
WOS记录号	WOS:000926123600003
出版者	BMC
WOS关键词	IN-VITRO ; ALTERNATIVE PROMOTERS ; ARABIDOPSIS-THALIANA ; CHLORELLA-VULGARIS ; MOLECULAR-CLONING ; MARINE MACROALGAE ; BROWN-ALGAE ; LIGHT ; IDENTIFICATION ; EXPRESSION
引用统计	被引频次：1[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
条目标识符	http://ir.qdio.ac.cn/handle/337002/183471
专题	实验海洋生物学重点实验室
通讯作者	Jiang, Peng
作者单位	1.Chinese Acad Sci, Qingdao 266071, Peoples R China 2.Chinese Acad Sci, Inst Oceanol, Ctr Ocean Mega Sci, Shandong Prov Key Lab Expt Marine Biol, Qingdao 266071, Peoples R China 3.Qingdao Natl Lab Marine Sci & Technol, Lab Marine Biol & Biotechnol, Qingdao 266237, Peoples R China 4.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
第一作者单位	中国科学院海洋研究所
通讯作者单位	中国科学院海洋研究所
推荐引用方式 GB/T 7714	Wang, Zhenghua,Wu, Chunhui,Jiang, Peng. Cloning and characterization of nitrate reductase gene in kelp Saccharina japonica (Laminariales, Phaeophyta)[J]. BMC PLANT BIOLOGY,2023,23(1):14.
APA	Wang, Zhenghua,Wu, Chunhui,&Jiang, Peng.(2023).Cloning and characterization of nitrate reductase gene in kelp Saccharina japonica (Laminariales, Phaeophyta).BMC PLANT BIOLOGY,23(1),14.
MLA	Wang, Zhenghua,et al."Cloning and characterization of nitrate reductase gene in kelp Saccharina japonica (Laminariales, Phaeophyta)".BMC PLANT BIOLOGY 23.1(2023):14.