Cloning and Expression Analysis of Chloroplast-targeted Ferredoxin-NADP+ Oxidoreductase Gene of Tea Plant (Camellia sinensis) cv.Huangjinya
1 State Key Laboratory of Crop Biology, Tai’an, 271018
2 College of Horticulture Science and Engineering, Shandong Agricultural University, Tai’an, 271018
Journal of Tea Science Research, 2020, Vol. 10, No. 1 doi: 10.5376/jtsr.2020.10.0001
Received: 01 Jul., 2020 Accepted: 06 Jul., 2020 Published: 24 Jul., 2020
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This article was first published in Molecular Plant Breeding in Chinese, and here was authorized to translate and publish the paper in English under the terms of Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Preferred citation for this article:
Zhao X.X., Fan Y.G., Tian Y.Y., Wang H.Y., Zhang L.X., and Li M., 2020, Cloning and expression analysis of Chloroplast-targeted Ferredoxin-NADP+ oxidoreductase gene of tea plant (Camellia sinensis) cv. Huangjinya, Journal of Tea Science Research, 10(1): 1-11 (doi: 10.5376/jtsr.2020.10.0001)
The chloroplast-targeted ferredoxin-NADP+ oxidoreductase (LFNR) is a hydrophilic flavin protein located at the end of the photosynthesis electron delivery chain in higher plants. It’s a hub for linking photosynthesis electronic transportation with chloroplast redox metabolism. In order to explore the relationship between LFNR gene and yellowing of ‘Huangjinya’controlled by light, we took separately one bud two leaves of ‘Huangjinya’ ‘Shuchazao’（CK） as the material to clone CsLFNR gene and then we obtained the sequence with a similarity of 100%, named CsLFNR1.1 (GenBank accession no. MT311318). Throw bioinformatics analysis, we know its cDNA gene length is 1095bp,coding 364 amino acids, The protein molecular weight is 40.623KDa, the theoretical isoelectric point is 8.86.CsLFNR1.1 is a alkaline protein without transmembrane structure and signal peptide, it has a chloroplast transport peptide(cTP), the secondary structure contains 23.35% alpha helix, 5.49% beta-corner, 28.85% extension chain and 42.31% irregular curling, It locates in the chloroplast. Through the Nucleotide BLAST, Protein BLAST and DNAMAN software, we found that CsLFNR1.1 and its translated amino acid sequences had 4 bases and 3 amino acid differences with NCBI ‘Shuchazao’ Gene CsLFNR1(XM_028233617.1) and Protein (XP_028089418.1), the similarity reached 99.63% and 99.18%. CsLFNR1.1 and CsLFNR1 have small differences in the physical and chemical properties and secondary structure, the tertiary structure prediction template of them is consistent. In addition, they have the same conserved domain and active site. We took different shade ‘Huangjinya’ blades as the material for qRT-PCR, the result showed that the expression of CsLFNR1.1 gene response to the light intensity, its expression increased with the increase of light intensity. This study provides a theoretical basis and scientific basis for further exploring the role of CsLFNR1.1 gene in the light regulation process of the new shoots and leaves of ‘Huangjinya’.
Camellia sinensis; FNR; Gene clone; Bioinformatics analysis; Expression analysis