Cloning and expression analysis of β-carotene hydroxylase gene from kale (Brassica oleracea L. var. acephala)

doi:10.3969/j.issn.1004-1524.2019.01.11

›› 2019, Vol. 31 ›› Issue (1): 80-85.DOI: 10.3969/j.issn.1004-1524.2019.01.11

• Horticultural Science • Previous Articles Next Articles

Cloning and expression analysis of β-carotene hydroxylase gene from kale (Brassica oleracea L. var. acephala)

WANG Yushu¹, WANG Huan², GUO Yu¹, ZHOU Minghui¹, CHEN Lu¹, CHEN Yang¹

1.Key Laboratory of Resistance Gene Engineering and Preservation of Biodiversity in Cold Areas, College of Life Sciences, Agriculture and Forestry, Qiqihar University, Qiqihar 161006, China;
2. College of Chemistry and Chemical Engineering, Qiqihar University, Qiqihar 161006, China

Received:2018-04-03 Online:2019-01-25 Published:2019-04-09

Abstract

Abstract: The β-carotene hydroxylase gene from kale (Brassica oleracea L. var. acephala) was cloned by homology-based cloning and RT-PCR strategies, named as BoBCH(GenBank accession number: MH016242). Sequence analysis showed that the full-length of BoBCH cDNA was 906 bp, which encoded a protein with 301 amino acids, an estimated molecular weight was 33.8 ku and an isoelectric point was 9.67. The results of conservative structural domain analysis showed that BoBCH belonged to the FA_hydroxylase protein superfamily. Phylogenetic tree analysis revealed that the kale had the closest evolutionary relationship with Brassica oleracea. Subcellular localization analysis of TMHMM and Wolf-Psort showed that it might be targeted to the chloroplast with four transmembrane regions. The qRT-PCR test results indicated that the highest was leaf, the next was stem, and the lowest level was in root; in the different periods of leaves, the expression level in the ornamental period was high, and the expression level was lower in seedling stage and rosette stage.

Key words: kale, β-carotene hydroxylase gene, cloning, expression analysis

CLC Number:

S635.9

WANG Yushu, WANG Huan, GUO Yu, ZHOU Minghui, CHEN Lu, CHEN Yang. Cloning and expression analysis of β-carotene hydroxylase gene from kale (Brassica oleracea L. var. acephala)[J]. , 2019, 31(1): 80-85.

References

[1] FRANK H A, COGDELL R J.Carotenoids in photosynthesis[J]. Photochemistry and Photobiology, 1996, 63(3): 257-264.
[2] 吴疆. 枸杞β-胡萝卜素羟化酶基因及其启动子的克隆和研究[D]. 天津: 天津大学, 2015.
WU J.Cloning and research of β-carotenoid hydroxylase gene and its promoter of Lycium barbarum[D]. Tianjin: Tianjin University, 2015. (in Chinese with English abstract)
[3] KATO M, IKOMA Y, MATSUMOTO H, et al.Accumulation of carotenoids and expression of carotenoid biosynthetic genes during maturation in citrus fruit[J]. Plant Physiology, 2004, 134(2): 824-837.
[4] PEGUERO-PINA J J, GIL-PELEGTINIL E, MORALES F. Three pools of zeaxanthin in Quercus coccifera leaves during light transitions with different roles in rapidly reversible photoprotective energy dissipation and photoprotection[J].Journal of Experimental Botany, 2013, 64(6): 1649-1661.
[5] POGSON B J, RISSLER H M.Genetic manipulation of carotenoid biosynthesis and photoprotection[J]. Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences, 2000, 355(1402):1395-1403.
[6] TIAN L, MAGALLANES-LUNDBACK M, MUSETTI V, et al.Functional analysis of β-and ɛ-ring carotenoid hydroxylases in Arabidopsis[J]. Plant Cell, 2003, 15(6): 1320-1332.
[7] TIAN L, DELLAPENNA D.Characterization of a second carotenoid β-hydroxylase gene from Arabidopsis and its relationship to the LUT1 locus[J]. Plant Molecular Biology, 2001, 47(3):379-388.
[8] 焦芳婵, 曾建敏, 吴兴富, 等. 烟草β-胡萝卜素羟化酶基因的特征分析[J].分子植物育种, 2015, 13(8):1831-1837.
JIAO F C, ZENG J M, WU X F, et al.Characteristic analysis of β-carotene hydroxylase gene in Nicotiana tabacum L.[J].Molecular Plant Breeding, 2015, 13(8):1831-1837. (in Chinese with English abstract)
[9] 冯唐锴, 李思光, 汪艳璐, 等. 南丰蜜橘β-胡萝卜素羟化酶基因的克隆和序列分析[J]. 西北植物学报, 2007, 27(2):238-243.
FENG T K, LI S G, WANG Y L, et al.Cloning and sequence analysis of β-carotene hydroxylase gene from Nanfeng orange (Citrus reticulate Blanco var. kinokuni (Tanaka) H. H. Hu)[J]. Acta Botanica Boreali-Occidentalia Sinica, 2007, 27(2): 238-243. (in Chinese with English abstract)
[10] BOUVIER F, KELLER Y, HARLINGUE A D,et al.Xanthophyll biosynthesis: molecular and functional characterization of carotenoid hydroxylases from pepper fruits (Capsicum annuum L.)[J]. Biochimica et Biophysica Acta, 1998, 1391(3):320-328.
[11] 王雅琼, 郑伟尉, 臧运祥, 等. 不同品种羽衣甘蓝生长期色素含量变化规律研究[J]. 中国农学通报, 2013, 29(10):154-161.
WANG Y Q, ZHENG W W, ZANG Y X, et al.Studies on change of pigment contents during different developmental stages among several flowering kale cultivars[J]. Chinese Agricultural Science Bulletin, 2013, 29(10):154-161. (in Chinese with English abstract)
[12] LIVAK K J, SCHMITTGEN T D.Analysis of relative gene expression data using real-time quantitative PCR and the 2^-ΔΔCt[J]. Method, 2001, 25(4): 402-408.
[13] JIANG W, JING J, GANG W, et al.Cloning and characterization of a novelβ-carotene hydroxylase gene from Lycium barbarum and its expression in Escherichia coli[J].Biotechnology and Applied Biochemistry, 2014, 61(6): 637-645.
[14] DAVISION P A, HUNTER C N, HORTON P.Overexpression of beta-carotene hydroxylase enhances stress tolerance in Arabidopsis[J].Nature, 2002, 418(6894): 203-206.
[15] 冯唐锴,李思光,罗玉萍,等. 植物β-胡萝卜素羟化酶研究进展[J].生物技术通报,2007(l): 54-58.
FENG T K, LI S G, LUO Y P, et al.The research of β-carotene hydroxylase[J]. Biotechnology Bulletin, 2007(l): 54-58. (in Chinese with English abstract)
[16] YAN J, KANDIANIS C B, HARJES C E, et al.Rare genetic variation at Zea mays Crtrb1 increases beta-carotene in maize grain[J]. Nature Genetics, 2010, 42(4): 322-327.

Cloning and expression analysis of β-carotene hydroxylase gene from kale (Brassica oleracea L. var. acephala)

RichHTML

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	HE Jiaqi, ZHAI Ying, ZHANG Jun, QIU Shuang, LI Mingyang, ZHAO Yan, ZHANG Meijuan, MA Tianyi. Cloning and expression analysis of GmDof1.5 in soybean under abiotic stress [J]. Acta Agriculturae Zhejiangensis, 2021, 33(1): 1-7.
[2]	WANG Weike, LU Na, YAN Jing, SONG Jiling, YUAN Weidong, ZHOU Zufa. Cloning and expression analysis of PpSAMS gene of Pleurotus pulmonarius [J]. Acta Agriculturae Zhejiangensis, 2021, 33(1): 62-68.
[3]	SUN Ruiping, WANG Feng, CHAO Zhe, LIU Hailong, XING Manping, LIU Quanwei, HUANG Lili, ZHENG Xinli, WEI Limin. Cloning and tissue expression analysis of PDK4 gene in Tunchang pig [J]. , 2020, 32(6): 978-985.
[4]	WANG Weike, SONG Jiling, LU Na, YUAN Weidong, YAN Jing, CHEN Guanping. Cloning and expression of the PpFBD1 involved in primordium formation of Pleurotus pulmonarius [J]. , 2020, 32(1): 93-97.
[5]	ZHANG Aiju, LIU Shili, LIU Jindian, ZHANG Genfang, ZHOU Zhiming. Cloning, characterization, and expression patterns of one sarco/endoplasmic reticulum calcium ATPase isoform from freshwater mussel Hyriopsis cumingii [J]. , 2019, 31(4): 545-555.
[6]	WANG Jing, PENG Shuang, HU Sheng, ZHUO Wei, CHEN Qian, LI Liqin. Cloning expression vector construction and expression analysis of tobacco ATPase4 gene [J]. , 2019, 31(2): 173-181.
[7]	GONG Shasha, MENG Qingling, QIAO Jun, ZHONG Wenqiang, HUANG Yunfu, ZHANG Guowu, CHEN Ying, CAI Xuepeng. Gene cloning and bioactivity analysis of chitinase gene AO-483 from Arthrobotrys oligospora XJ-A1 [J]. , 2019, 31(2): 222-228.
[8]	CHEN Guanju, CHAI Yiqiu, LI Xiaola, FANG Ming, LIU Yougao, JIN Yiwei. Cloning of sublitisin-like protease gene cDNA from Isaria cicadae and characterization of its sequence and deduced protein [J]. , 2019, 31(10): 1663-1670.
[9]	XU Qingsong, ZHAO Jia, WEI Yunmin, HAN Rongrong, LIU Lusheng, JIANG Caode, YU Yong-xiong. Cloning and expression analysis of oxalate oxidase gene MsOXO from Medicago sativa [J]. , 2019, 31(1): 11-19.
[10]	ZHANG Gaofeng, YANG Kankan, ZHANG Qian, WANG Yuanhong, YU Zhaorong, ZHANG Xueqi, LU Zhimin, LIU Zimin, LI Yongdong, WANG Yong. Prokaryotic expression and bioinformatics analysis of orf virus Anhui strain 023 gene [J]. , 2019, 31(1): 30-38.
[11]	YANG Zhou, LYU Ke, LYU Shan, WANG Junjie, ZHANG Di. Cloning and sequence analysis of two ARF genes and two Aux/IAA genes in Agapanthus praecox ssp. orientalis [J]. , 2019, 31(1): 86-97.
[12]	TANG Xiao, DENG Mengsheng, ZOU Xue, Li Liqin, ZHU Yuanzhi, WANG Xiyao. Cloning and expression analysis of StDWF1 in Solanum tuberosum [J]. , 2018, 30(6): 909-917.
[13]	ZHANG Yang, YAO Wencheng, WANG Bin, LU Lizhi, WANG Zhaoshan, XU Qi, CHANG Guobin, CHEN Guohong. Expression analysis of crest phenotype related genes from crested white duck [J]. , 2018, 30(5): 707-710.
[14]	XUE Shengling, JIANG Min, CHANG Jiaqi, LIU Yang, WEI Lin, ZHOU Jiankun, ZHANG Fen, SUN Bo. Cloning and prokaryotic expression of 1-deoxy-D-xylulose-5-phosphate synthase encoding gene (BaDXS1) in Brassica alboglabra [J]. , 2018, 30(5): 771-777.
[15]	LIANG Minhua, YANG Zhenfeng, SU Xinguo, SONG Chunbo. Cloning and expression analysis of phytoene synthase gene from peach fruit [J]. , 2018, 30(3): 399-405.