Isolation and expression analysis of a SKIP gene from Brassica oleracea var. italica

doi:10.3969/j.issn.1004-1524.2022.05.11

Acta Agriculturae Zhejiangensis ›› 2022, Vol. 34 ›› Issue (5): 966-973.DOI: 10.3969/j.issn.1004-1524.2022.05.11

• Horticultural Science • Previous Articles Next Articles

Isolation and expression analysis of a SKIP gene from Brassica oleracea var. italica

WEI Haizhong¹(), PAN Liqin¹, TANG Ziyi², TIAN Shengye², HE Haiye², YIN Longfei², ZHENG Dewei², ZHANG Huijuan², JIANG Ming²^,^*()

1. Taizhou Vocational College of Science and Technology, Taizhou 318020, Zhejiang, China
2. College of Life Science, Taizhou University, Taizhou 318000, Zhejiang, China

Received:2021-12-17 Online:2022-05-25 Published:2022-06-06
Contact: JIANG Ming

Abstract

Abstract:

SKIP (SKI-interacting protein) plays an important role in plant growth, development and stress responses. SKIP gene from Brassica oleracea var. italic was isolated and its sequence was analyzed by using bioinformatic tools. Besides, its expression patterns under biotic stresses (Hyaloperonospora parasitica and Xanthomonas campestris pv. campestris) and abiotic stresses (NaCl and PEG6000) were obtained by quantitative real-time PCR. The results indicated that broccoli SKIP was 1 836 bp in length with no intron identified. BoiSKIP encoded 611 amino acids and contained one SNW/SKI domain, which was located at 188-347 of the deduced protein sequences. The molecular weight and theoretical isoelectric point were 69.22 ku and 9.15, respectively, with a grand average of hydropathy index of -1.12, and BoiSKIP was identified as a hydrophilic protein. Phylogenetic analysis indicated that SKIPs from Brassica plants were grouped to the same clade, with a support rate of 100%. The closest relationship was found between BoiSKIP and cabbage SKIP, and they were gathered in the same branch. Expression analysis results indicated that BoiSKIP was induced by both NaCl and PEG6000, and the highest expression levels were observed at 24 h and 12 h, respectively. BoiSKIP was also induced by both H. parasitica and X. campestris pv. campestris, and the highest expression levels were found at 24 h and 72 h, respectively. Isolation and expression analysis of broccoli SKIP gene provided an insight into future study about its function on stress resistance responses.

Key words: Brassica oleracea var. italica, phylogenetic analysis, downy mildew, black rot

CLC Number:

S635

WEI Haizhong, PAN Liqin, TANG Ziyi, TIAN Shengye, HE Haiye, YIN Longfei, ZHENG Dewei, ZHANG Huijuan, JIANG Ming. Isolation and expression analysis of a SKIP gene from Brassica oleracea var. italica[J]. Acta Agriculturae Zhejiangensis, 2022, 34(5): 966-973.

Figures/Tables 6

Fig.1 Sequence of broccoli BoiSKIP gene and its deduced protein The SNW/SKI domain was highlighted.

Fig.2 Hydrophobic/hydrophilic character predication of BoiSKIP

Fig.3 Comparisons of SNW/SKI domains of SKIP from different plants XP_013688403, Brassica napus; XP_018456548, Raphanus sativus; CAA7013451, Microthlaspi erraticum; KAF2606023, Brassica cretica; OAP19162, Arabidopsis thaliana; RID56290, Brassica rapa; XP_006301006, Capsella rubella; XP_006390116, Eutrema salsugineum; XP_010471904, Camelina sativa; XP_013591577, Brassica oleracea var. oleracea; XP_020890221, A. lyrata subsp. lyrata; XP_031736674, Cucumis sativus.

Fig.4 Phylogenetic tree of BoiSKIP and its homologous sequences

Fig.5 Expression of BoiSKIP under abiotic stresses

Fig.6 Expression of BoiSKIP challenged by pathogens A, Hyaloperonospora parasitica ; B, Xanthomonas campestris pv. campestris.

References 26

[1]	OWIS A I. Broccoli, the green beauty: a review[J]. Journal of Pharmaceutical Sciences & Research, 2015, 7(9): 696-703.
[2]	LATTÉ K P, APPEL K E, LAMPEN A. Health benefits and possible risks of broccoli: an overview[J]. Food and Chemical Toxicology, 2011, 49(12): 3287-3309. DOI URL
[3]	WANG J S, GU H H, YU H F, et al. Genotypic variation of glucosinolates in broccoli (Brassica oleracea var. italica) florets from China[J]. Food Chemistry, 2012, 133(3): 735-741. DOI URL
[4]	高庆生, 陈永生, 管春松, 等. 西兰花生产现状、存在的问题及建议[J]. 蔬菜, 2020(11): 29-31.
	GAO Q S, CHEN Y S, GUAN C S, et al. Production status, problems and suggestions of broccoli[J]. Vegetables, 2020(11): 29-31. (in Chinese with English abstract)
[5]	VARALAKSHMI B, GANESHAN G, GOPALAKRISHNAN C, et al. Identification of sources of resistance to Alternaria leaf spot (Alternaria brassicicola), black rot (Xanthomonas campestris) and downy mildew (Peronospora parasitica) in cauliflower (Brassica oleraceae)[J]. Indian Journal of Agricultural Sciences, 2009, 79(6): 482-483.
[6]	COELHO P S, MONTEIRO A A. Inheritance of downy mildew resistance in mature broccoli plants[J]. Euphytica, 2003, 131(1): 65-69. DOI URL
[7]	NAGAI H, MIYAKE N, KATO S, et al. Improved control of black rot of broccoli caused by Xanthomonas campestris pv. campestris using a bacteriophage and a nonpathogenic Xanthomonas sp. strain[J]. Journal of General Plant Pathology, 2017, 83(6): 373-381. DOI URL
[8]	GUPTA M, VIKRAM A, BHARAT N. Black rot-A devastating disease of crucifers: a review[J]. Agricultural Reviews, 2013, 34(4): 269. DOI URL
[9]	SINGH D, DHAR S, YADAVA D K. Genetic and pathogenic variability of Indian strains of Xanthomonas campestris pv. campestris causing black rot disease in Crucifers[J]. Current Microbiology, 2011, 63(6): 551-560. DOI URL
[10]	MURIES B, CARVAJAL M, MARTÍNEZ-BALLESTA M. Response of three broccoli cultivars to salt stress, in relation to water status and expression of two leaf aquaporins[J]. Planta, 2013, 237(5): 1297-1310. DOI URL
[11]	KIM Y N, KHAN M A, KANG S M, et al. Enhancement of drought-stress tolerance of Brassica oleracea var. italica L. by newly isolated Variovorax sp. YNA59[J]. Journal of Microbiology and Biotechnology, 2020, 30(10): 1500-1509. DOI URL
[12]	MORA A A, EARLE E D. Resistance to Alternaria brassicicola in transgenic broccoli expressing a Trichoderma harzianum endochitinase gene[J]. Molecular Breeding, 2001, 8(1): 1-9. DOI URL
[13]	LI Z S, MEI Y J, LIU Y M, et al. The evolution of genetic diversity of broccoli cultivars in China since 1980[J]. Scientia Horticulturae, 2019, 250: 69-80. DOI URL
[14]	CIANCALEONI S, NEGRI V. A method for obtaining flexible broccoli varieties for sustainable agriculture[J]. BMC Genetics, 2020, 21(1): 51. DOI URL
[15]	LI Y, XIA C C, FENG J L, et al. The SNW domain of SKIP is required for its integration into the spliceosome and its interaction with the Paf1 complex in Arabidopsis[J]. Molecular Plant, 2016, 9(7): 1040-1050. DOI URL
[16]	ZHANG D Q, QIAO X J, WANG L M, et al. Skip is essential for Notch signaling to induce Sox2 in cerebral arteriovenous malformations[J]. Cellular Signalling, 2020, 68: 109537. DOI URL
[17]	LIM G H, ZHANG X, CHUNG M S, et al. A putative novel transcription factor, AtSKIP, is involved in abscisic acid signalling and confers salt and osmotic tolerance in Arabidopsis[J]. New Phytologist, 2010, 185(1): 103-113. DOI URL
[18]	WANG X X, WU F M, XIE Q G, et al. SKIP is a component of the spliceosome linking alternative splicing and the circadian clock in Arabidopsis[J]. Plant Cell, 2012, 24(8): 3278-3295. DOI URL
[19]	CUI Z B, TONG A Z, HUO Y Q, et al. SKIP controls flowering time via the alternative splicing of SEF pre-mRNA in Arabidopsis[J]. BMC Biology, 2017, 15(1): 80. DOI URL
[20]	LI Y, YANG J, SHANG X D, et al. SKIP regulates environmental fitness and floral transition by forming two distinct complexes in Arabidopsis[J]. New Phytologist, 2019, 224(1): 321-335. DOI URL
[21]	ZHANG H J, YIN L F, SONG F M, et al. SKIP silencing decreased disease resistance against Botrytis cinerea and Pseudomonas syringae pv. tomato DC3000 in tomato[J]. Frontiers in Plant Science, 2020, 11: 593267. DOI URL
[22]	FRASCH M, SAUMWEBER H. Two proteins from Drosophila nuclei are bound to chromatin and are detected in a series of puffs on polytene chromosomes[J]. Chromosoma, 1989, 97(4): 272-281. DOI URL
[23]	WIELAND C, MANN S, VON BESSER H, et al. The Drosophila nuclear protein Bx42, which is found in many puffs on polytene chromosomes, is highly charged[J]. Chromosoma, 1992, 101(8): 517-525. DOI URL
[24]	DAHL R, WANI B, HAYMAN M J. The Ski oncoprotein interacts with Skip, the human homolog of Drosophila Bx42[J]. Oncogene, 1998, 16 (12): 1579-1586. DOI URL
[25]	HOU X, XIE K B, YAO J L, et al. A homolog of human ski-interacting protein in rice positively regulates cell viability and stress tolerance[J]. Proceedings of the National Academy of Sciences, 2009, 106(15): 6410-6415. DOI URL
[26]	WANG X M, LI Z G, YAN F, et al. ZmSKIP, a homologue of SKIP in maize, is involved in response to abiotic stress in tobacco[J]. Plant Cell, Tissue and Organ Culture, 2013, 112(2): 203-216.

Isolation and expression analysis of a SKIP gene from Brassica oleracea var. italica

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