Garlic Chitinase Gene AsCHI1: Identification and Its Response to Salt Stress

doi:10.11924/j.issn.1000-6850.casb2021-0275

Abstract

Abstract:

The study aims to investigate the sequence characteristics of garlic chitinase gene and its response to salt stress, and to identify its function in resistance to stresses. The AsCHI1 gene was isolated by RT-PCR technology using garlic cultivar ‘Cangshan siliuban’ as the research material. BioXM 2.6, ProtParam, DNAMAN, SignalP 5.0, SOPMA, SWISS-MODEL, NCBI and MEGA5 were adopted to analyze the sequence characteristics of nucleotides and its encoded amino acids. Fluorescent quantitative PCR technology was introduced to detect its expression in different tissues and under salt stress. The open reading frame of the gene was 933 bp in length encoding 310 amino acids. The protein encoded by AsCHI1 gene belonged to chitinase Class I and GH19 family. Amino acid sequence analysis showed that AsCHI1 contained a signal peptide region at the N-terminus possessed by most chitinases, whereas the C-terminus was highly consistent with the amino acid sequences of CHI from other species. In terms of genetic relationship, it was relatively close to Lilium longiflorum LlCHI (QBZ68892.1) in the Liliaceae family and tea CsCHI (XP_028075045.1) in the Theaceae family. Real-time fluorescent quantitative PCR analysis showed that the AsCHI1 gene was expressed in garlic roots, garlic cloves and leaves, but the highest expression was observed in the roots. In addition, salt stress could significantly induce the expression of AsCHI1 gene in various tissues. This gene may play an important role in the resistance to salt stress in garlic plants.

Key words: Allium sativum, salt stress, AsCHI1 gene, gene cloning, response

CLC Number:

S633.4

ZHANG Yuyang, ZHOU Xue, LIU Lingyi, XU Wujun, REN Xuqin, WANG Guanglong, XIONG Aisheng. Garlic Chitinase Gene AsCHI1: Identification and Its Response to Salt Stress[J]. Chinese Agricultural Science Bulletin, 2022, 38(5): 23-29.

Figures/Tables 8

References 27

[1]	ALI S, GANAI B A, KAMILI A N, et al. Pathogenesis-related proteins and peptides as promising tools for engineering plants with multiple stress tolerance[J]. Microbiologica research, 2018, 212-213:29-37. doi: 10.1016/j.micres.2018.04.008 URL
[2]	COLLINGE D B, KRAGH K M, MIKKELSEN J D, et al. Plant chitinases[J]. Plant journal, 1993, 3(1):31-40. doi: 10.1046/j.1365-313X.1993.t01-1-00999.x URL
[3]	OYELEYE A, NORMI Y M. Chitinase: diversity, limitations, and trends in engineering for suitable applications[J]. Bioscience reports, 2018, 38(4): BSR2018032300.
[4]	MATHEW G M, MADHAVAN A, ARUN K B, et al. Thermophilic chitinases: structural, functional and engineering attributes for industrial applications[J]. Applied biochemistry and biotechnology, 2021, 193(1):142-164. doi: 10.1007/s12010-020-03416-5 URL
[5]	郭林霞, 董旋, 赵德刚. 转杜仲几丁质酶基因EuCHIT1番茄提高对灰霉病的抗性[J]. 植物生理学报, 2016, 52(5):703-714.
[6]	ZHANG C W, HUANG M Y, SANG X C, et al. Association between sheath blight resistance and chitinase activity in transgenic rice plants expressing McCHIT1 from bitter melon[J]. Transgenic research, 2019, 28(3-4):381-390. doi: 10.1007/s11248-019-00158-x URL
[7]	DING X, GOPALAKRISHNAN B, JOHNSON L B, et al. Insect resistance of transgenic tobacco expressing an insect chitinase gene[J]. Transgenic research, 1998, 7(2):77-84. doi: 10.1023/A:1008820507262 URL
[8]	罗亮. 棉花几丁质酶基因Ghchi6的抗蚜功能研究[D]. 荆州:长江大学, 2017:24-41.
[9]	王媛. 兴安落叶松几丁质酶基因的功能分析[D]. 呼和浩特:内蒙古农业大学, 2018:12-26.
[10]	AHMED N U, PARK J I, JUNG H J, et al. Molecular characterization of stress resistance-related chitinase genes of Brassica rapa[J]. Plant physiology and biochemistry, 2012, 58:106-115. doi: 10.1016/j.plaphy.2012.06.015 URL
[11]	KONG Q, MOSTAFA H H A, YANG W, et al. Comparative transcriptome profiling reveals that brassinosteroid-mediated lignification plays an important role in garlic adaption to salt stress[J]. Plant physiology and biochemistry, 2021, 158:34-42. doi: 10.1016/j.plaphy.2020.11.033 URL
[12]	QIU Z, ZHENG Z, ZHANG B, et al. Formation, nutritional value, and enhancement of characteristic components in black garlic: A review for maximizing the goodness to humans[J]. Comprehensive reviews in food science and food safety, 2020, 19(2):801-834. doi: 10.1111/crf3.v19.2 URL
[13]	梁志乐, 尚珂含, 王立辉, 等. 大蒜谷胱甘肽硫转移酶基因AsGST的克隆及其对盐胁迫的响应[J]. 核农学报, 2019, 33(6):1088-1095.
[14]	WANG G L, REN X Q, LIU J X, et al. Transcript profiling reveals an important role of cell wall remodeling and hormone signaling under salt stress in garlic[J]. Plant physiology and biochemistry, 2019, 135:87-98. doi: 10.1016/j.plaphy.2018.11.033 URL
[15]	WANG G, TIAN C, WANG Y, et al. Selection of reliable reference genes for quantitative RT-PCR in garlic under salt stress[J]. PeerJ, 2019, 7:e7319. doi: 10.7717/peerj.7319 URL
[16]	LIVAK K J, SCHMITTGEM T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2^-ΔΔCT method[J]. Methods, 2001, 25:402-408. doi: 10.1006/meth.2001.1262 URL
[17]	ASAKURA H, YAMAKAWA T, TAMURA T, et al. Transcriptomic and metabolomic analyses provide insights into the upregulation of fatty acid and phospholipid metabolism in tomato fruit under drought stress[J]. Journal of agricultural and food chemistry, 2021, 69(9):2894-2905. doi: 10.1021/acs.jafc.0c06168 URL
[18]	JIN J, LI K, QIN J, et al. The response mechanism to salt stress in Arabidopsis transgenic lines over-expressing of GmG6PD[J]. Plant physiology and biochemistry, 2021, 162:74-85. doi: 10.1016/j.plaphy.2021.02.021 URL
[19]	WANG W, DU J, CHEN L, et al. Transcriptomic, proteomic, and physiological comparative analyses of flooding mitigation of the damage induced by low-temperature stress in direct seeded early indica rice at the seedling stage[J]. BMC genomics, 2021, 22(1):176. doi: 10.1186/s12864-021-07458-9 URL
[20]	KESARI P, PATIL D N, KUMAR P, et al. Structural and functional evolution of chitinase-like proteins from plants[J]. Proteomics, 2015, 15(10):1693-1705. doi: 10.1002/pmic.201400421 URL
[21]	BHATTACHARYA D, NAGPURE A, Gupta R K. Bacterial chitinases: properties and potential[J]. Critical reviews in biotechnology, 2007, 27(1):21-28. doi: 10.1080/07388550601168223 URL
[22]	HAN L B, LI Y B, WANG F X, et al. The cotton apoplastic protein CRR1 stabilizes chitinase 28 to facilitate defense against the fungal pathogen Verticillium dahlia[J]. Plant cell, 2019, 31(2):520-536. doi: 10.1105/tpc.18.00390 URL
[23]	NAVARRO-GONZález S S, Ramírez-Trujillo J A, Peña-Chora G, et al. Enhanced tolerance against a fungal pathogen and insect resistance in transgenic tobacco plants overexpressing an endochitinase gene from Serratia marcescens[J]. International journal of molecular sciences, 2019, 20(14):3482.
[24]	YANG X, YANG J, LI H, et al. Overexpression of the chitinase gene CmCH1 from Coniothyrium minitans renders enhanced resistance to Sclerotinia sclerotiorum in soybean[J]. Transgenic research, 2020, 29(2):187-198. doi: 10.1007/s11248-020-00190-2 URL
[25]	王亚, 贺绥欢, 裴越琳, 等. 草莓几丁质酶基因FaChi1-FaChi4的转录特性及其对干旱胁迫、外施脱落酸及灰霉菌的响应[J]. 中国农业大学学报, 2015, 20(6):108-116.
[26]	KASHYAP P, DESWAL R. A novel class I Chitinase from Hippophae rhamnoides: Indications for participating in ICE-CBF cold stress signaling pathway[J]. Plant science, 2017, 259:62-70. doi: 10.1016/j.plantsci.2017.03.004 URL
[27]	周洁, 黄婧. 柳树几丁质酶基因SlChi的克隆和功能验证[J]. 分子植物育种, 2018, 16(24):8013-8021.