Please wait a minute...
Journal of Integrative Agriculture  2017, Vol. 16 Issue (04): 892-899    DOI: 10.1016/S2095-3119(16)61480-6
Plant Protection Advanced Online Publication | Current Issue | Archive | Adv Search |
Identification of the key chitinase genes in Tetranychus cinnabarinus (Boisduval) based on the expression and sequence characteristic analysis
XU Hao-ran, HE Lin, XIAO Wei, SHEN Guang-mao

Key Laboratory of Entomology and Pest Control Engineering, College of Plant Protection, Southwest University,  Chongqing 400715, P.R.China

Download:  PDF in ScienceDirect  
Export:  BibTeX | EndNote (RIS)      
Abstract  Chitin is an important content in the exoskeletons of arthropods, and its hydrolyzation is catalyzed by chitinases during the process of molting, thus, the chitinases are considered as ideal target to interfere the growth of arthropods.  This study intends to clarify the characteristic of the chitinases during the development of Tetranychus cinnabarinus, and screen out important genes as potential control targets.  The results showed that the total enzyme concentration of chitinases was significantly higher in larva, the first and second nymph than that in egg and adult.  Base on the transcriptome data, six unigenes encoding chitinases were identified and their expression patterns in different developmental stages were detected. The expressions of TcCHIT1 and TcCHIT10 showed high abundance during the molting process and their expression change during the developmental stages was consistent with the enzyme concentration.  The full-length of these two genes were further cloned, and the structural characteristics of their proteins were analyzed by constructing the three-dimensional structure model.  The results provide basic information to understand the characteristic of chitinases in T. cinnabarinus and might be considered as target for control.
Keywords:  Tetranychus cinnabarinus      chitinase      gene expression      protein structure  
Received: 16 May 2016   Accepted:
Fund: 

This project was supported by the National Natural Sciences Foundation of China (31401748), the Fundamental Research Funds for the Central Universities of China (XDJK2014C096, 2362015xk04), and the National Student’s Program for Innovation and Entrepreneurship Training Program, China (201510635023).

Corresponding Authors:  SHEN Guang-mao, E-mail: blackaet@163.com    
About author:  XU Hao-ran, E-mail: zgcxhr123@163.com

Cite this article: 

XU Hao-ran, HE Lin, XIAO Wei, SHEN Guang-mao. 2017. Identification of the key chitinase genes in Tetranychus cinnabarinus (Boisduval) based on the expression and sequence characteristic analysis. Journal of Integrative Agriculture, 16(04): 892-899.

Alvarenga E S L, Mansur J F, Justi S A, Figueira-Mansur J, dos Santos V M, Lopez S G, Masuda H, Lara F A, Melo A C A, Moreira M F. 2016. Chitin is a component of the Rhodnius prolixus midgut. Insect Biochemistry and Molecular Biology, 69, 61–70.
Arakane Y, Muthukrishnan S. 2010. Insect chitinase and chitinase-like proteins. Cellular and Molecular Life Sciences, 67, 201–216.
Arnold K, Bordoli L, Kopp J, Schwede T. 2006. The SWISS-MODEL workspace: A web-based environment for protein structure homology modelling. Bioinformatics, 22, 195–201.
Auger P, Migeon A, Ueckermann E A, Tiedt L, Navajas M. 2013. Evidence for synonymy between Tetranychus urticae and Tetranychus cinnabaribus (Acari, prostigmata, Tetranychidae): Review and new data. Acarologia, 53, 383–415.
Bradford M M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72, 248–254.
Chen Q S, Zhao S, Zou J, Shi L, He L. 2012. Monitoring of acaricide resistance in Tetranychus cinnabarinus. Chinese Journal of Applied Entomology, 49, 364–369. (in Chinese)
Cheong S P S, Huang J, Bendena W G, Tobe S S, Hui J H L. 2015. Evolution of ecdysis and metamorphosis in Arthropods: The rise of regulation of juvenile hormone. Integrative and Comparative Biology, 55, 878–890.
Grbi? M, Van Leeuwen T, Clark R M, Rombauts S, Rouzé P, Grbi? V, Osborne E J, Dermauw W, Ngoc P C T, Ortego F. 2011. The genome of Tetranychus urticae reveals herbivorous pest adaptations. Nature, 479, 487–492.
Guo F, Zhang Z, Zhao Z M. 1998. Pesticide resistance of Tetranychus cinnabarinus (Acari: Tetranychidae) in China: A review. Systematic and Applied Acarology, 3, 3–7.
Kariu T, Smith A, Yang X L, Pal U. 2013. A chitin deacetylase-like protein is a predominant constituent of tick peritrophic membrane that influences the persistence of lyme disease pathogens within the vector. PLOS ONE, 8, e78376.
Khajuria C, Buschman L L, Chen M S, Muthukrishnan S, Zhu K Y. 2010. A gut-specific chitinase gene essential for regulation of chitin content of peritrophic matrix and growth of Ostrinia nubilalis larvae. Insect Biochemistry and Molecular Biology, 40, 621–629.
Van Leeuwen T, Demaeght P, Osborne E J, Dermauw W, Gohlke S, Nauen R, Grbic M, Tirry L, Merzendorfer H, Clark R M. 2012. Population bulk segregant mapping uncovers resistance mutations and the mode of action of a chitin synthesis inhibitor in arthropods. Proceedings of the National Academy of Sciences of the United States of America, 109, 4407–4412.
Van Leeuwen T, Dermauw W. 2016. The molecular evolution of xenobiotic metabolism and resistance in chelicerate mites. Annual Review of Entomology, 61, 475–498.
Li D Q, Zhang J Q, Wang Y, Liu X J, Ma E B, Sun Y, Li S, Zhu K Y, Zhang J Z. 2015. Two chitinase 5 genes from Locusta migratoria: Molecular characteristics and functional differentiation. Insect Biochemistry and Molecular Biology, 58, 46–54.
Merzendorfer H, Zimoch L. 2003. Chitin metabolism in insects: structure, function and regulation of chitin synthases and chitinases. Journal of Experimental Biology, 206, 4393–4412.
Muthukrishnan S, Merzendorfer H, Arakane Y, Kramer K J. 2012. Chitin metabolism in insects. In: Gilbert L I, ed., Insect Molecular Biology and Biochemistry. Academic Press, San Diego. pp. 193–235.
Von Ohlen T, Luce-Fedrow A, Ortega M T, Ganta R R, Chapes S K. 2012. Identification of critical host mitochondrion-associated genes during Ehrlichia chaffeensis infections. Infection and Immunity, 80, 3576–3586.
Osman G H, Assem S K, Alreedy R M, El-Ghareeb D K, Basry M A, Rastogi A, Kalaji H M. 2016. Development of insect resistant maize plants expressing a chitinase gene from the cotton leaf worm, Spodoptera littoralis. Scitific Reports, 5, 18067.
Pesch Y Y, Riedel D, Patil K R, Loch G, Behr M. 2016. Chitinases and imaginal disc growth factors organize the extracellular matrix formation at barrier tissues in insects. Scientific Report, 6, 18340.
Reynolds S E, Samuels R I. 1996. Physiology and biochemistry of insect molting fluid. Annual Review of Insect Physiology, 26, 157–232.
Schmittgen T D, Livak K J. 2008. Analyzing real-time PCR data by the comparative CT method. Nature Protocols, 3, 1101–1108.
Shen G M, Shi L, Xu Z F, He L. 2014. Inducible expression of mu-class glutathione S-transferases is associated with fenpropathrin resistance in Tetranychus cinnabarinus. International Journal of Molecular Sciences, 15, 22626–22641.
Shi L, Xu Z F, Shen G M, Song C G, Wang Y, Peng J F, Zhang J, He L. 2015a. Expression characteristics of two novel cytochrome P450 genes involved in fenpropathrin resistance in Tetranychus cinnabarinus (Boisduval). Pesticide Biochemistry and Physiology, 119, 33–41.
Shi L, Zhang J, Shen G, Xu Z, Wei P, Zhang Y, Xu Q, He L. 2015b. Silencing NADPH-cytochrome P450 reductase results in reduced acaricide resistance in Tetranychus cinnabarinus (Boisduval). Scientific Reports, 5, 15581.
Sun W, Jin Y, He L, Lu W C, Li M. 2010. Suitable reference gene selection for different strains and developmental stages of the carmine spider mite, Tetranychus cinnabarinus, using quantitative real-time PCR. Journal of Insect Science, 10, 208.
Tamura K, Dudley J, Nei M, Kumar S. 2007. MEGA4: Molecular evolutionary genetics analysis (MEGA) software version 4.0. Molecular Biology and Evolution, 24, 1596–1599.
Tetreau G, Cao X L, Chen Y R, Muthukrishnan S, Jiang H B, Blissard G W, Kanost M R, Wang P. 2015. Overview of chitin metabolism enzymes in Manduca sexta: Identification, domain organization, phylogenetic analysis and gene expression. Insect Biochemistry and Molecular Biology, 62, 114–126.
Wang Y, Zhao S, Shi L, Xu Z, He L. 2014. Resistance selection and biochemical mechanism of resistance against cyflumetofen in Tetranychus cinnabarinus (Boisduval). Pesticide Biochemistry and Physiology, 111, 24–30.
Xi Y, Pan P L, Ye Y X, Yu B, Xu H J, Zhang C X. 2015. Chitinase-like gene family in the brown planthopper, Nilaparvata lugens. Insect Molecular Biology, 24, 29–40.
Xi Y, Pan P L, Ye Y X, Yu B, Zhang C X. 2014. Chitin deacetylase family genes in the brown planthopper, Nilaparvata lugens (Hemiptera: Delphacidae). Insect Molecular Biology, 23, 695–705.
Xia W K, Ding T B, Niu J Z, Liao C Y, Zhong R, Yang W J, Liu B, Dou W, Wang J J. 2014. Exposure to diflubenzuron results in an up-regulation of a chitin synthase 1 gene in citrus red mite, Panonychus citri (Acari: Tetranychidae). International Journal of Molecular Sciences, 15, 3711–3728.
Xu Z F, Zhu W Y, Liu Y C, Liu X, Chen Q S, Peng M, Wang X Z, Shen G M, He L. 2014. Analysis ofinsecticide resistance-related genes of the carmine spider mite Tetranychus cinnabarinus based on a de novo assembled transcriptome. PLOS ONE, 9, e94779.
Zhang J, Zhang X, Arakane Y, Muthukrishnan S, Kramer K J, Ma E, Zhu K Y. 2011. Comparative genomic analysis of chitinase and chitinase-like genes in the african malaria mosquito (Anopheles gambiae), PLoS ONE, 6, e19899.
Zhu K Y, Merzendorfer H, Zhang W, Zhang J, Muthukrishnan S. 2016. Biosynthesis, turnover, and functions of chitin in insects. Annual Review of Entomology, 61, 177–196.
Zhu Q S, Arakane Y, Banerjee D, Beeman R W, Kramer K J, Muthukrishnan S. 2008a. Domain organization and phylogenetic analysis of the chitinase-like family of proteins in three species of insects. Insect Biochemistry and Molecular Biology, 38, 452–466.
Zhu Q S, Arakane Y, Beeman R W, Kramer K J, Muthukrishnan S. 2008b. Functional specialization among insect chitinase family genes revealed by RNA interference. Proceedings of the National Academy of Sciences of the United States of America, 105, 6650–6655.
[1] ZHAO Shu-ping, DENG Kang-ming, ZHU Ya-mei, JIANG Tao, WU Peng, FENG Kai, LI Liang-jun.

Optimization of slow-release fertilizer application improves lotus rhizome quality by affecting the physicochemical properties of starch [J]. >Journal of Integrative Agriculture, 2023, 22(4): 1045-1057.

[2] ZHANG Yan-mei, AO De, LEI Kai-wen, XI Lin, Jerry W SPEARS, SHI Hai-tao, HUANG Yan-ling, YANG Fa-long. Dietary copper supplementation modulates performance and lipid metabolism in meat goat kids[J]. >Journal of Integrative Agriculture, 2023, 22(1): 214-221.
[3] JIANG Yong, MA Xin-yan, XIE Ming, ZHOU Zheng-kui, TANG Jing, CHANG Guo-bin, CHEN Guo-hong, HOU Shui-sheng. Dietary threonine deficiency affects expression of genes involved in lipid metabolism in adipose tissues of Pekin ducks in a genotype-dependent manner[J]. >Journal of Integrative Agriculture, 2022, 21(9): 2691-2699.
[4] RONG Hao, YANG Wen-jing, XIE Tao, WANG Yue, WANG Xia-qin, JIANG Jin-jin, WANG You-ping. Transcriptional profiling between yellow- and black-seeded Brassica napus reveals molecular modulations on flavonoid and fatty acid content[J]. >Journal of Integrative Agriculture, 2022, 21(8): 2211-2226.
[5] AN Feng, ZHANG Kang, ZHANG Ling-kui, LI Xing, CHEN Shu-min, WANG Hua-sen, CHENG Feng. Genome-wide identification, evolutionary selection, and genetic variation of DNA methylation-related genes in Brassica rapa and Brassica oleracea[J]. >Journal of Integrative Agriculture, 2022, 21(6): 1620-1632.
[6] FAN Xiao-xue, BIAN Zhong-hua, SONG Bo, XU Hai. Transcriptome analysis reveals the differential regulatory effects of red and blue light on nitrate metabolism in pakchoi (Brassica campestris L.)[J]. >Journal of Integrative Agriculture, 2022, 21(4): 1015-1027.
[7] LIU Cong, LI De-xiong, HUANG Xian-biao, Zhang Fu-qiong, Xie Zong-zhou, Zhang Hong-yan, Liu Ji-hong. Manual thinning increases fruit size and sugar content of Citrus reticulata Blanco and affects hormone synthesis and sugar transporter activity[J]. >Journal of Integrative Agriculture, 2022, 21(3): 725-735.
[8] DUAN Yao-ke, HAN Rong, SU Yan, WANG Ai-ying, LI Shuang, SUN Hao, GONG Hai-jun. Transcriptional search to identify and assess reference genes for expression analysis in Solanum lycopersicum under stress and hormone treatment conditions[J]. >Journal of Integrative Agriculture, 2022, 21(11): 3216-3229.
[9] Kashif NOOR, Hafiza Masooma Naseer CHEEMA, Asif Ali KHAN, Rao Sohail Ahmad KHAN. Expression profiling of transgenes (Cry1Ac and Cry2A) in cotton genotypes under different genetic backgrounds[J]. >Journal of Integrative Agriculture, 2022, 21(10): 2818-2832.
[10] WANG Pei-pei, WANG Zhao-ke, GUAN Le, Muhammad Salman HAIDER, Maazullah NASIM, YUAN Yong-bing, LIU Geng-sen, LENG Xiang-peng. Versatile physiological functions of the Nudix hydrolase family in berry development and stress response in grapevine[J]. >Journal of Integrative Agriculture, 2022, 21(1): 91-112.
[11] GUO Bing-bing, LI Jia-ming, LIU Xing, QIAO Xin, Musana Rwalinda FABRICE, WANG Peng, ZHANG Shao-ling, WU Ju-you. Identification and expression analysis of the PbrMLO gene family in pear, and functional verification of PbrMLO23[J]. >Journal of Integrative Agriculture, 2021, 20(9): 2410-2423.
[12] JI Xiao-hao, WANG Bao-liang, WANG Xiao-di, WANG Xiao-long, LIU Feng-zhi, WANG Hai-bo. Differences of aroma development and metabolic pathway gene expression between Kyoho and 87-1 grapes[J]. >Journal of Integrative Agriculture, 2021, 20(6): 1525-1539.
[13] CHEN Chang-long, YUAN Fang, LI Xiao-ying, MA Rong-cai, XIE Hua. Jasmonic acid and ethylene signaling pathways participate in the defense response of Chinese cabbage to Pectobacterium carotovorum infection[J]. >Journal of Integrative Agriculture, 2021, 20(5): 1314-1326.
[14] WANG Lu-lu, ZHAO Chun-fang, LIU Chang-jun, ZHANG Hao, LIAN Ling. Analysis of DNA methylation of CD79B in MDV-infected chicken spleen[J]. >Journal of Integrative Agriculture, 2021, 20(11): 2995-3002.
[15] WANG Xi-cheng, WU Wei-min, ZHOU Bei-bei, WANG Zhuang-wei, QIAN Ya-ming, WANG Bo, YAN Li-chun. Genome-wide analysis of the SCPL gene family in grape (Vitis vinifera L.)[J]. >Journal of Integrative Agriculture, 2021, 20(10): 2666-2679.
No Suggested Reading articles found!