鸭梨,PbChiIV,表达模式,SA,梨轮纹病菌," /> 鸭梨,PbChiIV,表达模式,SA,梨轮纹病菌,"/> Cloning and Expression of Chitinase <em>PbChiIV</em> Genes from <em>Pyrus bretschneideri</em> ‘Yali’

Scientia Agricultura Sinica ›› 2015, Vol. 48 ›› Issue (11): 2279-2286.doi: 10.3864/j.issn.0578-1752.2015.11.019

• RESEARCH NOTES • Previous Articles    

Cloning and Expression of Chitinase PbChiIV Genes from Pyrus bretschneideri ‘Yali’

LI Peng-peng1, LIU Ning2, ZHANG Yu-xing1, PENG Jian-ying1, DONG Jin-gao2   

  1. 1College of Horticulture, Agricultural University of Hebei, Baoding 071001, Hebei
    2College of Life Science, Agricultural University of Hebei/The Lab of Mycotoxin & Molecular Plant Pathology, Baoding 071001, Hebei
  • Received:2014-10-19 Online:2015-06-01 Published:2015-06-01

RichHTML

1

PDF

680

Abstract: 【Objective】This study aimed to clone a full length cDNA of PbChiIV，fruit and the expression after treatment with the salicylic acid(SA) and Botryosphaeria berengeriana de. Not. f. sp. piricola (Nose) Koganecawa et. Sokwwa.【Method】The full lengthof PbChiIV was obtained from Pyrus bretschneideri ‘Yali’ using specific primers. The similarity of the nucleotide sequence and deduced amino acid sequence was analyzed by using BLAST on NCBI. BiotEdit software was used to compare the amino acid sequence. Phylogenetic tree was constructed by using MEGA6.0. Quantitative real-time PCR (qRT-PCR) was performed to determine the expression pattern of PbChiIV in different tissues and the expression of this gene after treatment with SA and B.berengeriana. f. sp. piricola.【Result】The cDNA lengths of PbChiIV genes was 819 bp (GenBank accession number: KJ872676), encoding 272 amino acids. The deduced amino acid sequence shares 100%, 79%, 73%, 73%, 72%, 72%, 72%, 69%, 68%, 67%, 67%, 65%, 67% and 62% identity with P. pyrifolia ACM45716.1, Populus trichocarpa XP_006376418.1, Vitis vinifera NP_001268173.1, Arabidopsis thaliana CAA74930.1, Medicago sativa ACL36992.1, Medicago truncatula AAR87869.1, Vigna unguiculata CAA61281.1, Corylus heterophylla AEM97876.1, Galega orientalis AAP03085.1, Vitis vinifera NP_001268075.1, Vitis pseudoreticulata ABY66958.1, Vitis vinifera AAB65777.1, Nicotiana tabacum BAF44533.1 and Gossypium barbadense AER29902.1. This gene belongs to IV chitinase gene. Expression analysis showed that the expression of PbChiIV in root was the highest, which was 4.32 and 2.96 times compared with stem and leaf, respectively. Followed by the expression in fruit, which was 2.48 and 1.70 times compared to that of stem and leaf, respectively. In the young fruit and mature fruit, the expression of the gene could be induced by SA and B.berengeriana. f. sp. piricola. The highest expression of the gene was 2.83 and 3.80 times compared with controls after treatment with SA, respectively. The highest expression of the gene was 1.82 and 1.66 times as compared with controls after treatment with pathogen, respectively. The highest expression of the gene was 2.49 and 3.43 times as compared with controls after treatment with SA and pathogen. In addition, the expression was up to the peak at 72, 24 and 72 h after treatment with SA and pathogen, respectively.【Conclusion】PbChiIV may be involved in SA-mediated basic signaling pathway of plant defense responses and disease resistance signal transduction network. It is speculated that PbChiIV is involved in defense responses mediated by B.berengeriana. f. sp. piricola. It might play an important role in disease resistance. from Pyrus bretschneideri ‘Yali’ and detect the expression characteristics of PbChiIV in root, stem, leaf

Key words: Pyrus bretschneideri ‘Yali’, PbChiIV de. Not. f. sp. piricola (Nose) Koganecawa et. Sokwwa), expression pattern, SA, pear ring rot pathogen (Botryosphaeria berengeriana

CLC Number: 

  • 鸭梨
[1]    Leon-Reyes A, Spoel S H, De Lange E S, Abe H, Kobayashi M, Tsuda S, Millenaar F F, Welschen R A M, Ritsema T, Pieterse C M J. Ethylene modulates the role of nonexpression pathogenesis-related genes 1 in cross talk between salicylate and jasmonate signaling. Plant Physiology, 2009, 149: 1797-1809.
[2]    蒋选利, 李振岐, 康振生, 闫海林. 几丁质酶与植物的抗病性. 西北农业学报, 2002, 11(3): 71-75. 
Jiang X L, Li Z Q, Kang Z S, Yan H L. Chitinase and plant disease resistance. Acta Agriculturae Boreali-occidentalis Sinica, 2002, 11(3): 71-75. (in Chinese)
[3]    张福丽, 王占斌, 王志英. 几丁质酶在植物抗真菌病害中的作用. 林业科技, 2006, 31(3): 24-27.
Zhang F L, Wang Z B, Wang Z Y. Chitinase and its function in fungal resistance. Forestry Science Technology, 2006, 31(3): 24-27. (in Chinese)
[4]    Guan Y, Ramalingam S, Nagegowda D, Taylor P W J, Chye M L. Brassica juncea chitinase BjCHI1 inhibits growth of fungal phytopathogens and agglutinates gram-negative bacteria. Journal of Experimental Botany, 2008, 59(12): 3475-3484. 
[5]    张志忠, 吴菁华, 吕柳新, 林义章. 植物几丁质酶及其应用研究进展. 福建农林大学学报: 自然科学版, 2005, 34(4): 494-499.
Zhang Z Z, Wu J H, Lü L X, Lin Y Z. Progress of plant chitinase and its application. Journal of Fujian Agriculture and Forestry University: Natural Science Edition, 2005, 34(4): 494-499. (in Chinese)
[6]    Xiao Y H, Li X B, Yang X Y, Luo M, Hou L, Guo S H, Luo X Y, Pei Y. Cloning and characterization of a balsam pear class Ⅰ chitinase gene (Mcchit1) and its ectopic expression enhances fungal resistance in transgenic plants. Biotechnology & Biochemistry, 2007, 71(5): 1211-1219.
[7]    刘栋峰, 张朝红, 王跃进. 黄冠梨几丁质酶基因家族cDNA全长序列的克隆与分析. 农业生物技术学报, 2009, 17(6): 1042-1049.
Liu D F, Zhang C H, Wang Y J. Full-length cDNA clone and sequence analysis of the chitinase family gene from pear. Journal of Agricultural Biotechnology, 2009, 17(6): 1042-1049. (in Chinese)
[8]    Broglie K E, Biddle P. Cressman R, Broglie R. Functional analysis of DNA sequences responsible for ethylene regulation of a bean chitinase gene in transgenic tobacco. The Plant Cell, 1989, 1: 599-607.
[9]    Bartnicki-Garcia S. Cell wall chemistry, morphogenesis, and taxonomy of fungi. Annual Review Microbiology, 1968, 22: 87-108.
[10]   Brokaert W F, Parijs J V, Allen A K, Peumans W J. Comparison of some molecular, enzymatic and antifungal properties of chitinases from thorn-apple, tobacco and wheat. Physiological and Molecular Plant Pathology, 1988, 33: 319-331.
[11]   Schlumbaum A, Mauch F, Vögeli U, Boller T. Plant chitinases are potent inhibitors of fungal growth. Nature, 1986, 324: 365-367.  
[12]   蔡斌华, 张计育, 高志红, 渠慎春, 佟兆国, 靡林, 乔玉山, 章镇. 一种改良的提取草莓属叶片总RNA的方法. 江苏农业学报, 2008, 24(6): 875-877.
Cai B H, Zhang J Y, Gao Z H, Qu S C, Tong Z G, Mi L, Qiao Y S, Zhang Z. An improved method for isolation of total RNA from the leaves of Fragaria spp. Jiangsu Journal of Agricultural Sciences, 2008, 24(6): 875-877. (in Chinese)
[13]   Livak K J, Schmittgen T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods, 2001, 25: 402-408.
[14]   Samac D A, Hironaka C M, Yallay P E, Shah D M. Isolation and characterization of genes encoding basic and acidic chitinase in Arabidopsis thaliana. Plant Physiology, 1990, 93: 907-914. 
[15]   Lotan T, Ori N, Fluhr R. Pathogenesis-related proteins are developmentally regulated in tobacco flowers. The Plant Cell, 1989, 1: 881-887.
[16]   Memelink J, Linthorst H J M, Schilperoot R A, Hoge J H C. Tobacco genes encoding acidic and basic isoforms of pathogenesis-related proteins display different expression patterns. Plant Molecular Biology, 1990, 14: 119-126.
[17]   Neale A D, Wahleithner J A, Lund M, Bonnett H T, Kelly A, Meeks-Wagner D R, Peacock W J, Dennis E S. Chitinase, β-1,3-glucanase, osmotin, and extensin are expressed in tobacco explants during flower formation. The Plant Cell, 1990, 2: 673-684.
[18]   Creelman R A, Mullet J E. Biosynthesis and action of jasmonates in plants. Annual Review Plant Physiology, 1997, 48: 355-381.
[19]   Dempsey D A, Shah J, Klessing D F. Salicylic acid and disease resistance in plants. Critical Reviews in Plant Sciences, 1999, 18(4): 547-575.
[20]   Van Loon L C, Rep M, Pieterse C M J. Significance of inducible defense-related proteins in infected plants. Annual Review of Phytopathology, 2006, 44: 135-162.
[21]   Von Dahl C C, Baldwin I T. Deciphering the role of ethylene in plant herbivore interactions. Journal of Plant Growth Regulation, 2007, 26: 201-209.
[22]   Glazebrook J. Genes controlling expression of defense responses in Arabidopsis-2001 status. Current Opinion in Plant Biology, 2001, 4: 301-308.        
[23]   Lamb C J, Lawton M A, Dixon R A. Signal and transduction mechanisms for activation of plant defenses against microbial attack. Cell, 1989, 56: 215-224.
[24]   Mohamed F, Lydia F, Masumi I, Hideo I. Expression of potential defense responses of Asian and European pears to infection with Venturia nashicola. Physiological and Molecular Plant Pathology, 2004, 64: 319-330.  
[25]   程曦, 田彩娟, 李爱宁, 邱金龙. 植物与病原微生物互作分子基础的研究进展. 遗传, 2012, 34(2): 134-144.
Cheng X, Tian C J, Li A N, Qiu J L. Advances onmolecular mechanisms of plant-pathogen interactions. Hereditas, 2012, 34(2): 134-144. (in Chinese)
[26]   Dodds P N, Rathjen J P. Plant immunity: Towards an integrated view of plant-pathogen interactions. Nature Review Genetics, 2010, 11: 539-548.
[1] LU XueLi, GILLANI SyedaWajeeha, MENG Chen, LI XiaoBin, SONG YiRu, BAI Yu, WANG JuYing, FENG XiaoFei, LIU ChenChen, LI YiQiang, XU ZongChang. Effects of Different Types of Salt Stress on Seed Germination of Pennisetum alopecuroides and Study on Sodium-Regulated Transcriptome [J]. Scientia Agricultura Sinica, 2026, 59(7): 1400-1419.
[2] ZHANG DongMei, ZHOU XinXin, XIAO GuiLin, ZENG XiangGuo, WANG ChunYan, WANG ZeXian, HAN YongChao. Phenotypic Characteristics of Strawberry Floral Organs in Response to Botrytis cinerea Infection and Methods for Gray Mold Resistance Evaluation [J]. Scientia Agricultura Sinica, 2026, 59(7): 1456-1466.
[3] YUE XiaoYu, ZHAO ShiChen, WANG Qin. The miR-362-3p Regulates the Proliferation and Steroid Hormone Synthesis of Mare Follicular Granulosa Cells by Targeting BMPR2 [J]. Scientia Agricultura Sinica, 2026, 59(7): 1564-1575.
[4] ZHANG ZhiLin, LIU Rong, ZONG XuXiao, HAO XiaoPeng, YANG Tao. Integrated Multi-Stage Evaluation of Salt Tolerance in Vicia faba L. and Itaconic Acid-Mediated Alleviation of Germination-Stage Salt Stress [J]. Scientia Agricultura Sinica, 2026, 59(6): 1172-1188.
[5] LIU Jia, ZHOU ShiQi, ZHOU Yan, SHEN GuangMao. Analysis of the Lethal Activity of Serine Carboxypeptidase DmSCBP1 from Dionaea muscipula Against Bactrocera dorsalis [J]. Scientia Agricultura Sinica, 2026, 59(6): 1244-1254.
[6] CAO HaiShun, ZHOU DongYuan, WANG Rui, SHI ZhaoWan, WU TingQuan, ZHANG ChangYuan. Identification of Short Hypocotyl Cucumber Germplasm Under Low Light Stress and QTL Mapping of the Trait [J]. Scientia Agricultura Sinica, 2026, 59(6): 1286-1301.
[7] ZHAO QingYao, WANG XiaoMing, XING Tong, LI LingYun, XU XingLian, ZHAO Xue. Extraction Optimization, Structural Characterization, and Anticoagulant Activity of Intestinal Polysaccharides from Yellow-Feathered Chickens [J]. Scientia Agricultura Sinica, 2026, 59(6): 1317-1332.
[8] CHEN YaRu, WANG Lei, FU Ming, HUANG Tao, ZHANG Hao, LIANG ZhenHua, PI JinSong, WU Yan. Molecular Mechanism of USP18 Facilitating Ferroptosis by Suppressing GPX4 Ubiquitination and Degradation in Duck Granulosa Cells [J]. Scientia Agricultura Sinica, 2026, 59(5): 1128-1140.
[9] DONG Yu, WU Qian, FENG Xuan, ZHENG YinYing, CUI BaiMing. A Novel Plasmid pEA60 of Erwinia amylovora Enhances the Pathogenicity of Strains by Regulating the Synthesis of Virulence Factors [J]. Scientia Agricultura Sinica, 2026, 59(5): 996-1007.
[10] CHEN Min, JIAO ZiLan, QIAO ChengBin, XU Hao, ZHANG Bi, MA DongHua, KONG WeiRu, WANG JingWen, SONG JiaWei, LUO ChengKe, LI PeiFu, TIAN Lei. Morpho-Physiological Responses and Adaptive Strategies of Rice Germplasm Accessions from Different Subspecies Under Salt Stress [J]. Scientia Agricultura Sinica, 2026, 59(4): 705-722.
[11] CUI ShiYou, CHEN PengJun, MIAO YuanQing, HAN JiJun, SHEN JunMing. Development and Field Evaluation of Glyphosate-Resistant Wheat Germplasm Generated Through EMS Mutagenesis [J]. Scientia Agricultura Sinica, 2026, 59(4): 723-733.
[12] LUO ZhengYing, HU SiZhen, LIN XiuQin, HU Xin, ZHANG Min, XU ChaoHua, LIU XinLong, ZENG QianChun. Identification and Functional Characterization of the PEBP Gene Family in Regulating Flowering Time in Saccharum spontaneum and Saccharum officinarum [J]. Scientia Agricultura Sinica, 2026, 59(4): 734-749.
[13] GUO FuCheng, TANG HaiJiang, HAO XinYi, MA GuoLin, YANG JiuJu, HUANG LinFeng, TIAN Lei, WANG Bin, LUO ChengKe. Effects of Different Irrigation Methods on Water-Salt Transport, Rice Yield, and Water Use Efficiency in Saline Soil in Ningxia [J]. Scientia Agricultura Sinica, 2026, 59(4): 750-764.
[14] WANG YongSheng, NIU Li, WANG ChangJie, MA LiHua, LIAN XiaoXiao, MENG YaXiong, MA XiaoLe, YAO LiRong, ZHANG Hong, YANG Ke, LI BaoChun, WANG HuaJun, SI ErJing, WANG JunCheng. Genome-Wide Association Study and Candidate Gene Identification for Thousand Grain Weight in Winter Wheat [J]. Scientia Agricultura Sinica, 2026, 59(3): 499-514.
[15] PEI HongXia, WANG LuYao, JIANG YaPing, LI ShengMei, GAO JingXia. Identification of Nuclear Male Sterility Genes in Pepper Based on BSA-Seq and Proteomics [J]. Scientia Agricultura Sinica, 2026, 59(3): 637-654.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
京ICP备09089781号-30
Copyright 2018 © Editorial office of Scientia Agricultura Sinica
Tel: 86-010-82106279    E-mail: zgnykx@mail.caas.net.cn
Supported by Beijing Magtech Co.Ltd