CHEN Jing,LU Xiumei,REN Qinqin,et al.Identification of circRNA and their target genes related to resistance to gummy stem blight in melon[J].Journal of Nanjing Agricultural University,2020,43(4):629-636.[doi:10.7685/jnau.201912037]





Identification of circRNA and their target genes related to resistance to gummy stem blight in melon
陈静1 鲁秀梅1 任琴琴1 孙亚亭1 包卫红2 陈雯倩2 钱春桃1
1. 南京农业大学园艺学院/作物遗传与种质创新国家重点实验室, 江苏 南京 210095;
2. 海门市农业科学研究所, 江苏 南通 226100
CHEN Jing1 LU Xiumei1 REN Qinqin1 SUN Yating1 BAO Weihong2 CHEN Wenqian2 QIAN Chuntao1
1. College of Horticulture/State Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095, China;
2. Haimen Agricultural Science Research Institute, Nantong 226100, China
melongummy stem blightcircRNAtarget genes
[目的] 本文旨在通过甜瓜接种蔓枯病菌前、后circRNA的研究及靶基因的筛选验证,探究circRNA在甜瓜抗蔓枯病过程中的作用。[方法] 利用感病品种‘白皮脆’和抗病材料PI420145,分别在接种前、后取样,构建circRNA文库,通过RNA-Seq、Real-time PCR验证以及基因结构分析,探索circRNA在抗蔓枯病过程中的作用。[结果] 12个circRNA文库中共鉴定出3 136个circRNA,过滤后共获得1 511个circRNA。聚类分析结果表明,circRNA的种类和表达具有品种差异性,且接种蔓枯病菌会引起circRNA的变化。接种蔓枯病菌前、后,在PI420145和‘白皮脆’中分别筛选到4个和3个差异表达的circRNA,其中1个circRNA在2种材料中都有差异表达;对6个靶标基因的结构域分析及RT-qPCR分析结果表明,MELO3C022310MELO3C002560MELO3C010763基因与甜瓜蔓枯病抗性相关。[结论] circRNA参与了甜瓜材料PI420145对蔓枯病入侵的防御反应。
[Objectives] The purpose of this article was to investigate the role of circRNA involved in the resistance of melon to gummy stem blight through the study of circRNA and screening of target genes before and after the inoculation of Didymella bryoniae.[Methods] The circRNA libraries were constructed by sampling the susceptible variety ‘Baipicui’ and disease-resistant material PI420145 before and after inoculation,respectively. The RNA-Seq,real-time PCR and gene structure analysis were combined to explore the role of circRNA in the resistance to gummy stem blight.[Results] A total of 3 136 circRNA were identified from 12 libraries,and 1 511 circRNA were resultantly obtained after filtering. Through cluster analysis,it was concluded that the types and expression of circRNA were various among different cultivars and could be changed by the inoculation of D.bryoniae. Before and after inoculation,4 and 3 differentially expressed circRNA were screened in PI420145 and ‘Baipicui’,respectively,and one of which was differentially expressed in both materials. The results of domain analysis and RT-qPCR of the 6 target genes showed that the genes of MELO3C022310,MELO3C002560 and MELO3C010763 were related to resistance of melon gummy stem blight.[Conclusions] The circRNA was involved in the defense response of melon source PI420145 to the invasion of D.bryoniae.


[1] Ye C Y,Chen L,Liu C,et al. Widespread noncoding circular RNAs in plants[J]. New Phytologist,2015,208(1):88-95.
[2] Morris K V,Mattick J S. The rise of regulatory RNA[J]. Nature Reviews Genetics,2014,15(6):423-437.
[3] Wei J,Wei W,Xu H,et al. Circular RNA hsa_circRNA_102958 may serve as a diagnostic marker for gastric cancer[J]. Cancer Biomarkers,2020,27(2):139-145.
[4] Ruan Z B,Chen G C,Zhang R,et al. Circular RNA expression profiles during the differentiation of human umbilical cord-derived mesenchymal stem cells into cardiomyocyte-like cells[J]. Journal of Cellular Physiology,2019,234:16412-16423.
[5] Dou Y,Li S,Yang W,et al. Genome-wide discovery of circular RNAs in the leaf and seedling tissues of Arabidopsis thaliana[J]. Current Genomics,2017,18(4):360-365.
[6] Zhou R,Xu L,Zhao L,et al. Genome-wide identification of circRNAs involved in tomato fruit coloration[J]. Biochemical and Biophysical Research Communications,2018,499(3):466-469.
[7] Cheng J,Zhang Y,Li Z,et al. A lariat-derived circular RNA is required for plant development in Arabidopsis[J]. Science China Life Sciences,2018,61(2):204-213.
[8] Ye C Y,Zhang X,Chu Q,et al. Full-length sequence assembly reveals circular RNAs with diverse non-GT/AG splicing signals in rice[J]. RNA Biology,2017,14(8):1055-1063.
[9] Ghorbani A,Izadpanah K,Peters J R,et al. Detection and profiling of circular RNAs in uninfected and maize Iranian mosaic virus-infected maize[J]. Plant Science,2018,274:402-409.
[10] Wang Z P,Liu Y F,Li D W,et al. Identification of circular RNAs in kiwifruit and their species-specific response to bacterial canker pathogen invasion[J]. Frontiers in Plant Science,2017,8:413.
[11] Wang J Y,Yang Y W,Jin L M,et al. Re-analysis of long non-coding RNAs and prediction of circRNAs reveal their novel roles in susceptible tomato following TYLCV infection[J]. BMC Plant Biology,2018,18(1):104.
[12] 李冰冰,王哲,曹亚兵,等. 丛枝病对白花泡桐环状RNA表达谱变化的影响[J]. 河南农业大学学报,2018,52(3):327-334. Li B B,Wang Z,Cao Y B,et al. Effects on the changes of the expression profiles of circRNA related to paulownia witches’ broom in Paulownia fortunei[J]. Journal of Henan Agricultural University,2018,52(3):327-334(in Chinese with English abstract).
[13] 刘文睿,张永兵,周晓慧,等. 甜瓜抗蔓枯病基因Gsb-1的分子标记及其与抗源PI420145中抗病基因的关系[J]. 中国瓜菜,2009,22(5):1-4. Liu W R,Zhang Y B,Zhou X H,et al. SSR marker linked to gummy stem blight resistance gene Gsb-1 in melon and its allelism with resistance gene from PI420145[J]. China Cucurbits and Vegetables,2009,22(5):1-4(in Chinese with English abstract).
[14] 李英.瓜类蔓枯病菌的生物学特性和黄瓜抗病资源的筛选[D]. 南京:南京农业大学,2007:39-47. Li Y. Study on biology characteristics of Didymella bryoniae and screening of resistance germplasm of cucumber[D]. Nanjing:Nanjing Agricultural University,2007:39-47(in Chinese with English abstract).
[15] Zhang Y P,Kyle M,Anagnostou K,et al. Screening melon(Cucumis melo)for resistance to gummy stem blight in the greenhouse and field[J]. HortScience,1997,32(1):117-121.
[16] Xiang L X,Cai C W,Cheng J R,et al. Identification of circularRNAs and their targets in Gossypium under Verticillium wilt stress based on RNA-seq[J]. Peer J,2018,6(3):e4500.
[17] 张敏.番茄晚疫病密切相关circRNA及其功能初探[D]. 大连:大连理工大学,2018:21-31. Zhang M. The preliminary study of circRNA which is closely related to tomato late blight[D]. Dalian:Dalian University of Technology,2018:21-31(in Chinese with English abstract).
[18] Tugizimana F,Steenkamp P A,Piater L A,et al. Unravelling the metabolic reconfiguration of the post-challenge primed state in Sorghum bicolor responding to Colletotrichum sublineolum infection[J]. Metabolites,2019,9(10):194.
[19] Zhang Y,Zhang F,Huang X Z. Characterization of an Arabidopsis thaliana DUF761-containing protein with a potential role in development and defense responses[J]. Theoretical and Experimental Plant Physiology,2019,31(2):303-316.
[20] Santos-Silva L K,Soares-Costa A,Gerald L T S,et al. Recombinant expression and biochemical characterization of sugarcane legumain[J]. Plant Physiology and Biochemistry,2012,57:181-192.
[21] Hu P,Meng Y,Wise R P. Functional contribution of chorismate synthase,anthranilate synthase,and chorismate mutase to penetration resistance in barley-powdery mildew interactions[J]. Molecular Plant-Microbe Interactions,2009,22(3):311-320.
[22] Sun X Y,Wang L,Ding J C,et al. Integrative analysis of Arabidopsis thaliana transcriptomics reveals intuitive splicing mechanism for circular RNA[J]. FEBS Letters,2016,590(20):3510-3516.
[23] 齐凤坤. 番茄芝麻斑病侵染机理初步探讨[D]. 哈尔滨:东北农业大学,2010:25-26. Qi F K. Tomato Helminthosporium carposaprum infection mechanism of preliminary study[D]. Harbin:Northeast Agricultural University,2010:25-26(in Chinese with English abstract).
[24] Robert-Seilaniantz A,Navarro L,Bari R,et al. Pathological hormone imbalances[J]. Current Opinion in Plant Biology,2007,10(4):372-379.


 MIAO Yongmei,WANG Wanyang,YANG Hailin,et al.Physiological effects of exogenous Ca2+, SA and ABA in alleviating low temperature stress of melon seedlings[J].Journal of Nanjing Agricultural University,2013,36(4):25.[doi:10.7685/j.issn.1000-2030.2013.04.005]
 MA Hua,CHENG Chunyan,XU Jian,et al.Verification of cucumber-sour cucumber introgression lines and screening resistant to gummy stem blight[J].Journal of Nanjing Agricultural University,2015,38(4):369.[doi:10.7685/j.issn.1000-2030.2015.03.003]
 XU Binghua,QIAN Chuntao,WANG Hongying,et al.The expression analysis of defense genes in the genes pyramided melon(Cucumis melo L.)resistance to gummy stem blight[J].Journal of Nanjing Agricultural University,2014,37(4):63.[doi:10.7685/j.issn.1000-2030.2014.05.010]
 BI Yanfei,XU Binghua,GUO Jing,et al.Pyramiding disease resistance genes by marker-assisted selection in melon(Cucumis melo L.)and’Baipicui’breed improvement[J].Journal of Nanjing Agricultural University,2015,38(4):375.[doi:10.7685/j.issn.1000-2030.2015.03.004]
 LU Xiumei,ZHANG Ning,XIA Meiling,et al.Changes of endogenous hormone contents and expression analysis of related genes in melon with different resistance after inoculated with Didymella bryoniae[J].Journal of Nanjing Agricultural University,2018,41(4):248.[doi:10.7685/jnau.201706036]


更新日期/Last Update: 1900-01-01