白花泡桐和毛泡桐丛枝病发生与蛋白质表达谱变化
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摘要
为深入了解泡桐丛枝病发生与蛋白质变化的关系,以白花泡桐和毛泡桐健康、患丛枝病组培幼苗以及经15mg·L~(-1)和75mg·L~(-1)硫酸二甲酯(DMS)处理的患病组培幼苗为材料,采用iTRAQ技术研究这2个品种、8种材料的蛋白质表达谱变化。结果表明,12种蛋白质与泡桐丛枝病发生密切相关,其主要定位于叶绿体(7个,58.33%)和线粒体(3个,25%),功能与光合作用、蛋白质翻译、抗逆性相关。实时荧光定量PCR验证结果显示部分蛋白质的基因在mRNA的表达水平与iTRAQ测试的蛋白质翻译水平结果一致。研究结果为深入揭示泡桐丛枝病发生机理奠定基础。
In order to understand the relationship between the occurrence of paulownia witches'broom and the protein profile changes of the host,in this study,the technology of isobaric tags for relative and absolute quantitation(iTRAQ)was adopted to analyze the protein changes in the healthy,infected,and 15 mg·L~(-1),75 mg·L~(-1) dimethyl sulfate treated infected tissue culture plantlets of two Paulownia species(P.fortunei and P.tomentosa).A total of 12 differentially abundant proteins closely related to the witches'broom were obtained.Subcellular localization showed that 7 proteins were located in chloroplasts and 3 located in mitochondria,with functions related to photosynthesis,protein translation,and stress resistance.Then the quantitative real-time PCR(qRT-PCR)showed the expression level of the differentially abundant proteins coding genes were consistent with the generated by iTRAQ results.The results would lay the foundation for revealing the mechanism of paulownia witches'broom.
In order to understand the relationship between the occurrence of paulownia witches'broom and the protein profile changes of the host,in this study,the technology of isobaric tags for relative and absolute quantitation(iTRAQ)was adopted to analyze the protein changes in the healthy,infected,and 15 mg·L~(-1),75 mg·L~(-1) dimethyl sulfate treated infected tissue culture plantlets of two Paulownia species(P.fortunei and P.tomentosa).A total of 12 differentially abundant proteins closely related to the witches'broom were obtained.Subcellular localization showed that 7 proteins were located in chloroplasts and 3 located in mitochondria,with functions related to photosynthesis,protein translation,and stress resistance.Then the quantitative real-time PCR(qRT-PCR)showed the expression level of the differentially abundant proteins coding genes were consistent with the generated by iTRAQ results.The results would lay the foundation for revealing the mechanism of paulownia witches'broom.
引文
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[2]姚焕英,高志勇,王君龙.泡桐属植物生物学及药用价值研究[J].渭南师范学院学报,2016,31(16):29-34.YAO H Y,GAO Z Y,WANG J L.The study on the biology and medicinal value of plants in Paulownia[J].Journal of Weinan Normal University,2016,31(16):29-34.(in Chinese)
[3]朱志文,李鹏,黄卫华,等.间作类型对新造泡桐林生长及其土壤养分含量的影响[J].湖南林业科技,2017,44(2):15-19.ZHU Z W,LI P,HUANG W H,et al.Effects on the growth of the newPaulownia fortunei plantation and soil chemical properties responded by different intercropping patterns[J].Hunan Forestry Science&Technology,2017,44(2):15-19.(in Chinese)
[4]陈占宽,杨艳坤,叶金山,等.白花泡桐羟基肉桂酰辅酶A还原酶mRNA全序列克隆及序列分析[J].西北林学院学报,2011,26(4):99-103.CHEN Z K,YANG Y K,YE J S,et al.Cloning and analysis of cinnamoyl-CoA reductase mRNA of Paulownia fortunei[J].Journal of Northwest Forestry University,2011,26(4):99-103.(in Chinese)
[5]胡佳续,唐芳,张莹,等.天津滨海新区泡桐丛枝病植原体16SrDNA基因序列分析[J].中国植保导刊,2016,36(1):5-9.HU J X,TANG F,ZHANG Y,et al.Sequence analysis of 16SrDNA of phytoplasma causing Paulownia witches’-broom in Binhai new district of Tianjin[J].China Plant Protection,2016,36(1):5-9.(in Chinese)
[6]TAWIDIAN P,JAWHARI M,SOBH H,et al.The potential of grafting with selected stone fruit varieties for management of almond witches’broom[J].Phytopathologia Mediterranea,2018,56(3):458-469.
[7]郑建伟,毕会涛,寇首道,等.泡桐丛枝病发生过程中物质变化研究现状[J].河南科学,2000,18(4):405-407.
[8]雷启义,董志,周江菊,等.泡桐丛枝病的分子生物学研究进展[J].安徽农业科学,2008,36(22):9609-9610.LEI Q Y,DONG Z,ZHOU J J,et al.Advance of the molecular research on Paulownia witches’broom disease[J].Journal of Anhui Agricultural Sciences,2008,36(22):9609-9564.(in Chinese)
[9]LIU R,DONG Y P,FAN G Q,et al.Discovery of genes related to witches’broom disease in Paulownia tomentosa×Paulownia fortunei by a de novo assembled transcriptome[J].Plos One,2013,8(11):e80238.
[10]MOU H Q,LU J,ZHU S F,et al.Transcriptomic analysis of Paulowniainfected by Paulownia witches-broom phytoplasma[J].Plos One,2013,8(10):e77217.
[11]FAN G Q,DONG Y P,DENG M J,et al.Plant-pathogen interaction,circadian rhythm,and hormone-related gene expression provide indicators of phytoplasma infection in Paulownia fortunei[J].International Journal of Molecular Sciences,2014,15(12):23141-23162.
[12]CAO X B,Fan G Q,Deng M J,et al.Identification of genes related to Paulownia witches’broom by AFLP and MSAP[J].International Journal of Molecular Sciences,2014,15(8):14669-14683.
[13]FAN G Q,NIU S Y,ZHAO Z L,et al.Identification of microRNAs and their targets in Paulownia fortunei,plants free from phytoplasma pathogen after methyl methane sulfonate treatment[J].Biochimie,2016,127:271-280.
[14]FAN G Q,CAO Y B,DENG M J,et al.Identification and dynamic expression profiling of microRNAs and target genes of Paulownia tomentosa,in response to Paulownia,witches’broom disease[J].Acta Physiologiae Plantarum,2017,39(1):28.
[15]曹亚兵,翟晓巧,邓敏捷,等.泡桐丛枝病发生与代谢组变化的关系[J].林业科学,2017,53(6):85-93.CAO Y B,ZHAI X Q,DENG M J,et al.Relationship between metabolites variation and Paulownia witches’broom[J].Scientia Silvae Sinicae,2017,53(6):85-93.(in Chinese)
[16]WANG Z,ZHAI X Q,CAO Y B,et al.Long non-coding RNAs responsive to witches’broom disease in Paulownia tomentosa[J].Forests,2017,8(9).
[17]FAN G Q,CAO Y B,WANG Z.Regulation of long non-coding RNAs responsive to phytoplasma infection in Paulownia tomentosa[J].International Journal of Genomics,2018,2018(22):1-16.
[18]CAO Y B,FAN G Q,ZHAI X Q,et al.Genome-wide analysis of lncRNAs in Paulownia tomentosa,infected with phytoplasmas[J].Acta Physiologiae Plantarum,2018,40(3):49.
[19]范国强,赵改丽,翟晓巧,等.硫酸二甲酯处理豫杂一号泡桐丛枝病幼苗的形态变化及其SSR分析[J].东北林业大学学报,2011,39(5):30-33.FAN G Q,ZHAO G L,ZHAI X Q,et al.Morphological changes of Paulownia tomentosa×P.fortunei seedlings with witches’broom treated with dimethyl sulphonate and their SSRanalyses[J].Journal of Northeast Forestry University,2011,39(5):30-33.(in Chinese)
[20]赵改丽,赵振利,范国强,等.硫酸二甲酯对白花泡桐丛枝病幼苗形态变化及DNA碱基序列的影响[J].河南农业大学学报,2011,45(3):287-291.ZHAO G L,ZHAO Z L,FAN G Q,et al.Effects of dimethyl sulphonate on the morphological changes of Paulownia fortunei seedlings with witches’broom and their DNA base sequences[J].Journal of Henan Agricultural University,2011,45(3):287-291.(in Chinese)
[21]范国强,赵改丽,翟晓巧,等.硫酸二甲酯对毛泡桐丛枝病幼苗植原体及SSR扩增位点的影响[J].南京林业大学学报:自然科学版,2012,36(3):5-8.FAN G Q,ZHAO G L,ZHAI X Q,et al.Effect of dimethyl sulphate on phytoplasma of Paulownia tomentosa seedling infected by witches’broom and its DNA loci at SSR level[J].Journal of Nanjing Forestry University:Nat.Sci.Edi.,2012,36(3):78-84.(in Chinese)
[22]周凯凯,张胜,赵忠.不同树龄银杏叶片差异蛋白组学研究[J].西北林学院学报,2018,33(1):105-112.ZHOU K K,ZHANG S,ZHAO Z,et al.Proteomic analysis on Ginkgoleaves with different ages[J].Journal of Northwest Forestry University,2018,33(1):105-112.(in Chinese)
[23]张艺萍,瞿素萍,杨秀梅,等.百合抗病无性系抗枯萎病的蛋白质组学分析[J].园艺学报,2018,45(3):530-540.ZHANG Y P,ZHAI S P,YANG X M,et al.Proteomics analysis of lily blight disease-resistant clones[J].Acta Horticulturae Sinica,2018,45(03):530-540.(in Chinese)
[24]ZHANG M,CHENG S T,WANG H Y,et al.iTRAQ-based proteomic analysis of defence responses triggered by the necrotrophic pathogen Rhizoctonia solani in cotton[J].Journal of Proteomics,2016,152:226-235.
[25]NI W C,ZHU L M,SHA R H,et al.Comparative iTRAQproteomic profiling of susceptible and resistant apple cultivars infected by Alternaria alternata apple pathotype[J].Tree Genetics&Genomes,2017,13(1):23.
[26]LIU J,SUI Y,CHEN H Z,et al.Proteomic analysis of Kiwifruit in response to the postharvest pathogen,Botrytis cinerea[J].Frontiers in Plant Science,2018,9:1-15.
[27]范国强,李有,郑建伟,等.泡桐丛枝病发生相关蛋白质的电泳分析[J].林业科学,2003,29(2):119-122.FAN G Q,LI Y,ZHENG J W,et al.SDS-PAGE of proteins related to Paulownia witche’s broom[J].Scientia Silvae Sinicae,2003,29(2):119-122.(in Chinese)
[28]王鹏伟,史俊华.5-氮胞苷和利福平对丛枝病泡桐幼苗形态和蛋白质变化的协同影响[J].河南林业科技,2010,30(3):4-7.WANG P W,SHI J H.Effects of 5-Aza and other extraneous substances on the changes of morphology and protein of Paulowniaseedlings with witche’s broom[J].Journal of Henan Forestry Science and Technology,2010,30(3):4-7.(in Chinese)
[29]WEI Z,WANG Z,LI X Y,et al.Comparative proteomic analysis of Paulownia fortunei response to phytoplasma infection with dimethyl sulfate treatment[J].International Journal of Genomics,2017,2017(3):1-11.
[30]WANG Z,LIU W S,FAN G Q,et al.Quantitative proteomelevel analysis of Paulownia witches’broom disease with methyl methane sulfonate assistance reveals diverse metabolic changes during the infection and recovery processes[J].Peerj,2017,5(7):e3495.
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