杨树受溃疡病菌侵染初期的转录组
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摘要
欧美杨细菌性溃疡病是一种对杨树人工林造成严重损害的新型枝干病害。为探讨杨树抵御该病害的分子机制,分别以染病初期和未染病的中林46杨(Populus×euramericana‘Zhonglin 46’)树皮组织为材料开展了转录组测定和分析。基因差异表达结果表明,2 597个基因在中林46杨受到溃疡病菌侵染初期发生变化,其中1 412个基因表达上调,1 185个基因表达下调。表达量变化最大的前20个基因包含了编码锌指蛋白、漆酶、内切几丁质酶、胰蛋白与蛋白酶抑制剂等与抗病相关的基因。差异表达基因的GO注释和KEGG通路富集性分析进一步表明,类黄酮和苯丙烷类物质在杨树抵御病原菌侵染的初期起了一定作用。有趣的是,发现维生素B6代谢、有机含硒化合物代谢和苯并恶嗪生物合成通路等抗病相关通路的差异表达基因在染病初期全部或绝大部分受到诱导。
Lonsdalea canker of poplar is a new type of canker disease that causes severe damage to poplar plantation.To explore the molecular defense mechanism of poplar in response to Lonsdalea canker,this investigation performed transcriptome sequencing of the bark of Populus euramericana‘Zhonglin 46'infected or not with Lonsdalea quercina subsp.populi.The results showed that 2 597 genes of poplars were significantly changed at the early stage of infection of poplars.Among these genes,1 412 genes were up-regulated while 1 185 genes were down-regulated.Twenty genes with the most significant differential expression abundance included genes encoding zinc finger proteins,laccase,endochitinase,trypsin and protease inhibitor and other genes related with disease resistance.GO annotation and the enrichment analysis of the KEGG pathway of the differentially expressed genes(DEGs) showed that flavonoid and phenylpropanoid should be involved in the defense response of poplar to pathogen infection at the early stage.Interestingly,we found that all or most of the DEGs involved in resistance-related pathways such as Vitamin B6 metabolism,selenocompound metabolism and benzoxazinoid biosynthesis pathways of poplar were induced at the early stage of the infection.
Lonsdalea canker of poplar is a new type of canker disease that causes severe damage to poplar plantation.To explore the molecular defense mechanism of poplar in response to Lonsdalea canker,this investigation performed transcriptome sequencing of the bark of Populus euramericana‘Zhonglin 46'infected or not with Lonsdalea quercina subsp.populi.The results showed that 2 597 genes of poplars were significantly changed at the early stage of infection of poplars.Among these genes,1 412 genes were up-regulated while 1 185 genes were down-regulated.Twenty genes with the most significant differential expression abundance included genes encoding zinc finger proteins,laccase,endochitinase,trypsin and protease inhibitor and other genes related with disease resistance.GO annotation and the enrichment analysis of the KEGG pathway of the differentially expressed genes(DEGs) showed that flavonoid and phenylpropanoid should be involved in the defense response of poplar to pathogen infection at the early stage.Interestingly,we found that all or most of the DEGs involved in resistance-related pathways such as Vitamin B6 metabolism,selenocompound metabolism and benzoxazinoid biosynthesis pathways of poplar were induced at the early stage of the infection.
引文
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[8] FANG Z, ZHOU D, YE X, et al. Identification of candidate anthocyanin-related genes by transcriptomic analysis of ‘Furongli’ plum (Prunus salicina Lindl.) during fruit ripening using RNA-Seq[J]. Frontiers in Plant Science,2016,7.doi:10.3389/fpls.2016.01338.
[9] XIE R, PAN X, ZHANG J, et al. Effect of salt-stress on gene expression in citrus roots revealed by RNA-seq[J]. Functional & Integrative Genomics,2018,18(2):155-173.
[10] 侯佳,孙丰硕,吴秋明,等.一种高效提取杨树发病树皮总RNA的方法及应用[J].植物生理学报,2014,50(2):223-228.
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[16] SUGANO S, KAMINAKA H, RYBKA Z, et al. Stress-responsive zinc finger gene ZPT2-3 plays a role in drought tolerance in petunia[J]. The Plant Journal,2003,36(6):830-841.
[17] IIDA A, KAZUOKA T, TORIKAI S, et al. A zinc finger protein RHL41 mediates the light acclimatization response in Arabidopsis[J]. The Plant Journal,2000,24(2):191-203.
[18] MUKHOPADHYAY A, VIJ S, TYAGI A K. Overexpression of a zinc-finger protein gene from rice confers tolerance to cold, dehydration, and salt stress in transgenic tobacco[J]. Proceedings of the National Academy of Sciences,2004,101(16):6309-6314.
[19] LIU H, ZHANG H, YANG Y, et al. Functional analysis reveals pleiotropic effects of rice RING-H2 finger protein gene OsBIRF1 on regulation of growth and defense responses against abiotic and biotic stresses[J]. Plant Molecular Biology,2008,68(1/2):17-30.
[20] LI L, STEFFENS J. Overexpression of polyphenol oxidase in transgenic tomato plants results in enhanced bacterial disease resistance[J]. Planta,2002,215(2):239-247.
[21] SCHLUMBAUM A, MAUCH F, V?GELI U, et al. Plant chitinases are potent inhibitors of fungal growth[J]. Nature,1986,324:365-367.
[22] DING X, GOPALAKRISHNAN B, JOHNSON L B, et al. Insect resistance of transgenic tobacco expressing an insect chitinase gene[J]. Transgenic Res,1998,7(2):77-84.
[23] HILDER V A, GATEHOUSE A M R, SHEERMAN S E, et al. A novel mechanism of insect resistance engineered into tobacco[J]. Nature,1987,330:160-163.
[24] GRATIVOL C, HEMERLY A S, FERREIRA P C G. Genetic and epigenetic regulation of stress responses in natural plant populations[J]. Biochimica et Biophysica Acta,2012,1819(2):176-185.
[25] MIRANDA M, RALPH S G, MELLWAY R, et al. The transcriptional response of hybrid poplar (Populus trichocarpa×P. deltoides) to infection by Melampsora medusae leaf rust involves induction of flavonoid pathway genes leading to the accumulation of proanthocyanidins[J]. MPMI,2007,20(7):816-831.
[26] CARLSEN S C K, UNDERSTRUP A, FOMSGAARD I S, et al. Flavonoids in roots of white clover: interaction of arbuscular mycorrhizal fungi and a pathogenic fungus[J]. Plant and Soil,2008,302(1/2):33-43.
[27] VOGT T. Phenylpropanoid biosynthesis[J]. Molecular Plant,2010,3(1):2-20.
[28] DENSLOW S A, RUESCHHOFF E E, DAUB M E. Regulation of the Arabidopsis thaliana vitamin B6 biosynthesis genes by abiotic stress[J]. Plant Physiology and Biochemistry,2007,45(2):152-161.
[29] HAVAUX M, KSAS B, SZEWCZYK A, et al. Vitamin B6 deficient plants display increased sensitivity to high light and photo-oxidative stress[J]. BMC Plant Biol,2009,9.doi:10.1186/1471-2229-9-130.
[30] DANON A, MIERSCH O, FELIX G, et al. Concurrent activation of cell death-regulating signaling pathways by singlet oxygen in Arabidopsis thaliana[J]. The Plant Journal,2005,41(1):68-80.
[31] TITIZ O, TAMBASCO-STUDART M, WARZYCH E, et al. PDX1 is essential for vitamin B6 biosynthesis, development and stress tolerance in Arabidopsis[J]. The Plant Journal,2006,48(6):933-946.
[32] KUMAR M, BIJO A J, BAGHEL R S, et al. Selenium and spermine alleviate cadmium induced toxicity in the red seaweed Gracilaria dura by regulating antioxidants and DNA methylation[J]. Plant Physiology and Biochemistry,2012,51:129-138.
[33] NIEMEYER H M. Hydroxamic acids (4-hydroxy-1,4-benzoxazin-3-ones), defence chemicals in the gramineae[J]. Phytochemistry,1988,27(11):3349-3358.
[34] 刘小红,李健强,周立刚,等.丁布对小麦赤霉病菌和玉米小斑病菌的抑制作用[J].菌物学报,2004,23(1):109-114.
[35] SINGH S K, ELAND C, HARHOLT J, et al. Cell adhesion in Arabidopsis thaliana is mediated by ectopically parting cells 1-a glycosyltransferase (GT64) related to the animal exostosins[J]. The Plant Journal,2005,43(3):384-397.
[36] O’DONOGHUE E M, SOMERFIELD S D, WATSON L M, et al. Galactose metabolism in cell walls of opening and senescing petunia petals[J]. Planta,2008,229(3):709-721.
[2] LI Y, HE W, REN F, et al. A canker disease of Populus×euramericana in China caused by Lonsdalea quercina subsp. populi[J]. Plant Disease,2013,98(3):368-378.
[3] YANG R, DENG C, WEI J, et al. A large-scale mutational analysis of two-component signaling systems of Lonsdalea quercina revealed that KdpD-KdpE regulates bacterial virulence against host poplar trees[J]. Molecular Plant-Microbe Interactions,2018,31(7): 724-736.
[4] 刘振阳,苏衍修,左涛,等.溃疡病不同抗性杨树SA和JA信号转导相关基因的差异表达分析[J].中国农学通报,2015,31(14):156-163.
[5] 左涛,赵树堂,卢孟柱,等.杨树二氢黄酮醇-4-还原酶基因(DFR)的克隆及反义表达对儿茶素合成的影响[J].东北林业大学学报,2016,44(10):49-55.
[6] 左涛,包海,陈慧,等.杨树LAR基因的克隆及表达对儿茶素合成的影响[J].中南林业科技大学学报,2016,36(12):121-128.
[7] SOCQUET-JUGLARD D, KAMBER T, POTHIER J F, et al. Comparative RNA-seq analysis of early-infected peach leaves by the invasive phytopathogen Xanthomonas arboricola pv. pruni[J]. PLoS One,2013,8(1).doi:10.1371/journal.pone.0054196.
[8] FANG Z, ZHOU D, YE X, et al. Identification of candidate anthocyanin-related genes by transcriptomic analysis of ‘Furongli’ plum (Prunus salicina Lindl.) during fruit ripening using RNA-Seq[J]. Frontiers in Plant Science,2016,7.doi:10.3389/fpls.2016.01338.
[9] XIE R, PAN X, ZHANG J, et al. Effect of salt-stress on gene expression in citrus roots revealed by RNA-seq[J]. Functional & Integrative Genomics,2018,18(2):155-173.
[10] 侯佳,孙丰硕,吴秋明,等.一种高效提取杨树发病树皮总RNA的方法及应用[J].植物生理学报,2014,50(2):223-228.
[11] LI R, YU C, LI Y, et al. SOAP2: an improved ultrafast tool for short read alignment[J]. Bioinformatics,2009,25(15):1966-1967.
[12] MORTAZAVI A, WILLIAMS B A, MCCUE K, et al. Mapping and quantifying mammalian transcriptomes by RNA-Seq[J]. Nature Methods,2008,5(7):621-628.
[13] AUDIC S, CLAVERIE J M. The significance of digital gene expression profiles[J]. Genome Res,1997,7(10):986-995.
[14] CONESA A, GOTZ S, GARCIA-GOMEZ J M, et al. Blast2GO: a universal tool for annotation, visualization and analysis in functional genomics research[J]. Bioinformatics,2005,21(18):3674-3676.
[15] KANEHISA M, ARAKI M, GOTO S, et al. KEGG for linking genomes to life and the environment[J]. Nucleic Acids Research,2008,36:D480-D484.doi:10.1093/nar/gkm882.
[16] SUGANO S, KAMINAKA H, RYBKA Z, et al. Stress-responsive zinc finger gene ZPT2-3 plays a role in drought tolerance in petunia[J]. The Plant Journal,2003,36(6):830-841.
[17] IIDA A, KAZUOKA T, TORIKAI S, et al. A zinc finger protein RHL41 mediates the light acclimatization response in Arabidopsis[J]. The Plant Journal,2000,24(2):191-203.
[18] MUKHOPADHYAY A, VIJ S, TYAGI A K. Overexpression of a zinc-finger protein gene from rice confers tolerance to cold, dehydration, and salt stress in transgenic tobacco[J]. Proceedings of the National Academy of Sciences,2004,101(16):6309-6314.
[19] LIU H, ZHANG H, YANG Y, et al. Functional analysis reveals pleiotropic effects of rice RING-H2 finger protein gene OsBIRF1 on regulation of growth and defense responses against abiotic and biotic stresses[J]. Plant Molecular Biology,2008,68(1/2):17-30.
[20] LI L, STEFFENS J. Overexpression of polyphenol oxidase in transgenic tomato plants results in enhanced bacterial disease resistance[J]. Planta,2002,215(2):239-247.
[21] SCHLUMBAUM A, MAUCH F, V?GELI U, et al. Plant chitinases are potent inhibitors of fungal growth[J]. Nature,1986,324:365-367.
[22] DING X, GOPALAKRISHNAN B, JOHNSON L B, et al. Insect resistance of transgenic tobacco expressing an insect chitinase gene[J]. Transgenic Res,1998,7(2):77-84.
[23] HILDER V A, GATEHOUSE A M R, SHEERMAN S E, et al. A novel mechanism of insect resistance engineered into tobacco[J]. Nature,1987,330:160-163.
[24] GRATIVOL C, HEMERLY A S, FERREIRA P C G. Genetic and epigenetic regulation of stress responses in natural plant populations[J]. Biochimica et Biophysica Acta,2012,1819(2):176-185.
[25] MIRANDA M, RALPH S G, MELLWAY R, et al. The transcriptional response of hybrid poplar (Populus trichocarpa×P. deltoides) to infection by Melampsora medusae leaf rust involves induction of flavonoid pathway genes leading to the accumulation of proanthocyanidins[J]. MPMI,2007,20(7):816-831.
[26] CARLSEN S C K, UNDERSTRUP A, FOMSGAARD I S, et al. Flavonoids in roots of white clover: interaction of arbuscular mycorrhizal fungi and a pathogenic fungus[J]. Plant and Soil,2008,302(1/2):33-43.
[27] VOGT T. Phenylpropanoid biosynthesis[J]. Molecular Plant,2010,3(1):2-20.
[28] DENSLOW S A, RUESCHHOFF E E, DAUB M E. Regulation of the Arabidopsis thaliana vitamin B6 biosynthesis genes by abiotic stress[J]. Plant Physiology and Biochemistry,2007,45(2):152-161.
[29] HAVAUX M, KSAS B, SZEWCZYK A, et al. Vitamin B6 deficient plants display increased sensitivity to high light and photo-oxidative stress[J]. BMC Plant Biol,2009,9.doi:10.1186/1471-2229-9-130.
[30] DANON A, MIERSCH O, FELIX G, et al. Concurrent activation of cell death-regulating signaling pathways by singlet oxygen in Arabidopsis thaliana[J]. The Plant Journal,2005,41(1):68-80.
[31] TITIZ O, TAMBASCO-STUDART M, WARZYCH E, et al. PDX1 is essential for vitamin B6 biosynthesis, development and stress tolerance in Arabidopsis[J]. The Plant Journal,2006,48(6):933-946.
[32] KUMAR M, BIJO A J, BAGHEL R S, et al. Selenium and spermine alleviate cadmium induced toxicity in the red seaweed Gracilaria dura by regulating antioxidants and DNA methylation[J]. Plant Physiology and Biochemistry,2012,51:129-138.
[33] NIEMEYER H M. Hydroxamic acids (4-hydroxy-1,4-benzoxazin-3-ones), defence chemicals in the gramineae[J]. Phytochemistry,1988,27(11):3349-3358.
[34] 刘小红,李健强,周立刚,等.丁布对小麦赤霉病菌和玉米小斑病菌的抑制作用[J].菌物学报,2004,23(1):109-114.
[35] SINGH S K, ELAND C, HARHOLT J, et al. Cell adhesion in Arabidopsis thaliana is mediated by ectopically parting cells 1-a glycosyltransferase (GT64) related to the animal exostosins[J]. The Plant Journal,2005,43(3):384-397.
[36] O’DONOGHUE E M, SOMERFIELD S D, WATSON L M, et al. Galactose metabolism in cell walls of opening and senescing petunia petals[J]. Planta,2008,229(3):709-721.
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