菜心BrAGL24基因mRNA在嫁接体中的长距离运输分析
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摘要
为研究结球甘蓝/菜心嫁接体中开花调控基因的mRNA是否从砧木向接穗进行了运输,本试验以参与开花调控的菜心AGL24基因为研究对象,构建结球甘蓝菜心异源嫁接体,并从接穗结球甘蓝中鉴定菜心AGL24的序列。结果表明,利用结球甘蓝和菜心的转录组测序数据,拼接获得了结球甘蓝和菜心的AGL24基因序列(BoAGL24和BrAGL24)。利用菜心BrAGL24基因中的种间差异序列(C1～C7),在异源嫁接(结球甘蓝/菜心嫁接)的接穗结球甘蓝茎尖的转录组测序文库(T2)中筛选得到2条来自砧木菜心BrAGL24的reads。利用结球甘蓝BoAGL24的种间差异序列(G1～G7),分析结球甘蓝内源BoAGL24基因的转录表达量,发现异源嫁接的结球甘蓝茎尖中AGL24的表达量总体上高于同源嫁接的结球甘蓝茎尖中AGL24的表达量。异源嫁接使砧木菜心BrAGL24基因的mRNA运输到接穗结球甘蓝中,且增加了接穗结球甘蓝茎尖中内源BoAGL24基因的转录表达水平。本研究鉴定了菜心BrAGL24基因的mRNA运输特性,为进一步研究AGL24基因的mRNA运输与结球甘蓝/菜心嫁接启动接穗结球甘蓝开花机制奠定了一定的理论基础。
To investigate whether mRNAs of flowering-regulated genes transport from rootstock to scion in head cabbageChinese flowering cabbage heterograft,flowering-regulated AGL24 was selected to analyze mRNA transport between rootstock and scion. Head cabbage-Chinese flowing cabbage heterograft was constructed,and AGL24 of Chinese flowing cabbage was indentified from shoot pex from head cabbage scion. Based on transcriptome sequencing data of head cabbage and Chinese flowering cabbage,AGL24 gene sequences of head cabbage and Chinese flowering cabbage were obtained and named BoAGL24 and BrAGL24,respectively. With interspecific differential sequences of BrAGL24 of Chinese flowering cabbage( C1-C7) two BrAGL24 reads of Chinese flowering cabbage were identified from shoot apex RNA-seq library from heterograft( T2). The endogenous transcriptional expression level of BoAGL24 was analyzed based on interspecific differential sequences of BoAGL24( G1-G7),results indicated the expression level of BoAGL24 was higher in T2 than that in T1. Heterograft not only caused mRNA transport of the BrAGL24 gene from rootstock to scion but also increased the transcriptional expression of the endogenous BoAGL24 in stem apex. It is the first study that identify mRNA transport of BrAGL24 gene of Chinese flowering cabbage,which lays the foundation for further study of molecule mechanism related to AGL24 mRNA transport and promoting scion flowering caused by head cabbage-Chinese flowering cabbage heterograft.
To investigate whether mRNAs of flowering-regulated genes transport from rootstock to scion in head cabbageChinese flowering cabbage heterograft,flowering-regulated AGL24 was selected to analyze mRNA transport between rootstock and scion. Head cabbage-Chinese flowing cabbage heterograft was constructed,and AGL24 of Chinese flowing cabbage was indentified from shoot pex from head cabbage scion. Based on transcriptome sequencing data of head cabbage and Chinese flowering cabbage,AGL24 gene sequences of head cabbage and Chinese flowering cabbage were obtained and named BoAGL24 and BrAGL24,respectively. With interspecific differential sequences of BrAGL24 of Chinese flowering cabbage( C1-C7) two BrAGL24 reads of Chinese flowering cabbage were identified from shoot apex RNA-seq library from heterograft( T2). The endogenous transcriptional expression level of BoAGL24 was analyzed based on interspecific differential sequences of BoAGL24( G1-G7),results indicated the expression level of BoAGL24 was higher in T2 than that in T1. Heterograft not only caused mRNA transport of the BrAGL24 gene from rootstock to scion but also increased the transcriptional expression of the endogenous BoAGL24 in stem apex. It is the first study that identify mRNA transport of BrAGL24 gene of Chinese flowering cabbage,which lays the foundation for further study of molecule mechanism related to AGL24 mRNA transport and promoting scion flowering caused by head cabbage-Chinese flowering cabbage heterograft.
引文
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[22] ParkinⅠA P,Koh C,Tang H,Robinson S J,Kagale S,Clarke WE,Town C D,Nixon J,KrishnakumarⅤ,Bidwell S L,DenoeudF,Belcram H,Links M G,Just J,Clarke C,Bender T,HuebertT,Mason A S,Pires J C,Barker G,Moore J,Walley P G,ManoliS,Batley J,Edwards D,Nelson M N,Wang X,Paterson A H,King G,BancroftⅠ,Chalhoub B,Sharpe A G. Transcriptome andmethylome profiling reveals relics of genome dominance in themesopolyploid Brassica oleracea[J]. Genome Biology,2014,15(6):R77
[23]肖旭峰,曹必好,王勇,陈国菊,雷建军.菜薹花芽分化及BrcuFLC基因的克隆与表达[J].园艺学报,2008,35(6):827-832
[24] Turnbull C G N,Lopez-Cobollo R M. Heavy traffic in the fast lane:long-distance signalling by macromolecules[J]. New Phytologist,2013,198(1):33-51
[25] Spiegelman Z,Golan G,Wolf S. Don’t kill the messenger:long-distance trafficking of mRNA molecules[J]. Plant Science,2013,213:1-8
[26] Lewsey M G,Hardcastle T J,Melnyk C W,Molnar A,Valli A,Urich M A,Nery J R,Baulcombe D C,Ecker J R. Mobile smallRNAs regulate genome-wide DNA methylation[J]. Proceedings ofthe National Academy of Sciences,2016,113(6):801-810
[27] Yang Y,Mao L,Jittayasothorn Y,Kang Y,Jiao C,Fei Z,Zhong GY. Messenger RNA exchange between scions and rootstocks in graftedgrapevines[J]. BMC Plant Biology,2015,15(1):251
[28] Yang H,Yu T. Arabidopsis floral regulators FVE and AGL24 arephloem-mobile RNAs[J]. Botanical Studies,2010,51(1):17-26
[29] Lough T J,Lucas W J. Integrative plant biology:role of phloemlong-distance macromolecular trafficking[J]. Annual Review PlantBiology,2006,57(1):203-232
[30]王文娇,王世峰,李梅兰,侯雷平.不同砧木嫁接黄瓜蜡质合成基因表达分析及其miRNA预测[J].核农学报,2017,31(10):1896-1903
[31] Li Y,Li C,Bai L,He C,Yu X. MicroRNA and target generesponses to salt stress in grafted cucumber seedlings[J]. ActaPhysiologiae Plantarum,2016,38(2):1-12
[2]马三梅,王永飞.菜心育种的研究进展[J].北方园艺,2006(3):40-41
[3]蒋欣梅,赵容秋,于锡宏.高温春化逆转对甘蓝碳、氮代谢的影响[J].园艺学报,2009,36(增刊):1967
[4]陶鹏,李必元,岳智臣,王五宏,雷娟利,赵彦婷,钟新民.结球甘蓝与菜心正反嫁接比较试验[J].浙江农业科学,2017,58(4):595-596
[5] Jung C,Müller A E. Flowering time control and applications in plantbreeding[J]. Trends Plant Science,2009,14(10):563-573
[6]程建徽,梅军霞,郑婷,魏灵珠,吴江.不同砧木对欧亚种葡萄红亚历山大产量和品质的影响[J].核农学报,2015,29(8):1607-1616
[7]马佩勇,贾赵东,边小峰,郭小丁,谢一芝.不同砧木与嫁接方式对甘薯开花结实的影响[J].浙江农业学报,2014,26(6):1425-1430
[8]韩霄.从作物的源流库理论展望新型育种技术[J].生物技术通报,2015,31(4):34-39
[9]王燕,谢辉,陈利萍.植物嫁接诱导的遗传变异机理的研究进展[J].遗传,2011,33(6):585-590
[10] Li X,Bian H,Song D,Ma S,Han N,Wang J,Zhu M. Floweringtime control in ornamental gloxinia(Sinningia speciosa)bymanipulation of miR159 expression[J]. Annals Botany,2013,111(5):791-799
[11] Huang N C, Yu T S. The sequences of Arabidopsis GA-INSENSITIVE RNA constitute the motifs that are necessary andsufficient for RNA long-distance trafficking[J]. Plant Journal,2009,59(6):921-929
[12] Kanehira A, Yamada K, Iwaya T, Tsuwamoto R, Kasai A,Nakazono M,Harada T. Apple phloem cells contain some mRNAstransported over long distances[J]. Tree Genetics Genomes,2010,6(5):635-642
[13] Zhang W,Thieme C J,Kollwig G,Apelt F,Yang L,Winter N,Andresen N,Walther D,Kragler F. tRNA-related sequences triggersystemic mRNA transport in plants[J]. Plant Cell,2016,28(6):1237-1249
[14] GregisⅤ,Sessa A,Colombo L,Kater M M. AGL24,SHORTVEGETATIVE PHASE, and APETALA1 redundantly controlAGAMOUS during early stages of flower development in Arabidopsis[J]. Plant Cell,2006,18(6):1373-1382
[15] GregisⅤ,Sessa A,Dorca-Fornell C,Kater M M. The Arabidopsisfloral meristem identity genes AP1,AGL24 and SVP directly repressclass B and C floral homeotic genes[J]. Plant Journal,2009,60(4):626-637
[16]蒋炜,周雯文,李朝闯,闫凯,王宇,王志敏,宋明,汤青林.青花菜开花促进因子AGL19与整合子AGL24和SOC1的互作研究[J].园艺学报,2017,44(10):1905-1913
[17] Liu C,Chen H,Er H L,Soo H M,Kumar P P,Han J H,Liou YC,Yu H. Direct interaction of AGL24 and SOC1 integrates floweringsignals in Arabidopsis[J]. Development,2008,135(8):1481-1491
[18] Lee J,Oh M,Park H,LeeⅠ. SOC1 translocated to the nucleus byinteraction with AGL24 directly regulates leafy[J]. Plant Journal,2008,55(5):832-843
[19] Lee J,LeeⅠ. Regulation and function of SOC1,a floweringpathway integrator[J]. Journal Experimental Botany,2010,61(9):2247-2254
[20] Torti S,Fornara F. AGL24 acts in concert with SOC1 and FULduring Arabidopsis floral transition[J]. Plant Signaling Behavior,7(10):1251-1254
[21] Wang X,Wang H,Wang J,Sun R,Wu J,Liu S,Bai Y,Mun J,BancroftⅠ,Cheng F,Huang S,Li X,Hua W,Wang J,Wang X,Freeling M,Pires J C,Paterson A H,Chalhoub B,Wang B,Hayward A,Sharpe A G,Park B,Weisshaar B,Liu B,Li B,LiuB,Tong C,Song C,Duran C,Peng C,Geng C,Koh C,Lin C,Edwards D,Mu D,Shen D,Soumpourou E,Li F,Fraser F,Conant G,Lassalle G,King G J,Bonnema G,Tang H,Wang H,Belcram H,Zhou H,Hirakawa H,Abe H,Guo H,Wang H,JinH,ParkinⅠA P,Batley J,Kim J,Just J,Li J,Xu J,Deng J,Kim JA,Li J,Yu J,Meng J,Wang J,Min J,Poulain J,Wang J,Hatakeyama K,Wu K,Wang L,Fang L,Trick M,Links M G,Zhao M,Jin M,Ramchiary N,Drou N,Berkman P J,Cai Q,Huang Q,Li R,Tabata S,Cheng S,Zhang S,Zhang S,Huang S,Sato S,Sun S,Kwon S,Choi S,Lee T,Fan W,Zhao X,Tan X,Xu X,Wang Y,Qiu Y,Yin Y,Li Y,Du Y,Liao Y,Lim Y,Narusaka Y,Wang Y,Wang Z,Li Z,Wang Z,Xiong Z,Zhang Z.The genome of the mesopolyploid crop species Brassica rapa[J].Nature Genetics,2011,43:1035-1039
[22] ParkinⅠA P,Koh C,Tang H,Robinson S J,Kagale S,Clarke WE,Town C D,Nixon J,KrishnakumarⅤ,Bidwell S L,DenoeudF,Belcram H,Links M G,Just J,Clarke C,Bender T,HuebertT,Mason A S,Pires J C,Barker G,Moore J,Walley P G,ManoliS,Batley J,Edwards D,Nelson M N,Wang X,Paterson A H,King G,BancroftⅠ,Chalhoub B,Sharpe A G. Transcriptome andmethylome profiling reveals relics of genome dominance in themesopolyploid Brassica oleracea[J]. Genome Biology,2014,15(6):R77
[23]肖旭峰,曹必好,王勇,陈国菊,雷建军.菜薹花芽分化及BrcuFLC基因的克隆与表达[J].园艺学报,2008,35(6):827-832
[24] Turnbull C G N,Lopez-Cobollo R M. Heavy traffic in the fast lane:long-distance signalling by macromolecules[J]. New Phytologist,2013,198(1):33-51
[25] Spiegelman Z,Golan G,Wolf S. Don’t kill the messenger:long-distance trafficking of mRNA molecules[J]. Plant Science,2013,213:1-8
[26] Lewsey M G,Hardcastle T J,Melnyk C W,Molnar A,Valli A,Urich M A,Nery J R,Baulcombe D C,Ecker J R. Mobile smallRNAs regulate genome-wide DNA methylation[J]. Proceedings ofthe National Academy of Sciences,2016,113(6):801-810
[27] Yang Y,Mao L,Jittayasothorn Y,Kang Y,Jiao C,Fei Z,Zhong GY. Messenger RNA exchange between scions and rootstocks in graftedgrapevines[J]. BMC Plant Biology,2015,15(1):251
[28] Yang H,Yu T. Arabidopsis floral regulators FVE and AGL24 arephloem-mobile RNAs[J]. Botanical Studies,2010,51(1):17-26
[29] Lough T J,Lucas W J. Integrative plant biology:role of phloemlong-distance macromolecular trafficking[J]. Annual Review PlantBiology,2006,57(1):203-232
[30]王文娇,王世峰,李梅兰,侯雷平.不同砧木嫁接黄瓜蜡质合成基因表达分析及其miRNA预测[J].核农学报,2017,31(10):1896-1903
[31] Li Y,Li C,Bai L,He C,Yu X. MicroRNA and target generesponses to salt stress in grafted cucumber seedlings[J]. ActaPhysiologiae Plantarum,2016,38(2):1-12