彭磊;吴艳;刘小翠;杨鵾;范付华;文晓鹏;

• 1:贵州大学农业生物工程研究院/生命科学学院·山地植物资源保护与种质创新教育部重点实验室
• 2:贵州省安顺市农业科学院
• 3:贵州大学林学院

摘要(Abstract)：

【目的】克隆Ty3-gypsy类反转录转座子反转录酶(RT)序列并分析其特性,为火龙果遗传变异和进化研究提供新的信息。【方法】根据Ty3-gypsy类反转录转座子RT保守区设计简并引物,从红肉和白肉火龙果及其突变体中扩增出430 bp左右的目标片段,回收、克隆、测序获得RT序列,并进行生物信息学分析;采用RT-PCR检测其转录活性。【结果】共获得82条RT序列,其中有55条序列发生终止密码子突变,5条序列发生移码突变;经RT序列对比,火龙果与水稻、拟南芥有较高的同源性;3条RT序列具有转录活性。【结论】火龙果Ty3-gypsy类反转录转座子RT序列具有高度异质性,部分序列具有转录活性,与其他物种存在反转录转座子的横向传递。

关键词(KeyWords)： 火龙果;Ty3-gypsy类反转录转座子;反转录酶;异质性;转录活性

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(31560549,31260464)

作者(Author): 彭磊;吴艳;刘小翠;杨鵾;范付华;文晓鹏;

Email:

参考文献(References)：

• [1]徐玲,杨静,刘林,李成云.植物反转录转座子的研究进展[J].江西农业学报,2012,24(6):42-46.XU Ling,YANG Jing,LIU Lin,LI Chengyun.Research progress in retrotransposons of plants[J].Acta Agriculturae Jiangxi,2012,24(6):42-46.
• [2]VELASCO R,ZHARKIKH A,AFFOURTIT J,……,VIOLA R.The genome of the domesticated apple(Malus×domestica Borkh.)[J].Nature Genetics,2010,42(10):833-839.
• [3]MA J X,DEVOS K M,BENNETZEN J L.Analyses of LTR-retrotransposon structures reveal recent and rapid genomic DNA loss in rice[J].Genome Research,2004,14(5):860-869.
• [4]CENCI A,COMBES M C,LASHERMES P.Genome evolution in diploid and tetraploid Coffea species as revealed by comparative analysis of orthologous genome segments[J].Plant Molecular Biology,2012,78(1/2):135-145.
• [5]HUO H,CONNER J A,OZIAS-AKINS P.Genetic mapping of the apospory-specific genomic region in Pennisetum squamulatum using retrotransposon-based molecular markers[J].Theoretical and Applied Genetics,2009,119(2):199-212.
• [6]LEE S,KIM J H,PARK K C,KIM N S.LTR-retrotransposons and inter-retrotransposon amplified polymorphism(IRAP)analysis in Lilium species[J].Genetica,2015,143(3):343-352.
• [7]周芬静,高燕会,童再康.石蒜Ty1-copia类反转录转座子反转录酶基因(RT)的克隆与分析[J].农业生物技术学报,2015,23(4):469-480.ZHOU Fenjing,GAO Yanhui,TONG Zaikang.Cloning and analysis of reverse transcriptase gene(RT)of Ty1-copia retrotransposons in Lycoris radiate[J].Journal of Agricultural Biotechnology,2015,23(4):469-480.
• [8]DIAS E S,HATT C,HAMON S,HAMON P,RIGOREAU M,CROUZILLAT D,CARARETO C M A,KOCHKO A,GUYOT R.Large distribution and high sequence identity of a Copia-type retrotransposon in angiosperm families[J].Plant Molecular Biology,2015,89(1):83-97.
• [9]XIONG T L,XIAO J H,LI Y X,BIAN S N,HUANG D W.Diversity and evolution of Ty1-copia retroelements within Chalcidoidea by reverse transcriptase domain analysis[J].Insect Molecular Biology,2015,24(5):503-516.
• [10]ZUCCOLO A,SCOFIELD D G,DEPAOLI E,MORGANTE M.The Ty1-copia LTR retroelement family PARTC is highly conserved in conifers over 200 MY of evolution[J].Gene,2015,568(1):89-99.
• [11]孙俊,房经贵,孙其宝,章镇.苹果Ty1-copia类逆转座子家族鉴定及特性分析[J].西北植物学报,2009,29(12):2373-2377.SUN Jun,FANG Jinggui,SUN Qibao,ZHANG Zhen.Identification and characteristics of apple Ty1-copia-like retrotransposons[J].Acta Botanica Boreali-Occidentalia Sinica,2009,29(12):2373-2377.
• [12]徐玲,杨静,刘林,李成云.水稻gypsy类逆转座子对不同胁迫条件的响应[J].湖南农业大学学报(自然科学版),2012,38(6):591-596.XU Ling,YANG Jing,LIU Lin,LI Chengyun.Response of rice gypsy retrotransposons to different stress conditions[J].Journal of Hunan Agricultural University(Natural Sciences),2012,38(6):591-596.
• [13]张文波,陈凌,李雪辉,白玉娥,林晓飞.兴安落叶松Ty3-gypsy类逆转座子逆转录酶的多样性分析[J].分子植物育种,2016,14(5):1098-1106.ZHANG Wenbo,CHEN Ling,LI Xuehui,BAI Yu’e,LIN Xiaofei.Sequence diversity analysis of reverse transcriptases of Ty3-gypsy-like retrotransposons in Larix gmelinii[J].Molecular Plant Breeding,2016,14(5):1098-1106.
• [14]侯小改,郭大龙,黄燕梅,张曦.牡丹Ty3-gypsy类反转录转座子反转录酶序列的克隆及分析[J].园艺学报,2013,40(1):98-106.HOU Xiaogai,GUO Dalong,HUANG Yanmei,ZHANG Xi.Cloning and analysis of reverse transcriptase of Ty3-gypsy-like retrotransposons in tree peony(Paeonia)[J].Acta Horticulturae Sinica,2013,40(1):98-106.
• [15]申世辉,马玉华,蔡永强.火龙果研究进展[J].中国热带农业,2015(1):48-52.SHEN Shihui,MA Yuhua,CAI Yongqiang.Research progress of pitaya[J].China Tropical Agriculture,2015(1):48-52.
• [16]TEL-ZUR N,ABBO S,BAR-ZVI D,MIZRAHI Y.Genetic relationships among Hylocereus and Selenicereus Vine Cacti(Cactaceae):evidence from hybridization and cytological studies[J].Annals of Botany,2004,94(4):527-534.
• [17]张冰雪,范付华,乔光,宋莎,文晓鹏,刘涛.贵州地方火龙果芽变种质DNA指纹图谱及遗传多样性的ISSR分析[J].果树学报,2013,30(4):573-577.ZHANG Bingxue,FAN Fuhua,QIAO Guang,SONG Sha,WEN Xiaopeng,LIU Tao.DNA fingerprint construction and genetic relationship analysis of dragon fruit germplasm in Guizhou by ISSR markers[J].Journal of Fruit Science,2013,30(4):573-577.
• [18]袁亚芳,陈明贤,陈清西,庄卫东,郑金水.福建地区火龙果种质资源调查及ISSR分析[J].中国农学通报,2013,29(34):216-220.YUAN Yafang,CHEN Mingxian,CHEN Qingxi,ZHUANG Weidong,ZHENG Jinshui.Investigate and ISSR analysis of pitaya germplasm resources in Fujian province[J].Chinese Agricultural Science Bulletin,2013,29(34):216-220.
• [19]GBADEGESIN M A,BEECHING J R.Highly heterogeneous Ty3/Gypsy-like retrotransposon sequences in the genome of cassava(Manihot esculenta Crantz)[J].African Journal of Biotechnology,2011,10(20):3951-3963.
• [20]ZUCCOLO A,AMMIRAJU J S S,KIM H,SANYAL A,JACKSON S,WING R A.Rapid and differential proliferation of the Ty3-gypsy LTR retrotransposon Atlantys in the genus Oryza[J].Rice,2008,1(1):85-99.
• [21]JEUNG J U,CHOB S K,LEE S J,SHIN J S.Characterization of Ty3-gypsy-like elements in Hibiscus syriacus[J].Molecules and Cells,2005,19(3):318-327.
• [22]KOBAYASHI S,GOTO-YAMAMOTO N,HIROCHIKA H.Retrotransposon-induced mutations in grape skin color[J].Science,2004,304(5673):982.
• [23]ERDMANN P M,LEE R K,BASSETT M J,MCCLEAN P E.A molecular marker tightly linked to P,a gene required for flower and seedcoat color in commonbean(Phaseolus vulgaris L.),contains the Ty3-gypsy retrotransposon Tpv3g[J].Genome,2002,45(4):728-736.
• [24]范付华,乔光,郑思成,文晓鹏.火龙果Ty1-copia类反转录转座子反转录酶序列的克隆及分析[J].园艺学报,2012,39(2):265-272.FAN Fuhua,QIAO Guang,ZHENG Sicheng,WEN Xiaopeng.Cloning and analysis of reverse transcriptase of Ty1-copia retrotransposons in Hylocereus undatu[J].Acta Horticulturae Sinica,2012,39(2):265-272.
• [25]KUMEKAWA N,OHTSUBO E,OHTSUBO H.Identification and phylogenetic analysis of gypsy-type retrotransposons in the plant kingdom[J].Genes&Genetic Systems,1999,74(6):299-307.
• [26]FAN F H,WEN X P,DING G J,CUI B W.Isolation,identification and characterization of genomic LTR retrotransposon sequences from masson pine(Pinus massoniana)[J].Tree Genetics&Genomes,2013,9(5):1237-1246.
• [27]蒋爽,滕元文,宗宇,蔡丹英.植物LTR反转录转座子的研究进展[J].西北植物学报,2013,33(11):2354-2360.JIANG Shuang,TENG Yuanwen,ZONG Yu,CAI Danying.Review of retrotransposons in plants[J].Acta Botanica Boreali-Occidentalia Sinica,2013,33(11):2354-2360.
• [28]周鹏,仇宗浩,翟锐,梁东,徐凌飞.早酥梨及其红色芽变Ty1-copia反转录转座子逆转录酶的克隆与分析[J].西北农林科技大学学报(自然科学版),2014,42(5):168-182.ZHOU Peng,QIU Zonghao,ZHAI Rui,LIANG Dong,XU Lingfei.Cloning and comparative analysis of reverse transcriptase of Ty1-copia retrotranspon in‘Zaosu’(Pyrus bretschneideri Rehd.)pear and its red mutant[J].Journal of Northwest A&F University(Natural Science Edition),2014,42(5):168-182.
• [29]TANG Y M,MA Y Z,LI L C,YE X G.Identification and characterization of reverse transcriptase domain of transcriptionally active retrotransposons in wheat genomes[J].Journal of Integrative Plant Biology Formerly Acta Botanica Sinica,2005,47(5):604-612.
• [30]李聪,斯金平,高燕会,朱玉球,蒋园.脱落酸对铁皮石斛Ty1-copia类反转录转座子转录活性的影响[J].中国中药杂志,2014,39(10):1788-1794.LI Cong,SI Jinping,GAO Yanhui,ZHU Yuqiu,JIANG Yuan.Effect of ABA on transcriptionally active Ty1-copia retrotransposons in Dendrobium officinale[J].China Journal of Chinese Materia Medica,2014,39(10):1788-1794.
• [31]POUTEAU S,GRANDBASTIEN M A,BOCCARA M.Microbial elicitors of plant defence responses activate transcription of a retrotransposon[J].The Plant Journal,1994,5(4):535-542.
• [32]TAKEDA S,SUGIMOTO K,OTSUKI H,HIROCHIKA H.Transcriptional activation of the tobacco retrotransposon Tto1 by wounding and methyl jasmonate[J].Plant Molecular Biology,1998,36(3):365-376.
• [33]MHIRI C,MOREL J B,VERNHETTES S,CASACUBERTA J M,LUCAS H,GRANDBASTIEN M A.The promoter of the tobacco Tnt1 retrotransposon is induced by wounding and by abiotic stress[J].Plant Molecular Biology,1997,33(2):257-266.
• [34]HIROCHIKA H,SUGIMOTO K,OTSUKIT Y,TSUGAWAT H,KANDA M.Retrotransposons of rice involved in mutations induced by tissue culture[J].Proceeding of the National Academy of Sciences,1996,93(15):7783-7788.
• [35]KIMURA Y,TOSA Y,SHIMADA S,SOGO R,KUSABA M,SUNAGA T,BETSUYAKU S,ETO Y,NAKAYASHIKI H,MAYAMA S.OARE-1,a Ty1-copia retrotransposon in oat activated by abiotic and biotic stresses[J].Plant Cell Physiology,2001,42(12):1345-1354.
• [36]HIROCHIKA H.Activation of tobacco retrotransposons during tissue culture[J].The EMBO Journal,1993,12(6):2521-2528.
• [37]蒋爽,蔡丹英,滕元文.梨反转录转座子逆转录酶序列预测及其进化和转录分析[J].园艺学报,2014,41(11):2196-2207.JIANG Shuang,CAI Danying,TENG Yuanwen.Prediction,evolution and expression analysis of reverse transcriptase of LTR retrotransposons in pear[J].Acta Horticulturae Sinica,2014,41(11):2196-2207.
• [38]RAJPUT M K,UPADHYAYA K C.Characterization of heterogeneity in Ty1-copia group retrotransposons in chickpea(Cicer arietinum L.)[J].Molekuliarnaia Biologiia,2010,44(4):601-607.
• [39]LAO N T,MALONEY A P,ATKINS J F,KAVANAGH T A.Versatile dual reporter gene systems for investigating stop codon read through in plants[J].Plo S One,2009,4(10):e7354.
• [40]GRANDBASTIEN M A,LUCAS H,MOREL J B,MHIRI C,VERNHETTES S,CASACUBERTA J M.The expression of the tobacco Tnt1 retrotransposon is linked to plant defense responses[J].Genetica,1997,100(1/3):241-252.
• [41]HARADA T,SUNAKO T,WAKASA Y,SOEJIMA J,SATOH T,NIIZEKI M.An allele of the 1-aminocyclopropane-1-carboxylate synthase gene(Md-ACS1)accounts for the low level of ethylene production in climacteric fruits of some apple cultivars[J].Theoretical and Applied Genetics,2000,101(5/6):742-746.
• [42]BISWAS M K,BAIG M N R,CHENG Y J,DENG X X.Retrotransposon based genetic similarity within the genus Citrus and its relatives[J].Genetic Resources and Crop Evolution,2010,57(7):963-972.
• [43]YAO J L,DONG Y H,MORRIS B A M.Parthenocarpic apple fruit production conferred by transposon insertion mutations in a MADS-box transcription factor[J].Proceedings of the National Academy of Sciences,2001,98(3):1306-1311.