WANG Wencheng,MA Chunhui,WANG Ran,et al.Cloning and Expression Analysis of Adventitious Rooting Related Gene PbARRO-1 of Pyrus betulifolia Bunge.[J].Northern Horticulture,2017,41(12):97-102.[doi:10.11937/bfyy.201712024]
杜梨不定根形成相关基因PbARRO-1的克隆及定量表达分析
- Title:
- Cloning and Expression Analysis of Adventitious Rooting Related Gene PbARRO-1 of Pyrus betulifolia Bunge.
- 文献标志码:
- A
- 摘要:
- 以杜梨叶片为试材，采用同源序列和RACE技术克隆了梨ARRO（adventitious rooting related oxygenase）基因，并进行了基因的生物信息学和时空表达分析，以期对梨属植物不定根产生研究提供帮助。结果表明：PbARRO-1基因开放读码框全长831 bp，编码276个氨基酸，相对分子量为30.535 9 kD，等电点pI为5.53；推导氨基酸序列同源性分析显示该基因与苹果、梅花、毛果杨等具有较高同源性，其中苹果达到92%。荧光实时定量PCR分析表明，该基因在秋子梨根、茎、叶中均有表达，叶片中表达量初期较高，后期较低；茎中表达量则呈现先升高后降低的趋势，在第15天达到峰值，推测PbARRO-1基因与茎上不定根的形成密切相关。
- Abstract:
- The leaves of Pyrus betulifolia were used as the materials，a novel adventitious rooting related oxygenase gene (ARRO gene) was cloned by Homology-based and RACE technology，and then analyzed with bioinformatics methods and quantitative real-time RT-PCR technology.The study laid a foundation for adventitious root formation studying.The results showed that the open reading frame of PbARRO-1 was 831 bp and encodes 276 amino acids，the molecular weight and isoelectric point of the protein was 30.535 9 kD and 5.53 respectively.Amino acid sequence homology analysis showed that the deduced amino acid sequence of PbARRO-1 gene had 92% homologies with Malus domestica and 80% homologies with Prunus mume.Quantitative real-time RT-PCR analysis indicated that PbARRO-1 gene expressed in roots，stems，leaves of Pyrus ussuriensis，in leaves its expression level was higher at early stage and lower in later stage;in stems its expression level increased at first and then decreased，and reached peak value at 15 days.The conclusion inferred that PbARRO-1 gene was closely related to root induction.
参考文献/References:
[1]李志霞,聂继云,李静,等.梨产业发展分析与建议[J].中国南方果树,2014,43(5):144-147. [2]张绍铃.梨学[M].北京:中国农业出版社,2013. [3]沙守峰,张绍铃,李俊才.梨矮化砧木的选育及其应用研究进展[J].北方园艺,2009(8):140-143. [4]薛明超,马春晖,李鼎立,等.不同梨砧木嫩枝扦插繁殖技术研究[J].中国果树,2015(3):227-227. [5]徐凌飞,刘亚杰,李慧,等.梨矮化砧木云南榅桲硬枝扦插繁殖试验[J].北方园艺,2011(17):54-55. [6]王德芬,张梅,李鼎立,等.秋子梨叶片高效再生体系的构建[J].北方园艺,2016(4):97-101. [7]周佳红,邢才华,曹慧,等.秋子梨茎段薄层细胞培养再生初探[J].北方园艺,2015(6):95-98. [8]王乔春,SVENSSON M.培养基种类对梨试管苗茎增殖的影响[J].四川农业大学学报,1993(1):77-81. [9]杨芳.秋子梨组织培养与离体保存的研究[D].北京:中国农业大学,2007. [10]付镇芳,孟颢光,张朝红,等.‘砀山酥’梨叶片再生体系的建立[J].北方园艺,2011(14):98-101. [11]SMOLKA A,WELANDER M,OLSSON P,et al.Involvement of the ARRO-1，gene in adventitious root formation in apple[J].Plant Science,2009,177(6):710-715. [12]贺丹,李睿,纪思羽,等.牡丹不定根形成相关基因PsARRO-1的克隆及表达分析[J].植物生理学报,2014(8):1151-1158. [13]BUTLER E D,GALLAGHER T F.Characterization of auxin-induced ARRO-1 expression in the primary root of Malus domestica[J].Journal of Experimental Botany,2000,51(351):1765-1766. [14]LI T Y,WANG Y,ZHANG X Z,et al.Isolation and characterization of ARRO-1, genes from apple rootstocks in response to auxin treatment[J].Plant Molecular Biology Reporter,2012,30(6):1408-1414. [15]SEDIRA M,BUTLER E,GALLAGHER T,et al.Verification of auxin-induced gene expression during adventitious rooting in rol B-transformed and untransformed apple Jork[J].Plant Science,2005,168(5):1193-1198. [16]LIVAK K J,SCHMITTGEN T D.Analysis of relative gene expression data using real-time quantitative PCR and the 2-⊿⊿Ct method[J].Method,2001,25(4):402-408. [17]吴静.牡丹不定根形成相关基因PSARRO-1的克隆与序列分析[D].郑州:河南农业大学,2010.
相似文献/References:
[1]梁玉勇,廖玲,左北梅,等.太子参不定根诱导及增殖培养研究[J].北方园艺,2013,37(02):147.
LIANG Yu-yong,LIAO Ling,ZUO Bei-mei,et al.Study on Adventitious Roots Induction and Proliferation Culture of Pseudostellaria heterophylla[J].Northern Horticulture,2013,37(12):147.
[2]马玲玲,王鹏,张振宇,等.梓属植物嫩枝扦插生根能力的评价[J].北方园艺,2014,38(15):72.
MA Ling-ling,WANG Peng,ZHANG Zhen-yu,et al.Evaluation on Rooting Ability of Softwood Cuttings on Catalpa[J].Northern Horticulture,2014,38(12):72.
[3]秦嘉泽,全雪丽,田海丽,等.长白山野生膜荚黄芪不定根总黄酮积累规律研究（Ⅰ）[J].北方园艺,2014,38(16):158.
QIN Jia-ze,QUAN Xue-li,TIAN Hai-li,et al.Study on the Accumulation Pattern of Total Flavonoids for Adventitious Roots of Wild Astragalus membranaceus in Changbai Mountain[J].Northern Horticulture,2014,38(12):158.
[4]赵兴宇,李丽丽,杨洪一.蓝莓茎段离体快繁研究[J].北方园艺,2015,39(01):103.[doi:10.11937/bfyy.201501028]
ZHAO Xing-yu,LI Li-li,YANG Hong-yi.Study on Rapid Proliferation of Stem Segments in vitro of Blueberry[J].Northern Horticulture,2015,39(12):103.[doi:10.11937/bfyy.201501028]
[5]焦延静,邢瑞肖,杜国强,等.杜梨根癌病原菌质粒类型鉴定及枯草芽孢杆菌对其抑制研究[J].北方园艺,2015,39(18):129.[doi:10.11937/bfyy.201518034]
JIAO Yanjing,XING Ruixiao,DU Guoqiang,et al.Identification of Bacteria Plasmid Type Collected From Crown Gall and Efficacy of Bacillus subtilis to Their Control in Pyrus betulaefolia Bge[J].Northern Horticulture,2015,39(12):129.[doi:10.11937/bfyy.201518034]
[6]李 丹 丹,王 忆,韩 振 海,等.应用正交设计对组培杜梨快速繁殖的研究[J].北方园艺,2009,33(01):0.[doi:10.11937/bfyy.200901012]
LI Dan -dan,W ANG Yi,HAN Zhen -hai,et al.Studies on Rapid Propagation Tissue Culture in Pyrusbetulifolia Bunge by Applying Orthogonal Design[J].Northern Horticulture,2009,33(12):0.[doi:10.11937/bfyy.200901012]
[7]李军霞,吴翠云.杜梨种子休眠与萌发过程中酶活性变化[J].北方园艺,2011,35(11):0.[doi:10.11937/bfyy.2011011006]
LI Jun-xia,WU Cui-yun.The Change of Enzyme Activity in the Dornmny and Germination Process of Pyrus betulaefolia[J].Northern Horticulture,2011,35(12):0.[doi:10.11937/bfyy.2011011006]
[8]臧忠婧,吴 瑕.草原樱桃扦插繁殖生根解剖学分析[J].北方园艺,2010,34(03):0.[doi:10.11937/bfyy.201003078]
ZANG Zhong-jingI,WU Xia.The Anatomical Observation on Rooting of the Cutting of Ground Cherry[J].Northern Horticulture,2010,34(12):0.[doi:10.11937/bfyy.201003078]
[9]付爽,汪洋,吕爽,等.低温对膜荚黄芪不定根总黄酮积累的影响[J].北方园艺,2017,41(22):154.[doi:10.11937/bfyy.20170244]
FU Shuang,WANG Yang,LYU Shuang,et al.Effects of Low Temperature on Accumulation of Total Flavonoids inAdventitious Roots of Astragalus membranaceus[J].Northern Horticulture,2017,41(12):154.[doi:10.11937/bfyy.20170244]
[10]于斌,朱进,周国林,等.淹水胁迫对黄瓜幼苗生长及不定根解剖结构的影响[J].北方园艺,2018,42(03):61.[doi:10.11937/bfyy.20172604]
YU Bin,ZHU Jin,ZHOU Guolin,et al.Effects of Waterlogging Stress on Growth and Adventitious Roots Anatomical Structure in Cucumber Seedling[J].Northern Horticulture,2018,42(12):61.[doi:10.11937/bfyy.20172604]
备注/Memo
第一作者简介：王文成（1994-），男，硕士研究生，研究方向为果树种质资源与分子育种。E-mail：2542633288@qq.com.责任作者：李鼎立（1979-），男，博士，副教授，研究方向为果树种质资源与分子育种。E-mail：qauldl@163.com.基金项目：山东省高等学校科技计划资助项目（J14LF05）；山东省优秀中青年科学家科研奖励基金资助项目（BS2011NY007）；国家自然科学基金青年基金资助项目（31401832）；现代农业产业技术体系建设专项资金资助项目（CARS-29）。