苹果生长素输入载体MdAUX1的基因克隆及在拟南芥和烟草中的遗传转化

doi:10.7525/j.issn.1673-5102.2016.06.014

植物研究 ›› 2016, Vol. 36 ›› Issue (6): 902-908.doi: 10.7525/j.issn.1673-5102.2016.06.014

苹果生长素输入载体MdAUX1的基因克隆及在拟南芥和烟草中的遗传转化

安建平, 王小非, 刘鑫, 李浩浩, 由春香, 郝玉金

山东农业大学园艺科学与工程学院, 作物生物学国家重点实验室, 泰安 271018

收稿日期:2016-09-05 出版日期:2016-11-15 发布日期:2016-11-16
通讯作者: 郝玉金,E-mail:haoyujin@sdau.edu.cn E-mail:haoyujin@sdau.edu.cn
作者简介:安建平(1990-),男,硕士研究生,主要从事果树生物技术的研究。
基金资助:
国家自然科学基金（31430074）；山东省农业工程和产业体系（SDAIT-03-033-03）

Molecular Cloning and Genetic Transformation Analysis of an Apple Auxin Influx Transporter Gene MdAUX1 in Arabidopsis thaliana and Tobacco

AN Jian-Ping, WANG Xiao-Fei, LIU Xin, LI Hao-Hao, YOU Chun-Xiang, HAO Yu-Jin

College of Horticulture Science and Engineering, Shandong Agricultural University, State Key Laboratory of Crop Biology, Tai'an 271018

Received:2016-09-05 Online:2016-11-15 Published:2016-11-16
Supported by:
The National Natural Science Foundation of China(31430074);Shandong Agricultural Engineering and industrial System(SDAIT-03-033-03)

摘要/Abstract

摘要： 生长素的极性运输对调节植物生长发育起重要作用。生长素转运蛋白调控生长素的极性运输。本研究从‘嘎啦’（Malus×domestic‘Royal Gala’）苹果中克隆了生长素输入载体基因MdAUX1（基因序列号：MDP0000384437）。序列分析表明，该基因包含长为1 443 bp完整的开放阅读框，编码含有480个氨基酸的蛋白。氨基酸序列比对和系统进化树分析表明，AUX1在不同物种间具有高度的序列保守性，其中，苹果MdAUX1与欧洲甜樱桃PaAUX1亲缘关系最近。半定量PCR分析显示，MdAUX1在苹果的茎中表达量较高。通过农杆菌介导的遗传转化获得转MdAUX1基因的拟南芥和烟草。转基因拟南芥根系表型观察结果显示，MdAUX1基因在拟南芥中异位表达后，会抑制主根伸长，促进侧根形成；在外源NAA处理下，转基因拟南芥对主根伸长的抑制作用得到部分恢复。在烟草中过量表达MdAUX1基因能够显著促进植株地上部分生长。以上结果说明苹果MdAUX1基因在调节植物生长发育中发挥重要作用。

关键词: 苹果, 生长素输入载体, MdAUX1, 遗传转化

Abstract: Auxin has been identified to play critical roles in regulating plant growth and development. The polar transport of auxin is regulated by auxin transporters. In this study, an auxin influx transporter gene named MdAUX1 was cloned from Malus×domestic‘Royal Gala’. Sequence analysis showed that the length of MdAUX1 gene was 1 443 bp, which encoded 480 amino acids. Amino acid sequence alignment and phylogenetic tree analysis showed that the AUX1 had a high sequence conservation among different species, and apple MdAUX1 exhibited the highest sequence similarity with Prunus avium PaAUX1. RT-PCR analysis demonstrated the MdAUX1 gene was mainly expressed in stem. We generated transgenic Arabidopsis and tobacco plants expressing the apple MdAUX1 by Agrobacterium-mediated genetic transformation. Ectopic expression of MdAUX1 in Arabidopsis exhibited the phenotype of inhibition of primary root elongation and increase lateral root number. Furthermore, there was no significant inhibition on primary root elongation phenotype when treated with the NAA. Overexpressing MdAUX1 in tobacco could significantly promote the growth of aerial parts of plant. These results suggested that MdAUX1 played a vital role in plant growth and development.

Key words: apple, auxin influx transporters, MdAUX1, genetic transformation

中图分类号:

S661.1

安建平, 王小非, 刘鑫, 李浩浩, 由春香, 郝玉金. 苹果生长素输入载体MdAUX1的基因克隆及在拟南芥和烟草中的遗传转化[J]. 植物研究, 2016, 36(6): 902-908.

AN Jian-Ping, WANG Xiao-Fei, LIU Xin, LI Hao-Hao, YOU Chun-Xiang, HAO Yu-Jin. Molecular Cloning and Genetic Transformation Analysis of an Apple Auxin Influx Transporter Gene MdAUX1 in Arabidopsis thaliana and Tobacco[J]. Bulletin of Botanical Research, 2016, 36(6): 902-908.

参考文献

1. 倪为民,陈晓亚,许智宏,等.生长素极性运输研究进展[J].植物学报,2000,42(3):221-228. Ni W M,Chen X Y,Xu Z H,et al.Advances in study of polar auxin transport[J].Acta Botanica Sinica,2000,42(3):221-228.
2. Kepinski S,Leyser O.Plant development:auxin in loops[J].Current Biology,2005,15(6):R208-R210.
3. 刘进平.生长素受体与信号转导机制研究进展[J].生物技术通报,2007,3:22. Liu J P.Advances in research on auxin receptors and auxin signaling mechanism[J].Biotechnology Bulletin,2007,3:22.
4. 王凤娇,宋晓婕,王万军.植物生长素的极性运输[J].现代农业科技,2009,3:11-14. Wang F J,Song X J,Wang W J.Auxin polar transport[J].Modern Agricultural Science and Technology,2009,3:11-14
5. Parry G,Marchant A,May S,et al.Quick on the uptake:characterization of a family of plant auxin influx carriers[J].Journal of Plant Growth Regulation,2001,20(3):217-225.
6. 董秀春.毛白杨生长素信号转导因子基因的分离与功能的初步分析[D]. 泰安:山东农业大学,2008. Dong X C.Isolation and functional characterization of gene encoding auxin signaling components in Populus tomentosa Carr.[D].Tai'an:Shandong Agricultural University,2008.
7. Muller A,Guan C,Gaweiler L,et al.AtPIN2 defines a locus of Arabidopsis for root gravitropism control[J].EMBO Journal,1998,17:6903-6911.
8. 任怡怡,戴绍军,刘炜.花生生长素输入载体基因PNAUX1的克隆及表达模式分析[J].山东农业科学,2012,03:1-6. Ren Y Y,Dai S J,Liu W.Cloning of auxin influx carrier gene PNAUX1 from peanut and analysis of its expression pattern[J].Shandong Agricultural Sciences,2012,03:1-6.
9. 闫辉.毛白杨PtAUX1和PtPIN1基因的克隆及遗传转化[D].泰安:山东农业大学,2013. Yan H.Cloning of PtAUX1 and PtPIN1 from Populus tomentosa Carr and transformation.[D].Tai'an:Shandong Agricultural University,2013.
10. Zhao H,Ma T,Wang X,et al.OsAUX1 controls lateral root initiation in rice(Oryza sativa L.)[J].Plant,Cell & Environment,2015,38(11):2208-2222.
11. Yu C,Sun C,Shen C,et al.The auxin transporter,OsAUX1,is involved in primary root and root hair elongation and in Cd stress responses in rice(Oryza sativa L.)[J].The Plant Journal,2015,83(5):818-830.
12. Liu W,Li R J,Han T T,et al.Salt stress reduces root meristem size by nitric oxide-mediated modulation of auxin accumulation and signaling in Arabidopsis[J].Plant Physiology,2015,168(1):343-356.
13. Li J,Xu H H,Liu W C,et al.Ethylene inhibits root elongation during alkaline stress through AUXIN1 and associated changes in auxin accumulation[J].Plant Physiology,2015,168(4):1777-1791.
14. Li G,Song H,Li B,et al.Auxin resistant1 and PIN-FORMED2 protect lateral root formation in Arabidopsis under iron stress[J].Plant Physiology,2015,169(4):2608-2623.
15. Guilfoyle T J,Hagen G.Auxin response factors[J].Current Opinion in Plant Biology,2007,10(5):453-460.
16. Clough S J,Bent A F.Floral dip:a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana[J].The Plant Journal,1998,16(6):735-743.
17. 李运合,孙光明,吴蓓.植物生长素的极性运输载体研究进展[J].西北植物学报,2009,08:1714-1722. Li Y H,Sun G M,Wu B.Advances on carriers of plant polar auxin transport[J].Acta Botanica Boreali-Occidentalia,2009,08:1714-1722.
18. 曲桂芹,张贤泽,霍俊伟.影响大豆体细胞胚诱导因素的研究[J].植物研究,2001,21:210-214. Qu G Q,Zhang X Z,Huo J W.Study on somatic embryos induction in soybean(Glycine Max L.)[J].Bulletin of Botanical Research,2001,21:210-214.
19. Reed J W.Roles and activities of Aux/IAA proteins in Arabidopsis[J].Trends in Plant Science,2001,6(9):420-425.
20. Yang Y,Hammes U Z,Taylor C G,et al.High-affinity auxin transport by the AUX1 influx carrier protein[J].Current Biology,2006,16(11):1123-1127.
21. 张碧玉,苏良辰,孙叙卓.拟南芥生长素输入载体AUX1研究[J].亚热带植物科学,2010,39(1):88-91. Zhang B Y,Su L C,Sun X Z.Research on auxin influx transporter AUX1 of Arabidopsis[J].Subtropical Plant Science,2010,39(1):88-91.
22. Nemhauser J L,Mockler T C,Chory J.Interdependency of brassinosteroid and auxin signaling in Arabidopsis[J].PLoS Biology,2004,2(9):e258.
23. Carrier D J,Bakar N T A,Swarup R,et al.The binding of auxin to the Arabidopsis auxin influx transporter AUX1[J].Plant Physiology,2008,148(1):529-535.

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苹果生长素输入载体MdAUX1的基因克隆及在拟南芥和烟草中的遗传转化

Molecular Cloning and Genetic Transformation Analysis of an Apple Auxin Influx Transporter Gene MdAUX1 in Arabidopsis thaliana and Tobacco

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