Rabdosinate调节生长素极性运输蛋白PIN1、PIN3和PIN4抑制拟南芥幼苗根生长

doi:10.7525/j.issn.1673-5102.2019.06.014

摘要/Abstract

摘要： 利用3种拟南芥生长素极性运输外运载体突变体及4种转基因株系研究了二萜rabdosinate抑制拟南芥幼苗主根及侧根生长的作用机制。结果显示，60~80 μmol·L^-1的rabdosinate显著抑制野生型拟南芥幼苗主根生长及侧根形成，而对突变体pin1、pin2和pin3主根未显示明显的抑制效应，对侧根的抑制减弱；发现rabdosinate （60~80 μmol·L^-1）引起生长素报告株系根尖DR5活性升高，并增加融合蛋白PIN1-GFP丰度以及减少PIN3-GFP和PIN4-GFP的丰度。推断rabdosinate可通过增加PIN1丰度促进了根部生长素向顶运输，而减少PIN3丰度降低根尖部生长素的横向转运，引起了生长素在根尖部的累积及生长素浓度梯度的改变，进而抑制幼苗主根生长及侧根发育。

关键词: rabdosinate, 二萜, 拟南芥, 生长素极性运输, 根生长

Abstract: This paper studied the mechanism of inhibition of primary root growth and lateral root development regulated by rabdosinate using polar auxin carriers reporter lines(PIN1::PIN1:GFP,PIN3::PIN3:GFP,PIN4::PIN4:GFP) and auxin reporters line(DR5::DR5:GFP). The results showed that the length of primary root and the number of lateral root in wild-type Arabidopsis thaliana seedlings were significantly reduced by 60~80 μmol·L^-1 rabdosinate. However, the mutants pin1, pin3 and pin4 showed a lower level of sensitivity to rabdosinate in terms of the inhibition of root elongation and lateral root formation compared with the WT seedlings.Rabdosinate(60~80 μmol·L^-1)induced a dose-dependent increase in DR5 activity and the abundance of PIN1-GFP fusion proteins in the root tips of A.thaliana, and significantly decreased the abundance of PIN3-GFP and PIN4-GFP at their respective cellular locations. These results suggest that rabdosinate may promote acropetal auxin transport activities by increasing the abundance of PIN1 and block transverse auxin transport activities by decreasing the abundance of PIN3-GFP and PIN4-GFP, resulting in auxin accumulation in the root meristematic cells and the inhibition of root growth and lateral root development.

Key words: rabdosinate, diterpene, Arabidopsis thaliana, polar auxin transport, root growth

中图分类号:

Q949.748.3

李芃, 郇兆蔚, 丁兰. Rabdosinate调节生长素极性运输蛋白PIN1、PIN3和PIN4抑制拟南芥幼苗根生长[J]. 植物研究, 2019, 39(6): 908-916.

LI Peng, HUAN Zhao-Wei, DING Lan. Rabdosinate Regulates Polar Auxin Carriers PIN1,PIN3 and PIN4 and Inhibits Root Growth of Arabidopsis thaliana Seedlings[J]. Bulletin of Botanical Research, 2019, 39(6): 908-916.

参考文献

1. Ashour M,Wink M and Gershenzon J. Biochemistry of Terpenoids:monoterpenes sesquiterpenes and diterpenes[J]. Annual Plant Reviews,2010(40):258-303.
2. Withers S T,Keasling J D. Biosynthesis and engineering of isoprenoid small molecules[J]. Applied Microbiology & Biotechnology,2007,73(5):980-990.
3. Tholl D. Biosynthesis and biological functions of terpenoids in plants[J]. Advances in Biochemical Engineering/biotechnology,2015,148:63-106.
4. 蒲洁莹,何莉,吴思宇,等. 甘草属植物中三萜类化合物的抗病毒作用研究进展[J]. 病毒学报. 2013,29(6):673-679. Pu J Y,He L,Wu S Y,et al. Anti-virus research of triterpenoids in Licorice[J]. Chinese Journal of Virology,2013,29(6):673-679.
5. 李国华,李晔,梅锡玲,等. 灵芝三萜类化合物研究进展[J]. 中草药,2015,46(12):1858-1862. Li G H,Li H,Mei X L,et al. Study progress on triterpenoids in Ganoderma lucidum[J]. Chinese Traditional and Herbal Drugs,2015,46(12):1858-1862
6. Weir T L,Park S W,Vivanco J M. Biochemical and physiological mechanisms mediated by allelochemicals[J]. Current Opinion in Plant Biology,2004,7(4):472-479.
7. 陈圣宾,李振基. 外来植物入侵的化感作用机制探讨[J]. 生态科学,2005,24(1):69-74. Chen S B,Li Z J. Discussion on allelopathy mechanism of exotic plant invasion[J]. Ecological Science,2005,24(4):69-74.
8. Kato-Noguchi,Hisashi,Peters,et al. The role of momilactones in rice allelopathy[J]. Journal of Chemical Ecology,2013,39(2):175-185.
9. 王建花,陈婷,林文雄. 植物化感作用类型及其在农业中的应用[J]. 中国生态农业学报,2013,21(10):1173-1183. Wang J H,Chen T,Lin W X,et al. Plant aiielopathy types and their and application in agriculture[J]. Chinese Journal of Eco-Agriculture,2013,21(10):1173-1183.
10. Ding L,Jing H W,Qin B,et al. Regulation of cell division and growth in roots of Lactuca sativa L. seedlings by the ent-kaurene diterpenoid rabdosin B[J]. Journal of Chemical Ecology,2010,36(5):553-563.
11. Ding L,Jing H W,Wang T,et al. Regulation of root growth in Lactuca sativa L. seedlings by the ent-kaurane diterpenoid epinodosin[J]. Journal of Plant Growth Regulation,2010,29(4):419-427.
12. Hu Y F,Yang L J,Na X F,et al. Narciclasine inhibits the responses of Arabidopsis roots to auxin[J]. Planta,2012,236(2):597-612.
13. Hu Y F,Na X F,Li J L,et al. Narciclasine,a potential allelochemical,affects subcellular trafficking of auxin transporter proteins and actin cytoskeleton dynamics in Arabidopsis roots[J]. Planta,2015,242(6):1349-1360.
14. Abrahim D,Francischini A C,Pergo E M,et al. Effects of α-pinene on the mitochondrial respiration of maize seedlings[J]. Plant Physiology and Biochemistry,2003,41(11-12):985-991.
15. Eva B,Jan H. Hormone interactions at the root apical meristem[J]. Plant Molecular Biology,2009,69(4):383-396.
16. Sun H D,Huang S X,Han Q B. Diterpenoids from Isodon species and their biological activities[J]. Cheminform,2006,38(5):673-698.
17. Ding L,Qi L L,Jing H W,et al. Phytotoxic effects of Leukamenin E(an ent-kaurene diterpenoid) on root growth and root hair development in Lactuca sativa L. seedlings[J]. Journal of Chemical Ecology,2008,34(11):1492-1500.
18. 程蔚玲,丁兰,李金平,等. Leukamenin E调节拟南芥幼苗生长发育的模式及其机制[J]. 生态学杂志,2017,36(3):676-686. Cheng W L,Ding L,LI J P,et al. The pattern and mechanism of seedling growth and development regulated by Leukamenin E[J]. Chinese Journal of Ecology,2017,36(3):676-686.
19. 丁兰,祁林林,景宏伟,等. 四种对映-贝壳杉烷二萜化合物化感作用潜能的初步评价[J]. 西北师范大学学报:自然科学版,2008,44(6):74-78. Ding L,Qi L L,Jing H W,et al. Preliminary evaluation of the allelopathic potential of four ent-kaurene diterpene[J]. Journal of Northwest Normal University:Natural Science,2008,44(6):74-78.
20. 丁兰,景宏伟,王涛,等. 甘肃产毛叶香茶菜二萜成分及其化感潜能评估[J]. 西北师范大学学报:自然科学版,2010,46(2):88-93. Ding L,Jing H W,Wang T,et al. Chemical constituent s and allelopathic potential of Isodon japonica(Burman f1) Hara[J]. Journal of Northwest Normal University:Natural Science,2010,46(2):88-93.
21. 丁兰,王炜,汪涛,等. 甘肃产蓝萼香茶菜二萜化学成分研究[J]. 广西植物. 2008,28(2):265-268. Ding L,Wang W,Wang T,et al. Chemical components of diterpenoids from Isodon japonica var. galaucocalyx from Gansu Prpvince,China[J]. Guihaia,2008,28(2):265-268.
22. Zhang H,Jennings A,Barlow P W et al. Dual pathways for regulation of root branching by nitrate[J]. Proceedings of the National Academy of Sciences of the United States of America,1999,96(11):6529-6534.
23. Araniti F,Bruno L,Sunseri F,et al. The allelochemical farnesene affects Arabidopsis thaliana root meristem altering auxin distribution[J]. Plant Physiology and Biochemistry,2017,121:14-20.
24. Benková E,Michniewicz M,Sauer M,et al. Local,efflux-dependent auxin gradients as a common module for plant organ formation[J]. Cell,2003,115:591-602.
25. Casimiro I,Marchant A,Bhalerao R P,et al. Auxin transport promotes Arabidopsis lateral root initiation[J]. Plant Cell,2001,13(4):843-852.
26. 邹纯雪,门淑珍. 生长素的外输载体PIN蛋白家族研究进展[J]. 中国细胞生物学学报,2013(5):574-582. Zou C X,Men S Z. Research advances in auxin efflux carrier PIN proteins[J]. Chinese Journal of Cell Biology,2013(5):574-582.
27. Bradow J M,Connick W J. Volatile seed germination inhibitors from plant residues[J]. Journal of Chemical Ecology,1990,16(3):645-666.
28. Asplund R O. Monoterpenes:relationship between structure and inhibition of germination[J]. Phytochemistry,1968,7(11):1995-1997.
29. 李俊华,种康. 植物生长素极性运输调控机理的研究进展[J]. 植物学通报,2006,23(5):466-477. Li J H,Zhong K. Current research advances on polar auxin transport in plant[J]. Chinese Bulletin of Botany. 2006,23(5):466-477.
30. Ding L,Hou Q,Zhou Q,et al. Structure-activity relationships of eight ent-kaurene diterpenoids from three Isodon plants[J]. Research on Chemical Intermediates,2010,36(4):443-452.
31. Vilches-Barro A,Maizel A. Talking through walls:mechanisms of lateral root emergence in Arabidopsis thaliana[J]. Current Opinion in Plant Biology,2015,23:31-38.
32. Blilou I,Xu J,Wildwater M,et al. The PIN auxin efflux facilitator network controls growth and patterning in Arabidopsis roots[J]. Nature,2005,433(7021):39-44.
33. Friml J,Wisniewska J N,Benková E,et al. Lateral relocation of auxin efflux regulator PIN3 mediates tropism in Arabidopsis[J]. Nature,2002,415:806-809.
34. Friml J,Benková E,Blilou I,et al. AtPIN4 mediates sink-driven auxin gradients and root patterning in Arabidopsis[J]. Cell,2002,108(5):661-673.

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