水稻SUMO化E3连接酶SIZ1调控缺磷条件下根的发育和根构型形成

doi:10.3969/j.issn.1001-7216.2015.01.005

摘要/Abstract

摘要：

植物应对磷胁迫的方法之一是改变根系的构型。以水稻SUMO化E3连接酶SIZ1突变体ossiz1为供试材料,研究了OsSIZ1在水稻根发育中的作用以及其与磷胁迫、生长素之间的关系。与野生型相比,OsSIZ1抑制ossiz1种子根和不定根的伸长,促进侧根密度的增加和根毛的增多。缺磷时,突变体ossiz1的反应更强烈,即不定根伸长、侧根密度增大和根毛增多的趋势更加明显。说明OsSIZ1参与调控水稻根构型的改变,低磷时效果更明显。ossiz1地上部和地下部的总磷浓度显著高于野生型,说明OsSIZ1在水稻中负调控磷素的吸收利用。定量RT-PCR结果显示,ossiz1中OsYUCCA1和OsPIN1a/1b的相对表达量显著高于野生型,说明OsSIZ1负调控根中生长素的合成与极性运输,并且缺磷时负调控作用减弱。结果表明,SUMO化E3连接酶OsSIZ1调控缺磷条件下根构型的形成,而且这一过程可能是通过调控生长素分布完成的。

关键词: 水稻, 磷, OsSIZ1, 根构型

Abstract:

Plants cope with the low P environment by altering their root architecture. Here, we studied the role of SUMO E3 ligase SIZ1 on the root development and architecture, and the relationship among OsSIZ1, phosphate stress and auxin using ossiz1 mutants. Compared with wild-type (WT), the ossiz1 mutation inhibited the elongation of seminal roots and adventitious roots, promoted lateral roots formation and root hair proliferation. However, the response of the ossiz1 mutation was stronger than WT under Pi deficiency, namely, the trend of root elongation and the increase of the lateral root density and the root hair number of ossiz1 were more obvious relative to WT seedlings. This suggests that OsSIZ1 regulates the root architecture, and the effect is more obvious under Pi deficiency. The total P concentration of shoots and roots in ossiz1 is higher than WT, this suggests that OsSIZ1 negatively regulates phosphorus acquisition and use. And relative expression of OsYUCCA1 and OsPIN1a/1b in ossiz1 roots is markedly up-regulated compared with WT. This suggests that OsSIZ1 negatively regulates the auxin biosynthesis and polar auxin transport, which is weakened under Pi-deficient conditions. Together, these results suggest that OsSIZ1 regulates Pi starvation-induced root architecture remodeling possibly through the control of auxin patterning.

Key words: rice, phosphate, OsSIZ1, root architecture

中图分类号:

周红敏, 王化敦, 孙瑞, 裴文霞, 吴学能, 曹越, 孙雅菲, 徐国华, 孙淑斌. 水稻SUMO化E3连接酶SIZ1调控缺磷条件下根的发育和根构型形成[J]. 中国水稻科学, 2015, 29(1): 35-44.

Hong-min ZHOU, Hua-dun WANG, Rui SUN, Wen-xia PEI, Xue-neng WU, Yue CAO, Ya-fei SUN, Guo-hua XU, Shu-bin SUN. OsSIZ1 Regulates the Development and Architecture of the Roots under Phosphate Starvation Conditions in Rice[J]. Chinese Journal OF Rice Science, 2015, 29(1): 35-44.

图/表 7

表1 生长素生物合成基因OsYUCCA1和生长素外流蛋白基因OsPINs的序列

Table 1 Primers used to amplify the OsYUCCA1 and OsPINs cDNA fragments.

基因 Gene	引物序列 Primer sequences (5'-3')
OsActin	F: GGAACTGGTATGGTCAAGGC
	R: AGTCTCATGGATAACCGCAG
OsPIN1a	F: TCATCTGGTCGCTCGTCTGC
	R: CGAACGTCGCCACCTTGTTC
OsPIN1b	F: TGCACCCTAGCATTCTCAGCA
	R: CCCTCCTCCCAAATTCTACTT
OsYUCCA1	F: TCATCGGACGCCCTCAACGTCGC
	R: GGCAGAGCAAGATTATCAGTC
OsPIN2	F:CAACACCTACTCCAGCCTC
	R:TGGACCAGTCAAGAACCTC

图1 突变体ossiz1及其野生型种子根的生长情况 A-发芽7 d后水稻幼苗种子根的表型,白色线条表示1 cm; B-发芽7 d后水稻幼苗种子根不同部位的根毛生长,白色线条表示2 cm; C和D-营养液开始处理后7 d内水稻幼苗种子根的根长和生长速率。WT-野生型。

Fig.1. Growth performances of seminal roots of ossiz1 and wild type. A, Root phenotype of rice seedlings 7 d after germination, bar=1 cm; B, The root hair proliferation on the seminal roots of rice seedlings 7 d after germination, bar=2 cm; C,D, The length and the growth rate of seminal root of rice seedlings for 7 d after transfer to the nutrient solution. WT, Wild type.

图2 正常供磷(+P)和缺磷(-P)固体培养基中野生型和ossiz1根系的表型分析 A、C中白色线条分别表示1 cm,2 cm。

Fig. 2. Root phenotypic analysis of wild type(WT) and ossiz1 seedlings in the Pi-sufficient(+P) and Pi-deficient(-P) solid culture medium. In A, Bar=1 cm; In C, Bar=2 cm.

图3 正常供磷(+P)和缺磷(-P)条件下ossiz1突变体及其野生型根系的表型分析 A、C中白色线条分别表示1 cm; 2 cm。

Fig. 3. Root phenotype of wild type (WT) and ossiz1 seedlings under Pi-sufficient(+P) and Pi-deficient(-P) conditions. In A, Bar=1 cm; In C; Bar=2 cm.

图4 正常供磷和缺磷条件下ossiz1突变体及其野生型根系中不定根的统计分析 A-不定根的扫描图; B-每株水稻苗根系中无成簇大侧根发生的不定根(a, c, e, g)和有成簇大侧根发生的不定根(b, d, f, h); C-有成簇大侧根发生的不定根占总不定根数的百分比。

Fig. 4. Statistical analysis of adventitious roots of ossiz1 and its wild type (WT) seedlings under Pi-sufficient (+P) and Pi-deficient (-P) conditions. A,Root scanning; B, Adventitious roots without large lateral root formation in clumps (a, c, e, g) and adventitious roots with large lateral root formation in clumps (b, d, f, h); C,The ratio of adventitious roots with large lateral root formation in clumps to the total adventitious roots.

图5 水培正常供磷和缺磷条件下ossiz1及其野生型地上部和地下部总磷浓度

Fig. 5. Total P concentration in the shoots and roots of ossiz1 and its wild type (WT) seedlings under Pi-sufficient (+P) and Pi-deficient (-P) conditions in hydroponics culture.

图6 水培试验时WT和ossiz1根中生长素途径中相关基因的表达 A-正常供磷和缺磷处理2周后生长素生物合成基因OsYUCCA1的相对表达量; B-正常供磷和缺磷处理2周后生长素外流基因OsPINs的相对表达量。

Fig. 6. Expression of auxin genes in roots of WT and ossiz1 in hydroponics experiment. A,Relative expression of auxin biosynthesis gene OsYUCCA1 subjected to +P and -P conditions for 2 weeks; B, Relative expression of auxin efflux transporters OsPINs subjected to +P and -P conditions for 2 weeks.

参考文献 53

[1]	杨辉霞, 童依平, 王道文. 拟南芥低磷胁迫反应分子机理研究的最新进展. 植物学通报, 2007, 24: 726-734.
[2]	Chiou T J, Lin S I.Signaling network in sensing phosphate availability in plants.Annu Rev Plant Biol, 2011, 62: 185-206.
[3]	Grime J P, Crick J C, Rincon J E.The ecological significance of plasticity.Symp Soc Exp Biol, 1986, 40: 5-29.
[4]	Price A H, Tomos A D, Virk D S.Genetic dissection of root growth in rice (Oryza sativa L.): Ⅰ. A hydrophonic screen.Theor Appl Genet, 1997, 95: 132-142.
[5]	Narang R A, Bruene A, Altmann T.Analysis of phosphate acquisition efficiency in different Arabidopsis accessions.Plant Physiol, 2000, 124: 1786-1799.
[6]	Malamy J E.Intrinsic and environmental response pathways that regulate root system architecture.Plant Cell Environ, 2005, 28: 67-77.
[7]	López-Bucio J, Cruz-Ramírez A, Herrera-Estrella L.The role of nutrient availability in regulating root architecture.Curr Opin Plant Biol, 2003, 6: 280-287.
[8]	Raghothanma K G.Phosphate acquisition.Annu Rev Plant Physiol Plant Mol Biol, 1999, 50: 665-693.
[9]	黄荣, 孙虎威, 刘尚俊, 等. 低磷胁迫下水稻根系的发生及生长素的响应. 中国水稻科学, 2012, 26(5): 563-568.
[10]	Friml J, Vieten A, Sauer M, et al.Efflux-dependent auxin gradients establish the apical-basal axis ofArabidopsis. Nature, 2003, 426: 47-53.
[11]	Benkova E, Michniewicz M, Sauer M, et al.Local, efflux-dependent auxin gradients as a common module for plant organ formation.Cell, 2003, 115: 591-602.
[12]	Mattsson J, Ckurshumova W, Berleth T.Auxin signaling in Arabidopsis leaf vascular development.Plant Physiol, 2003, 131: 1327-1339.
[13]	Blakeslee J J, Bandyopadhyay A, Peer W A, et al.Relocalization of the PIN1 auxin efflux facilitator plays a role in phototropic responses.Plant Physiol, 2004, 134: 28-31.
[14]	Palme K, Dovzhenko A, Ditengou F A.Auxin transport and gravitational research: Perspectives.Protoplasma, 2006, 229: 175-181.
[15]	Vanneste S, Friml J.Auxin: A trigger for change in plant development.Cell, 2009, 136: 1005-1016.
[16]	Zhi G E, Ge L, Wang L.Molecular mechanism of adventitious root formation in rice.Plant Growth Regul, 2012, 68: 325-331.
[17]	Yamamoto Y, Kamiya N, Morinaka Y, et al.Auxin biosynthesis by the YUCCA genes in rice.Plant Physiol, 2007, 143: 1362-1371.
[18]	Morita Y, Kyozuka J.Characterization of OsPID, the rice ortholog of PINOID, and its possible involvement in the control of polar auxin transport.Plant Cell Physiol, 2007, 48: 540-549.
[19]	Dong L, Wang L, Zhang Y, et al.An auxin-inducible F-box protein CEGENDUO negatively regulates auxin-mediated lateral root formation in Arabidopsis.Plant Mol Biol, 2006, 60: 599-615.
[20]	Lee S H, Cho H T.PINOID positively regulates auxin efflux in Arabidopsis root hair cells and tobacco cells.Plant Cell, 2006, 18: 1604-1616.
[21]	Duan Q H, Kita D, Li C, et al.FERONIA receptor-like kinase regulates RHO GTPase signaling of root hair development.PNAS, 2010, 107(41): 17821-17826.
[22]	Shen C J, Wang S K, Zhang S N, et al.OsARF16, a transcription factor, is required for auxin and phosphate starvation response in rice (Oryza sativa L.). Plant, Cell,Environ, 2013, 36: 607-620.
[23]	Lopez-Bucio J, Hernandez-Abreu E, Sanchez-Calderon L, et al.An auxin transport independent pathway is involved in phosphate stress-induced root architectural alterations in Arabidopsis.Plant Physiol, 2005, 137: 681-691.
[24]	Lopez-Bucio J, Hernandez-Abreu E, Sanchez-Calderon L, et al.Phosphate availability alters architecture and causes changes in hormone sensitivity in the Arabidopsis root system.Plant Physiol, 2002, 129: 244-256.
[25]	Pérez-Torres C A, Lo’pez-Bucio J, Cruz-Ramı’rez A, et al. Phosphate availability alters lateral root development in Arabidopsis by modulating auxin sensitivity via a mechanism involving the TIR1 auxin receptor.Plant Cell, 2008, 20: 3258-3272.
[26]	Nacry P, Canivenc G, Muller B, et al.A role for auxin redistribution in the responses of the root system architecture to phosphate starvation inArabidopsis. Plant Physiol, 2005, 138: 2061-2074.
[27]	Chen Y N, Fan X R, Song W J, et al.Over-experssion of OsPIN2 leads to increased tiller numbers, angle and shorter plant height through suppression of OsLAZY1.Plant Biol, 2012, 10: 139-149.
[28]	陈泉, 施蕴渝. 小泛素相关修饰SUMO研究进展. 生命科学, 2004, 16(1): 1-6.
[29]	徐庞连, 曾棉炜, 黄丽霞,等. 植物SUMO化修饰及其生物学功能.植物学通报, 2008, 25(5): 608-615.
[30]	Miura K, Rus A, Sharkhuu A, et al.Arabidopsis SUMO E3 ligase SIZl controls phosphate deficiency responses.PNAS, 2005, 102: 7760-7765.
[31]	Miura K, Jin J B, Hasegawa P M.Sumoylation, a post-translational regulatory process in plants.Curr Opin Plant Biol, 2007, 10: 495-502.
[32]	Yoo C Y, Miura K, Jin J B, et al.SIZl small ubiquitin-like modifier E3 ligase facilitates basal thermotolerance in Arabidopsis independent of salicylic acid.Plant Physiol, 2006, 142: 1548-1558.
[33]	Catala R, Ouyang J, Abreu I A, et al.The Arabidopsis E3 SUMO ligase SIZ1 regulates plant growth and drought responses.Plant Cell, 2007, 19: 2952-2966.
[34]	Saminathan T, Guo C L, Chuang M H.Rice SIZ1, a SUMO E3 ligase, controls spikelet fertility through regulation of anther dehiscence.New Phytol, 2011, 189(3): 869-882.
[35]	Miura K, Rus A, Sharkhuu A, et al.The Arabidopsis SUMO E3 ligase SIZ1 controls phosphate deficiency responses.PNAS, 2005, 102: 7760-7765.
[36]	Miura K, Lee J, Gong Q.SIZ1 Regulation of phosphate starvation-induced root architecture remodeling involves the control of auxin accumulation.Plant Physiol, 2011, 155: 1000-1012.
[37]	Wang H D, Makeen K, Yan Y.OsSIZ1 regulates the vegetative growth and reproductive development in rice.Plant Mol Biol Rep, 2010, 29: 411-417.
[38]	鲍士旦. 土壤农化分析.北京:中国农业出版社, 1999.
[39]	Beemster G T, De Vusser K, De Tavernier E, et al.Variation in growth rate between Arabidopsis ecotypes is correlated with cell division and A-typecyclin-dependent kinase activity.Plant Physiol, 2002, 129: 854-864.
[40]	Mouchel C F, Briggs G C, Hardtke C S.Natural genetic variation in Arabidopsis identifies BREVIS RADIX, a novel regulator of cell proliferation and elongation in the root.Genes Dev, 2004, 18: 700-714.
[41]	Zhu Y, Dong A, Meyer D, et al.Arabidopsis NRP1 and NRP2 encode histone chaperones and are required for maintaining postembryonic root growth.Plant Cell, 2006, 18: 2879-2892.
[42]	Jia L, Zhang B, Mao C, et al.OsCYT-INV1 for alkaline/neutral invertase is involved in root cell development and reproductivity in rice (Oryza sativa L.).Planta, 2008, 228: 51-59.
[43]	Miura K, Lee J, Miura T, Hasegawa PM.SIZ1 controls cell growth and plant development in Arabidopsis through salicylic acid.Plant Cell Physiol, 2010, 51: 103-113.
[44]	Huang L, Yang S, Zhang S, et al.The Arabidopsis SUMO E3 ligase AtMMS21, a homologue of NSE2/MMS21, regulates cell proliferation in the root.Plant J, 2009, 60: 666-678.
[45]	郭朝晖, 张杨珠, 黄见良, 等.磷对杂交水稻根系特性与养分吸收的影响. 湖南农业大学学报.自然科学版, 2001, 27(5):350-354.
[46]	郭再华, 贺立源, 徐才国.水稻耐低磷特性研究.应用与环境生物学报, 2004, 10(6):681-685.
[47]	郭再华, 贺立源, 徐才国.磷水平对不同耐低磷水稻苗根系生长及氮、磷、钾吸收的影响. 应用与环境生物学报, 2006, 12(4):449-452.
[48]	Sasaki O, Yamazaki K, Kawata S.The relationship between the diameters and the structures of lateral roots in rice plants.Jpn J Crop Sci, 1984, 53: 169-175.
[49]	Rebouillat J, Dievart A, Verdeil J L.Molecular genetics of rice root development.Rice, 2009, 2: 15-34.
[50]	Zhu Z X, Liu Y, Liu S J, et al.A gain-of-function mutation in OsIAA11 affects lateral root development in rice.Mol Plant, 2012, 5: 154-161.
[51]	Grierson C, Schiefelbein J.Root Hairs// The Arabidopsis Book. Roekville MD: Ameriean Society of Plant Biologists, 2002.
[52]	Peret B, De Rybel B, Casimiro I, et al.Arabidopsis lateral root development: An emerging story.Trends Plant Sci, 2009, 14: 399-408.
[53]	Benkova E, Bielach A.Lateral root organogenesis-from cell to organ.Curr Opin Plant Biol, 2010, 13: 677-683.