中国农业科学 ›› 2021, Vol. 54 ›› Issue (16): 3369-3380.doi: 10.3864/j.issn.0578-1752.2021.16.001
张雅文1(),包淑慧1,唐振家1,王小文1,2,杨芳3,张德春4(),胡一兵1()
收稿日期:
2020-12-18
接受日期:
2021-02-12
出版日期:
2021-08-16
发布日期:
2021-08-24
通讯作者:
张德春,胡一兵
作者简介:
张雅文,E-mail: 基金资助:
ZHANG YaWen1(),BAO ShuHui1,TANG ZhenJia1,WANG XiaoWen1,2,YANG Fang3,ZHANG DeChun4(),HU YiBing1()
Received:
2020-12-18
Accepted:
2021-02-12
Online:
2021-08-16
Published:
2021-08-24
Contact:
DeChun ZHANG,YiBing HU
摘要:
【目的】蔗糖是植物体内光合产物运输的主要形式。蔗糖-质子同向运输蛋白(sucrose transporter/sucrose carrier,SUT/SUC)在植物细胞之间的蔗糖跨膜运输以及植物组织和器官之间的蔗糖分配中具有重要作用。水稻SUT家族共有5个成员。已有研究表明,敲除OsSUT1、OsSUT2、OsSUT3、OsSUT4对水稻的生长发育产生显著影响,说明这些基因不可替代。然而,OsSUT5的功能还未见报道。阐明OsSUT5在水稻生长发育中的作用,将为全面了解SUT蛋白在模式植物水稻中的生理功能提供新的依据。【方法】通过对水稻OsSUT5的时空表达特性进行实时荧光定量PCR分析,同时利用OsSUT5的推测启动子驱动GUS在水稻体内表达,获得其基因编码蛋白的组织定位信息,以及在烟草叶片表皮细胞内进行OsSUT5-GFP融合蛋白瞬时表达,对蛋白进行亚细胞定位,比较基因编辑技术(CRISPR-Cas9)创制的OsSUT5不同纯合突变株系与野生型对照水稻的形态和生理特征,获得OsSUT5的功能信息。【结果】转录水平上,OsSUT5在水稻茎、叶、花序和颖果中均有表达,并且该基因编码蛋白在营养器官中的表达集中于维管组织。在生殖器官中,OsSUT5的表达主要位于花药及发育的颖果内。该基因编码蛋白在水稻发育的颖果特别是盾片和胚根鞘中高表达。烟草叶片表皮细胞内的瞬时表达显示OsSUT5-GFP融合蛋白定位于细胞质膜。与野生型水稻相比,3个OsSUT5纯合突变株系均表现为花粉活性以及离体萌发率明显降低。与此相吻合的是,OsSUT5突变株系未授粉的小穗比例相对于野生型显著增加,同时突变株系结实率显著下降。通过对OsSUT5突变株系颖果的观察和比较显示,OsSUT5突变体水稻颖果的垩白相对于野生型明显增多,其颖果长度相对于野生型对照也有一定程度的增加,但统计结果表明OsSUT5突变体水稻的千粒重与野生型无显著区别。【结论】OsSUT5在水稻花粉发育甚至受精过程中可能发挥重要作用;敲除OsSUT5对水稻的结实率、籽粒的形态和品质有明显影响。推断OsSUT5在水稻开花结实中的作用(包括对花粉活性和颖果内胚乳发育的影响)与其蔗糖运输功能密切相关。
张雅文, 包淑慧, 唐振家, 王小文, 杨芳, 张德春, 胡一兵. 蔗糖转运蛋白OsSUT5在水稻花粉发育及结实中的作用[J]. 中国农业科学, 2021, 54(16): 3369-3380.
ZHANG YaWen, BAO ShuHui, TANG ZhenJia, WANG XiaoWen, YANG Fang, ZHANG DeChun, HU YiBing. Function of Sucrose Transporter OsSUT5 in Rice Pollen Development and Seed Setting[J]. Scientia Agricultura Sinica, 2021, 54(16): 3369-3380.
[1] |
KHUSH G S. Green revolution: The way forward. Nature Review Genetics, 2001, 2(10):815-822.
doi: 10.1038/35093585 |
[2] | LEMOINE R, LA CAMERA S, ATANASSOVA R, DÉDALDÉCHAMP F, ALLARIO T, POURTAU N, BONNEMAIN J L, LALOI M, COUTOS-THÉVENOT P, MAUROUSSET L, FAUCHER M, GIROUSSE C, LEMONNIER P, PARRILLA J, DURAND M. Source-to-sink transport of sugar and regulation by environmental factors. Frontiers in Plant Science, 2013, 4:272. |
[3] | BIHMIDINE S, HUNTER C T 3RD, JOHNS C E, KOCH K E, BRAUN D M. Regulation of assimilate import into sink organs: update on molecular drivers of sink strength. Frontiers in Plant Science, 2013, 4:177. |
[4] | YADAV U P, AYRE B G, BUSH D R. Transgenic approaches to altering carbon and nitrogen partitioning in whole plants: Assessing the potential to improve crop yields and nutritional quality. Frontiers in Plant Science, 2015, 6:275. |
[5] |
SAUER N. Molecular physiology of higher plant sucrose transporters. FEBS Letters, 2007, 581(12):2309-2317.
doi: 10.1016/j.febslet.2007.03.048 |
[6] | LEMOINE R. Sucrose transporters in plants: Update on function and structure. Biochimica et Biophysica Acta, 2000, 1465(1/2):246-262. |
[7] |
ABRAMSON J, SMIRNOVA I, KASHO V, VERNER G, KABACK H R, IWATA S. Structure and mechanism of the lactose permease of Escherichia coli. Science, 2003, 301(5633):610-615.
doi: 10.1126/science.1088196 |
[8] |
RIESMEIER J W, WILLMITZER L, FROMMER W B. Isolation and characterization of a sucrose carrier cDNA from spinach by functional expression in yeast. The EMBO Journal, 1992, 11(13):4705-4713.
doi: 10.1002/embj.1992.11.issue-13 |
[9] |
HIROSE T, IMAIZUMI N, SCOFIELD G N, FURBANK R T, OHSUGI R. cDNA cloning and tissue-specific expression of a gene for sucrose transporter from rice (Oryza sativa L.). Plant and Cell Physiology, 1997, 38(12):1389-1396.
doi: 10.1093/oxfordjournals.pcp.a029134 |
[10] |
AOKI N, HIROSE T, SCOFIELD G N, WHITFELD P R, FURBANK R T. The sucrose transporter gene family in rice. Plant and Cell Physiology, 2003, 44(3):223-232.
doi: 10.1093/pcp/pcg030 |
[11] |
LALONDE S, WIPF D, FROMMER W B. Transport mechanisms for organic forms of carbon and nitrogen between source and sink. Annual Review of Plant Biology, 2004, 55:341-372.
doi: 10.1146/annurev.arplant.55.031903.141758 |
[12] |
AYRE B G. Membrane-transport systems for sucrose in relation to whole-plant carbon partitioning. Molecular Plant, 2011, 4(3):377-394.
doi: 10.1093/mp/ssr014 |
[35] | 杜琳. OsSUT对水稻灌浆生理的分子调控[D]. 福州: 福建农林大学, 2010. |
DU L. Molecular regulation of OsSUT on rice filling physiology[D]. Fuzhou: Fujian Agricultural & Forestry University, 2010. (in Chinese) | |
[13] | DUO P, GU X, XUE L J, LEEBENS-MACK J H, TSAI C J. Bayesian phylogeny of sucrose transporters: Ancient origins, differential expansion and convergent evolution in monocots and dicots. Frontiers in Plant Science, 2014, 5:615. |
[14] |
ZHANG C K, HAN L, SLEWINSKI T L, SUN J L, ZHANG J, WANG Z Y, TURGEON R. Symplastic phloem loading in poplar. Plant Physiology, 2014, 166(1):306-313.
doi: 10.1104/pp.114.245845 |
[36] |
LIVAK K J, SCHMITTGEN T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)). Methods, 2001, 25:402-408.
doi: 10.1006/meth.2001.1262 |
[37] |
MA L, ZHANG D C, MIAO Q, YANG J, XUAN Y H, HU Y B. Essential role of sugar transporter OsSWEET11 during the early stage of rice grain filling. Plant and Cell Physiology, 2017, 58(5):863-873.
doi: 10.1093/pcp/pcx040 |
[15] |
KÜHN C, GROF C P. Sucrose transporters of higher plants. Current Opinion in Plant Biology, 2010, 13(3):287-297.
doi: 10.1016/j.pbi.2010.02.001 |
[16] |
HACKEL A, SCHAUER N, CARRARI F, FERNIE A R, GRIMM B C, KÜHN C. Sucrose transporter LeSUT1 and LeSUT2 inhibition affects tomato fruit development in different ways. The Plant Journal, 2006, 45(2):180-192.
doi: 10.1111/tpj.2006.45.issue-2 |
[17] |
SIVITZ A B, REINDERS A, WARD J M. Arabidopsis sucrose transporter AtSUC1 is important for pollen germination and sucrose-induced anthocyanin accumulation. Plant Physiology, 2008, 147(1):92-100.
doi: 10.1104/pp.108.118992 |
[18] |
PAYYAVULA R S, TAY K H C, TSAI C J, HARDING S A. The sucrose transporter family in Populus: The importance of a tonoplast PtaSUT4 to biomass and carbon partitioning. The Plant Journal, 2011, 65(5):757-770.
doi: 10.1111/tpj.2011.65.issue-5 |
[19] | CHINCINKA I, GIER K, KRÜGEL U, LIESCHE J, HE H, GRIMM B, HARREN F J M, CRISTESCU S M, KÜHN C. Photoperiodic regulation of the sucrose transporter StSUT4 affects the expression of circadian-regulated genes and ethylene production. Frontiers in Plant Science, 2013, 4:26. |
[20] | FURBANK R T, SCOFIELD G N, HIROSE T, WANG X D, PATRICK J, OFFLER C E. Cellular localization and function of a sucrose transporter OsSUT1 in developing rice seeds. Australian Journal of Plant Physiology, 2001, 28(12):1187-1196. |
[21] |
HIROSE T, TAKANO M, TERAO T. Cell wall invertase in developing rice caryopsis: Molecular cloning of OsCIN1 and analysis of its expression in relation to its role in grain filling. Plant and Cell Physiology, 2002, 43(4):452-459.
doi: 10.1093/pcp/pcf055 |
[22] |
HIROSE T, ZHANG Z, MIYAO A, HIROCHIKA H, OHSUGI R, TERAO T. Disruption of a gene for rice sucrose transporter, OsSUT1 impairs pollen function but pollen maturation is unaffected. Journal of Experimental Botany, 2010, 61(13):3639-3646.
doi: 10.1093/jxb/erq175 |
[23] |
ISHIMARU K, HIROSE T, AOKI N, TAKAHASHI S, ONO K, YAMAMOTO S, WU J, SAJI S, BABA T, UGAKI M, MATSUMOTO T, OHSUGI R. Antisense expression of a rice sucrose transporter OsSUT1 in rice (Oryza sativa L.). Plant and Cell Physiology, 2001, 42(10):1181-1185.
doi: 10.1093/pcp/pce148 |
[24] |
ISHIBASHI Y, OKAMURA K, MIYAZAKI M, PHAN T, YUASA T, IWAYA-INOUE M. Expression of rice sucrose transporter gene OsSUT1 in sink and source organs shaded during grain filling may affect grain yield and quality. Environmental and Experimental Botany, 2014, 97:49-54.
doi: 10.1016/j.envexpbot.2013.08.005 |
[25] |
SCOFIELD G N, HIROSE T, AOKI N, FURBANK R. Involvement of the sucrose transporter, OsSUT1, in the long- distance pathway for assimilate transport in rice. Journal of Experimental Botany, 2007, 58(12):3155-3169.
doi: 10.1093/jxb/erm153 |
[26] |
SCOFIELD G N, HIROSE T, GAUDRON J A, FURBANK R T. Antisense suppression of the rice transporter gene, OsSUT1, leads to impaired grain filling and germination but does not affect photosynthesis. Functional Plant Biology, 2002, 29(7):815-826.
doi: 10.1071/PP01204 |
[27] |
ZHANG J, LI D, XU X, ZISKA L H, ZHU J, LIU G, ZHU C. The potential role of sucrose transport gene expression in the photosynthetic and yield response of rice cultivars to future CO2 concentration. Physiologia Plantarum, 2020, 168:218-226.
doi: 10.1111/ppl.v168.1 |
[28] |
EOM J S, CHO J I, REINDERS A, LEE S W, YOO Y, TUAN P Q, CHOI S B, BANG G, PARK Y I, CHO M H, BHOO S H, AN G, HAHN T R, WARD J M, JEON J S. Impaired function of the tonoplast-localized sucrose transporter in rice, OsSUT2, limits the transport of vacuolar reserve sucrose and affects plant growth. Plant Physiology, 2011, 157(1):109-119.
doi: 10.1104/pp.111.176982 |
[29] | 李孟珠, 王高鹏, 巫月, 任怡, 李刚华, 刘正辉, 丁艳锋, 陈琳. 水稻蔗糖转运蛋白OsSUT4参与蔗糖转运的功能研究. 中国水稻科学, 2020, 34(6):491-498. |
LI M Z, WANG G P, WU Y, REN Y, LI G H, LIU Z H, DING Y F, CHEN L. Function analysis of sucrose transporter OsSUT4 in sucrose transport in rice. Chinese Journal of Rice Science, 2020, 34(6):491-498. (in Chinese) | |
[30] | 岳萌萌. 水稻蔗糖转运蛋白OsSUT4的功能分析[D]. 泰安: 山东农业大学, 2020. |
YUE M M. Functional analysis of a rice sucrose transporter OsSUT4[D]. Taian: Shandong Agricultural University, 2020. (in Chinese) | |
[31] |
SUN Y, REINDERS A, LAFLEUR K R, MORI T, WARD J M. Transport activity of rice sucrose transporters OsSUT1 and OsSUT5. Plant and Cell Physiology, 2010, 51(1):114-122.
doi: 10.1093/pcp/pcp172 |
[32] |
BAUD S, WUILLÈME S, LEMOINE R, KRONENBERGER J, CABOCHE M, LEPINIEC L, ROCHAT C. The AtSUC5 sucrose transporter specifically expressed in the endosperm is involved in early seed development in Arabidopsis. The Plant Journal, 2005, 43(6):824-836.
doi: 10.1111/tpj.2005.43.issue-6 |
[33] | LUDWIG A, STOLZ J, SAUER N. Plant sucrose-H + symporters mediate the transport of vitamin H. The Plant Journal, 2000, 24(4):503-509. |
[34] | 张武君, 管其龙, 付艳萍, 苏军. 反义抑制水稻蔗糖转运蛋白基因(OsSUT5)的表达降低其愈伤组织诱导和植株再生频率. 农业生物技术学报, 2014, 22:825-831. |
ZHANG W J, GUAN Q L, FU Y P, SU J. Antisense suppression of rice sucrose transporter gene (OsSUT5) expression reduces its callus induction and plant regeneration frequency. Journal of Agricultural Biotechnology, 2014, 22:825-831. (in Chinese) | |
[38] |
MIAO J, GUO D, ZHANG J, HUANG Q, QIN G, ZHANG X, WAN J, GU H, QU L J. Targeted mutagenesis in rice using CRISPR-Cas system. Cell Research, 2013, 23:1233-1236.
doi: 10.1038/cr.2013.123 |
[39] | 戚继艳, 阳江华, 唐朝荣. 植物蔗糖转运蛋白的基因与功能. 植物学通报, 2007, 24:532-543. |
QI J Y, YANG J H, TANG C R. Sucrose transporter genes and their functions in plants. Chinese Bulletin of Botany, 2007, 24:532-543. (in Chinese) | |
[40] | 陈钢, 母天燕, 曹光球, 林强, 郑宏, 曹世江. 杉木蔗糖转运蛋白SUT基因的克隆和功能分析. 西北林学院学报, 2020, 35(2):8-14. |
CHEN G, MU T Y, CAO G Q, LIN Q, ZHENG H, CAO S J. Cloning and functional analysis of Chinese Fir sucrose transporter SUT gene. Journal of Northwest Forestry University, 2020, 35(2):8-14. (in Chinese) | |
[41] | 康爽, 贺红霞, 王铭, 郭嘉, 薛晶. 杏蔗糖转运蛋白基因PaSUC4的获得及其生物信息学分析. 生物技术进展, 2016, 6:105-112. |
KANG S, HE H X, WANG M, GUO J, XUE J. The acquisition and bioinformatic analysis of sucrose transporter protein gene PaSUC4 from apricot. Current Biotechnology, 2016, 6:105-112. (in Chinese) | |
[42] | 齐素坤, 侯夫云, 秦桢, 李爱贤, 王庆美, 张立明. 植物蔗糖转运蛋白研究进展. 山东农业科学, 2016, 48(2):149-153. |
QI S K, HOU F Y, QIN Z, LI A X, WANG Q M, ZHANG L M. Research progress of sucrose transporters in plant. Shandong Agricultural Science, 2016, 48(2):149-153. (in Chinese) | |
[43] | 张清, 胡伟长, 张积森. 植物蔗糖转运蛋白研究进展. 热带作物学报, 2016, 37(1):193-202. |
ZHANG Q, HU W C, ZHANG J S. Sucrose transporters in plants. Chinese Journal of Tropical Crops, 2016, 37(1):193-202. (in Chinese) | |
[44] | 张立军, 李晓宇, 阮燕晔. 双子叶与单子叶植物蔗糖转运蛋白的研究进展. 沈阳农业大学学报, 2008, 39(3):259-264. |
ZHANG L J, LI X Y, RUAN Y Y. Progress on sucrose transporters in dicotyledons and monocotyledons. Journal of Shenyang Agricultural University, 2008, 39(3):259-264. (in Chinese) | |
[45] | 王忠. 水稻的开花与结实. 北京: 科学出版社, 2014: 91-109. |
WANG Z. The Flowering and Fruiting of Rice. Beijing: Science Press, 2014: 91-109. (in Chinese) | |
[46] | 杨宏远. 水稻生殖生物学. 杭州: 浙江大学出版社, 2005: 91-98. |
YANG H Y. Rice Reproductive Biology. Hangzhou: Zhejiang University Press, 2005: 91-98. (in Chinese) | |
[47] | 周立军, 江玲, 翟虎渠, 万建民. 水稻垩白的研究现状与改良策略. 遗传, 2009, 31(6):563-572. |
ZHOU L J, JIANG L, ZHAI H Q, WAN J M. Research status and improvement strategies of rice chalkiness. Heredity, 2009, 31(6):563-572. (in Chinese) | |
[48] |
LI D, XU R, LV D, ZHANG C, YANG H, ZHANG J, WEN J, LI C, TAN X. Identification of the core pollen-specific regulation in the rice OsSUT3 promoter. International Journal of Molecular Sciences, 2020, 21(6):1909.
doi: 10.3390/ijms21061909 |
[49] | 李丹丹, 李娟, 栾一方, 张春龙, 徐汝聪, 吕东, 谭亚玲, 谭学林. 水稻OsSUT3基因启动子的克隆及表达分析. 分子植物育种, 2018, 16:7225-7233. |
LI D D, LI J, LUAN Y F, ZHANG C L, XU R C, LÜ D, TAN Y L, TAN X L. Cloning and expression analysis of OsSUT3 gene promoter from Oryza sativa. Molecular Plant Breeding. 2018, 16:7225-7233. (in Chinese) |
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