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Acta Agronomica Sinica ›› 2021, Vol. 47 ›› Issue (3): 472-480.doi: 10.3724/SP.J.1006.2021.03027

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Functional analysis of plasma membrane intrinsic protein ZmPIP1;1 involved in drought tolerance and photosynthesis in maize

ZHOU Lian, LIU Chao-Xian, XIONG Yu-Han, ZHOU Jing, CAI Yi-Lin*()   

  1. Maize Research Institute / Academy of Agricultural Sciences / State Cultivation Base of Crop Stress Biology for Southern Mountainous Land, Southwest University, Chongqing 400715, China
  • Received:2020-05-14 Accepted:2020-09-13 Online:2021-03-12 Published:2020-10-06
  • Contact: CAI Yi-Lin E-mail:caiyilin1789@163.com
  • Supported by:
    National Natural Science Foundation of China(31601312)

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Abstract:

Plasma membrane intrinsic protein (PIP) is one of the main subfamily of aquaporin regulates diverse physiology functions during plant growth and development. In previous research, the expression of ZmPIP1;1 was induced by osmosis or salt stress. However, the biological function of ZmPIP1;1 was still unclear in maize. In this study, ZmPIP1;1 overexpressed transgenic plants were obtained and exhibited less water loss rate and enhanced drought tolerance compared to wild type. Transcriptome sequencing indicated significant changes in the expression levels of genes involved in ABA biosynthesis and its signaling pathways. Photosynthetic activity, kernel width and kernel weight was increased in ZmPIP1;1 overexpression maize, but not growth under normal condition compared with wild type. Moreover, interaction between ZmPIP1;1 and ZmPIP2;6 was observed by bimolecular fluorescence complementation (BiFC) experiment, resulting in re-localized on plasma membrane and chloroplast in maize mesophyll protoplast. Our study laid an important foundation for understanding the molecular mechanism of ZmPIP1;1, and provided a new method of molecular breeding for high photosynthetic efficiency.

Key words: maize, ZmPIP, drought stress, photosynthesis, re-localization

Fig. 1

Identification of ZmPIP1;1 overexpressed transgenic maize A: schematic illustration of T-DNA sequence of ZmPIP1;1 overexpression vector. B: PCR detection of T1 generation of ZmPIP1;1 overexpression transgenic lines; 1-5: five independent transgenic lines; MK: DNA marker; PC: positive control; WT: wild type. C: qRT-PCR analysis of four representative ZmPIP1;1 overexpressed transgenic lines."

Fig. 2

Phenotypes and water loss rate of ZmPIP1;1 overexpressed transgenic lines under drought treatment A: phenotype of WT and ZmPIP1;1 overexpressed transgenic maize under drought stress treatment for 15 days. B: relative shoot biomass of different genotypes grown in soil with the indicated treatments, compared with normal conditions. Values are means of four biological replicates with error bars indicating standard derivations (SD). C: determination of water loss from detached leaves of WT and ZmPIP1;1 overexpressed transgenic line. Values followed by * are significantly different at P < 0.01."

Fig. 3

Clusters of orthologous groups (COG) function classification of consensus sequences T and G columns represent the functions of “signal transduction mechanisms” and “carbohydrate transport and metabolism”, respectively."

Fig. 4

Relative expression level of key genes involved in ABA biosynthesis and its signaling pathway The relative expression level of (A) ZmNCED9-1, ZmNCED9-2, ZmABA1, and ZmAAO3 involved in ABA biosynthesis (B) ZmABF1, ZmPYL2, ZmABI5, and ZmSnRK3 involved in ABA signaling pathway in leaves of the WT and ZmPIP1;1 overexpressed lines under normal and drought stress treatments. Data are means of three biological replicates with error bars indicating standard deviations (SD). Values followed by asterisk are significantly different at P < 0.01. ZmACT1 was used as the internal control."

Fig. 5

AN and Ci of WT and ZmPIP1;1 overexpressing transgenic maize plants All data are means ± standard deviations (SD) (n = 4). Values followed by asterisk are significantly different at P < 0.01."

Fig. 6

Phenotypic characterization of ZmPIP1;1 overexpressed transgenic maize kernel Phenotypic determination (A) and photograph (B) of ZmPIP1;1 overexpressed transgenic maize kernel. Values followed by asterisk are significantly different at P < 0.01. Bar = 1 cm."

Fig. 7

Observation of BiFC experiment of ZmPIP1;1 and ZmPIP2;6 in maize protoplasts pCAMBIA-ZmPIP2;6-GFP, pDOE-ZmPIP1;1-ZmPIP2;6, and pDOE- ZmPIP2;6-ZmPIP1;1 were co-localized with a mCherry-labeled plasma membrane marker (mCherry-PM; CD3-1007). Bar = 20 μm."

[1] Yu C. China’s water crisis needs more than words. Nature, 2011,470:307.
pmid: 21331001
[2] Long S P, Zhu X G, Naidu S L, Ort D R. Can improvement in photosynthesis increase crop yields? Plant Cell Environ, 2006,29:315-330.
doi: 10.1111/j.1365-3040.2005.01493.x pmid: 17080588
[3] Nowicka B, Ciura J, Szymanska R, Kruk J. Improving photosynthesis, plant productivity and abiotic stress tolerance- current trends and future perspectives. J Plant Physiol, 2018,231:415-433.
[4] Chaumont F, Moshelion M, Daniels M J. Regulation of plant aquaporin activity. Biol Cell, 2005,97:749-764.
[5] Maurel C. Plant aquaporins: novel functions and regulation properties. FEBS Lett, 2007,581:2227-2236.
pmid: 17382935
[6] Kaldenhoff R, Ribas-Carbo M, Sans J F, Lovisolo C, Heckwolf M, Uehlein N. Aquaporins and plant water balance. Plant Cell Environ, 2008,31:658-666.
pmid: 18266903
[7] Chaumont F, Tyerman S D. Aquaporins: highly regulated channels controlling plant water relations. Plant Physiol, 2014,164:1600-1618.
pmid: 24449709
[8] Chaumont F, Barrieu F, Wojcik E, Chrispeels M J, Jung R. Aquaporins constitute a large and highly divergent protein family in maize. Plant Physiol, 2001,125:1206-1215.
[9] Johanson U, Karlsson M, Johansson I, Gustavsson S, Sjovall S, Fraysse L, Weig A R, Kjellbom P. The complete set of genes encoding major intrinsic proteins inArabidopsis provides a framework for a new nomenclature for major intrinsic proteins in plants. Plant Physiol, 2001,126:1358-1369.
doi: 10.1104/pp.126.4.1358 pmid: 11500536
[10] Ishibashi K. Aquaporin superfamily with unusual npa boxes: S-aquaporins (superfamily, sip-like and subcellular-aquaporins). Cell Mol Biol (Noisy-le-grand), 2006,52:20-27.
[11] Maurel C, Verdoucq L, Luu D T, Santoni V. Plant aquaporins: membrane channels with multiple integrated functions. Annu Rev Plant Biol, 2008,59:595-624.
pmid: 18444909
[12] Danielson J A, Johanson U. Unexpected complexity of the aquaporin gene family in the moss Physcomitrella patens. BMC Plant Biol, 2008,8:45-59.
doi: 10.1186/1471-2229-8-45 pmid: 18430224
[13] Bienert G P, Bienert M D, Jahn T P, Boutry M, Chaumont F. Solanaceae XIPs are plasma membrane aquaporins that facilitate the transport of many uncharged substrates. Plant J, 2011,66:306-317.
pmid: 21241387
[14] Kammerloher W, Fischer U, Piechottka G P, Schaffner A R. Water channels in the plant plasma membrane cloned by immunoselection from a mammalian expression system. Plant J, 1994,6:187-199.
doi: 10.1046/j.1365-313x.1994.6020187.x pmid: 7920711
[15] Murata K, Mitsuoka K, Hirai T, Walz T, Agre P, Heymann J B, Engel A, Fujiyoshi Y. Structural determinants of water permeation through aquaporin-1. Nature, 2000,407:599-605.
[16] Fetter K, Van Wilder V, Moshelion M, Chaumont F. Interactions between plasma membrane aquaporins modulate their water channel activity. Plant Cell, 2004,16:215-228.
pmid: 14671024
[17] Zelazny E, Borst J W, Muylaert M, Batoko H, Hemminga M A, Chaumont F. FRET imaging in living maize cells reveals that plasma membrane aquaporins interact to regulate their subcellular localization. Proc Natl Acad Sci USA, 2007,104:12359-12364.
[18] Vandeleur R K, Mayo G, Shelden M C, Gilliham M, Kaiser B N, Tyerman S D. The role of plasma membrane intrinsic protein aquaporins in water transport through roots: diurnal and drought stress responses reveal different strategies between isohydric and anisohydric cultivars of grapevine. Plant Physiol, 2009,149:445-460.
pmid: 18987216
[19] Chen W, Yin X, Wang L, Tian J, Yang R, Liu D, Yu Z, Ma N, Gao J. Involvement of rose aquaporin RhPIP1;1 in ethylene-regulated petal expansion through interaction with RhPIP2;1. Plant Mol Biol, 2013,83:219-233.
[20] Yaneff A, Sigaut L, Marquez M, Alleva K, Pietrasanta L I, Amodeo G. Heteromerization of PIP aquaporins affects their intrinsic permeability. Proc Natl Acad Sci USA, 2014,111:231-236.
[21] Afzal Z, Howton T C, Sun Y, Mukhtar M S. The roles of aquaporins in plant stress responses. J Dev Biol, 2016,4:9-30.
[22] Alexandersson E, Danielson J A, Rade J, Moparthi V K, Fontes M, Kjellbom P, Johanson U. Transcriptional regulation of aquaporins in accessions ofArabidopsis in response to drought stress. Plant J, 2010,61:650-660.
[23] Bae E K, Lee H, Lee J S, Noh E W. Drought, salt and wounding stress induce the expression of the plasma membrane intrinsic protein 1 gene in poplar (Populus alba × P. tremula var. glandulosa). Gene, 2011,483:43-48.
[24] Peng Y, Lin W, Cai W, Arora R. Overexpression of aPanax ginseng tonoplast aquaporin alters salt tolerance, drought tolerance and cold acclimation ability in transgenic Arabidopsis plants. Planta, 2007,226:729-740.
[25] Zhou S, Hu W, Deng X, Ma Z, Chen L, Huang C, Wang C, Wang J, He Y, Yang G, He G. Overexpression of the wheat aquaporin gene,TaAQP7, enhances drought tolerance in transgenic tobacco. PLoS One, 2012,7:e52439.
pmid: 23285044
[26] Sreedharan S, Shekhawat U K, Ganapathi T R. Transgenic banana plants overexpressing a native plasma membrane aquaporin MusaPIP1;2 display high tolerance levels to different abiotic stresses. Plant Biotechnol J, 2013,11:942-952.
[27] Zhou L, Zhou J, Xiong Y, Liu C, Wang J, Wang G, Cai Y. Overexpression of a maize plasma membrane intrinsic protein ZmPIP1;1 confers drought and salt tolerance in Arabidopsis. PLoS One, 2018,13:e0198639.
[28] Uehlein N, Otto B, Hanson D T, Fischer M, McDowell N, Kaldenhoff R. Function ofNicotiana tabacum aquaporins as chloroplast gas pores challenges the concept of membrane CO2 permeability. Plant Cell, 2008,20:648-657.
[29] Hanba Y T, Shibasaka M, Hayashi Y, Hayakawa T, Kasamo K, Terashima I, Katsuhara M. Overexpression of the barley aquaporin HvPIP2;1 increases internal CO2 conductance and CO2 assimilation in the leaves of transgenic rice plants. Plant Cell Physiol, 2004,45:521-529.
doi: 10.1093/pcp/pch070 pmid: 15169933
[30] Heckwolf M, Pater D, Hanson D T, Kaldenhoff R. TheArabidopsis thaliana aquaporin AtPIP1;2 is a physiologically relevant CO2 transport facilitator. Plant J, 2011,67:795-804.
pmid: 21564354
[31] 周练, 熊雨涵, 洪祥德, 周京, 刘朝显, 王久光, 王国强, 蔡一林. 玉米质膜内在蛋白ZmPIP2;6响应渗透、盐和干旱胁迫的功能鉴定. 中国农业科学, 53:461-473.
Zhou L, Xiong Y H, Hong X D, Zhou J, Liu C X, Wang J G, Wang G Q, Cai Y L. Functional characterization of a maize plasma membrane intrinsic protein ZmPIP2;6 responses to osmotic, salt and drought stress. Sci Agric Sin, 2020,53:461-473 (in Chinese with English abstract).
[32] Gookin T E, Assmann S M. Significant reduction of BiFC non-specific assembly facilitates in planta assessment of heterotrimeric G-protein interactors. Plant J, 2014,80:553-567.
[33] Nelson B K, Cai X, Nebenfuhr A. A multicolored set ofin vivo organelle markers for co-localization studies in Arabidopsis and other plants. Plant J, 2007,51:1126-1136.
doi: 10.1111/j.1365-313X.2007.03212.x pmid: 17666025
[34] Bart R, Chern M, Park C J, Bartley L, Ronald P C. A novel system for gene silencing using siRNAs in rice leaf and stem-derived protoplasts. Plant Methods, 2006,2:13-21.
[35] Nakashima K, Yamaguchi-Shinozaki K. ABA signaling in stress-response and seed development. Plant Cell Rep, 2013,32:959-970.
pmid: 23535869
[36] Wang Z Y, Xiong L, Li W, Zhu J K, Zhu J. The plant cuticle is required for osmotic stress regulation of abscisic acid biosynthesis and osmotic stress tolerance in Arabidopsis. Plant Cell, 2011,23:1971-1984.
doi: 10.1105/tpc.110.081943 pmid: 21610183
[37] Lee S C, Lan W, Buchanan B B, Luan S. A protein kinase-phosphatase pair interacts with an ion channel to regulate ABA signaling in plant guard cells. Proc Natl Acad Sci USA, 2009,106:21419-21424.
[38] Chater C, Peng K, Movahedi M, Dunn J A, Walker H J, Liang Y K, McLachlan D H, Casson S, Isner J C, Wilson I, Neill S J, Hedrich R, Gray J E, Hetherington A M. Elevated CO2-induced responses in stomata require ABA and ABA signaling. Curr Biol, 2015,25:2709-2716.
[39] Sorrentino G, Haworth M, Wahbi S, Mahmood T, Zuomin S, Centritto M. Abscisic acid induces rapid reductions in mesophyll conductance to carbon dioxide. PLoS One, 2016,11:e0148554.
[40] Flexas J, Ribas-Carbo M, Hanson D T, Bota J, Otto B, Cifre J, McDowell N, Medrano H, Kaldenhoff R. Tobacco aquaporin NtAQP1 is involved in mesophyll conductance to CO2 in vivo. Plant J, 2006,48:427-439.
pmid: 17010114
[41] Ding L, Gao L, Liu W, Wang M, Gu M, Ren B, Xu G, Shen Q, Guo S. Aquaporin plays an important role in mediating chloroplastic CO2 concentration under high-N supply in rice (Oryza sativa) plants. Physiol Plant, 2016,156:215-226.
[42] Heinen R B, Bienert G P, Cohen D, Chevalier A S, Uehlein N, Hachez C, Kaldenhoff R, Le Thiec D, Chaumont F. Expression and characterization of plasma membrane aquaporins in stomatal complexes of Zea mays. Plant Mol Biol, 2014,86:335-350.
doi: 10.1007/s11103-014-0232-7 pmid: 25082269
[43] Lin W, Peng Y, Li G, Arora R, Tang Z, Su W, Cai W. Isolation and functional characterization ofPgTIP1, a hormone-autotrophic cells-specific tonoplast aquaporin in ginseng. J Exp Bot, 2007,58:947-956.
pmid: 17237160
[44] Zhou L, Wang C, Liu R, Han Q, Vandeleur R K, Du J, Tyerman S D, Shou H. Constitutive overexpression of soybean plasma membrane intrinsic protein GmPIP1;6 confers salt tolerance. BMC Plant Biol, 2014,14:181-193.
pmid: 24998596
[45] Patrick J W, Zhang W, Tyerman S D, Offler C E, Walker N A. Role of membrane transport in phloem translocation of assimilates and water. Funct Plant Biol, 2001,28:697-709.
[46] Liu F, Vantoai T, Moy L P, Bock G, Linford L D, Quackenbush J. Global transcription profiling reveals comprehensive insights into hypoxic response inArabidopsis. Plant Physiol, 2005,137:1115-1129.
doi: 10.1104/pp.104.055475 pmid: 15734912
[47] Zhou Y, Setz N, Niemietz C, Qu H, Offler C E, Tyerman S D, Patrick J W. Aquaporins and unloading of phloem-imported water in coats of developing bean seeds. Plant Cell Environ, 2007,30:1566-1577.
doi: 10.1111/j.1365-3040.2007.01732.x pmid: 17927694
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