Skip to main content

Advertisement

SpringerLink
Log in

Cloning and characterisation of a pepper aquaporin, CaAQP, which reduces chilling stress in transgenic tobacco plants

  • Original Paper
  • Published:
Plant Cell, Tissue and Organ Culture (PCTOC) Aims and scope Submit manuscript

Abstract

Ubiquitous cell membrane proteins called aquaporins are members of major intrinsic proteins (MIPs), which control the specific transport of water molecules across cell membranes. A pepper aquaporin gene (CaAQP), which exhibits the structural features of tonoplast intrinsic proteins of the MIP subfamily, was isolated from the leaves of chilling-treated seedlings of pepper (Capsicum annuum L.) cv. P70. Assays indicated high levels of expression in young seeds, green fruits and flower buds and low levels of expression in the stems, leaves and roots of pepper. The expression patterns were strongly and rapidly induced by HgCl2, low temperature, abscisic acid, fluridone and osmotic stresses. The responsiveness of pepper seedlings pretreated with abscisic acid at low temperatures demonstrated up-regulation of CaAQP by chilling, which is potentially involved in ABA signalling. Our results indicated that overexpression of CaAQP decreased chilling stress in transgenic plants, likely by increasing the stomatal aperture under stress, increasing the rate of membrane damage during the recovery stage, thereby affecting the intercellular CO2 concentration with lower stomatal conductance and transpiration rates. VIGS of CaAQP in pepper plants caused significant growth retardation. These results suggested that CaAQP played a crucial role in the plant response to abiotic stresses.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9

Similar content being viewed by others

Abbreviations

CaMV:

Cauliflower mosaic virus

Ci:

Intercellular CO2 concentrations

GIPs:

GlpF-like intrinsic proteins

Gs:

Stomatal conductance

HIPs:

Hybrid intrinsic proteins

L0 :

Root hydraulic conductance

MIPs:

Major intrinsic proteins

NIPs:

Nodulin26-like intrinsic proteins

NtL25 :

L25 ribosomal protein

PIPs:

Plasma membrane intrinsic proteins

Pn:

Net photosynthetic rate

PSV:

Protein storage vacuoles

RACE:

Rapid amplification of cDNA ends

RCBD:

Randomized complete block design

REL:

Relative electrolyte leakage

RWC:

Relative water content

SA:

Salicylic acid

SIPs:

Small basic intrinsic proteins

SE:

Standard error

TDFs:

Transcript-derived fragments

TIPs:

Tonoplast intrinsic proteins

Tr:

Transpiration rate

TMHs:

Transmembrane helices

UTR:

Non-coding region

XIPs:

Uncharacterized X intrinsic proteins

References

  • Allen DJ, Ort DR (2001) Impacts of chilling temperatures on photosynthesis in warm-climate plants. Trends Plant Sci 6:36–42

    Article  CAS  PubMed  Google Scholar 

  • Aroca R, Amodeo G, Fernandez-Illescas S, Herman EM, Chaumont F, Chrispeels MJ (2005) The role of aquaporins and membrane damage in chilling and hydrogen peroxide induced changes in the hydraulic conductance of maize roots. Plant Physiol 137:341–353

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Atkin OK, Tjoelker MG (2003) Thermal acclimation and the dynamic response of plant respiration to temperature. Trends Plant Sci 8:343–351

    Article  CAS  PubMed  Google Scholar 

  • Baaziz KB, Lopez D, Bouzid S, Cochard H, Venisse JS, Sakr S (2012) Early gene expression in the walnut tree occurring during stimulation of leaf hydraulic conductance by irradiance. Biol Plant 56:657–666

    Article  Google Scholar 

  • Benfey PN, Ren L, Chua NH (1989) The CaMV 35S enhancer contains at least two domains which can confer different developmental and tissue-specific expression patterns. EMBO J 8:2195–2202

    CAS  PubMed Central  PubMed  Google Scholar 

  • Boursiac Y, Chen S, Luu DT, Sorieul M, van den Dries N, Maurel C (2005) Early effects of salinity on water transport in Arabidopsis roots. Molecular and cellular features of aquaporin expression. Plant Physiol 139:790–805

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Chaumont F, Barrieu F, Wojcik E, Chrispeels MJ, Jung R (2001) Aquaporins constitute a large and highly divergent protein family in maize. Plant Physiol 125:1206–1215

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Cruz-Huerta N, Williamson JG, Darnell RL (2011) Low night temperature increases ovary size in sweet pepper cultivars. HortScience 46:396–401

    Google Scholar 

  • Daniels MJ, Chaumont F, Mirkov TE, Chrispeels MJ (1996) Characterization of a new vacuolar membrane aquaporin sensitive to mercury at a unique site. Plant Cell 8:587–599

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Danielson JÅH, Johanson U (2008) Unexpected complexity of the Aquaporin gene family in the moss Physcomitrella patens. BMC Plant Biol. doi:10.1186/1471-2229-8-45

  • Del Amor FM, Rubio JS (2009) Effects of antitranspirant spray and potassium: calcium: magnesium ratio on photosynthesis, nutrient and water uptake, growth, and yield of sweet pepper. J Plant Nutr 32:97–111

    Article  Google Scholar 

  • Deng MH, Wen JF, Zhu HS, Zou XX (2009) The hottest pepper variety in China. Genet Resour Crop Evol 56:605–608

    Article  Google Scholar 

  • Ermawati N, Liang YS, Cha JY, Shin D, Jung MH, Lee JJ, Lee BH, Han CD, Lee KH, Son D (2009) A new tip homolog, ShTIP, from Salicornia shows a different involvement in salt stress compared to that of TIP from Arabidopsis. Biol Plant 53:271–277

    Article  CAS  Google Scholar 

  • Faize M, Burgos L, Faize L, Piqueras A, Nicolas E, Barba-Espin G, Clemente-Moreno MJ, Alcobendas R, Artlip T, Hernandez JA (2011) Involvement of cytosolic ascorbate peroxidase and Cu/Zn-superoxide dismutase for improved tolerance against drought stress. J Exp Bot 62:2599–2613

    Article  CAS  PubMed  Google Scholar 

  • Forrest KL, Bhave M (2007) Major intrinsic proteins (MIPs) in plants: a complex gene family with major impacts on plant phenotype. Funct Integr Genomics 7:263–289

    Article  CAS  PubMed  Google Scholar 

  • Forrest KL, Bhave M (2008) The PIP and TIP aquaporins in wheat form a large and diverse family with unique gene structures and functionally important features. Funct Integr Genomics 8:115–133

    Article  CAS  PubMed  Google Scholar 

  • Gattolin S, Sorieul M, Hunter PR, Khonsari RH, Frigerio L (2009) In vivo imaging of the tonoplast intrinsic protein family in Arabidopsis roots. BMC Plant Biol. doi:10.1186/1471-2229-9-133

  • Gattolin S, Sorieul M, Frigerio L (2011) Mapping of tonoplast intrinsic proteins in maturing and germinating Arabidopsis seeds reveals dual localization of embryonic TIPs to the tonoplast and plasma membrane. Mol Plant 4:180–189

    Article  CAS  PubMed  Google Scholar 

  • Godínez-Hernández Y, Anaya-López JL, Díaz-Plaza R, González-Chavira M, Torres-Pacheco I, Rivera-Bustamante RF, Guevara-González RG (2001) Characterization of resistance to pepper huasteco geminivirus in chili peppers from Yucatán, México. HortScience 36:139–142

    Google Scholar 

  • Guo WL, Chen RG, Gong ZH, Yin YX, Ahmed SS, He YM (2012) Exogenous abscisic acid increases antioxidant enzymes and related gene expression in pepper (Capsicum annuum) leaves subjected to chilling stress. Genet Mol Res 11:4063–4080

    Article  CAS  PubMed  Google Scholar 

  • Gupta AB, Sankararamakrishnan R (2009) Genome-wide analysis of major intrinsic proteins in the tree plant Populus trichocarpa: characterization of XIP subfamily of aquaporins from evolutionary perspective. BMC Plant Biol. doi:10.1186/1471-2229-9-134

  • Hunter PR, Craddock CP, Di Benedetto S, Roberts LM, Frigerio L (2007) Fluorescent reporter proteins for the tonoplast and the vacuolar lumen identify a single vacuolar compartment in Arabidopsis cells. Plant Physiol 145:1371–1382

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Javot H, Maurel C (2002) The role of aquaporins in root water uptake. Ann Bot 90:301–313

    Article  CAS  PubMed  Google Scholar 

  • Johanson U, Karlsson M, Johansson I, Gustavsson S, Sjovall S, Fraysse L, Weig AR, Kjellbom P (2001) The complete set of genes encoding major intrinsic proteins in Arabidopsis provides a framework for a new nomenclature for major intrinsic proteins in plants. Plant Physiol 126:1358–1369

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Korkmaz A, Korkmaz Y, Demirkiran AR (2010) Enhancing chilling stress tolerance of pepper seedlings by exogenous application of 5-aminolevulinic acid. Environ Exp Bot 67:495–501

    Article  CAS  Google Scholar 

  • Lee YH, Jung M, Shin SH, Lee JH, Choi SH, Her NH, Harn CH (2009) Transgenic peppers that are highly tolerant to a new CMV pathotype. Plant Cell Rep 28:223–232

    Article  CAS  PubMed  Google Scholar 

  • Li DD, Wu YJ, Ruan XM, Li B, Zhu L, Wang H, Li XB (2009) Expressions of three cotton genes encoding the PIP proteins are regulated in root development and in response to stresses. Plant Cell Rep 28:291–300

    Article  CAS  PubMed  Google Scholar 

  • Li Z, Wang S, Tao Q, Pan J, Si L, Gong Z, Cai R (2012) A putative positive feedback regulation mechanism in CsACS2 expression suggests a modified model for sex determination in cucumber (Cucumis sativus L.). J Exp Bot 63:4475–4484

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Lukatkin AS, Brazaityte A, Bobinas C, Duchovskis P (2012) Chilling injury in chilling-sensitive plants: a review. Zemdirbyste 99:111–124

    Google Scholar 

  • Ma S, Quist TM, Ulanov A, Joly R, Bohnert HJ (2004) Loss of TIP1;1 aquaporin in Arabidopsis leads to cell and plant death. Plant J 40:845–859

    Article  CAS  PubMed  Google Scholar 

  • Mahdieh M, Mostajeran A, Horie T, Katsuhara M (2008) Drought stress alters water relations and expression of PIP-type aquaporin genes in Nicotiana tabacum plants. Plant Cell Physiol 49:801–813

    CAS  PubMed  Google Scholar 

  • Martinez-Ballesta MC, Diaz R, Martinez V, Carvajal M (2003) Different blocking effects of HgCl2 and NaCl on aquaporins of pepper plants. J Plant Physiol 160:1487–1492

    Article  CAS  PubMed  Google Scholar 

  • Park W, Scheffler BE, Bauer PJ, Campbell BT (2010) Identification of the family of aquaporin genes and their expression in upland cotton (Gossypium hirsutum L.). BMC Plant Biol. doi:10.1186/1471-2229-10-142

  • Peng YH, Lin WL, Cai WM, Arora R (2007) Overexpression of a Panax ginseng tonoplast aquaporin alters salt tolerance, drought tolerance and cold acclimation ability in transgenic Arabidopsis plants. Planta 226:729–740

    Article  CAS  PubMed  Google Scholar 

  • Reuscher, S, Akiyama M, Mori C, Aoki K, Shibata D, Shiratake K (2013) Genome-wide identification and expression analysis of aquaporins in tomato. PLoS One. doi:10.1371/journal.pone.0079052

  • Ruiz-Lozano JM, del Mar Alguacil M, Barzana G, Vernieri P, Aroca R (2009) Exogenous ABA accentuates the differences in root hydraulic properties between mycorrhizal and non mycorrhizal maize plants through regulation of PIP aquaporins. Plant Mol Biol 70:565–579

    Article  CAS  PubMed  Google Scholar 

  • Ryu MY, Cho SK, Kim WT (2010) The Arabidopsis C3H2C3-type RING E3 ubiquitin ligase AtAIRP1 is a positive regulator of an abscisic acid-dependent response to drought stress. Plant Physiol 154:1983–1997

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Sade N, Vinocur BJ, Diber A, Shatil A, Ronen G, Nissan H, Wallach R, Karchi H, Moshelion M (2009) Improving plant stress tolerance and yield production: is the tonoplast aquaporin SlTIP2;2 a key to isohydric to anisohydric conversion? New Phytol 181:651–661

    Article  CAS  PubMed  Google Scholar 

  • Sade N, Gebretsadik M, Seligmann R, Schwartz A, Wallach R, Moshelion M (2010) The role of tobacco Aquaporin1 in improving water use efficiency, hydraulic conductivity, and yield production under salt stress. Plant Physiol 152:245–254

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Schmidt GW, Delaney SK (2010) Stable internal reference genes for normalization of real-time RT-PCR in tobacco (Nicotiana tabacum) during development and abiotic stress. Mol Genet Genomics 283:233–241

    CAS  PubMed  Google Scholar 

  • Schussler MD, Alexandersson E, Bienert GP, Kichey T, Laursen KH, Johanson U, Kjellbom P, Schjoerring JK, Jahn TP (2008) The effects of the loss of TIP1;1 and TIP1;2 aquaporins in Arabidopsis thaliana. Plant J 56:756–767

    Article  CAS  PubMed  Google Scholar 

  • Schwarz D, Rouphael Y, Colla G, Venema JH (2010) Grafting as a tool to improve tolerance of vegetables to abiotic stresses: thermal stress, water stress and organic pollutants. Sci Hortic 127:162–171

    Article  CAS  Google Scholar 

  • Smart LB, Moskal WA, Cameron KD, Bennett AB (2001) MIP genes are down-regulated under drought stress in Nicotiana glauca. Plant Cell Physiol 42:686–693

    CAS  PubMed  Google Scholar 

  • Soto G, Alleva K, Amodeo G, Muschietti J, Ayub ND (2012) New insight into the evolution of aquaporins from flowering plants and vertebrates: orthologous identification and functional transfer is possible. Gene 503:165–176

    Article  CAS  PubMed  Google Scholar 

  • Sreedharan S, Shekhawat UK, Ganapathi TR (2013) Transgenic banana plants overexpressing a native plasma membrane aquaporin MusaPIP1; 2 display high tolerance levels to different abiotic stresses. Plant Biotechnol J 11:942–952

    Article  CAS  PubMed  Google Scholar 

  • Subramanyam K, Sailaja KV, Subramanyam K, Rao DM, Lakshmidevi K (2011) Ectopic expression of an osmotin gene leads to enhanced salt tolerance in transgenic chilli pepper (Capsicum annum L.). Plant Cell Tissue Organ Cult 105:181–192

    Article  CAS  Google Scholar 

  • Tyerman SD, Niemietz CM, Bramley H (2002) Plant aquaporins: multifunctional water and solute channels with expanding roles. Plant Cell Environ 25:173–194

    Article  CAS  PubMed  Google Scholar 

  • Venkatesh J, Yu JW, Park SW (2013) Genome-wide analysis and expression profiling of the Solanum tuberosum aquaporins. Plant Physiol Biochem 73:392–404

    CAS  PubMed  Google Scholar 

  • Wan HJ, Yuan W, Ruan MY, Ruan MY, Ye QJ, Wang RQ, Li ZM, Zhou GZ, Yao ZP, Zhao J, Liu SJ, Yang YJ (2011) Identification of reference genes for reverse transcription quantitative real-time PCR normalization in pepper (Capsicum annuum L.). Biochem Biophys Res Commun 416:24–30

    Article  CAS  PubMed  Google Scholar 

  • Wang JE, Li DW, Gong ZH, Zhang YL (2013a) Optimization of virus-induced gene silencing in pepper (Capsicum annum L.). Genet Mol Res 12:2492–2506

    Article  PubMed  Google Scholar 

  • Wang JE, Liu KK, Li DW, Zhang YL, Zhao Q, He YM, Gong ZH (2013b) A novel peroxidase CanPOD gene of pepper is involved in defense responses to Phytophtora capsici infection as well as abiotic stress tolerance. Int J Mol Sci 14:3158–3177

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Zeng YW, Wang JJ, Yang ZY, Shen SQ, Wu LH, Chen XY, Meng JG (2001) The diversity and sustainable development of crop genetic resources in the Lancang River Valley. Genet Resour Crop Evol 48:297–306

    Article  Google Scholar 

  • Zhou S, Hu W, Deng XM, Ma ZB, Chen LH, Huang C, Wang C, Wang J, He YZ, Yang GX, He GY (2012) Overexpression of the wheat aquaporin gene, TaAQP7, enhances drought tolerance in transgenic tobacco. PLoS One. doi:10.1371/journal.pone.0052439

Download references

Acknowledgments

This work was supported by the National Natural Science Foundation of China (Nos. 31201615, 31272163), “The Twelfth Five-Year” Plan of National Science and Technology in Rural Areas (No. 2011BAD12B03) and the Shaanxi Provincial Science and Technology Coordinating Innovative Engineering Project (No. 2012KTCL02-09). Language help was provided by Elsevier Webshop language services.

Author information

Authors and Affiliations

  1. College of Horticulture, Northwest A&F University, Yangling, 712100, Shaanxi, People’s Republic of China

    Yan-Xu Yin, Wei-Li Guo, Ying-Li Zhang, Jiao-Jiao Ji, Huai-Juan Xiao, Fei Yan, Yan-Yan Zhao, Wen-Chao Zhu, Ru-Gang Chen & Zhen-Hui Gong

  2. Guizhou Academy of Agricultural Sciences, Guiyang, 550006, Guizhou, People’s Republic of China

    Wen-Chao Zhu

  3. Institute of Vegetables, Hangzhou Academy of Agricultural Sciences, Hangzhou, 311104, Zhejiang, People’s Republic of China

    Wei-Guo Chai

Authors
  1. Yan-Xu Yin
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Wei-Li Guo
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ying-Li Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jiao-Jiao Ji
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Huai-Juan Xiao
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Fei Yan
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Yan-Yan Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Wen-Chao Zhu
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Ru-Gang Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Wei-Guo Chai
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Zhen-Hui Gong
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Zhen-Hui Gong.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOC 399 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Yin, YX., Guo, WL., Zhang, YL. et al. Cloning and characterisation of a pepper aquaporin, CaAQP, which reduces chilling stress in transgenic tobacco plants. Plant Cell Tiss Organ Cult 118, 431–444 (2014). https://doi.org/10.1007/s11240-014-0495-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11240-014-0495-3

Keywords

Search

Navigation