Skip to main content
SpringerLink
Log in

Physiological, structural, and proteomic analysis of chloroplasts during natural senescence of Ginkgo leaves

  • Original Paper
  • Published:
Plant Growth Regulation Aims and scope Submit manuscript

Abstract

A proteomic analysis was conducted to study the changes in chloroplast proteins during leaf senescence in Ginkgo under natural conditions. We also examined the physiological changes in senescing chloroplasts, including changes in photosynthesis and alterations in chloroplast ultrastructure. Decreased photosynthetic activity was observed during leaf senescence, while chloroplast structures were damaged with grana thylakoid unstacking and large accumulation of osmiophilic granules. Chloroplast proteins were extracted from Ginkgo leaves collected at an early stage (June) and late stage (October) of leaf senescence, and separated by two-dimensional gel electrophoresis. More than 850 protein spots were reproducibly detected, including 27 that were up-regulated and 21 that were down-regulated during senescence. Mass spectrometry analysis and database searches identified 17 spots, which represented 15 different proteins. Fifteen of these spots were chloroplast proteins, and the other two spots were proteins of unknown localization in Ginkgo leaves. Five of the proteins were previously reported as senescence-related proteins, and 10 were novel; (E)-4-hydroxy-3-methyl-but-2-enyl diphosphate synthase, Ycf4, putative 97B2-like cytochrome P450, limonene synthase, light-independent protochlorophyllide reductase subunit B, taxadiene synthase, geranylgeranyl diphosphate synthase, ribosomal protein L2, RNA polymerase alpha chain, and RNA polymerase IV second largest subunit. These proteins are involved in photosynthesis, transcription and translation, secondary metabolism, and reactive oxygen species scavenging. The results indicated that the decline of photosynthesis was mainly due to the decrease in stomatal conductance and the degradation of the photosynthetic apparatus during leaf senescence.

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

Similar content being viewed by others

Abbreviations

2-DE:

Two-dimensional gel electrophoresis

CBB:

Coomassie brilliant blue

Chl:

Chlorophyll

DPOR:

Light-independent protochlorophyllide reductase

gs :

Stomatal conductance

IEF:

Isoelectric focusing

MS:

Mass spectrometry

PMF:

Peptide mass fingerprinting

Pn:

Net photosynthetic rate

PSI:

Photosystem I

PSII:

Photosystem II

RbcL:

Rubisco large subunit

ROS:

Reactive oxygen species

SOD:

Superoxide dismutase

TCA:

Trichloracetic acid

References

  • Adamo A, Pinney JW, Kunova A, Westhead DR, Meyer P (2008) Heat stress enhances the accumulation of polyadenylated mitochondrial transcripts in Arabidopsis thaliana. PLoS ONE 3:e2889

    Article  PubMed  Google Scholar 

  • Agarwal GK, Rakwal R, Yonekura M, Kubo A, Saji H (2002) Proteome analysis of differentially displayed proteins as a tool for investigating ozone stress in rice (Oryza sativa L.) seedlings. Proteomics 2:947–959

    Article  Google Scholar 

  • Ananieva K, Ananiev ED, Mishev K, Georgieva K, Tzvetkova N, Staden JV (2008) Changes in photosynthetic capacity and polypeptide patterns during natural senescence and rejuvenation of Cucurbita pepo L. (zucchini) cotyledons. Plant Growth Regul 54:23–29

    Article  CAS  Google Scholar 

  • Andaluz S, Lopez-Millan AF, Rivas JD, Aro EM, Abadia J, Abadia A (2006) Proteomic profiles of thylakoid membranes and changes in response to iron deficiency. Photosynth Res 89:141–155

    Article  PubMed  CAS  Google Scholar 

  • Asada K (2006) Production and scavenging of reactive oxygen species in chloroplasts and their functions. Plant Physiol 141:391–396

    Article  PubMed  CAS  Google Scholar 

  • Blodner C, Majcherczyk A, Kues U, Polle A (2007) Early drought-induced changes to the needle proteome of Norway spruce. Tree Physiol 27:1423–1431

    Article  PubMed  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  PubMed  CAS  Google Scholar 

  • Bricker TM, Frankel LK (1998) The structure and function of the 33 kDa extrinsic protein of photosystem II: a critical assessment. Photosynth Res 56:157–173

    Article  CAS  Google Scholar 

  • Delieu T, Walker DA (1972) An improved cathode for the measurement of photosynthetic oxygen evolution by isolated chloroplasts. New Phytol 71:201–225

    Article  CAS  Google Scholar 

  • Friso G, Giacomelli L, Ytterberg AJ, Peltier JB, Rudella A, Sun Q, van Wijk KJ (2004) In-depth analysis of the thylakoid membrane proteome of Arabidopsis thaliana chloroplasts: new proteins, new functions, and a plastid proteome database. Plant Cell 16:478–499

    Article  PubMed  CAS  Google Scholar 

  • Gan S, Amasino RM (1997) Making sense of senescence: molecular genetic regulation and manipulation of leaf senescence. Plant Physiol 113:313–319

    PubMed  CAS  Google Scholar 

  • He XY, Ruan YN, Chen W, Lu T (2006) Responses of anti-oxidative system in leaves of Ginkgo biloba to elevated ozone concentration in urban area. Bot Stud 47:409–416

    CAS  Google Scholar 

  • He XY, Fu SL, Chen W, Zhao TH, Xu S, Tuba Z (2007) Changes in effects of ozone exposure on growth, photosynthesis, and respiration of Ginkgo biloba in Shenyang urban area. Photosynthetica 45:555–561

    Article  CAS  Google Scholar 

  • Hirao T, Watanabe A, Kurita M, Kondo T, Takata K (2008) Complete nucleotide sequence of the Cryptomeria japonica D. Don. chloroplast genome and comparative chloroplast genomics: diversified genomic structure of coniferous species. BMC Plant Biol 8:70

    Article  PubMed  Google Scholar 

  • Hirschberg J (2001) Carotenoid biosynthesis in flowering plants. Curr Opin Plant Biol 4:210–218

    Article  PubMed  CAS  Google Scholar 

  • Kahlau S, Bock R (2008) Plastid transcriptomics and translatomics of tomato fruit development and chloroplast-to-chromoplast differentiation: chromoplast gene expression largely serves the production of a single protein. Plant Cell 20:856–874

    Article  PubMed  CAS  Google Scholar 

  • Krupinska K (2007) Fate and activities of plastids during leaf senescence. In: Wise RR, Hoober JK (eds) The structure and function of plastids. Springer, The Netherlands, pp 433–449

    Chapter  Google Scholar 

  • Kudla J, Bock R (1999) RNA editing in an untranslated region of the Ginkgo chloroplast genome. Gene 234:81–86

    Article  PubMed  CAS  Google Scholar 

  • Lichtenthaler HK, Wellburn AR (1983) Determinations of total carotenoids and chlorophylls a and b of leaf extracts in different solvents. Biochem Soc Trans 11:591–592

    CAS  Google Scholar 

  • Lim PO, Kim HJ, Nam HG (2007) Leaf senescence. Annu Rev Plant Biol 58:115–136

    Article  PubMed  CAS  Google Scholar 

  • Lovdal T, Lillo CR (2009) Reference gene selection for quantitative real-time PCR normalization in tomato subjected to nitrogen, cold, and light stress. Anal Biochem 387:238–242

    Article  PubMed  CAS  Google Scholar 

  • Lu T, He XY, Chen W, Yan K, Zhao TH (2009) Effects of elevated O3 and/or elevated CO2 on lipid peroxidation and antioxidant systems in Ginkgo biloba leaves. Bull Environ Contam Toxicol 83:92–96

    Article  PubMed  CAS  Google Scholar 

  • Naver H, Boudreau E, Rochaix JD (2001) Functional studies of Ycf3. Its role in assembly of photosystem I and interactions with some of its subunits. Plant Cell 13:2731–2746

    PubMed  CAS  Google Scholar 

  • Overdieck D, Strassemeyer J (2005) Gas exchange of Ginkgo biloba leaves at different CO2 concentration levels. Flora 200:159–167

    Article  Google Scholar 

  • Ozawa S, Nield J, Terao A, Stauber EJ, Hippler M, Koike H, Rochaix JD, Takahashi Y (2009) Biochemical and structural studies of the large Ycf4-photosystem I assembly complex of the green alga Chlamydomonas reinhardtii. Plant Cell 21:2424–2442

    Article  PubMed  CAS  Google Scholar 

  • Pandey S, Kumar S, Nagar PK (2003) Photosynthetic performance of Ginkgo biloba L. grown under high and low irradiance. Photosynthetica 41:505–511

    Article  CAS  Google Scholar 

  • Peng L, Shimizu H, Shikanai T (2008) The chloroplast NAD(P)H dehydrogenase complex interacts with photosystem I in Arabidopsis. J Biol Chem 283:34873–34879

    Article  PubMed  CAS  Google Scholar 

  • Pflieger D, Rossier J (2008) Purification and proteomic analysis of chloroplasts and their sub-organellar compartments. Organelle Proteomics Methods Mol Biol 432:19–36

    Article  Google Scholar 

  • Quirino BF, Noh YS, Himelblau E, Amasino RM (2000) Molecular aspects of leaf senescence. Trends Plant Sci 5:278–282

    Article  PubMed  CAS  Google Scholar 

  • Sanderson MJ, Wojciechowski MF, Hu JM, Khan TS, Brady SG (2000) Error, bias, and long-branch attraction in data for two chloroplast photosystem genes in seed plants. Mol Biol Evol 17:782–797

    Article  PubMed  CAS  Google Scholar 

  • Sarijeva G, Knapp M, Lichtenthaler HK (2007) Differences in photosynthetic activity, chlorophyll and carotenoid levels, and in chlorophyll fluorescence parameters in green sun and shade leaves of Ginkgo and Fagus. J Plant Physiol 164:950–955

    Article  PubMed  CAS  Google Scholar 

  • Skribanek A, Solymosi K, Hideg E, Boddi B (2008) Light and temperature regulation of greening in dark-grown ginkgo (Ginkgo biloba). Physiol Plant 134:649–659

    Article  PubMed  CAS  Google Scholar 

  • Song CP, Mei HS (1991) Studies on the generation of superoxide anion in the chloroplasts during senescence. Acta Biophys Sinica 7:161–168

    Google Scholar 

  • Tang L, Okazawa A, Fukusaki E, Kobayashi A (2000) Removal of magnesium by Mg-dechelatase is a major step in the chlorophyll-degrading pathway in Ginkgo biloba in the process of autumnal tints. Z Naturforsch 55c:923–926

    Google Scholar 

  • Tang L, Okazawa A, Itoh Y, Fukusaki E, Kobayashi A (2004) Expression of chlorophyllase is not induced during autumnal yellowing in Ginkgo biloba. Z Naturforsch 59c:415–420

    Google Scholar 

  • Terry AC, Quick WP, Beerling DJ (2000) Long-term growth of ginkgo with CO2 enrichment increases leaf ice nucleation temperatures and limits recovery of the photosynthetic system from freezing. Plant Physiol 124:183–190

    Article  PubMed  CAS  Google Scholar 

  • Tholl D (2006) Terpene synthases and the regulation, diversity and biological roles of terpene metabolism. Curr Opin Plant Biol 9:1–8

    Article  Google Scholar 

  • Wilson KA, McManus MT, Gordon ME, Jordan TW (2002) The proteomics of senescence in leaves of white clover, Trifolium repens (L.). Proteomics 2:1114–1122

    Article  PubMed  CAS  Google Scholar 

  • Zhang CJ, Chu HJ, Chen GX, Shi DW, Zuo M, Wang J, Lu CG, Wang P, Chen L (2007) Photosynthetic and biochemical activities in flag leaves of a newly developed superhigh-yield hybrid rice (Oryza sativa) and its parents during the reproductive stage. J Plant Res 120:209–217

    Article  PubMed  CAS  Google Scholar 

  • Zhao CF, Wang JQ, Cao ML, Zhao K, Shao JM, Lei TT, Yin JN, Hill GG, Xu NZ, Liu SQ (2005) Proteomic changes in rice leaves during development of field-grown rice plants. Proteomics 5:961–972

    Article  PubMed  CAS  Google Scholar 

  • Zimmermann P, Zentgraf U (2005) The correlation between oxidative stress and leaf senescence during plant development. Cell Mol Biol Lett 10:515–534

    PubMed  CAS  Google Scholar 

Download references

Acknowledgments

Financial support was provided by the National Natural Sciences Foundation of China (No. 31271621) and the Natural Science Foundation of the Jiangsu Higher Education Institutions of China (No. 11KJA180001). We also gratefully acknowledge both two anonymous reviewers and the editors of this Journal for critical suggestions.

Author information

Authors and Affiliations

  1. College of Life Sciences, Nanjing Normal University, 1 Wenyuan Road, Nanjing, 210046, China

    Xiao-Dong Wei, Da-Wei Shi & Guo-Xiang Chen

  2. Institute of Food and Crops, Jiangsu Academy of Agricultural Sciences, Nanjing, China

    Xiao-Dong Wei

  3. College of Forest Resources and Environment, Nanjing Forestry University, Nanjing, China

    Da-Wei Shi

Authors
  1. Xiao-Dong Wei
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Da-Wei Shi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Guo-Xiang Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Guo-Xiang Chen.

Additional information

Xiao-Dong Wei, Da-Wei Shi contributed equally to this work.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wei, XD., Shi, DW. & Chen, GX. Physiological, structural, and proteomic analysis of chloroplasts during natural senescence of Ginkgo leaves. Plant Growth Regul 69, 191–201 (2013). https://doi.org/10.1007/s10725-012-9761-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10725-012-9761-8

Keywords

Search

Navigation