Skip to main content
SpringerLink
Log in

Physiological and transcriptional analysis reveals pathways involved in iron deficiency chlorosis in fragrant citrus

  • Original Article
  • Published:
Tree Genetics & Genomes Aims and scope Submit manuscript

Abstract

Iron (Fe) deficiency chlorosis is a yield-limiting problem in citrus production regions with calcareous soils. Physiological and transcriptional analyses of fragrant citrus (Citrus junos Sieb. ex Tanaka) leaves from Fe-sufficient (IS) and Fe-deficient (ID) plants were investigated in this study. The physiological results showed that Fe, potassium, and nitrogen levels decreased by 12, 15, and 41% in ID leaves, respectively. However, zinc and copper levels increased by 49 and 35% in ID leaves, respectively. The chlorophyll (Chl) content, photosynthesis rate, stomatal conductance, and transpiration rate in ID leaves decreased by 55, 33, 38, and 42%, respectively, compared with IS leaves. Moreover, transcriptional profiling analysis showed that genes associated with Chl metabolism, photosynthesis, and nitrogen metabolism were dramatically downregulated by Fe deficiency. The expression of glutamyl-tRNA reductase 1, chlorophyll(ide) b reductase, and geranylgeranyl diphosphate reductase in ID leaves was 0.26–0.37 times that in IS leaves. The expression levels of 16 photosynthesis-related genes were severely downregulated by Fe deficiency. In addition, the transcription levels of nitrate transporter, nitrate reductase, and ferredoxin-nitrite reductase genes in ID leaves were 0.38–0.45 times those in IS leaves. Taken together, these results indicated that the block of Chl biosynthesis, the reduction of photosynthesis, and the repression of nitrogen absorption resulted in the chlorosis symptoms observed in fragrant citrus leaves.

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

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

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

Similar content being viewed by others

References

  • Abadia J, Lopez-Millan AF, Rombola A, Abadia A (2002) Organic acids and Fe deficiency: a review. Plant Soil 241:75–86

    Article  CAS  Google Scholar 

  • Agnolon F, Santi S, Varanini Z, Pinton R (2002) Enzymatic response of cucumber roots to different levels of Fe supply. Plant Soil 241:35–41

    Article  CAS  Google Scholar 

  • Belkhodja R, Morales F, Sanz M, Abadía A, Abadía J (1998) Iron deficiency in peach trees: effects on leaf chlorophyll and nutrient concentrations in flowers and leaves. Plant Soil 203:257–268

    Article  CAS  Google Scholar 

  • Borlotti A, Vigani G, Zocchi G (2012) Iron deficiency affects nitrogen metabolism in cucumber (Cucumis sativus L.) plants. BMC Plant Biol 12:1

    Article  Google Scholar 

  • Briat J-F, Curie C, Gaymard F (2007) Iron utilization and metabolism in plants. Curr Opin Plant Biol 10:276–282

    Article  CAS  PubMed  Google Scholar 

  • Chen L, Smith BR, Cheng L (2004a) CO2 assimilation, photosynthetic enzymes, and carbohydrates of ‘Concord’ grape leaves in response to iron supply. J Am Soc Hortic Sci 129:738–744

  • Chen Y, Shi J, Tian G, Zheng S, Lin Q (2004b) Fe deficiency induces Cu uptake and accumulation in Commelina communis. Plant Sci 166:1371–1377

    Article  CAS  Google Scholar 

  • Cimen B, Yesiloglu T, Incesu M, Yilmaz B (2014) Growth and photosynthetic response of young ‘Navelina’ trees budded on to eight citrus rootstocks in response to iron deficiency New Zealand. Journal of Crop and Horticultural Science 42:170–182

    Article  CAS  Google Scholar 

  • Curie C, Panaviene Z, Loulergue C, Dellaporta SL, Briat J-F, Walker EL (2001) Maize yellow stripe1 encodes a membrane protein directly involved in Fe(III) uptake. Nature 409:346–349

    Article  CAS  PubMed  Google Scholar 

  • Eberhard S, Finazzi G, Wollman F-A (2008) The dynamics of photosynthesis. Annu Rev Genet 42:463–515

    Article  CAS  PubMed  Google Scholar 

  • Eggink LL, LoBrutto R, Brune DC, Brusslan J, Yamasato A, Tanaka A, Hoober JK (2004) Synthesis of chlorophyll b: localization of chlorophyllide a oxygenase and discovery of a stable radical in the catalytic subunit. BMC Plant Biol 4:1–16

  • Graziano M, Lamattina L (2007) Nitric oxide accumulation is required for molecular and physiological responses to iron deficiency in tomato roots. Plant J 52:949–960

    Article  CAS  PubMed  Google Scholar 

  • Hänsch R, Mendel RR (2009) Physiological functions of mineral micronutrients (Cu, Zn, Mn, Fe, Ni, Mo, B, Cl). Curr Opin Plant Biol 12:259–266

    Article  PubMed  Google Scholar 

  • Haydon MJ, Cobbett CS (2007) Transporters of ligands for essential metal ions in plants. New Phytol 174:499–506

    Article  CAS  PubMed  Google Scholar 

  • Holm G (1954) Chlorophyll mutations in barley. Acta Agric Scand 4:457–471

    Article  Google Scholar 

  • Jack R, Lancaster JMV, Kamin H, Nanette R, Orme-Johnson WH, Johnson O, Krueger RJ, Siegel LM (1978) Identification of the iron-sulfur center of spinach ferredoxin-nitrite reductase as a tetranuclear center, and preliminary EPR studies of mechanism. J Biol Chem 254:1268–1272

    Google Scholar 

  • Kim D, Pertea G, Trapnell C, Pimentel H, Kelley R, Salzberg SL (2013) TopHat2: accurate alignment of transcriptomes in the presence of insertions, deletions and gene fusions. Genome Biol 14:1

    Article  Google Scholar 

  • Kobayashi T, Nishizawa NK (2012) Iron uptake, translocation, and regulation in higher plants. Annu Rev Plant Biol 63:131–152

    Article  CAS  PubMed  Google Scholar 

  • Kramer D, Römheld V, Landsberg E, Marschner H (1980) Induction of transfer-cell formation by iron deficiency in the root epidermis of Helianthus annuus L. Planta 147:335–339

    Article  CAS  PubMed  Google Scholar 

  • Landsberg E-C (1989) Proton efflux and transfer cell formation as responses to Fe deficiency of soybean in nutrient solution culture. Plant Soil 114:53–61

    Article  CAS  Google Scholar 

  • Langmead B, Salzberg SL (2012) Fast gapped-read alignment with Bowtie 2. Nat Methods 9:357–359

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Larbi A, Abadía A, Abadía J, Morales F (2006) Down co-regulation of light absorption, photochemistry, and carboxylation in Fe-deficient plants growing in different environments. Photosynth Res 89:113–126

    Article  CAS  PubMed  Google Scholar 

  • Lee S, Chiecko JC, Kim SA, Walker EL, Lee Y, Guerinot ML, An G (2009) Disruption of OsYSL15 leads to iron inefficiency in rice plants. Plant Physiol 150:786–800

  • Li B, Dewey CN (2011) RSEM: accurate transcript quantification from RNA-Seq data with or without a reference genome. BMC Bioinformatics 12:1

    Article  Google Scholar 

  • Lin S, Baumer JS, Ivers D, de Cianzo SR, Shoemaker RC (1998) Field and nutrient solution tests measure similar mechanisms controlling iron deficiency chlorosis in soybean. Crop Sci 38:254–259

    Article  CAS  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408

  • Marschner P (2012) Marschner’s mineral nutrition of higher plants, 3rd edn. Academic Press, San Diego

    Google Scholar 

  • Martínez-Cuenca M-R, Forner-Giner MÁ, Iglesias DJ, Primo-Millo E, Legaz F (2013) Strategy I responses to Fe-deficiency of two Citrus rootstocks differing in their tolerance to iron chlorosis. Sci Hortic 153:56–63

    Article  Google Scholar 

  • Martinez-Cuenca M-R, Quinones A, Angeles Forner-Giner M (2016) Screening of ‘King’ mandarin (Citrus nobilis Lour) x Poncirus trifoliata ((L.) Raf.) hybrids as citrus rootstocks tolerants to iron chlorosis. Sci Hortic 198:61–69

    Article  Google Scholar 

  • Miller G, Pushnik J, Welkie G (1984) Iron chlorosis, a world wide problem, the relation of chlorophyll biosynthesis to iron. J Plant Nutr 7:1–22

    Article  CAS  Google Scholar 

  • Morales F, Abadía A, Abadía J (1990) Characterization of the xanthophyll cycle and other photosynthetic pigment changes induced by iron deficiency in sugar beet (Beta vulgaris L.) Plant Physiol 2:607–613

    Article  Google Scholar 

  • Morales F, Abadía A, Abadía J (1991) Chlorophyll fluorescence and photon yield of oxygen evolution in iron-deficient sugar beet (Beta vulgaris L.) leaves. Plant Physiol 97:886–893

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Mori S (1999) Iron acquisition by plants. Curr Opin Plant Biol 2:250–253

    Article  CAS  PubMed  Google Scholar 

  • Murata Y, Ma JF, Yamaji N, Ueno D, Nomoto K, Iwashita T (2006) A specific transporter for iron(III)-phytosiderophore in barley roots.1. Plant J 46:563–572

    Article  CAS  PubMed  Google Scholar 

  • Nouet C, Motte P, Hanikenne M (2011) Chloroplastic and mitochondrial metal homeostasis. Trends Plant Sci 16:395–404

    Article  CAS  PubMed  Google Scholar 

  • Pontoppidan B, Kannangara CG (1994) Purification and partial characterisation of barley glutamyl-tRNAGlu reductase, the enzyme that directs glutamate to chlorophyll biosynthesis. Eur J Biochem 225:529–537

    Article  CAS  PubMed  Google Scholar 

  • Robinson NJ, Procter CM, Connolly EL, Guerinot ML (1999) A ferric-chelate reductase for iron uptake from soils. Nature 397:694–697

    Article  CAS  PubMed  Google Scholar 

  • Santi S, Schmidt W (2009) Dissecting iron deficiency-induced proton extrusion in Arabidopsis roots. New Phytol 183:1072–1084

    Article  CAS  PubMed  Google Scholar 

  • Scheumann V, Schoch S, Rüdiger W (1998) Chlorophyll a formation in the chlorophyll b reductase reaction requires reduced ferredoxin. J Biol Chem 273:35102–35108

  • Tagawa K, Tsujimoto H, Arnon DI (1963) Role of chloroplast ferredoxin in the energy conversion process of photosynthesis. Proc Natl Acad Sci 49:567–572

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Tagliavini M, Rombolà AD (2001) Iron deficiency and chlorosis in orchard and vineyard ecosystems. Eur J Agron 15:71–92

    Article  CAS  Google Scholar 

  • Tagliavini M, Abadía J, Rombolà AD, Abadía A, Tsipouridis C, Marangoni B (2000) Agronomic means for the control of iron deficiency chlorosis in deciduous fruit trees. J Plant Nutr 23:2007–2022

    Article  CAS  Google Scholar 

  • Takahashi M (2003) Overcoming Fe deficiency by a transgenic approach in rice. Plant Cell Tissue Org Cult 72:211–220

    Article  CAS  Google Scholar 

  • Tan F, Tu H, Liang W, Long J, Wu X, Zhang H, Guo W (2015) Comparative metabolic and transcriptional analysis of a doubled diploid and its diploid citrus rootstock (C. junos cv. Ziyang xiangcheng) suggests its potential value for stress resistance improvement. BMC Plant Biol 15:89

  • Thornber JP (1975) Chlorophyll-proteins: light-harvesting and reaction center components of plants. Annu Rev Plant Physiol 26:127–158

    Article  CAS  Google Scholar 

  • Varotto C, Maiwald D, Pesaresi P, Jahns P, Salamini F, Leister D (2002) The metal ion transporter IRT1 is necessary for iron homeostasis and efficient photosynthesis in Arabidopsis thaliana. Plant J 31:589–599

    Article  CAS  PubMed  Google Scholar 

  • Vert G, Grotz N, Dédaldéchamp F, Gaymard F, Guerinot ML, Briat J-F, Curie C (2002) IRT1, an Arabidopsis transporter essential for iron uptake from the soil and for plant growth. Plant Cell 14:1223–1233

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Walker EL, Connolly EL (2008) Time to pump iron: iron-deficiency-signaling mechanisms of higher plants. Curr Opin Plant Biol 11:530–535

    Article  CAS  PubMed  Google Scholar 

  • Wang Y, Hsu P-K, Tsay Y-F (2012) Uptake, allocation and signaling of nitrate. Trends Plant Sci 17:458–467

    Article  CAS  PubMed  Google Scholar 

  • Wang N, Yan T, Fu L, Zhou G, Liu Y, Peng S (2014) Differences in boron distribution and forms in four citrus scion–rootstock combinations with contrasting boron efficiency under boron-deficient conditions. Trees 28:1589–1598

    Article  Google Scholar 

  • Wulandari C, Muraki S, Hisamura A, Ono H, Honda K, Kashima T, Subandiyah S, Masaoka Y (2014) Effect of iron deficiency on root ferric chelate reductase, proton extrusion, biomass production and mineral absorption of citrus root stock orange jasmine (Murraya exotica L.) J Plant Nutr 37:50–64

    Article  CAS  Google Scholar 

  • Zhao W, Yang X, Yu H, Jiang W, Sun N, Liu X, Liu X, Zhang X, Wang Y, Gu X (2015) RNA-Seq-based transcriptome profiling of early nitrogen deficiency response in cucumber seedlings provides new insight into the putative nitrogen regulatory network. Plant Cell Physiol 56:455–467

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by the National Natural Science Foundation of China (Grant No. 31272121) and the earmarked fund for the China Agriculture Research System (CARS-27).

Author information

Authors and Affiliations

  1. Key Laboratory of Horticultural Plant Biology, Ministry of Education, Huazhong Agricultural University, Wuhan, 430070, People’s Republic of China

    Long-Fei Jin, Yong-Zhong Liu, Wei Du, Li-Na Fu, Syed Bilal Hussain & Shu-Ang Peng

  2. Key Laboratory of Horticultural Crop Biology and Genetic Improvement (Central Region), Ministry of Education, Wuhan, 430070, People’s Republic of China

    Long-Fei Jin, Yong-Zhong Liu, Wei Du, Li-Na Fu, Syed Bilal Hussain & Shu-Ang Peng

Authors
  1. Long-Fei Jin
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yong-Zhong Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Wei Du
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Li-Na Fu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Syed Bilal Hussain
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Shu-Ang Peng
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Yong-Zhong Liu or Shu-Ang Peng.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Communicated by W.-W. Guo

Electronic supplementary material

Table S1

(DOCX 12 kb).

Table S2

(DOCX 11 kb).

Fig. S1

Randomness assessments of the two libraries (GIF 5 kb).

High resolution image (TIFF 1373 kb).

Fig. S2

Sequencing saturation analysis of the two libraries (GIF 6 kb).

High resolution image (TIFF 1372 kb).

Fig. S3

Up- and downregulated genes in chlorosis leaves. (GIF 8 kb).

High resolution image (TIFF 29 kb).

Fig. S4

Gene classification of differently expressed genes (GIF 84 kb).

High resolution image (TIFF 333 kb).

Data archiving statement

The raw data of the RNA-Seq has been uploaded to the Sequence Read Archive (SRA, https://www.ncbi.nlm.nih.gov/sra/) with the accession numbers SRR4419844 and SRR4419845.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Jin, LF., Liu, YZ., Du, W. et al. Physiological and transcriptional analysis reveals pathways involved in iron deficiency chlorosis in fragrant citrus. Tree Genetics & Genomes 13, 51 (2017). https://doi.org/10.1007/s11295-017-1136-x

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s11295-017-1136-x

Keywords

Search

Navigation