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European Journal of Plant Pathology

, Volume 148, Issue 3, pp 533–549 | Cite as

Differential expressions of photosynthetic genes provide clues to the resistance mechanism during Fusarium oxysporum f.sp. ciceri race 1 (Foc1) infection in chickpea (Cicer arietinum L.)

  • Anirban Bhar
  • Sumanti Gupta
  • Moniya Chatterjee
  • Senjuti Sen
  • Sampa Das
Article

Abstract

Fusarium oxysporum f.sp. ciceri race 1 (Foc1), a root-invading pathogen causes vascular wilt in chickpea (Cicer arietinum L.). Foc1 is known to induce reactive oxygen species (ROS) mediated localized defense responses at the site of colonization in roots. However, the effect of this localized infection on distant shoot tissues is still unknown. In the present study, the effect of Foc1 on shoot tissues of both susceptible and resistant chickpea plants was studied. Total pigment content and fluorescence of chlorophyll was measured. Occurrence of oxidative damage in shoots was confirmed by both biochemical and lipid peroxidation assays. Expression pattern of some redox responsive transcripts were also analyzed. Additionally, transcriptional accumulations of some key genes related to light reaction, carbon reduction and photosystem II (PSII) of photosynthesis were analyzed at different time points post infection. Expressional status of stress induced sugar metabolism related genes (sucrose synthase, β amylase and invertase) were also investigated. Finally, gene networks were constructed showing interconnection of the photosynthetic genes, sugar metabolism-related genes and redox responsive transcripts with other metabolic and stress related pathways. The results demonstrate that the infection in root tissues of chickpea by Foc1 dramatically increases the ROS levels in shoot tissues of susceptible plants. The oxidative outburst in shoot tissues of susceptible plants also hampers the photosynthetic stability by down-regulating the key photosynthetic genes. On the contrary, resistant chickpea lines are grossly devoid of such instances with few behavioral irregularities at later time points.

Keywords

Biotic stress Cicer arietinum Fusarium oxysporum f.sp. ciceri race 1 Oxidative burst Photosynthetic genes Wilt disease 

Notes

Acknowledgments

This work was supported by the grant provided to A. Bhar by Council of Scientific and Industrial Research, India (09/015(0378) /2009-EMR-1), to S. Gupta by Department of Biotechnology, Government of India (BT/PR9593/AGR/02/444/ 2007), to M. Chatterjee by Department of Biotechnology, Government of India (BT/01/COE/06/03/2006) and S. Sen by Indian Council of Agricultural Research (NFBSFARA/PB-2010/2010-11). S.Das was supported by funds provided by Bose Institute, Department of Science and Technology, Government of India. The funding organizations had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Authors thank Dr. S.C. Pande (ICRISAT, Patancheru) for providing fungal culture and Dr. S.K. Chaturvedi (IIPR, Kanpur) for providing chickpea seeds. Patient assistance of Mr. Swarnava Das for chlorophyll isolation and microscopic experiments is greatly acknowledged. Special thanks are reserved for Mr. Arup Kumar Dey for his help in green house experiments. Mr. Sudipta Basu is duly acknowledged for plant maintenance. Finally, authors acknowledge the Director, Bose Institute for infrastructural facilities.

Supplementary material

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ESM 1 (DOC 28 kb)
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Table S1 (DOC 58 kb)
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Table S2 (DOC 120 kb)
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Table S3 (XLS 60 kb)
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Table S4 (DOC 96 kb)
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Fig. S1:

External morphology and percentage of disease incidence in chickpea upon Foc1 infection. (a) Control JG62 plant. (b) Control WR315 plant. (c) Infected JG62 plant at 2 DPI. (d) Infected WR315 plant at 2 DPI. (e) Infected JG62 plant at 4 DPI. (f) Infected WR315 plant at 4 DPI. (g) Infected JG62 plant at 12 DPI. (h) Infected WR315 plant at 12 DPI. (i) Percentage of plants infected with Foc1 in both JG62 and WR315 plants. (GIF 163 kb)

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Fig. S2:

Graphical representation of DAB staining assay showing differential accumulation of hydrogen peroxide in root tissues of chickpea post Foc1 inoculation. (a) Accumulation of hydrogen peroxide in root tissues of susceptible JG62 plants at different time points. (b) Accumulation of hydrogen peroxide in root tissues of resistant WR315 plants at different time points. Bars represent standard error (n = 3), stars represent statistical significance, ns represents non significant data. (GIF 17 kb)

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Fig. S3

Relative expression profile of photosynthetic light reaction related genes in response to Foc1 infection (a-b) represents relative expression profile of genes involved in chlorophyll biosynthesis and degradation. (a) Expression pattern in JG62 plants. (b) Expression pattern in WR315 plants. (c-d) Represents relative expression profile of genes involved in maintenance of photosynthetic apparatus. (c) Expression pattern in JG62 plants (d) Expression pattern in WR315 plants. Value represents mean ± S.E (n = 81) and significant differences were calculated by student’s t-test at P < 0.05 (GIF 83 kb)

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Fig. S4

Relative expression profile of photosynthetic carbon reduction cycle related genes in response to Foc1 infection. (a-b) Represents relative expression profile of genes involved in regulation of RuBisCO activity. (a) Expression pattern in JG62 plants. (b) Expression pattern in WR315 plants. (c-d) Represents relative expression profile of genes involved in regulation of carbon reduction cycle. (c) Expression pattern in JG62 plants. (d) Expression pattern in WR315 plants. Value represents mean ± S.E (n = 81) and significant differences were calculated by student’s t-test at P < 0.05 (GIF 87 kb)

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Fig. S5

Light microscopic images of starch granules stained with Lugol’s iodine. (a) Panel represents susceptible plants at different post infection time points. (b) Panel represents resistant plants at different time points post infection. Bars represent 10 μm (GIF 41 kb)

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Fig. S6

Relative expression profile of sugar metabolism related genes in response to Foc1 infection. (a-b) Represents relative expression profile of sucrose synthase, invertase and β amylase. (a) Expression pattern in JG62 plants. (b) Expression pattern in WR315 plants. Value represents mean ± S.E (n = 81) and significant differences were calculated by student’s t-test at P < 0.05 (GIF 38 kb)

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Fig S7:

The network showing key interactors of hydrogen peroxide in plants. The network generated in Pathway Studio (Version 7.1) software taking H2O2 as small molecule central hub. (GIF 251 kb)

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Fig. S8

Network showing interacting partners of photosynthetic light reaction related genes. PORA, light-dependent NADPH:protochlorophyllide oxidoreductase A; G4, chlorophyll synthase; ATCLH1, chlorophyllase; LHCA3, PSI type III chlorophyll a/b-binding protein (Lhca3*1); LHCB5,the light-harvesting chlorophyll a/b binding protein CP26; HCF208, high chlorophyll fluorescence 208; CH1, chlorophyllide a oxygenase. (GIF 49 kb)

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Fig. S9

Network showing interacting partners of photosynthetic carbon reduction cycle related genes. RBCS1A, ribulose 1,5 bis phosphate carboxylase/oxygenase (RuBisCO) small subunit ; rbcL, ribulose 1,5 bis phosphate carboxylase/oxygenase (RuBisCO) large subunit; RCA, RuBisCO activase; CPN60B, chaperonin 60 beta; FBA1 (F3 K23.9), fructose bis phosphate aldolase 1; TIM, plastidic triose phosphate isomerase. (GIF 50 kb)

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Copyright information

© Koninklijke Nederlandse Planteziektenkundige Vereniging 2016

Authors and Affiliations

  • Anirban Bhar
    • 1
    • 2
  • Sumanti Gupta
    • 1
    • 3
  • Moniya Chatterjee
    • 1
  • Senjuti Sen
    • 1
  • Sampa Das
    • 1
  1. 1.Division of Plant BiologyBose InstituteKolkataIndia
  2. 2.Post Graduate Department of BotanyRamakrishna Mission Vivekananda Centenary CollegeKolkataIndia
  3. 3.Department of BotanyRabindra MahavidyalayaHooghlyIndia

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