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Molecular characterization and differential expression of β-1,3-glucanase during ripening in banana fruit in response to ethylene, auxin, ABA, wounding, cold and light–dark cycles

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Abstract

β-1,3-Glucanases (E.C. 3.2.1.39) are widely distributed enzyme among bacteria, fungi, and higher plants. Analyses of accumulation levels of β-1,3-glucanase protein in various tissues in banana have clearly indicated abundance of β-1,3-glucanase protein accumulation in ripe pulp tissue. After cloning of β-1,3-glucanase from banana pulp (cultivar Cavendish), we have carried out an in silico analysis to investigate the sequential, structural, and phylogenetic characteristics of the putative banana β-1,3-glucanase protein. As like other ripening specific genes, β-1,3-glucanase is regulated in response to a wide variety of factors. Therefore, we have analyzed the transcript accumulation pattern and protein levels of β-1,3-glucanase in response to ethylene, auxin, ABA, wounding and, low temperature in preclimacteric banana fruit. Expression profile analyses have indicated that whereas exogenous application of ethylene strongly stimulated β-1,3-glucanase transcript accumulation, ABA partially induced the expression of the gene. On the other hand, wound treatment did not induce β-1,3-glucanase expression. Conversely, auxin and cold treatment negatively regulated β-1,3-glucanase gene expression and thus inhibited glucanase activity. In addition, β-1,3-glucanase transcript level was markedly decreased by constant exposure to white light. Protein level and enzymatic activity of β-1,3-glucanase were substantially increased with considerable decrease in fruit firmness by ethylene treatment and reduced exposure to white light conditions as compared with other treatments. Together, the overall study of β-1,3-glucanase expression pattern, glucanase activity, and changes in fruit firmness during ripening in various conditions suggest the possible physiological function of β-1,3-glucanase in fruit pulp softening.

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Abbreviations

ABA:

Abscisic acid

β-1,3-gluc:

β-1,3-Glucanase

BSA:

Bovine serum albumin

cDNA:

Complementary DNA

DTT:

1,4-Dithiothreitol

IAA:

Indole acetic acid

ORF:

Open reading frame

RT-PCR:

Reverse transcriptase-polymerase chain reaction

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Acknowledgments

We thank Prof. Frederick Meins, FMIBR, Basel, Switzerland for providing the antibody of β-1,3-glucanase. We gratefully acknowledge the financial assistance from Council for Scientific and Industrial Research (CSIR). For technical assistance, we are thankful to Mr. Jadav Ghosh, Division of Plant Biology, Bose Institute.

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Authors and Affiliations

  1. Division of Plant Biology, Bose Institute, 93/1, Acharya Prafulla Chandra Road, Kolkata, West Bengal, 700 009, India

    Swarup Roy Choudhury, Sanjay Kumar Singh & Dibyendu N. Sengupta

  2. Department of Chemistry, Bose Institute, 93/1, Acharya Prafulla Chandra Road, Kolkata, West Bengal, 700 009, India

    Sujit Roy

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  1. Swarup Roy Choudhury
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  2. Sujit Roy
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Correspondence to Swarup Roy Choudhury.

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Communicated by P. Lakshmanan.

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Supplemental Figure 1

. Alignment of the deduced full-length amino acid sequence of β-1,3-glucanase with other β-1,3-glucanase sequences. Gaps are shown as dashes; letters on black background indicate identical amino acids. ClustalW (1.83) performed multiple sequence alignment (DOC 98 kb)

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Roy Choudhury, S., Roy, S., Singh, S.K. et al. Molecular characterization and differential expression of β-1,3-glucanase during ripening in banana fruit in response to ethylene, auxin, ABA, wounding, cold and light–dark cycles. Plant Cell Rep 29, 813–828 (2010). https://doi.org/10.1007/s00299-010-0866-0

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