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An effective in-gel assay protocol for the assessment of acid phosphatase (ACPase) isoform expression in the fungus Serendipita indica

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Abstract

The induction of acid phosphatase (ACPase) in the mycorrhizal fungi is an adaptive survival mechanism to cope in a low-phosphate environment. A mycorrhizal fungi Serendipita indica can induce the ACPase enzyme and enhance the phosphate (Pi) level to the host plant through Pi-solubilization mechanism, both intracellular and extracellular (media) levels. The spectrophotometer technique has been widely and commonly used to measure the ACPase enzyme activity in all microorganisms and plants using pNPP as a substrate. However, this technique cannot be useful when studying the involvement of ACPase isoforms in Pi-solubilization. In this article, we developed a single method to identify and express the ACPase isoforms of S. indica that contribute to the Pi-nutrition in the plant. This is native-PAGE electrophoresis with the in-gel assay and staining to detect the isoforms of the ACPase enzyme. The dark red-brown color developed after staining indicates the non-denatured (native) ACPase enzyme. This method utilized a modified minimal media for the de-repression of P-responsive genes such as ACPases with minimum salt contamination in the samples. This method will be helpful for the characterization of secretory and intracellular ACPases in fungi.

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All data related to this work will be available on request to the corresponding author.

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Funding

ASK is thankful to the UGC for providing Senior Research Fellowship to support her PhD research work. MK is thankful to the CSIR-Indian Institute of Toxicology Research, Lucknow, India for providing financial support (MLP002).

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Authors and Affiliations

  1. Environmental Toxicology Group, CSIR-Indian Institute of Toxicology Research (CSIR-IITR), Vishvigyan Bhavan, 31, Mahatma Gandhi Marg, Lucknow, Uttar Pradesh, 226001, India

    Aparna Singh Kushwaha & Manoj Kumar

  2. Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, Uttar Pradesh, 201002, India

    Aparna Singh Kushwaha & Manoj Kumar

Authors
  1. Aparna Singh Kushwaha
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  2. Manoj Kumar
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ASK: designing, protocol and experiment standardization, formal analysis, visualization, data analysis, literature survey and manuscript writing. MK: conception, data analyzing and manuscript editing.

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Correspondence to Manoj Kumar.

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All authors have seen, read, agreed and given consent for the communication and publication of this manuscript. The CSIR-IITR communication number for this manuscript is 3752.

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Kushwaha, A.S., Kumar, M. An effective in-gel assay protocol for the assessment of acid phosphatase (ACPase) isoform expression in the fungus Serendipita indica. 3 Biotech 12, 40 (2022). https://doi.org/10.1007/s13205-021-03103-0

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