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Development and validation of rapid and cost-effective protocol for estimation of amylose and amylopectin in maize kernels

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Abstract

Maize possesses wide variation in amylose and amylopectin which assumes significance as a part of both food-chain and different industrial applications. Estimation of amylose and amylopectin in maize kernels is important for developing suitable hybrids. The existing protocols for estimation of amylose and amylopectin in maize are elaborate and lengthy, and involve high cost. Here, we developed a rapid and cost-effective method for estimation of amylose and amylopectin in maize kernels. 10% toluene and 80% ethanol were used for removal of proteins (~ 10%) and lipids (~ 4%) from maize flour. The over-estimation of amylose was minimized using NaOH with KI to stop free KI to bind with amylopectin. Standards were improved by mixing amylose and amylopectin in different concentrations (0–100%), rather than using amylose or amylopectin alone. Standard curve generated regression equation of y = 90.436x + 0.8535 with R2 = 0.9989. Two types of samples viz., (1) protein, amylose and amylopectin (2) amylose and amylopectin, showed that starch fractions were highly comparable to expected values with correlation coefficient (r) of 0.9998 and mean standard deviation of 0.54. The protocol successfully estimated wide range of amylose (2.79–50.04%) and amylopectin (59.96–97.21%) among diverse maize inbreds including amylose extender1 (ae1) and waxy1 (wx1) mutants. Present protocol required 75% less time and 92.5% less cost compared to existing protocols. The newly developed method would be highly useful in developing maize hybrids high in amylose or amylopectin. This is the first report of rapid and cost-effective protocol for estimation of starch fractions in maize kernels.

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Acknowledgements

First author thanks Indian Council of Agricultural Research (ICAR), New Delhi for providing the fellowship during the M.Sc. program. Financial help received from Division of Genetics, ICAR-IARI, New Delhi, and ICAR-Consortia Research Platform on ‘Molecular breeding for improvement of tolerance to biotic and abiotic stresses, yield and quality traits in crops - Maize component’ (IARI Project Code No.: 12-143C) is thankfully acknowledged. We thank breeders of the national program and CIMMYT, Mexico for sharing the inbreds.

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Authors and Affiliations

  1. Division of Genetics, ICAR-Indian Agricultural Research Institute, New Delhi, 110012, India

    Shashidhar Bayappa Reddappa, Rashmi Chhabra, Zahirul Alam Talukder, Vignesh Muthusamy, Rajkumar Uttamrao Zunjare & Firoz Hossain

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  1. Shashidhar Bayappa Reddappa
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  2. Rashmi Chhabra
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  3. Zahirul Alam Talukder
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  4. Vignesh Muthusamy
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  5. Rajkumar Uttamrao Zunjare
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  6. Firoz Hossain
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Contributions

Conduct of experiment: SBR; development of protocol: RC, SBR; validation of protocol with maize inbreds: ZAT; development and maintenance of inbreds: VM; Statistical analysis: RUZ; Manuscript writing: SBR, RC and FH; Design of experiment: FH.

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Correspondence to Firoz Hossain.

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Reddappa, S.B., Chhabra, R., Talukder, Z.A. et al. Development and validation of rapid and cost-effective protocol for estimation of amylose and amylopectin in maize kernels. 3 Biotech 12, 62 (2022). https://doi.org/10.1007/s13205-022-03128-z

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  • DOI: https://doi.org/10.1007/s13205-022-03128-z

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