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Molecular mapping and candidate gene analysis of a new epicuticular wax locus in sorghum (Sorghum bicolor L. Moench)

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Abstract

Key message

A new epicuticular wax (bloom) locus has been identified and fine mapped to the 207.89 kb genomic region on chromosome 1. A putative candidate gene, Sobic.001G269200, annotated as GDSL-like lipase/acylhydrolase, is proposed as the most probable candidate gene involved in bloom synthesis/deposition.

Abstract

Deposition of epicuticular wax on plant aerial surface is one strategy that plants adapt to reduce non-transpiration water loss. Epicuticular wax (bloom)-less mutants in sorghum with their glossy phenotypes exhibit changes in the accumulation of epicuticular wax on leaf and culm surfaces. We report molecular mapping of a new sorghum locus, bloomless mutant (bm39), involved in epicuticular wax biosynthesis in sorghum. Inheritance studies involving a profusely bloom parent (BTx623) and a spontaneous bloomless mutant (RS647) indicated that the parents differed in a single gene for bloom synthesis. Bloomless was recessive to bloom deposition. Genetic mapping involving F2 and F7 mapping populations in diverse genetic backgrounds (BTx623 × RS647; 296A × RS647 and 27A × RS647) identified and validated the map location of bm39 to a region of 207.89 kb on chromosome 1. SSR markers, Sblm13 and Sblm16, flanked the bm39 locus to a map interval of 0.3 cM on either side. Nine candidate genes were identified, of which Sobic.001G269200 annotated for GDSL-like lipase/acylhydrolase is the most likely gene associated with epicuticular wax deposition. Gene expression analysis in parents, isogenic lines and sets of near isogenic lines also confirmed the reduced expression of the putative candidate gene. The study opens possibilities for a detailed molecular analysis of the gene, its role in epicuticular wax synthesis and deposition, and may help to understand its function in moisture stress tolerance and insect and pathogen resistance in sorghum.

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Abbreviations

CMS:

Cytoplasmic male sterility

CTAB:

Cetyltrimethylammonium bromide

dNTP:

Deoxynucleotide triphosphate

GBSS:

Granule-bound starch synthase

GDSL:

Glycine aspartic (glutamic) serine leucine

ORF:

Open reading frame

PCR:

Polymerase chain reaction

RT-PCR:

Reverse transcriptase polymerase chain reaction

SNP:

Single nucleotide polymorphism

SSRs:

Simple sequence repeats

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Acknowledgements

We sincerely thank the anonymous reviewers and the editor for their excellent comments and suggestions, which greatly improved the quality of the manuscript. We also thank the Director, ICAR-IIMR, Rajendranagar, Hyderabad, for the facilities for undertaking this study. We thank Dr. P. G. Padmaja (IIMR) for extending facilities for wax estimation, Dr. P. Rajendrakumar (IIMR) and Dr. M. Sheshu Madav (IIRR) for facilitating RNA expression studies and Dr. D. Sanjeev Rao (IIRR) for his suggestions in peptide analysis. The help of Mr. Jai Kishan, M. Bhaskar and M. Shankariah in conducting laboratory and field works is also gratefully acknowledged.

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Authors and Affiliations

  1. Marker-Assisted Selection Laboratory, ICAR-Indian Institute of Millet Research (IIMR), Rajendranagar, Hyderabad, 500 030, India

    G. Anurag Uttam, M. Praveen, Vilas A. Tonapi & R. Madhusudhana

  2. Department of Botany, Andhra University, Visakhapatnam, Andhra Pradesh, India

    Y. Venkateswara Rao

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  1. G. Anurag Uttam
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  2. M. Praveen
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  3. Y. Venkateswara Rao
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  4. Vilas A. Tonapi
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  5. R. Madhusudhana
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Correspondence to R. Madhusudhana.

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Communicated by Antonio Augusto Franco Garcia.

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Uttam, G.A., Praveen, M., Rao, Y.V. et al. Molecular mapping and candidate gene analysis of a new epicuticular wax locus in sorghum (Sorghum bicolor L. Moench). Theor Appl Genet 130, 2109–2125 (2017). https://doi.org/10.1007/s00122-017-2945-x

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