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Sugarcane DIRIGENT and O-METHYLTRANSFERASE promoters confer stem-regulated gene expression in diverse monocots

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Abstract

Transcription profiling analysis identified Saccharum hybrid DIRIGENT (SHDIR16) and Ο-METHYLTRANSFERASE (SHOMT), putative defense and fiber biosynthesis-related genes that are highly expressed in the stem of sugarcane, a major sucrose accumulator and biomass producer. Promoters (Pro) of these genes were isolated and fused to the β-glucuronidase (GUS) reporter gene. Transient and stable transgene expression analyses showed that both Pro DIR16 :GUS and Pro OMT :GUS retain the expression characteristics of their respective endogenous genes in sugarcane and function in orthologous monocot species, including rice, maize and sorghum. Furthermore, both promoters conferred stem-regulated expression, which was further enhanced in the stem and induced in the leaf and root by salicylic acid, jasmonic acid and methyl jasmonate, key regulators of biotic and abiotic stresses. Pro DIR16 and Pro OMT will enable functional gene analysis in monocots, and will facilitate engineering monocots for improved carbon metabolism, enhanced stress tolerance and bioenergy production.

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Abbreviations

aRNA:

Amplified RNA

BAC:

Bacterial artificial chromosome

CaMV35S:

Cauliflower mosaic virus 35S promoter

GUS:

β-Glucuronidase

JA:

Jasmonic acid

MeJA:

Methyl jasmonate

MS:

Murashige and Skoog

MU:

4-Methylumbelliferone

PCR:

Polymerase chain reaction

Pro:

Promoter

qRT-PCR:

Quantitative RT-PCR

SA:

Salicylic acid

SHDIR:

Saccharum hybrid dirigent

SHOMT:

Saccharum hybrid O-methyltransferase

UBI:

Ubiquitin

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Acknowledgments

We are grateful to members of the Thomas Laboratory at Texas A&M University for discussions of results, data analysis and support. We specifically acknowledge Dr. Andrew Tag and Rick Hammer for bioinformatics support. We also thank Dr. Madhurababu Kunta (Citrus Center, Texas A&M University-Kingsville) for assistance in photography, and Brennick Langston (Texas AgriLife Research, Texas A&M System) for sequence submission to NCBI and determination of the three-dimensional structure of sugarcane O-methyltransferase. This work was supported by grants from the Texas Grain and Grass Gene Initiative Program (No. 06-0001), the Rio Grande Valley Sugar Growers Cooperative and Texas AgriLife Research, Texas A&M System.

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Authors and Affiliations

  1. Department of Plant Pathology and Microbiology, Texas AgriLife Research, Texas A&M System, Weslaco, TX, 78596, USA

    Mona B. Damaj, Marco T. Buenrostro-Nava, Ian S. Curtis & T. Erik Mirkov

  2. Department of Trait Genetics and Technologies, DowAgroSciences LLC, 9330 Zionsville Road, Indianapolis, IN, 46268, USA

    Siva P. Kumpatla

  3. Laboratory for Crop Transformation, Institute for Plant Genomics and Biotechnology, Norman E. Borlaug Center for Southern Crop Improvement, Texas A&M University, College Station, TX, 77843-2123, USA

    Chandrakanth Emani & Keerti S. Rathore

  4. Laboratory for Functional Genomics, Department of Biology, Texas A&M University, College Station, TX, 77843-3258, USA

    Phillip D. Beremand & Terry L. Thomas

  5. Department of Molecular Biology and Traits, DowAgroSciences LLC, 9330 Zionsville Road, Indianapolis, IN, 46268, USA

    Avutu S. Reddy

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  1. Mona B. Damaj
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  2. Siva P. Kumpatla
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  9. Terry L. Thomas
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Correspondence to Terry L. Thomas or T. Erik Mirkov.

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Damaj, M.B., Kumpatla, S.P., Emani, C. et al. Sugarcane DIRIGENT and O-METHYLTRANSFERASE promoters confer stem-regulated gene expression in diverse monocots. Planta 231, 1439–1458 (2010). https://doi.org/10.1007/s00425-010-1138-5

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  • DOI: https://doi.org/10.1007/s00425-010-1138-5

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