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Nonsense-mediated mRNA degradation of CtFAD2-1 and development of a perfect molecular marker for olol mutation in high oleic safflower (Carthamus tinctorius L.)

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Abstract

There are two types of safflower oil, high oleic (HO) with 70–75 % oleic acid and high linoleic (HL) with about 70 % linoleic acid. The original HO trait in safflower, found in an introduction from India, is controlled by a partially recessive allele ol at a single locus (Knowles and Bill 1964). In the lipid biosynthesis pathway of developing safflower seeds, microsomal oleoyl phosphatidylcholine desaturase (FAD2) is largely responsible for the conversion of oleic acid to linoleic acid. In vitro microsomal assays indicated drastically reduced FAD2 enzyme activity in the HO genotype compared to conventional HL safflower. A previous study indicated that a single-nucleotide deletion was found in the coding region of CtFAD2-1 that causes premature termination of translation in the HO genotypes, and the expression of the mutant CtFAD2-1Δ was attenuated in the HO genotypes compared to conventional HL safflower (Guan et al. 2012). In this study, we hypothesise that down-regulation of CtFAD2-1 expression in the HO genotype may be explained by nonsense-mediated RNA decay (NMD). NMD phenomenon, indicated by gene-specific RNA degradation of defective CtFAD2-1Δ, was subsequently confirmed in Arabidopsis thaliana seed as well as in the transient expression system in Nicotiana benthamiana leaves. We have developed a perfect molecular marker corresponding to the olol mutation that can facilitate a rapid screening and early detection of genotypes carrying the olol mutation for use in marker-assisted selection for the management of the HO trait in safflower breeding programmes.

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Acknowledgments

This research project was financially supported by the Australian Grains Research and Development Corporation (GRDC). Shijiang Cao wishes to thank the China Scholarship Council (CSC) for providing a scholarship for studying in Australia. We also wish to thank Professor Sten Stymne of the Swedish University of Agricultural Sciences, Sweden, for teaching us the microsomal FAD2 enzyme assay. Excellent technical support from Anu Mathew, Luch Hac, Amratha Ashwin and Bei Dong are gratefully acknowledged. The authors are also grateful to Dr Phil Larkin for critically reviewing the manuscript.

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Authors and Affiliations

  1. Commonwealth Scientific and Industrial Research Organization Plant Industry, PO Box 1600, Canberra, ACT, 2601, Australia

    Qing Liu, Xue-Rong Zhou, Craig Wood, Allan Green & Surinder Singh

  2. School of Life Sciences, Northeast Normal University, Changchun, China

    Shijiang Cao

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  1. Qing Liu
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Correspondence to Qing Liu.

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Communicated by B. Hulke.

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Liu, Q., Cao, S., Zhou, XR. et al. Nonsense-mediated mRNA degradation of CtFAD2-1 and development of a perfect molecular marker for olol mutation in high oleic safflower (Carthamus tinctorius L.). Theor Appl Genet 126, 2219–2231 (2013). https://doi.org/10.1007/s00122-013-2129-2

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