Characterization of a sterile dwarf mutant and the cloning of zeaxanthin epoxidase in Asian cotton (Gossypium arboreum L.)
To document radiation damage incurred by nuclear DNA and understand the molecular mechanisms that control plant height, a sterile dwarf mutant (sd a ) derived from the Gossypium arboreum treated with 60Co γ-rays was characterized. A sequence analysis revealed that sd a included a partial deletion and a point mutation. The physiological changes and responses of the sd a mutant to exogenous hormones showed that the mutant’s chlorophyll contents, net photosynthetic rate, and abscisic acid levels were significantly reduced compared to those of wild-type. Using the sequence of a differentially expressed sequence tag encoding a zeaxanthin epoxidase (ZEP), a novel etiolation-associated gene, full-length cDNA was rapidly cloned using rapid amplification of cDNA ends. The full-length cDNA of this gene is 2414 bp, with a 2004-bp open reading frame encoding a putative ZEP of 667 amino acid residues that has a 73.421-kDa theoretical molecular mass and a 6.67 isoelectric point. The genomic sequence of GaZEP is 5233 bp, comprising 15 exons and 14 introns. The GaZEP promoter contains numerous cis-regulatory elements that are responsive to light, hormones, and stress. Real-time quantitative PCR indicated that GaZEP expression in the mutant was downregulated. The expression levels of GaZEP in roots and leaves were positively correlated with the root hair lengths and abscisic acid levels, respectively. Thus, in photosynthetic tissues, GaZEP mRNA levels might be regulated coordinately with variations in the abscisic acid levels. These results may assist in the further functional dissection of ZEP and in elucidating the molecular mechanism underlying sterile dwarfism in cotton.
KeywordsGossypium arboretum Sda Radiation mutation Deletion Abscisic acid Zeaxanthin epoxidase (ZEP) gene
We thank Prof. Baomin Wang (China Agricultural University, Beijing, China) for endogenous hormone measurements. This work was supported by grants from the China Agriculture Research System (No. CARS-34-YZ4), the National Natural Science Foundation of China (No. 30571184) and the National Key Technologies Research and Development (R&D) Program of China (No. 2004BA525B05-1).
- Anmarkrud JA, Kleven O, Augustin J, Bentz KH, Blomqvist D, Fernie KJ, Magrath MJ, Pärn H, Quinn JS, Robertson RJ, Szép T, Tarof S, Wagner RH, Lifjeld JT (2011) Factors affecting germline mutations in a hypervariable microsatellite: a comparative analysis of six species of swallows (Aves: Hirundinidae). Mutat Res 708(1–2):37–43CrossRefPubMedGoogle Scholar
- Audran C, Liotenberg S, Gonneau M, North H, Frey A, Tap-Waksman K, Vartanian N, Marion-Poll A (2001) Localisation and expression of zeaxanthin epoxidase mRNA in Arabidopsis in response to drought stress and during seed development. Aust J Plant Physiol 28:1161–1173Google Scholar
- Balmer LA, Beveridge DJ, Jazayeri JA, Thomson AM, Walker CE, Leedman PJ (2001) Identification of a novel AU–rich element in the 3ʹ untranslated region of epidermal growth factor receptor mRNA that is the target for regulated RNA–binding proteins. Mol Cell Biol 21:2070–2084CrossRefPubMedPubMedCentralGoogle Scholar
- Cheng WH, Endo A, Zhou L, Penney J, Chen HC, Arroyo A, Leon P, Nambara E, Asami T, Seo M, Koshiba T, Sheen J (2002) A unique short-chain dehydrogenase/reductase in Arabidopsis glucose signaling and abscisic acid biosynthesis and functions. Plant Cell 14(11):2723–2743CrossRefPubMedPubMedCentralGoogle Scholar
- Dharmadhikari AV, Gambin T, Szafranski P, Cao W, Probst FJ, Jin W, Fang P, Gogolewski K, Gambin A, George-Abraham JK, Golla S, Boidein F, Duban-Bedu B, Delobel B, Andrieux J, Becker K, Holinski-Feder E, Cheung SW, Stankiewicz P (2014) Molecular and clinical analyses of 16q24.1 duplications involving FOXF1 identify an evolutionarily unstable large minisatellite. BMC Med Genet 15:128CrossRefPubMedPubMedCentralGoogle Scholar
- Falchi R, Zanon L, De Marco F, Nonis A, Pfeiffer A, Vizzotto G (2013) Tissue-specific and developmental expression pattern of abscisic acid biosynthetic genes in peach fruit: possible role of the hormone in the coordinated growth of seed and mesocarp. J Plant Growth Regul 32:519–532CrossRefGoogle Scholar
- Li P, Dong H, Liu A, Liu J, Sun M, Wang G, Zhang S, Li Y, Mao S (2014) Diagnosis of premature senescence of cotton using SPAD value. Agric Sci 5:992–999Google Scholar
- Lind C, Dreyer I, López-Sanjurjo EJ, von Meyer K, Ishizaki K, Kohchi T, Lang D, Zhao Y, Kreuzer I, Al-Rasheid KA, Ronne H, Reski R, Zhu JK, Geiger D, Hedrich R (2015) Stomatal guard cells co-opted an ancient ABA-dependent desiccation survival system to regulate stomatal closure. Curr Biol 25(7):928–935CrossRefPubMedGoogle Scholar
- Wu C (2008) Isolation and characterization of a sterile-dwarf mutant and cloning of its related genes in asian cotton (Gossypium arboreum L.). Dissertation, Nanjing Agricultural University, pp 67–72Google Scholar
- Yang C, Zhu J, Jiang Y, Wang X, Gu M, Wang Y, Kang H, Fan X, Sha L, Zhang H, Xuan P, Zhou Y (2014) 100 Gy 60 Co γ-ray induced novel mutations in tetraploid wheat. Sci World J 2014:725813Google Scholar
- Zhang J, Wu YT, Guo WZ, Zhang TZ (2000) Fast screening of SSR markers in cotton with PAGE/siver staining. Cotton Sci Sin 12:267–269Google Scholar