Characterization of gibberellin overexpression lines in pea

  • Author / Creator
    Wickramarathna, Aruna
  • Abstract Gibberellins (GAs) are a class of plant hormones that regulate many aspects of plant growth and development including seed germination, stem elongation and fruit development. To investigate the regulation of GA biosynthesis and the impact of altered GA levels on plant growth and development, transgenic pea (Pisum sativum L. cv. Carneval) plants were generated to overexpress PsGA3ox1 (codes for GA 3β-hydroxylase which converts GA20 to bioactive GA1) under the control of the CaMV-35S promoter. Increased expression of the transgene PsGA3ox1 was correlated with altered plant phenotype including longer internodes, larger stipules and tendrils, and longer pods. Transgenic lines also showed upregulation of the GA catabolic genes PsGA2ox1 and/or PsGA2ox2, suggesting that GA1 substrate-induced feedback regulation also occurs to maintain GA homeostasis. Changes in endogenous GAs, quantified using an isotope dilution method, indicated that an increased flux in GA biosynthesis occurred in the expanding internodes, stipules and tendrils of the PsGA3ox1-overexpressor lines. Higher bioactive GA1 levels and growth were correlated with lower PsGA2ox1 transcript levels in elongating internodes, and oscillation of these parameters between adjacent elongating internodes in the PsGA3ox1-overexpression lines suggests that coordination of bioactive GA levels and growth occurs between adjacent internodes. During germination and early seedling growth, GA gene expression studies suggested that PsGA3ox1-overexpression increased the flux through to bioactive GA in the cotyledons, shoots and roots of pea seedlings, resulting in longer shoots but shorter roots. Auxins are a class of plant hormones involved in growth and differentiation of plants that can influence GA biosynthesis and action. The location and action of auxins is in part regulated by auxin carrier proteins. The expression patterns of the putative auxin efflux carrier genes PsPIN1 and PsPIN2 in elongating internodes were correlated with vascular re-patterning events in this tissue, and PsGA3ox1-overexpression appears to increase internode PsPIN1 and PsPIN2 transcript abundance and the formation of the vascular connections between the internode and the axillary buds. Overall, characterization of PsGA3ox1-overexpressor lines in pea demonstrated that bioactive GA levels are tightly regulated in pea tissues for the coordination of plant growth and development.

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  • Degree
    Doctor of Philosophy
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