Production of transgenic pineapple (Ananas cosmos (L.) Merr.) plants via adventitious bud regeneration

Autor: L. Xiong, Robert E. Paull, John S. Hu, C. Nagai, H. J. Atkinson, K. G. Rohrbach, X. He, G.-L. Nan, Paul H. Moore, M.-L. Wang, G. Uruu, Brent S. Sipes, K. T. Cheah
Rok vydání: 2009
Předmět:
Zdroj: In Vitro Cellular & Developmental Biology - Plant. 45:112-121
ISSN: 1475-2689
1054-5476
Popis: A new protocol for the production of transgenic pineapple plants was developed. Adventitious buds were induced directly from Agrobacterium-infected leaf bases and stem discs of in vitro plants, bypassing the establishment of callus cultures. Non-chimeric transgenic plants were obtained by multiple subculturing of primary transformants under increasing levels of selection. A total of 42 independent transgenic lines were produced from two cultivars with two different constructs: one containing a modified rice cystatin gene (Oc-IΔD86) and the other with the anti-sense gene to pineapple aminocyclopropane synthase (ACS). GUS histochemical staining provided the first evidence of the non-chimeric nature of the transformed plants. Their non-chimeric nature was further demonstrated by PCR analyses of the DNA extracted from individual leaves of a primary transformed plant and also from multiple plants propagated from a single transformation event. Southern hybridization confirmed random integration patterns of transgenes in the independent lines. For the Oc-IΔD86 gene, the expression at the mRNA level was detected via RT-PCR and its translation was detected by protein blot. Agronomic evaluation and bioassays of the transgenic plants will further validate the utility of this new tool for pineapple improvement.
Databáze: OpenAIRE
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