Authors: Wang, M.-L.¹; Uruu, G.²; Xiong, L.¹; He, X.¹; Nagai, C.¹; Cheah, K.²; Hu, J.³; Nan, G.-L.¹; Sipes, B.³; Atkinson, H.⁴; Moore, P.⁵; Rohrbach, K.³; Paull, R.⁶

Source: In Vitro Cellular and Development Biology - Plant, Volume 45, Number 2, April 2009 , pp. 112-121(10)

Publisher: Springer

Abstract:

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.

Keywords: Transgenic pineapple; Ananas comosus; Agrobacterium tumefaciens; Adventitious bud; Regeneration; Ethylene; Nematodes; Flowering

Document Type: Research article

DOI: http://dx.doi.org/10.1007/s11627-009-9208-8

Affiliations: 1: Hawaii Agriculture Research Center, P.O. Box 100, Kunia, HI, 96759, USA 2: Department of Tropical Plant and Soil Sciences, University of Hawaii at Manoa, 3190 Maile Way, Honolulu, HI, 96822, USA 3: Department of Plant and Environment Protection Sciences, University of Hawaii at Manoa, 3190 Maile Way, Honolulu, HI, 96822, USA 4: Centre for Plant Sciences, University of Leeds, Leeds, LS2 9JT, UK 5: USDA, ARS, PWA, Pacific Basin Agricultural Research Center, 64 Nowelo Street, Hilo, HI, 96720, USA 6: Department of Tropical Plant and Soil Sciences, University of Hawaii at Manoa, 3190 Maile Way, Honolulu, HI, 96822, USA, Email: paull@hawaii.edu

Publication date: 2009-04-01

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• In this Subject: Biology ,  Botany
• By this author: Wang, M.-L. ;  Uruu, G. ;  Xiong, L. ;  He, X. ;  Nagai, C. ;  Cheah, K. ;  Hu, J. ;  Nan, G.-L. ;  Sipes, B. ;  Atkinson, H. ;  Moore, P. ;  Rohrbach, K. ;  Paull, R.

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