The Introduction of Transgenes to Control Blackheart in Pineapple (Ananas Comosus L.) cv. Smooth Cayenne by Microprojectile Bombardment

Abstract

A transformation technique for the introduction of transgenes to control blackheart by particle bombardment has been developed for pineapple cv. Smooth Cayenne. Leaf callus cultures capable of high frequency organogenesis with a short regeneration time were used as explant material. Gus and gfp reporter genes were used to observe and determine transient and stable expression. The ppo gene, isolated from pineapple, was introduced to control blackheart. Co-transformation occurred with constructs containing the nptII gene conferring geneticin resistance. We have recovered 15 independent transgenic gus and gfp lines each from 8 separate experiments and 22 ppo lines from 11 experiments. Gus, gfp, ppo and nptII positive plants have been regenerated, which have been shown by Southern blot analysis to be stable transgenics containing multiple copies of the introduced genes. These results show that biolistic gene delivery in pineapple can be successfully achieved at an acceptable efficiency of 0.21-1.5% for genetic improvement of 'Smooth Cayenne', the industry standard throughout the world.

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

“…Other important agronomic problems, such as precocious flowering, have been tackled, and transgenic pineapples have been produced with delayed natural flowering (Trusov and Botella 2006). Transgenes have also been introduced to control chillinginduced internal fruit browning (Ko et al 2006).…”

“…Both Agrobacterium-mediated and biolistic transformation of pineapple have been previously reported (Graham et al 2001;Sripaoraya et al 2001;Espinosa et al 2002;Firoozabady et al 2006;Ko et al 2006;Yabor et al 2006). The present report describes a new method for Agrobacterium-mediated transformation of pineapple based on direct formation of adventitious buds from leaf bases and stem discs.…”

“…The present report describes a new method for Agrobacterium-mediated transformation of pineapple based on direct formation of adventitious buds from leaf bases and stem discs. Establishment and maintenance of undifferentiated cultures, either before or after the transformation events, were time-consuming integral steps in most of the other studies (Graham et al 2001;Espinosa et al 2002;Firoozabady et al 2006;Ko et al 2006;Yabor et al 2006). The use of undifferentiated cultures also results in a high frequency of somaclonal variations among regenerated pineapple plants (Wakasa 1979;Rangan 1984;Wakasa 1989;Firoozabady and Moy 2004).…”

Agrobacterium-mediated genetic transformation of pineapple var. Queen using a novel encapsulation-based antibiotic selection technique

“…The same technique was used to introduce a ppo (polyphenol oxidase) gene involved in the control of the bacterial-induced blackheart in pineapple cv. Smooth Cayenne though the transformation efficiency was less than 1.5% (Ko et al 2006).…”

Genomics of Pineapple, Crowning The King of Tropical Fruits

“…Another important agronomic problem such as natural flowering has also been tackled and transgenic pineapples have been produced with delayed natural flowering (Trusov and Botella 2006). Fruit quality issues have also been addressed with Ko et al (2006) introducing transgenes to control blackheart, an internal browning of fruit flesh as a result of chilling injury.…”