Abstract

<p><em>Dendrobium</em> orchid is one of the major export cut flowers not only in Thailand but also for several tropical countries. However, the production of ethylene by their flowers causes a shorter vase life. Flowers that contained lower levels of ethylene usually exhibited delayed senescence and consequently prolonged vase life. The transfer of antisense <em>ACC oxidase (ACO)</em> gene into orchid, in theory, may leads to decreased ethylene production because this gene can down regulates the ethylene biosynthesis pathway. This study focuses on the transformation and the existence and expression of the antisense <em>ACO</em>1 gene from papaya, namely (<em>CP-ACO</em>1), which was transferred in to <em>Dendrobium</em> 'Sonia Earsakul'. The successful stable transformation event obtained and the existence of the transferred gene was determined using PCR, dot blot hybridization and Southern blot hybridization techniques. The results revealed that antisense <em>CP-ACO</em>1 and <em>hygromycin phosphotransferase (hpt)</em> gene existed in all transgenic lines confirmed by PCR technique. The genomic dot blot confirmed the incorporation of the transgene in transgenic plant genome. Southern blot hybridization revealed the existed of one to four sets of the gene in transgenic lines. The expression of antisense <em>CP-ACO</em>1 gene was analyzed through the level of ACO enzyme activity and ethylene production in transgenic orchid. All of the transgenic lines had lower ACO enzyme activity and lower ethylene production than that of the non-transgenic orchid plants.<strong> </strong></p> <p> </p>

Highlights

  • Dendrobium orchid as one of the major cut flower in Thailand, have been export world-wide

  • The ethylene biosynthesis has two major enzymes: ACC synthase (ACS) and ACC oxidase (ACO) that are involved the rate of ethylene production

  • Regarding to the case that single Protocorm Like Bodies (PLBs) may form from multiple cells, each regenerated plantlet was considered to result from different origin of transformed cell

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Summary

Introduction

Dendrobium orchid as one of the major cut flower in Thailand, have been export world-wide. Transformation of antisense ACO gene from papaya (Carica papaya; dicotyledonous plant), that has 55% identity with ACO gene of Dendrobium orchid into Dendrobium ‘Sonia Earsakul’ (dicotyledonous plant) in this report ought to be beneficial research validating the decreased ethylene production in orchid. It is illustrate the effectiveness of antisense technology over the mono- and di-cotyledonous species

The Construction of Binary Vector Harboring Antisense CP-ACO1 Gene
Transplanting of Transgenic Plantlets
PCR Analysis
Dot Blot Analysis
Southern Blot Analysis
Analysis of ACC Oxidase Enzyme Activity
Analysis of Ethylene Production
Results and Discussion
Conclusion
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