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 Citrus and Other Subtropical Products Research: Fort Pierce, FL
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ARS Home Southeast Area Fort Pierce, Florida U.S. Horticultural Research Laboratory Citrus and Other Subtropical Products Research Research Publications at this Location





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Title:   Author  item
                                            Bai, Jinheitem
                                            Baldwin, Elizabeth

                                                         - *Liz*item
                                            LIAO, HUI-LING - University Of Floridaitem
                                            KOSTENYUK, IGOR - University Of Floridaitem
                                            BURNS, JACQUELINE - University Of Floridaitem
                                            IREY, MIKE - Southern Gardens Citrus
  Submitted to: Florida State Horticultural Society Meeting
                                    Publication Type: Abstract Only
                                    Publication Acceptance Date: 4/10/2012
                                    Publication  Date: N/A
                                    Citation: N/AInterpretive Summary:
                                Technical Abstract: Response of gene expression in the oxylipin pathway to chilling and heating in tomatoes at full ripe stage was investigated.  Total RNA was isolated from tomato pericarp tissue, and gene expression of Tomlox A-Tomlox D, HPL and ADH were determined by real-time Polymerase Chain Reaction (PCR) using the SYBR Green PCR kit, and expressed as ''CT in comparison to the reference controls, GAPGH and RPL8.  The widely used reference gene GAPGH was more stable than RPL8 in the postharvest stress research.  Primer PCR efficiencies of the genes were determined using a 100-fold dilution series of a bulked complementary DNA (cDNA) sample over three dilution points, and the results showed that primers designed using Primer Express 3.0 (Applied Biosystems) had higher efficiency over other primer design programs in the ABI PRISM 7500 Sequence Detection Fast System.  For orange research, total DNA was extracted from Huanglongbing infected fruit juice.  Citrus cytochrome oxidase (COX) was selected to represent host gene, and 16S rDNA was selected to represent the pathogen bacteria, Candidatus Liberibacter asiaticus (CLas).  Amplification of 16S rDNA was inhibited by unknown PCR enzyme inhibitors presented in the DNA extraction.  However, the inhibitors were removed by pathing through a silicon spin column. A multiplex qPCR of COX and CLas 16S DNA showed that the amplification of CLas 16S DNA was inhibited by COX in an ABI PRISM 7500 FAST sequence detection system.


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