Induced expression of selected plant defence related genes in pot azalea, Rhododendron simsii hybrid

A set of putative marker genes to study plant defense responses against Polyphagotarsonemus latus, a key pest in the production of Rhododendron simsii hybrids, was selected and validated. Genes belonged to the biosynthetic pathway of phytohormones jasmonic acid (JA) (RsLOX, RsAOS, RsAOC, RsOPR3 and...

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Bibliographic Details
Published in:Euphytica
Main Authors: Luypaert, Gil, Witters, Johan, Van Huylenbroeck, Johan, De Clercq, Patrick, De Riek, Jan, De Keyser, Ellen
Format: Article in Journal/Newspaper
Language:English
Published: 2017
Subjects:
Biology and Life Sciences
Jasmonic acid (JA)
Methyl jasmonate (MeJA)
Polyphagotarsonemus latus
Rhododendron simsii hybrids
RT-qPCR
RNA extraction quality
Salicylic acid (SA)
LATUS ACARI TARSONEMIDAE
TIME RT-PCR
ACID CARBOXYL METHYLTRANSFERASE
LEAF POLYPHENOL OXIDASE
HOT BORATE METHOD
HIGH-QUALITY RNA
JASMONIC-ACID
BROAD MITE
POLYPHAGOTARSONEMUS-LATUS
ARABIDOPSIS-THALIANA
Mite
Nordlicht
Online Access:https://biblio.ugent.be/publication/8533328
http://hdl.handle.net/1854/LU-8533328
https://doi.org/10.1007/s10681-017-2010-5
https://biblio.ugent.be/publication/8533328/file/8533331
Description
Summary:A set of putative marker genes to study plant defense responses against Polyphagotarsonemus latus, a key pest in the production of Rhododendron simsii hybrids, was selected and validated. Genes belonged to the biosynthetic pathway of phytohormones jasmonic acid (JA) (RsLOX, RsAOS, RsAOC, RsOPR3 and RsJMT) and salicylic acid (SA) (RsPAL and RsICS). Furthermore, RsPPO, a putative marker gene for oxidative stress response was successfully cloned from R. simsii. A CTAB-based extraction protocol was optimized to assure excellent RNA quality for subsequent RT-qPCR analysis. The RT-qPCR protocol was extensively tested and RsRG7 and RsRG14 were selected as reference genes from a geNorm pilot study. Validation of the marker genes was done after application with elicitors [methyl jasmonate (MeJA), coronatine, beta-aminobutyric acid and acibenzolar-Smethyl] or wounding. Both 100 mu M MeJA and 0.1 lM coronatine had a significant effect on the expression of all marker genes. Foliar application of MeJA on the shoots resulted in a significantly earlier response when compared to root application and subsequent sampling of the shoots. Expression patterns after MeJA treatment were generally the same in six R. simsii genotypes: 'Nordlicht', 'Elien', 'Aiko Pink', ' Michelle Marie', 'Mevrouw Gerard Kint' and 'Sachsenstern'. Wounding resulted in the same expression patterns as MeJA treatment except for RsJMT. None of the genotypes showed a significant induction of the latter gene 6 h upon wounding. Findings of these experiments indicated that the tolerant genotype 'Elien' has low basal expression levels of RsPPO. This might be the first step towards the breeding of mite-tolerant genotypes.

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