A community resource for high-throughput quantitative RT-PCR analysis of transcription factor gene expression in Medicago truncatula

Kakar, Klementina; Wandrey, Maren; Czechowski, Tomasz; Gaertner, Tanja; Scheible, Wolf‐Rüdiger; Stitt, Mark; Torres‐Jerez, Ivone; Xiao, Yongli; Redman, Julia C.; Wu, Hank; Cheung, Foo; Town, Christopher D.; Udvardi, Michael K.

doi:10.1186/1746-4811-4-18

Cited by 129 publications

(119 citation statements)

References 32 publications

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“…Three to four biological replicates and two technical replicates for each gene of interest were investigated. The cycle threshold values of quantitative PCR were used to normalize the expression levels of target genes with the expression level of the reference gene from M. truncatula (Kakar et al, 2008). The gene expression levels in leaves of AM-and P i -treated plants were compared with the control condition.…”

Section: Quantitative Rt-pcrmentioning

confidence: 99%

Enhanced Secondary- and Hormone Metabolism in Leaves of Arbuscular Mycorrhizal Medicago truncatula

et al. 2017

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Arbuscular mycorrhizas (AM) are the most common symbiotic associations between a plant's root compartment and fungi. They provide nutritional benefit (mostly inorganic phosphate [P i ]), leading to improved growth, and nonnutritional benefits, including defense responses to environmental cues throughout the host plant, which, in return, delivers carbohydrates to the symbiont. However, how transcriptional and metabolic changes occurring in leaves of AM plants differ from those induced by P i fertilization is poorly understood. We investigated systemic changes in the leaves of mycorrhized Medicago truncatula in conditions with no improved P i status and compared them with those induced by high-P i treatment in nonmycorrhized plants. Microarray-based genome-wide profiling indicated up-regulation by mycorrhization of genes involved in flavonoid, terpenoid, jasmonic acid (JA), and abscisic acid (ABA) biosynthesis as well as enhanced expression of MYC2, the master regulator of JA-dependent responses. Accordingly, total anthocyanins and flavonoids increased, and most flavonoid species were enriched in AM leaves. Both the AM and P i treatments corepressed iron homeostasis genes, resulting in lower levels of available iron in leaves. In addition, higher levels of cytokinins were found in leaves of AM-and P i -treated plants, whereas the level of ABA was increased specifically in AM leaves. Foliar treatment of nonmycorrhized plants with either ABA or JA induced the up-regulation of MYC2, but only JA also induced the up-regulation of flavonoid and terpenoid biosynthetic genes. Based on these results, we propose that mycorrhization and P i fertilization share cytokinin-mediated improved shoot growth, whereas enhanced ABA biosynthesis and JA-regulated flavonoid and terpenoid biosynthesis in leaves are specific to mycorrhization.

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Section: Quantitative Rt-pcrmentioning

confidence: 99%

Enhanced Secondary- and Hormone Metabolism in Leaves of Arbuscular Mycorrhizal Medicago truncatula

et al. 2017

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“…The reaction efficiency was 95-100%, as tested using a standard curve for each primer pair. Based on the existing bibliography (Kakar et al 2008;Mantiri et al 2008;Peréz et al 2015) selected 5 candidate reference genes and constructed onsite geNorm and NormFinder evaluation within them (supplement 1). For further analysis used ACTIN2 as reference gene.…”

Section: Quantitative Real-time Pcrmentioning

confidence: 99%

Identification of LEC1, L1L and Polycomb Repressive Complex 2 genes and their expression during the induction phase of Medicago truncatula Gaertn. somatic embryogenesis

Orłowska

Igielski

Łagowska

et al. 2016

Plant Cell Tiss Organ Cult

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However, the callus formation was observed in the nonembryogenic line as well, but the LEC1 and L1L genes were not expressed and the transcription of PRC2 genes was at stable level. It was only the SWN expression that decreased at the beginning of induction and did not change in the subsequent days in both lines. Our results indicate that LEC1 and L1L, known as marker genes of the late developmental stages, may be associated with the acquisition of embryogenic competency by somatic cells (prime events in the SE induction). The PRC2 complex genes are also expressed during embryogenic callus tissue formation on Medicago tuncatula leaf explants.

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“…As is the case for many other crops, useful resistance to R. solani in M. truncatula breeding populations has not yet been found, despite substantial screening efforts (Barbetti et al, 2006). However, we previously identified moderate resistance in A17 (the reference genotype for genome sequencing and the development of genomics tools; Kakar et al, 2008) and strong susceptibility in an ethylene-insensitive mutant of A17, sickle (skl; Penmetsa and Cook, 1997;Penmetsa et al, 2008). Here, we used promoter studies and gene expression profiling to further characterize the moderate resistance in A17.…”

mentioning

confidence: 99%

The B-3 Ethylene Response Factor MtERF1-1 Mediates Resistance to a Subset of Root Pathogens inMedicago truncatulawithout Adversely Affecting Symbiosis with Rhizobia

et al. 2010

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The fungal necrotrophic pathogen Rhizoctonia solani is a significant constraint to a range of crops as diverse as cereals, canola, and legumes. Despite wide-ranging germplasm screens in many of these crops, no strong genetic resistance has been identified, suggesting that alternative strategies to improve resistance are required. In this study, we characterize moderate resistance to R. solani anastomosis group 8 identified in Medicago truncatula. The activity of the ethylene-and jasmonateresponsive GCC box promoter element was associated with moderate resistance, as was the induction of the B-3 subgroup of ethylene response transcription factors (ERFs). Genes of the B-1 subgroup showed no significant response to R. solani infection. Overexpression of a B-3 ERF, MtERF1-1, in Medicago roots increased resistance to R. solani as well as an oomycete root pathogen, Phytophthora medicaginis, but not root knot nematode. These results indicate that targeting specific regulators of ethylene defense may enhance resistance to an important subset of root pathogens. We also demonstrate that overexpression of MtERF1-1 enhances disease resistance without apparent impact on nodulation in the A17 background, while overexpression in sickle reduced the hypernodulation phenotype. This suggests that under normal regulation of nodulation, enhanced resistance to root diseases can be uncoupled from symbiotic plant-microbe interactions in the same tissue and that ethylene/ERF regulation of nodule number is distinct from the defenses regulated by B-3 ERFs. Furthermore, unlike the stunted phenotype previously described for Arabidopsis (Arabidopsis thaliana) ubiquitously overexpressing B-3 ERFs, overexpression of MtERF1-1 in M. truncatula roots did not show adverse effects on plant development.

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A community resource for high-throughput quantitative RT-PCR analysis of transcription factor gene expression in Medicago truncatula

Cited by 129 publications

References 32 publications

Enhanced Secondary- and Hormone Metabolism in Leaves of Arbuscular Mycorrhizal Medicago truncatula

Enhanced Secondary- and Hormone Metabolism in Leaves of Arbuscular Mycorrhizal Medicago truncatula

Identification of LEC1, L1L and Polycomb Repressive Complex 2 genes and their expression during the induction phase of Medicago truncatula Gaertn. somatic embryogenesis

The B-3 Ethylene Response Factor MtERF1-1 Mediates Resistance to a Subset of Root Pathogens inMedicago truncatulawithout Adversely Affecting Symbiosis with Rhizobia

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