Helia Guerra Cardoso

Universidade de Évora - ICAAM (Portugal) · Laboratório Biologia Molecular

Citation: Campos, C.; Coito, J.L.; Cardoso, H.; Marques da Silva, J.; Pereira, H.S.; Viegas, W.; Nogales, A. Dynamic Regulation of Grapevine's microRNAs in Response to Mycorrhizal Symbiosis and High Temperature. Plants 2023, 12, 982. Abstract: MicroRNAs (miRNAs) are non-coding small RNAs that play crucial roles in plant development and stress respo...

In the last few years, the traceability and labeling of processed food and feeds have gained increasing importance due to the impact that mislabeling and product fraud may have on human/animal health or on the quality of final products, such as milk, cheese, and meat, as a consequence of animal dietary. The presence of contaminants or possible frau...

Organic agriculture, recognized as a more sustainable agricultural system, strongly depends on the use of highly resilient genotypes. Seed resilience, with increased tolerance to germination that provide vigorous seedlings under environmental stresses, currently represents one of the most important agronomical traits. Seed germination involves the...

The PIN-FORMED (PIN) proteins represent the most important polar auxin transporters in plants. Here, we characterized the PIN gene family in two olive genotypes, the Olea europaea subsp. europaea var. sylvestris and the var. europaea (cv. ‘Farga’). Twelve and 17 PIN genes were identified for vars. sylvestris and europaea, respectively, being distri...

Organic agriculture, recognized as a more sustainable agricultural system, strongly de-pends on the use of highly resilient genotypes. Resilient seeds, with increased tolerance to germinate and provide vigorous seedlings under environmental stresses, currently represent one of the most important agronomical traits. Seed germination involves the act...

Organic agriculture, recognized as a more sustainable agricultural system, strongly depends on the use of highly resilient genotypes. Resilient seeds, with increased tolerance to germinate and provide vigorous seedlings under environmental stresses, currently represent one of the most important agronomical traits. Seed germination involves the acti...

The availability of phenotyping tools to assist breeding programs in the selection of high-quality crop seeds is of obvious interest with consequences for both seed producers and consumers. Seed germination involves the activation of several metabolic pathways, such as cellular respiration to provide the required ATP and reducing power. This work t...

In vitro micropropagation is already a current multiplication tool for walnut self-rooted cultivars and rootstocks, but walnut grafting is still performed in the field or in greenhouses, mainly using seedlings as rootstocks. The present work describes a new approach to obtain clonal walnut-grafted plants, involving in vitro shoot production of ‘Par...

Alternative oxidase (AOX) is a key enzyme of the alternative respiration, known to be involved in plant development and in response to various stresses. To verify the role of DcAOX1 and DcAOX2a genes in carrot tap root growth and in response to cold stress, their expression was analyzed in two experiments: during root growth for 13 weeks and in res...

The author wishes to make an erratum to the published version of the paper [...]

Only a small portion of the stem cells participate in the process of adventitious root formation and the cells/tissues types involved in this process is species-dependent. In olive, it is still unclear which type of cells acquire competence for rooting. Regardless, the entire stem nodal segment (containing a mixture of distinct cell types) continue...

Heat stress negatively affects several physiological and biochemical processes in grapevine plants. In this work, two new methods, calorespirometry, which has been used to determine temperature adaptation in plants, and near-infrared (NIR) spectroscopy, which has been used to determine several grapevine-related traits and to discriminate among vari...

Several olive cultivars, characterized by high-quality olive oil show agronomical issues such as excessive vigor, high susceptibility to biotic and abiotic stresses, and low propagation ability. They are strong candidates for breeding based on new technologies to improve their performance in a short period of time. For this reason, the first step i...

Exogenous auxins supplementation plays a central role in the formation of adventitious roots (AR) for several plant species. However, the molecular mechanisms underlying the process of adventitious rooting are still not completely understood and many plants with economic value, including several olive cultivars, exhibit a recalcitrant behavior towa...

In the last decades, the interest on traditional Mediterranean fruits highly increased, not only due to the constant demand of consumers for new crop alternatives but also due to the identification in such species of molecules with important properties for human health (e.g., resveratrol from grapes and oleuropein from olives). Efforts to improve t...

Os trabalhos de transformação genética estão fortemente dependentes de protocolos eficientes de embriogénese somática, única forma de garantir a origem unicelular das plantas geneticamente modificadas. A Universidade de Évora tem em curso um trabalho de identificação dos genes relacionados com o enraizamento adventício em oliveira, sendo expectável...

Abstract: Propagation of some Olea europaea L. cultivars is strongly limited due to recalcitrant behavior in adventitious root formation by semi-hardwood cuttings. One example is the cultivar ”Galega vulgar”. The formation of adventitious roots is considered a morphological response to stress. Alternative oxidase (AOX) is the terminal oxidase of th...

Traceability of processed food and feed products has been gaining importance due to the impact that those products can have on human/animal health and to the associated economic and legal concerns, often related to adulterations and frauds as it can be the case for meat and milk. Despite mandatory traceability requirements for the analysis of feed...

Amplification plots from dilution series of gDNA from the selected plant species. gDNA dilutions ranged from final concentration of 0.04 ng to 80 ng, in a total of nine data points. The plant species were: maize, barley, soybean, wheat, flax, sunflower, peanut, rice and cotton. (PDF)

Sensitivity and linearity of the TaqMan qPCR assays. Serial dilutions of gDNA extracted from sunflower, rice, soybean, cotton, barley, flax, wheat, maize and peanut were tested by using a species-specific TaqMan assay. Data are expressed as the absolute target copy number (TCN) versus gDNA amount. Each data point is the mean of two technical replic...

List of the 31 species. (DOCX)

Results from end-point methodology applied to 7 feed samples (X8, X10, X13, X32, X42, Z2 and Z3) to identify the presence/absence of 9 plant species. The plant species were wheat, maize, soybean, barley, sunflower, cotton, flax, rice and peanut. (PDF)

Amplification plots to assess primers/probe specificity. Cross-reactivity in gDNA of 31 plant species, bulked into seven pools were performed. The nine plant species selected to design primers and probes were: maize, barley, soybean, wheat, flax, sunflower, peanut, rice and cotton. (PDF)

Composition of the pools used for cross reactivity tests. (DOCX)

“An alternative for olive cultivars recalcitrant to vegetative propagation is micropropagation through adventitious rooting, but the underlying molecular mechanisms of this process are still not well understood. Previous studies support that olive in vitro root induction is triggered by a strong and short oxidative shock. This shock may be enhanced...

A consortium of European enterprises and research institutions has been engaged in the Feed-Code Project withthe aim of addressing the requirements stated in European Union Regulation No. 767/2009, concerning market placement and use of feed of known and ascertained botanical composition. Accordingly, an interlaboratory trial was set up to compare...

In traditional plant breeding, selected elite breeding lines play an important role as a source for building-up and improving breeding populations. Currently, the focus for using wild relatives together with landraces in breeding programs is gaining more attention in the context of global climate changes. Differences in plant growth adaptation unde...

The plastid terminal oxidase (PTOX) gene is known to be implicated in the carotenoid biosynthesis pathway. In the present work we characterize the Daucus carota L. PTOX gene (DcPTOX) and describe the relation between DcPTOX transcript level and carotenoids accumulation during storage root development of white, yellow, orange, red, and purple carrot...

Alternative oxidase (AOX) protein is located in the inner mitochondrial membrane and is encoded in the nuclear genome being involved in plant response upon a diversity of environmental stresses and also in normal plant growth and development. Here we report the characterization of the AOX gene family of Hypericum perforatum L. Two AOX genes were id...

The nuclear-encoded plastid terminal oxidase gene (PTOX) is present in photosynthetic species and functions in the oxidation of the plastoquinone pool. It plays an important role on chlororespiration, chromorespiration and carotenoid biosynthesis. Here, we show short-term early response of carrot PTOX (DcPTOX) in leaves of two carrot inbred lines u...

Stress-adaptive cell plasticity in target tissues and cells for plant biomass growth is important for yield stability. In vitro systems with reproducible cell plasticity can help to identify relevant metabolic and molecular events during early cell reprogramming. In carrot, regulation of the central root meristem is a critical target for yield-dete...

(A) Growth curve of primary cultures system of D. carota L. cv. Rotin during 28 days in culture at 21°C in four individual plants. Data are shown as callus FW values and represented as mean ± SD. (B) Explants from the secondary phloem of carrot tap roots PCS. Aspect of the explants (1) before (T0), (2) 14 days, and (3) 28 days after in vitro inocul...

This review highlights a four-step rational for the hypothesis that mitochondria play an upstream central role for stress-induced somatic embryogenesis (SE): (1) Initiation of SE is linked to programmed cell death (PCD) (2) Mitochondria are crucially connected to cell death (3) SE is challenged by stress per se (4) Mitochondria are centrally linked...

Arbuscular mycorrhizal fungi (AMF) are root-inhabiting fungi that form mutualistic symbioses with their host plants. AMF symbiosis improves nutrient uptake and buffers the plant against a diversity of stresses. Rhizophagus irregularis is one of the most widespread AMF species in the world, and its application in agricultural systems for yield impro...

Carrot (Daucus carota L.) is a biennial plant that accumulates considerable amounts of carotenoid pigments in the storage root. To better understand the molecular mechanisms for carotenoid accumulation in developing storage roots, plastid terminal oxidase (PTOX) cDNA was isolated and selected for reverse-transcription quantitative polymerase chain...

The alternative oxidase (AOX) is a key enzyme of the alternative respiration pathway and is involved in plant response to a variety of environmental stresses. The genes encoding for this enzyme are highly polymorphic and thus likely good candidates for molecular marker development to assist plant breeding on stress tolerance. In carrot, DcAOX1 gene...

The development of new ‘deep phenotyping’ techniques for functional markers (FM) development on alternative oxisase (AOX) gene sequences are expected to greatly increase the efficiency of association studies between the candidate FM sequences and the desired phenotype. However, it is critical to perform these studies in the appropriate target tissu...

In angiosperms alternative oxidase (AOX) has been described as a small multigene family composed of three to five genes distributed in two different subfamilies, AOX1 and AOX2. Nevertheless, while AOX1 occurs in both monocots and eudicots, AOX2 occurs only in eudicots. Neimanis et al. recently suggested that the lack of AOX2 in monocots is due to a...

In angiosperms AOX has been described as a small multigene family composed of three to five genes distributed in two different subfamilies, AOX1 and AOX2 (Whelan, Millar and Day, 1996; Saisho et al., 1997; Vanlerberghe, 2013). Nevertheless, while AOX1 occurs in both monocots and eudicots, AOX2 occurs only in eudicots. Neimanis et al. (2013) recentl...

Plants live with a myriad of biotic and abiotic interactions with the soil and environment that determine their growth, productivity and life cycle, whereas the rhizosphere plays an interface role (Jeffries, Gianinazzi and Perotto, 2003). Arbuscular mycorrhizal fungi (AMF) establish the predominant mutualistic symbiotic relationship within the root...

Reverse transcription quantitative real time polymerase chain reaction (RT-qPCR) is a preferred method for rapid and accurate quantification of gene expression studies. Appropriate application of RT-qPCR requires accurate normalization through the use of suitable reference genes. This study aimed selecting robust and reliable reference genes which...

Reverse transcription-quantitative real-time PCR (RT-qPCR) is a widely used technique for gene expression analysis. The reliability of this method depends largely on the suitable selection of stable reference genes for accurate data normalization. Hypericum perforatum L. (St. John's wort) is a field growing plant that is frequently exposed to a var...

Functional marker (FM) development across plant species requires common allelic sequences that affect phenotypic variation in the same way. The identification of orthologous genes showing the same function across species could indicate conservation of functional polymorphisms. In this chapter we provide an overview over polymorphic sites described...

Key message The expression of carrot antifreeze protein enhanced chilling tolerance in heterologous host system tomato and AFP can be a potential gene candidate for producing chilling tolerant crop plants. Abstract In an attempt to improve chilling tolerance, the carrot gene encoding the antifreeze protein (AFP) was cloned under the control of cons...

Unlabelled: Alternative oxidase (AOX) has been proposed as a functional marker candidate in a number of events involving cell differentiation, including rooting efficiency in semi-hardwood shoot cuttings of olive (Olea europaea L.). To ascertain the general importance of AOX in olive rooting, the auxin-induced rooting process was studied in an in...

'Trincadeira' and 'Aragonez' are two of the most important Vitis vinifera L. cultivars used for red wine production in Portugal. Aiming to obtain a reproducible system for plant genetic transformation, the techniques of particle bombardment and transformation mediated by Agrobacterium tumefaciens were tested. In both systems putative transformed em...

Methyl-jasmonate (MJ)-treated hairy roots of Daucus carota L. were used to study the influence of alternative oxidase (AOX) in phenylpropanoid metabolism. Phenolic acid accumulation, as well as total flavonoids and lignin content of the MJ-treated hairy roots were decreased by treatment with salicylhydroxamic acid (SHAM), a known inhibitor of AOX....

Alternative oxidase (AOX) has been proposed as a promising functional marker candidate for multiple plant stress behaviour. The present paper describes natural polymorphism in AOX2b of Daucus carota L. (DcAOX2b). Exon-primed intron crossing-PCR (EPIC-PCR) revealed length variation (intron length polymorphisms, ILPs) in intron 1. Six fragment patter...

Olive (Olea europaea L.) trees are mainly propagated by adventitious rooting of semi-hardwood cuttings. However, efficient commercial propagation of valuable olive tree cultivars or landraces by semi-hardwood cuttings can often be restricted by a low rooting capacity.We hypothesize that root induction is a plant cell reaction linked to oxidative st...

Single nucleotide polymorphisms (SNPs) and insertion-deletions (InDels) are becoming important genetic markers for major crop species. In this study, we focus on variations at genomic level of the Daucus carota L. AOX2a gene. The use of gene-specific primers designed in exon regions on the boundaries of introns permitted to recognize intron length...

'Genomic design' refers to the structural organization of gene sequences. Recently, the role of intron sequences for gene regulation is being better understood. Further, introns possess high rates of polymorphism that are considered as the major source for speciation. In molecular breeding, the length of gene-specific introns is recognized as a too...

The present paper deals with the analysis of natural polymorphism in a selected alternative oxidase (AOX) gene of the medicinal plant, St John's wort. Four partial AOX gene sequences were isolated from the genomic DNA of a wild plant of Hypericum perforatum L. Three genes belong to the subfamily AOX1 (HpAOX1a, b and c) and one to the subfamily AOX2...

The gymnosperm Pinus pinea L. (stone pine) is a typical Mediterranean pine used for nuts and timber production, and as an ornamental around the world. Pine genomes are large in comparison to other species. The hypothesis that retrotransposons, such as gymny, made a large contribution to this alteration in genome size was recently confirmed. However...

Alternative oxidase (AOX) is a mitochondrial protein encoded by the nuclear genome. In higher plants AOX genes form a small multigene family mostly consisting of the two subfamilies AOX1 and AOX2. Daucus carota L. is characterized by a unique extension pattern of AOX genes. Different from other plant species studied so far it contains two genes in...

Plant alternative oxidase (AOX) is a mitochondrial inner membrane enzyme involved in alternative respiration. The critical importance of the enzyme during acclimation upon stress of plant cells is not fully understood and is still an issue of intensive research and discussion. Recently, a role of AOX was suggested for the ability of plant cells to...

The paper highlights Daucus carota L. as an ideal model to complement plant stress research on Arabidopsis thaliana L. Recently, alternative oxidase (AOX) is discussed as functional marker candidate for cell reprogramming upon stress. Carrot is the most studied species for cell reprogramming and our current research reveals that it is the only one...

The paper highlights Daucus carota L. as an ideal model to complement plant stress research on Arabidopsis thaliana L. Recently, alternative oxidase (AOX) is discussed as functional marker candidate for cell reprogramming upon stress. Carrot is the most studied species for cell reprogramming and our current research reveals that it is the only one...

With the aim to reduce fees of a micropropagation protocol for the Portuguese olive cultivar ‘Galega vulgar’, expensive Difco Bacto-Agar® was replaced by commercial agar-agar, and the analysed data revealed no significant differences between the two agars, for both the explants multiplication rate and the rooting capacity. The replacement of the OM...

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