Gema Ancillo - Academia.edu (original) (raw)

Papers by Gema Ancillo

XII International Citrus Congress - International Society of Citriculture, 2015

Background / Purpose: It is accepted that four basic taxa (Citrus medica, Citrus maxima, Citrus r... more Background / Purpose: It is accepted that four basic taxa (Citrus medica, Citrus maxima, Citrus reticulata and Citrus micrantha) have generated all cultivated Citrus species. The genomes of most of the modern Citrus cultivars, vegetatively propagated, are reported to be an interspecific mosaic of large DNA fragments issued from a limited number of inter-specific meiosis. Our main objective is to know how multilocus study of closely linked SNPs allows a phylogenetic assignation of DNA fragments of the main cultivated species. Main conclusion: Phylogenetic origin of specific DNA fragments can be assigned from multilocus analysis of closely linked SNPs.

PLOS ONE, 2015

Most cultivated Citrus species originated from interspecific hybridisation between four ancestral... more Most cultivated Citrus species originated from interspecific hybridisation between four ancestral taxa (C. reticulata, C. maxima, C. medica, and C. micrantha) with limited further interspecific recombination due to vegetative propagation. This evolution resulted in admixture genomes with frequent interspecific heterozygosity. Moreover, a major part of the phenotypic diversity of edible citrus results from the initial differentiation between these taxa. Deciphering the phylogenomic structure of citrus germplasm is therefore essential for an efficient utilization of citrus biodiversity in breeding schemes. The objective of this work was to develop a set of species-diagnostic single nucleotide polymorphism (SNP) markers for the four Citrus ancestral taxa covering the nine chromosomes, and to use these markers to infer the phylogenomic structure of secondary species and modern cultivars. Species-diagnostic SNPs were mined from 454 amplicon sequencing of 57 gene fragments from 26 genotypes of the four basic taxa. Of the 1,053 SNPs mined from 28,507 kb sequence, 273 were found to be highly diagnostic for a single basic taxon. Species-diagnostic SNP markers (105) were used to analyse the admixture structure of varieties and rootstocks. This revealed C. maxima introgressions in most of the old and in all recent selections of mandarins, and suggested that C. reticulata × C. maxima reticulation and introgression processes were important in edible mandarin domestication. The large range of phylogenomic constitutions between C. reticulata and C. maxima revealed in mandarins, tangelos, tangors, sweet oranges, sour oranges, grapefruits, and orangelos is favourable for genetic association studies based on phylogenomic structures of the germplasm. Inferred admixture structures were in agreement with previous hypotheses regarding the origin of several PLOS ONE |

Plant molecular biology, 1999

We have isolated cDNA clones encoding class I chitinase (ChtC) from potato leaves which share a h... more We have isolated cDNA clones encoding class I chitinase (ChtC) from potato leaves which share a high degree of nucleotide and amino acid sequence similarity to other, previously described basic (class I) chitinases (ChtB) from potato. Despite this similarity, characteristic features distinguish ChtC from ChtB, including an extended proline-rich linker region between the hevein and catalytic domains and presence of a potential glycosylation site (NDT) in the deduced protein. These differences are in accordance with the properties of purified chitinase C which is glycosylated and hence has a higher molecular mass in comparison to chitinase B. In contrast to the coding sequences, the 3'-untranslated regions of ChtC and ChtB exhibited a low degree of similarity, which allowed us to generate gene-specific probes to study the genomic organization and expression of both types of gene. Genomic DNA blots suggest that ChtC and ChtB are each encoded by one or two genes per haploid genome. ...

Applications in Plant Sciences, 2013

Annals of Botany, 2012

Aims Despite differences in morphology, the genera representing 'true citrus fruit trees' are sex... more Aims Despite differences in morphology, the genera representing 'true citrus fruit trees' are sexually compatible, and their phylogenetic relationships remain unclear. Most of the important commercial 'species' of Citrus are believed to be of interspecific origin. By studying polymorphisms of 27 nuclear genes, the average molecular differentiation between species was estimated and some phylogenetic relationships between 'true citrus fruit trees' were clarified. † Methods Sanger sequencing of PCR-amplified fragments from 18 genes involved in metabolite biosynthesis pathways and nine putative genes for salt tolerance was performed for 45 genotypes of Citrus and relatives of Citrus to mine single nucleotide polymorphisms (SNPs) and indel polymorphisms. Fifty nuclear simple sequence repeats (SSRs) were also analysed. † Key Results A total of 16 238 kb of DNA was sequenced for each genotype, and 1097 single nucleotide polymorphisms (SNPs) and 50 indels were identified. These polymorphisms were more valuable than SSRs for intertaxon differentiation. Nuclear phylogenetic analysis revealed that Citrus reticulata and Fortunella form a cluster that is differentiated from the clade that includes three other basic taxa of cultivated citrus (C. maxima, C. medica and C. micrantha). These results confirm the taxonomic subdivision between the subgenera Metacitrus and Archicitrus. A few genes displayed positive selection patterns within or between species, but most of them displayed neutral patterns. The phylogenetic inheritance patterns of the analysed genes were inferred for commercial Citrus spp. † Conclusions Numerous molecular polymorphisms (SNPs and indels), which are potentially useful for the analysis of interspecific genetic structures, have been identified. The nuclear phylogenetic network for Citrus and its sexually compatible relatives was consistent with the geographical origins of these genera. The positive selection observed for a few genes will help further works to analyse the molecular basis of the variability of the associated traits. This study presents new insights into the origin of C. sinensis.

BMC genetics, Jan 29, 2014

BackgroundThe most economically important Citrus species originated by natural interspecific hybr... more BackgroundThe most economically important Citrus species originated by natural interspecific hybridization between four ancestral taxa (Citrus reticulata, Citrus maxima, Citrus medica, and Citrus micrantha) and from limited subsequent interspecific recombination as a result of apomixis and vegetative propagation. Such reticulate evolution coupled with vegetative propagation results in mosaic genomes with large chromosome fragments from the basic taxa in frequent interspecific heterozygosity. Modern breeding of these species is hampered by their complex heterozygous genomic structures that determine species phenotype and are broken by sexual hybridisation. Nevertheless, a large amount of diversity is present in the citrus gene pool, and breeding to allow inclusion of desirable traits is of paramount importance. However, the efficient mobilization of citrus biodiversity in innovative breeding schemes requires previous understanding of Citrus origins and genomic structures. Haplotyping...

Virology, 2007

Changes in gene expression of Mexican lime plants in response to infection with a severe (T305) o... more Changes in gene expression of Mexican lime plants in response to infection with a severe (T305) or a mild (T385) isolate of Citrus tristeza virus (CTV) were analyzed using a cDNA microarray containing 12,672 probes to 6875 different citrus genes. Statistically significant (P b 0.01) expression changes of 334 genes were detected in response to infection with isolate T305, whereas infection with T385 induced no significant change. Induced genes included 145 without significant similarity with known sequences and 189 that were classified in seven functional categories. Genes related with response to stress and defense were the main category and included 28% of the genes induced. Selected transcription changes detected by microarray analysis were confirmed by quantitative real-time RT-PCR. Changes detected in the transcriptome upon infecting lime with T305 may be associated either with symptom expression, with a strain-specific defense mechanism, or with a general response to stress.

Planta, 2003

Chitinases are ubiquitous proteins that occur in all plants in multiple isoforms. We have isolate... more Chitinases are ubiquitous proteins that occur in all plants in multiple isoforms. We have isolated the ChtC2 gene encoding an unusual, basic (class I) chitinase from potato ( Solanum tuberosum L.). In contrast to other chitinase genes, ChtC2 is not activated by infection, but rather constitutively expressed in leaves and stems where it is restricted to epidermal cells. Sequence analysis revealed a number of potential regulatory elements in the promoter, but most striking was the presence of a 319-bp direct repeat located between -333 and -968 upstream of the transcription start site. For a functional analysis, a 1,322-bp promoter fragment and two 5&amp;#39; deletions of 782 bp and 162 bp in length were translationally fused to the beta-glucuronidase (GUS) reporter gene and used for transient expression studies by particle bombardment. All promoter constructs conferred expression of GUS activity in different epidermal cell types of potato leaves. Expression in parenchyma cells of the leaf mesophyll was not detectable with any of the ChtC2 gene promoter constructs, in contrast to the pattern observed with the 35S promoter from cauliflower mosaic virus. The epidermis-specific expression of the reporter gene was confirmed using transgenic potato plants containing the fusion of the entire ChtC2 promoter with the GUS reporter. Histochemical analysis indicated that the promoter was only active in epidermal cells of leaves.

Plant Physiology and Biochemistry, 2011

In this work, we studied the sequence of responses induced by flooding in citrus plants, with the... more In this work, we studied the sequence of responses induced by flooding in citrus plants, with the aim of identifying the signals that lead to stomatal closure. One-year-old seedlings of Carrizo citrange, grown in sand under greenhouse conditions, were waterlogged for 35 d and compared with normally watered well-drained plants. Significant decreases in stomatal conductance and transpiration were detected between flooded and control seedlings from a week after the beginning of the experiment. However ABA concentration in leaves only started to increase after three weeks of flooding, suggesting that stomata closed in the absence of a rise in foliar ABA. Therefore, stomatal closure in waterlogged seedlings does not appear to be induced by ABA, at least during the early stages of flood-stress. The low levels of ABA detected in roots and xylem sap from flooded seedlings indicated that it is very unlikely that the ABA increase in the leaves of these plants is due to ABA translocation from roots to shoots. We propose that ABA is produced in old leaves and transported to younger leaves. Flooding had no effect on water potential or the relative water content of leaves.

Plant Cell Reports, 2011

The production of cybrids, combining nucleus of a species with alien cytoplasmic organelles, is a... more The production of cybrids, combining nucleus of a species with alien cytoplasmic organelles, is a valuable method used for improvement of various crops. Several citrus cybrids have been created by somatic hybridization. These genotypes are interesting models to analyze the impact of cytoplasmic genome change on nuclear genome expression. Herein, we report genome-wide gene expression analysis in leaves of a citrus cybrid between C. reticulata cv 'Willowleaf mandarin' and C. limon cv 'Eureka lemon' compared with its lemon parent, using a Citrus 20K cDNA microarray. Molecular analysis showed that this cybrid possesses nuclear and chloroplast genomes of Eureka lemon plus mitochondria from Willowleaf mandarin and, therefore, can be considered as a lemon bearing foreign mitochondria. Mandarin mitochondria influenced the expression of a large set of lemon nuclear genes causing an over-expression of 480 of them and repression of 39 genes. Quantitative real-time RT-PCR further confirmed the credibility of microarray data. Genes over-expressed in cybrid leaves are predominantly attributed to the functional category ''cellular protein metabolism'' whereas in the down-regulated none functional category was enriched. Overall, mitochondria replacement affected different nuclear genes including particularly genes predicted to be involved in mitochondrial retrograde signaling. Mitochondria regulate all cell structures even chloroplast status. These results suggest that nuclear gene expression is modulated with respect to new information received from the foreign organelle, with the final objective to suit specific needs to ensure better cell physiological balance.

Plant Cell Reports, 2009

Allopolyploidy is known to induce novel patterns of gene expression and often gives rise to new p... more Allopolyploidy is known to induce novel patterns of gene expression and often gives rise to new phenotypes. Here we report on the first attempt to relate phenotypic inheritance in an allotetraploid somatic hybrid with gene expression. Carotenoid compounds in the fruit pulp of the two parental species and the hybrid were evaluated quantitatively by HPLC. Only very low levels of b-carotene and b-cryptoxanthin were observed in Citrus limon, while b-cryptoxanthin was a major component of C. reticulata, which also displayed high levels of phytoene, phytofluene, b-carotene, lutein, zeaxantin and violaxanthin. Total carotenoid content in mandarin juice sacs was 60 times greater than that in lemon. The allotetraploid hybrid produced all the same compounds as mandarin but at very low levels. Transgressive concentration of abscisic acid (ABA) was observed in the somatic hybrid. Real-time RT-PCR of total RNA from juice sacs was used to study expression of seven genes (CitDxs, CitPsy, CitPds, CitZds, CitLcy-b, CitChx-b, and CitZep) of the carotenoid biosynthetic pathway and two genes (CitNced1 and CitNced2) involved in abscisic acid synthesis from carotenoid. Gene expression was significantly higher for mandarin than lemon for seven of the nine genes analyzed. Lemon under expression was partially dominant in the somatic hybrid for three upstream steps of the biosynthetic pathway, particularly for CitDxs. Transgressive over expression was observed for the two CitNced genes. A limitation of the upstream steps of the pathway and a downstream higher consumption of carotenoids may explain the phenotype of the somatic hybrid.

Plant and Soil, 2011

Water deficit (WD) is a growing problem in agriculture. In citrus crops, genetically-determined c... more Water deficit (WD) is a growing problem in agriculture. In citrus crops, genetically-determined characteristics of the rootstock are important factors in plant responses to WD. Aquaporins are involved in regulating the water supply to the plant by mediating water flow through the cell membranes. Recent studies support a direct role for aquaporins in plant water relations and demonstrate their involvement in tolerance to WD. This study investigates the relationship between photosynthetic and water-balance parameters with levels of expression of aquaporins in conditions of moderate WD in the rootstocks Poncirus trifoliata (L.) Raf. (PT), Cleopatra Mandarin (Citrus reshni Hort. ex Tan.) (CM) and 030115 (a hybrid of the two former rootstocks). Under conditions of WD, the hybrid 030115 drastically reduced aquaporin expression, accompanied by a loss of plant vigour but without reducing the net CO2 assimilation (ACO2). PT maintained the same level of aquaporin expression under WD as under normal irrigation conditions, but suffered a sharp reduction in ACO2. CM, which has lower expression of aquaporins than PT under both normal irrigation conditions and WD, responded better to water stress conditions than PT. Thus, low levels of aquaporins, or repression of their expression, accompanied by decreased plant vigour resulted in a decrease in plasma membrane permeability, thereby facilitating water retention in the cells under conditions of water stress. This can induce water stress tolerance in citrus rootstocks.

Molecular Plant Pathology, 2012

Viroids are small (246-401 nucleotides), single-stranded, circular RNA molecules that infect seve... more Viroids are small (246-401 nucleotides), single-stranded, circular RNA molecules that infect several crop plants and can cause diseases of economic importance. Citrus are the hosts in which the largest number of viroids have been identified. Citrus exocortis viroid (CEVd), the causal agent of citrus exocortis disease, induces considerable losses in citrus crops. Changes in the gene expression profile during the early (pre-symptomatic) and late (postsymptomatic) stages of Etrog citron infected with CEVd were investigated using a citrus cDNA microarray. MaSigPro analysis was performed and, on the basis of gene expression profiles as a function of the time after infection, the differentially expressed genes were classified into five clusters. FatiScan analysis revealed significant enrichment of functional categories for each cluster, indicating that viroid infection triggers important changes in chloroplast, cell wall, peroxidase and symporter activities.

Journal of Experimental Botany, 2007

In recent years, class prediction experiments have been largely developed in cancer research with... more In recent years, class prediction experiments have been largely developed in cancer research with the aim of classifying unknown samples by examining their expression signature. In natural populations, a significant component of gene expression variability is also heritable. Citrus species are an ideal model to accomplish the study of these questions in plants, due to the existence of varieties derived from somatic mutations that are likely to differ from each other by one or a few point mutations but are phenotypically indistinguishable at early vegetative stages. The small genetic variability existing among these varieties makes molecular markers ineffective in distinguishing genotypes within a particular species. Gene expression profiles have been used to predict mandarin clementine varieties (Citrus clementina Hort. ex Tan.) by means of two independent supervised learning algorithms: Support Vector Machines and Prediction Analysis of Microarrays. The results show that transcriptional variation is variety-dependent in citrus, and supervised clustering methods may correctly assign blind samples to varieties when both training and test samples are under the same experimental conditions.

Journal of Experimental Botany, 2010

Differential gene expression analysis provides new insights into the molecular basis of iron defi... more Differential gene expression analysis provides new insights into the molecular basis of iron deficiency stress response in the citrus rootstock Poncirus trifoliata (L.) Raf.