Biology subjects

Garcia-Gil, M. R.

Publications and source records attributed to Garcia-Gil, M. R..

13 recordsLinked to original sources

bioRxiv2026

Uncovering bioactive metabolites from the Taxus wallichiana endophyte Annulohypoxylon purpureonitens using reverse metabolomics

Endophytic fungi associated with the Himalayan yew (Taxus wallichiana) represent an underexplored source of bioactive secondary metabolites. This study investigated the extracellular metabolites of Annulohypoxylon purpureonitens isolated from Nepalese T. wallichiana using bioactivity screening combined with LCMS/MS-based metabolomics. The fungal extract exhibited broad-spectrum antibacterial activity, showing the strongest inhibition against Staphylococcus aureusand Enterococcus faecalis (MIC = 500 ug/mL). It also displayed notable antioxidant capacity(DPPH, ABTS, TPC &TFC) and cytotoxicity against HeLa and MCF-7 cancer cell lines. Metabolite profiling via GNPS molecular networking, manual MS/MS validation, and MASST reverse metabolomics putatively identified diverse compounds, including hydroquinidine, chlorogenic acid, muramic acid, and cordycepin conjugates widely distributed across public microbial datasets. Overall, A. purpureonitens is a promising source of multifunctional metabolites, laying a foundation for future compound isolation and functional characterization.

pharmacology and toxicology↗

bioRxiv2024

In silico characterisation of PAL homologs and metabolomic profiling of shade response indicates potential presence of PTALs and Tyr as a probable precursor of lignin biosynthesis in conifers

Norway spruce and Scots pine show enhanced lignin synthesis under shade along with differential expression of defense-related genes that renders disease resilience. In general, phenylalanine (Phe) is the precursor for lignin synthesis in plants and tyrosine (Tyr) forms an additional lignin precursor in grasses. Phenylalanine ammonia-lyase (PAL) and tyrosine ammonia-lyase (TAL) from lignin biosynthesis pathway use either Phe or Tyr as precursors for lignin production, respectively. Grasses possess bifunctional phenylalanine/tyrosine ammonia-lyase (PTAL) that potentially can use both Phe and Tyr for lignin biosynthesis. Metabolomic profiles of seedlings revealed a relatively higher amount of Phe and Tyr under shade in Scots pine, while Norway spruce showed differential regulation of only Tyr under shade. Sequence analysis and phylogeny of PAL homologs in the two conifers coupled with correlation of up-regulation of precursors for lignin synthesis (Phe/Tyr) and enhanced lignin synthesis along with differential expression of PAL homologs under shade, suggests potential presence of PTALs in conifers, which is novel. Exome sequence analysis revealed latitudinal variation in allele frequencies of SNPs from coding regions of putative PAL and PTAL in Norway spruce, which may impact enzyme activity affecting lignin synthesis. Metabolomic analysis additionally identified metabolites involved in plant immunity, defense and stress response.

plant biology↗

bioRxiv2023

Breeding for growth has resulted in increased tolerance to drought at the cost of genetic diversity in Scots pine.

Climate change possess a threat to forests and forestry. Drought has been identified as a one of the main issues due to its interaction with other biotic and abiotic stresses. Few studies have been done regarding breeding effect on the adaptability to climate change. After a common garden experiment with seedling families of Scots pine from northern Sweden, we have found differences in drought tolerance between families of breeding and natural origin. We performed a high throughput analysis-based phenotyping on both canopy and root traits. Root architecture traits might be related to drought tolerance and show moderate to high heritability values. The heritability of root architecture traits can be useful not only for drought but also for adaptability to other abiotic stresses. Analysis on architecture traits show that, especially on canopy-traits, families from breeding origins show less phenotypic variance than the ones from natural origins. The methodology employed to evaluate drought tolerance and plant architecture might be useful for future research and forest management focused on climate change adaptability.

physiology↗

bioRxiv2022

QTL mapping of the narrow-branch "Pendula" phenotype in Norway spruce (Picea abies L. Karst.)

Pendula-phenotyped Norway spruce has a potential forestry interest for high density plantations. This phenotype is believed to be caused by a dominant single mutation. Despite the availability of RAPD markers linked to the trait, the nature of the mutation is yet unknown. We performed a Quantitative Trait Loci (QTL) mapping based on two different progenies of F1 crosses between pendula and normal crowned trees using NGS technologies. Approximately 25 % of all gene bearing scaffolds of Picea abies genome assembly v1.0 were mapped to 12 linkage groups and a single QTL, positioned near the center of LG VI, was found in both crosses. The closest probe-markers placed on the maps were positioned 0.82 cM and 0.48 cM away from the Pendula marker in two independent pendula-crowned x normal-crowned wildtype crosses, respectively. We have identified genes close to the QTL region with differential mutations on coding regions and discussed their potential role in changing branch architecture.

genomics↗

bioRxiv2022

Natural variation in PHY causes differential response to R/FR light following a latitudinal cline in Norway spruce

Detection of the genomic basis of local adaptation to environmental conditions is challenging in forest trees. Phytochromes (PHY) and cryptochromes (CRY) perceive the red (R)/far-red (FR) and blue light respectively and, play a fundamental role in regulating the light pathway in plants contributing to plant growth and development. Norway spruce shows an adaptive latitudinal cline for shade (low R:FR or FR-enriched light) tolerance and requirement of FR light for its growth, thus showing differential response to light quality. We analyzed the exome capture data that included a uniquely large data set of 1654 Norway spruce trees sampled from different latitudes across Sweden that differ in exposure to photoperiod and FR light received during the growing season. Statistically significant clinal variation was detected in allele and genotype frequencies of missense mutations in coding regions belonging to well-defined functional domains of PHYO (PAS-B), PHYP2 (PAS fold-2), CRY1 (CCT1) and CRY2 (CCT2) that strongly correlates with the latitudinal gradient in response to variable light quality in Norway spruce. Asn835Ser in PHYO displayed the steepest cline among all other polymorphisms. We propose that these variations represent signs of local adaptation to light quality in Norway spruce.

genomics↗

bioRxiv2022

Teasing apart the joint effect of demography and natural selection in the birth of a contact zone

Vast population movements induced by recurrent climatic cycles have shaped the genetic structure of plant species. This is especially true in Scandinavia that was repeatedly glaciated. During glacial periods trees were confined to refugia, south and east of the ice sheet, from which they recolonized Scandinavia as the ice melted away. This multi-pronged recolonization led to large contact zones in most species. We leverage large genomic data from 5000 trees to reconstruct the demographic history of Norway spruce (Picea abies) and test for the presence of natural selection during the recolonization process and the establishment of the contact zone. Sweden is today made up of two large genetic clusters, a southern one originating from the Baltics and a Northern one originating from Northern Russia. The contact zone delineating these two clusters closely matches the limit between two major climatic regions. This suggests that natural selection contributed to the establishment and the maintenance of the contact zone. To test this hypothesis we first used Approximate Bayesian Computation; an Isolation-with migration model with genome-wide linked selection fits the data better than a purely neutral one. Secondly, we identified loci characterized by both extreme allele frequency differences between geographic regions and association to the variables defining the climatic zones. These loci, many of which are related to phenology, form clusters present on all linkage groups. Altogether, the current genetic structure reflects the joint effect of climatic cycles, recolonization and selection on the establishment of strong local adaptation and contact zones. Significance StatementUnderstanding how past climatic events, human actions and evolutionary forces contributed to the present distribution of genetic diversity is crucial to predict their reaction to the current climate crisis. Vast distribution shifts induced by past environmental changes, local ecological processes, natural selection and human transfers contributed to the current distribution of Norway spruce across Northern Europe. Genome-wide polymorphisms from thousands of individuals show that Scandinavia was recolonized after the Last Glacial from both south and north. This two-pronged recolonization established a contact zone between two genetic clusters that matches the limit between two major climate zones. The contact zone is shaped and maintained by natural selection on a large number of loci that form blocks of co-adapted loci spread genome-wide.

evolutionary biology↗

bioRxiv2021

Distance-based exclusion method for parentage analysis using microsatellites (SSR) markers

Genotyping mistakes represent a challenge in parental assignment where even small errors can lead to significant amounts of unassigned siblings. Different parental assignment algorithms have been designed to approach this problem. The Exclusion method is the most applied for its reliability and biological meaning. However, the resolving power of this method is the lowest for data containing genotyping errors. We introduce a new distance-based approach which we coin as Distance-Based Exclusion (DBE). The DBE method calculates the distance between the offspring haplotype and haplotype of each of the potential fathers. The father with the lowest distance is then assigned as candidate father according to a distance ratio (). We have tested the Exclusion and DBE methods using a real dataset of 1230 offsprings subdivided into families of 25 individuals. Each family had six potential fathers and one known mother. Compared with the Exclusion method, the DBE method is able to solve 4.7% more individuals (64.4% Exclusion vs 69.1% DBE) using the most restrictive tested without errors. DBE method can also be used together with the Exclusion method for error calculation and to further solve unassigned individuals. Using a two-step approach, we were able to assign 98.1% of the offsprings with a total predicted error of 4.7%. Considering the results obtained, we propose the use of the DBE method in combination with the Exclusion method for parental assignment.

genetics↗

bioRxiv2020

Effect of number of annual rings and tree ages on genomic predictive ability for solid wood properties of Norway spruce

Genomic selection (GS) or genomic prediction is considered as a promising approach to accelerate tree breeding and increase genetic gain by shortening breeding cycle, but the efforts to develop routines for operational breeding are so far limited. We investigated the predictive ability (PA) of GS based on 484 progeny trees from 62 half-sib families in Norway spruce (Picea abies (L.) Karst.) for wood density, modulus of elasticity (MOE) and microfibril angle (MFA) measured with SilviScan, as well as for measurements on standing trees by Pilodyn and Hitman instruments. GS predictive abilities were comparable with those based on pedigree-based prediction. The highest PAs were reached with at least 80-90% of the dataset used as training set. Use of different statistical methods had no significant impact on the estimated PAs. We also compared the abilities to predict density, MFA and MOE of 19 year old trees with models trained on data from coring at different ages and to different depths into the stem. 78-95% of the maximal PAs obtained from coring to the pith at high age were reached by using data possible to obtain by drilling 3-5 rings towards the pith at tree age 10-12, thereby shortening the cycle and reducing the impact on the tree.

plant biology↗

bioRxiv2020

Evolutionary analysis of LP3 gene family in conifers: an ASR homolog

Drought has long been established as a major environmental stress for plants which have in turn developed several coping strategies, ranging from physiological to molecular mechanisms. LP3; a homolog of the Abscisic Acid, Stress and Ripening (ASR) gene was first detected in tomato; and has been shown to be present in four different isoforms in loblolly pine called LP3-0, LP3-1, LP3-2 and LP3-3. While ASR has already been extensively studied notably in tomato, the same cannot be said of LP3. Like ASR, the different LP3 isoforms have been shown to be upregulated in response to water deficit stress and to also act as transcription factors for genes likely involved in hexose transport. In this study we have investigated the evolutionary history of LP3 gene family, with the aim of relating it to that of ASR from a phylogenetic perspective and comparing the differences in selective pressure and codon usage. Phylogenetic analyses of different LP3 homologs compared to ASR show that LP3 is less divergent across species than ASR and that even when comparing the different sub-sections of the gene the divergence rate of LP3 is lower than that of ASR. Analysis of different gene parameters showed that there were differences in GC1% and GC2% but not in total or GC3% content. All genes had a relatively high CAI value associated with a low to moderate ENC value, which is indicative of high translation efficiency found in highly expressed genes. Analysis of codon usage also showed that LP3 preferentially uses different codons than ASR. Selective pressure analysis across most of the LP3 and ASR genes used in this study showed that these genes were principally undergoing purifying selection, with the exception of LP3-3 which seems to be undergoing diversifying selection most probably due to the fact that it likely recently diverged from LP3-0. This study thus provides insight in how ASR and LP3 have diverged from each other while remaining homologous.

evolutionary biology↗

bioRxiv2020

Molecular signatures of local adaptation to light in Norway Spruce: role of lignin mediated immunity

Study of natural variation is an efficient method to elucidate how plants adapt to local climatic conditions, a key process for the evolution of a species. However, it is challenging to determine the genetic basis of adaptive variation especially in forest trees which have large and complex genomes. Norway spruce is a shade tolerant conifer in which the requirement of far-red light for growth increases latitudinally northwards. In the current work, hypocotyl-length followed a latitudinal cline in response to SHADE (low red:far-red ratio). RNA-sequencing revealed differential gene expression in response to SHADE, between a southern and a northern natural population in Sweden. Exome capture included analysis of uniquely large data set (1654 trees) that revealed missense variations in coding regions of nine differentially expressed candidate genes, which followed a latitudinal cline in allele and genotype frequencies. These genes included five transcription factors involved in vital processes like bud-set/bud-flush, lignin pathway and cold acclimation, and other genes that take part in cell-wall remodeling, secondary cell-wall thickening, response to starvation and immunity. Findings from this work primarily suggests that the northern populations of Norway spruce are better adapted towards disease resistance under shade by up-regulation of lignin pathway that is linked to immunity and it forms concrete basis for local adaptation to light quality in Norway spruce, one of the most economically important conifer tree species in Sweden.

plant biology↗

bioRxiv2018

Increased prediction ability in Norway spruce trials using a marker x environment interaction and non-additive genomic selection model

A genomic selection (GS) study of growth and wood quality traits is reported based on control-pollinated Norway spruce families established in two Northern Swedish trials at two locations using exome capture as a genotyping platform. Non-additive effects including dominance and first-order epistatic interactions (including additive by additive, dominance by dominance, and additive by dominance) and marker-by-environment interaction (MxE) effects were dissected in genomic and phenotypic selection models. GS models partitioned additive and non-additive genetic variances more precisely compared with pedigree-based models. In addition, predictive ability (PA) in GS was substantially increased by including dominance and slightly increased by including MxE effects when these effects are significant. For velocity, response to GS (RGS) per year increased 91.3/43.7%, 86.9/82.9%, and 78.9/80.8% compared with response to phenotypic selection (RPS) per year when GS was based on 1) main marker effects (M), 2) M + MxE effects (A), and 3) A + dominance effects (AD) for site 1/site 2, respectively. This indicates that including MxE and dominance effects not only improves genetic parameter estimates but also may improve the genetic gain when they are significant. For tree height, Pilodyn, and modulus of elasticity (MOE), RGS per year improved up to 84.2%, 91.3%, and 92.6% compared with RPS per year, respectively.

genetics↗

bioRxiv2018

Functional Quantitative Trait Loci (QTL) analysis for adaptive traits in a three-generation Scots pine pedigree

In forest tree breeding, QTL identification aims to accelerate the breeding cycle and increase the genetic gain of traits with economical and ecological value. In our study, both phenotypic data and predicted breeding values were used in the identification QTL linked to the adaptive value in a three-generation pedigree population, for the first time in a conifer species (Pinus sylvestris L.). A total of 11 470 open pollinated F2-progeny trees established at three different locations, were measured for growth and adaptive traits. Breeding values were predicted for their 360 mothers, originating from a single cross of two parents. A multilevel LASSO association analysis was conducted to detect QTL using genotypes of the mothers with the corresponding phenotypes and estimated breeding values (EBVs). Different levels of genotype-by-environment (GxE) effects among sites and ages were detected for survival and height. Moderate-to-low narrow sense heritabilities and EBVs accuracies were found for all traits and all sites. We identified 18 AFLPs and 12 SNPs to be associated with QTL for one or more traits. 62 QTL were significant with percentages of variance explained ranging from 1.7 to 18.9%, mostly for traits based on phenotypic data. Two SNP-QTL showed pleiotropic effects for traits related with survival, seed and flower production. Furthermore, we detected several QTL with significant effects across multiple ages, which could be considered as strong candidate loci for early selection. The lack of reproducibility of some QTL detected across sites may be due to environmental heterogeneity and QTL-by-environment effects.

genetics↗

bioRxiv2018

Design and evaluation of a large sequence-capture probe set and associated SNPs for diploid and haploid samples of Norway spruce (Picea abies)

Massively parallel sequencing has revolutionized the field of genetics by providing comparatively high-resolution insights into whole genomes for large number of species so far. However, whole-genome resequencing of many conspecific individuals remains cost-prohibitive for most species. This is especially true for species with very large genomes with extensive genomic redundancy, such as the genomes of coniferous trees. The genome assembly for the conifer Norway spruce (Picea abies) was the first published draft genome assembly for any gymnosperm. Our goal was to develop a dense set of genome-wide SNP markers for Norway spruce to be used for assembly improvement and population studies. From 80,000 initial probe candidates, we developed two partially-overlapping sets of sequence capture probes: one developed against 56 haploid megagametophytes, to aid assembly improvement; and the other developed against 6 diploid needle samples, to aid population studies. We focused probe development within genes, as delineated via the annotation of ~67,000 gene models accompanying P. abies assembly version 1.0. The 31,277 probes developed against megagametophytes covered 19,268 gene models (mean 1.62 probes/model). The 40,018 probes developed against diploid tissue covered 26,219 gene modules (mean 1.53 probes/model). Analysis of read coverage and variant quality around probe sites showed that initial alignment of captured reads should be done against the whole genome sequence, rather than a subset of probe-containing scaffolds, to overcome occasional capture of sequences outside of designed regions. All three probe sets, anchored to the P. abies 1.0 genome assembly and annotation, are available for download.

genomics↗