Analyse comparative de la diversité génétique et de la structure des populations chez l'orge (Hordeum vulgare L.) à l'aide de marqueurs SSR, DArT et du pedigree

La diversité génétique de 92 cultivars d'orges canadiennes a été mesurée en utilisant deux types des marqueurs moléculaires (SSR et DArT) et le pedigree. Au total, 368 alleles ont été identifiés aux 50 locus SSR et utilisés pour créer une matrice de distance génétique. Le nombre d'allèles par locus variait de 2 à 13 (x = 7,36) et l'indice PIC variait entre 0,34 et 0,86 (x = 0,69). Pour les marqueurs bialléliques DArT, la matrice de distance génétique s'est basée sur 971 marqueurs dont les indices PIC variaient entre 0,06 et 0,50 (x = 0,39). Une troisième matrice de distance a été calculée basée sur le coefficient de parenté. Un groupement des génotypes a été effectué basé sur les trois matrices de distances génétiques et les dendrogrammes obtenus ont montré des relations génétiques parmi les cultivars d'orge. Une comparaison de la similarité topologique des trois dendrogrammes réalisée en utilisant l'indice de congruence a montré une très bonne concordance entre les trois dendrogrammes. Des analyses statistiques ont par ailleurs montré une corrélation fortement significative entre les matrices SSR et DArT (r = 0,80, p< 0,002) mais une plus faible corrélation positive du pedigree avec les deux types de marqueur (r = 0,46, p<0,002; r = 0,52, p< 0,002). Ces informations comparant les résultats de différentes méthodes d'estimation de la diversité génétique seront utiles pour l'amélioration et la conservation des ressources génétiques d'orge

Genetic architecture of wood properties based on association analysis and co-expression networks in white spruce

Association studies are widely utilized to analyze complex traits but their ability to disclose genetic architectures is often limited by statistical constraints, and functional insights are usually minimal in nonmodel organisms like forest trees. We developed an approach to integrate association mapping results with co-expression networks. We tested single nucleotide polymorphisms (SNPs) in 2652 candidate genes for statistical associations with wood density, stiffness, microfibril angle and ring width in a population of 1694 white spruce trees (Picea glauca). Associations mapping identified 229-292 genes per wood trait using a statistical significance level of P < 0.05 to maximize discovery. Over-representation of genes associated for nearly all traits was found in a xylem preferential co-expression group developed in independent experiments. A xylem co-expression network was reconstructed with 180 wood associated genes and several known MYB and NAC regulators were identified as network hubs. The network revealed a link between the gene PgNAC8, wood stiffness and microfibril angle, as well as considerable within-season variation for both genetic control of wood traits and gene expression. Trait associations were distributed throughout the network suggesting complex interactions and pleiotropic effects. Our findings indicate that integration of association mapping and co-expression networks enhances our understanding of complex wood traits

Modular organization of the white spruce (Picea glauca) transcriptome reveals functional organization and evolutionary signatures

Transcript profiling has shown the molecular bases of several biological processes in plants but few studies have developed an understanding of overall transcriptome variation. We investigated transcriptome structure in white spruce (Picea glauca), aiming to delineate its modular organization and associated functional and evolutionary attributes. Microarray analyses were used to: identify and functionally characterize groups of co-expressed genes; investigate expressional and functional diversity of vascular tissue preferential genes which were conserved among Picea species, and identify expression networks underlying wood formation. We classified 22 857 genes as variable (79%; 22 coexpression groups) or invariant (21%) by profiling across several vegetative tissues. Modular organization and complex transcriptome restructuring among vascular tissue preferential genes was revealed by their assignment to coexpression groups with partially overlapping profiles and partially distinct functions. Integrated analyses of tissue-based and temporally variable profiles identified secondary xylem gene networks, showed their remodelling over a growing season and identified PgNAC-7 (no apical meristerm (NAM), Arabidopsis transcription activation factor (ATAF) and cup-shaped cotyledon (CUC) transcription factor 007 in Picea glauca) as a major hub gene specific to earlywood formation. Reference profiling identified comprehensive, statistically robust coexpressed groups, revealing that modular organization underpins the evolutionary conservation of the transcriptome structure. © 2015 The Authors

Insect herbivory (Choristoneura fumiferana, Tortricidea) underlies tree population structure (Picea glauca, Pinaceae)

Variation in insect herbivory can lead to population structure in plant hosts as indicated by defence traits. In annual herbaceous, defence traits may vary between geographic areas but evidence of such patterns is lacking for long-lived species. This may result from the variety of selection pressures from herbivores, long distance gene flow, genome properties, and lack of research. We investigated the antagonistic interaction between white spruce (Picea glauca) and spruce budworm (SBW, Choristoneura fumiferana) the most devastating forest insect of eastern North America in common garden experiments. White spruces that are able to resist SBW attack were reported to accumulate the acetophenones piceol and pungenol constitutively in their foliage. We show that levels of these acetophenones and transcripts of the gene responsible for their release is highly heritable and that their accumulation is synchronized with the most devastating stage of SBW. Piceol and pungenol concentrations negatively correlate with rate of development in female SBW and follow a non-random geographic variation pattern that is partially explained by historical damage from SBW and temperature. Our results show that accumulation of acetophenones is an efficient resistance mechanism against SBW in white spruce and that insects can affect population structure of a long-lived plant

Clubroot resistance gene Rcr6 in Brassica nigra resides in a genomic region homologous to chromosome A08 in B. rapa

Background: Clubroot, caused by Plasmodiophora brassicae Woronin, is a very important disease of Brassica species. Management of clubroot relies heavily on genetic resistance. In a cross of Brassica nigra lines PI 219576 (highly resistant, R) × CR2748 (highly susceptible, S) to clubroot, all F1 plants were resistant to clubroot. There was a 1:1 ratio of R:S in the BC1 and 3R:1S in the F2, which indicated that a single dominant gene controlled clubroot resistance in PI 219576. This gene was designated Rcr6. Mapping of Rcr6 was performed using genome sequencing information from A-genome of B. rapa and B-genome of B. nigra though bulked segregant RNA sequencing (BSR-Seq) and further mapping with Kompetitive Allele Specific PCR (KASP) analysis. Results: Reads of R and S bulks from BSR-Seq were initially aligned onto B. rapa (A-genome; B. nigra has the B-genome) where Rcr6 was associated with chromosome A08. KASP analysis showed that Rcr6 was flanked by SNP markers homologous to the region of 14.8-15.4 Mb of chromosome A08. There were 190 genes annotated in this region, with five genes (Bra010552, Bra010588, Bra010589, Bra010590 and Bra010663) identified as encoding the toll-interleukin-1 receptor / nucleotide-binding site / leucine-rich-repeat (TIR-NBS-LRR; TNL) class of proteins. The reads from BSR-Seq were then aligned into a draft B-genome of B. nigra, where Rcr6 was mapped on chromosome B3. KASP analysis indicated that Rcr6 was located on chromosome B3 in a 0.5 Mb region from 6.1-6.6 Mb. Only one TNL gene homologous to the B. rapa gene Bra010663 was identified in the target region. This gene is a likely candidate for Rcr6. Subsequent analysis of the Rcr6 equivalent region based on a published B. nigra genome was performed. This gene is located into chromosome B7 of the published B-genome, homologous to BniB015819. Conclusion: Rcr6 was the first gene identified and mapped in the B-genome of Brassica species. It resides in a genomic region homologous to chromosome A08 of A-genome. Based on this finding, it could possibly integrate into A08 of B. napus using marker assisted selection with SNP markers tightly linked to Rcr6 developed in this study

Architecture génétique des caractères reliés au bois, à la croissance et à la résistance contre la tordeuse des bourgeons de l'épinette chez l'épinette blanche

Les analyses d’association visent à mieux comprendre l’architecture génétique des caractères quantitatifs comme ceux reliés à l’adaptation, la croissance et la structure du bois, et mettre en évidence les gènes qui les contrôlent. Ces connaissances sont utiles pour notamment mieux appréhender le potentiel de résilience dans le contexte de changements climatiques, ainsi que pour la conservation et l’amélioration génétique des arbres forestiers. Cette thèse porte sur l’épinette blanche (Picea glauca [Moench] Voss) avec deux objectifs principaux : 1) développer une compréhension plus intégrée des caractères quantitatifs, et ; 2) développer des stratégies permettant de mieux exploiter les analyses d’association face aux contraintes génétiques et statistiques liées à leur utilisation chez les arbres forestiers. La thèse s’appuie sur deux études d’association qui portent respectivement sur la croissance et les caractéristiques physiques du bois (chapitre 2), et la résistance contre la tordeuse des bourgeons de l’épinette (TBE, Choristoneura fumiferana Clemens) (chapitre 3). Nous avons d’abord analysé les caractéristiques du bois ainsi que la croissance en utilisant un seuil de significativité statistique permissif (P &lt; 0,05; sans correction pour tests multiples) pour maximiser la découverte de gènes comportant des SNPs significatifs. Selon les caractères étudiés, entre 229 et 292 gènes ont été identifiés, qui ont ensuite été soumis à des analyses complémentaires portant sur leurs annotations fonctionnelles et leur expression tissulaire. Les gènes associés aux caractères du bois se sont montrés surreprésentés dans un groupe de co-expression préférentiel au xylème secondaire. Une analyse de réseau de ce groupe de gènes a permis d’identifier certains facteurs de transcription de type MYB et NAC ayant un niveau de connectivité élevé, dont notamment PgNAC-7 qui était le gène ‘hub’ le plus connecté, suggérant un rôle dans la régulation des caractères physiques du bois. L’étude des caractères de défense contre la TBE s’est appuyée sur trois méthodes d’analyse d’association considérant soit un locus à la fois, plusieurs loci, ou encore plusieurs caractères simultanément. Les analyses ont permis d’identifier 33 gènes ayant des fonctions métaboliques variées et expliquant une proportion élevée de la variation phénotypique observée. Des corrélations phénotypiques faiblement positives ont également été observées entre la croissance et les facteurs de résistance contre la TBE, indiquant l’absence d’un compromis métabolique entre ces caractères. Nos résultats indiquent que les stratégies proposées dans la présente thèse pourraient être appliquées à d’autres caractères quantitatifs et permettre de développer des connaissances utiles pour mieux comprendre l'architecture génétique des caractères complexes chez les arbres forestiers.The general aim of association analyses is to decipher the genetic architecture of quantitative traits such as those related to adaptation, growth and wood structure, and to highlight the genes that control them. This knowledge is useful to better understand the potential for resilience toward climate change, and for the genetic conservation and improvement of forest trees. This thesis focuses on white spruce (Picea glauca [Moench] Voss) and has two main objectives: 1) develop a more integrated understanding of quantitative traits, and; 2) develop strategies to better exploit association analyses despite the genetic and statistical constraints related to their use in forest trees. The thesis is based on two association studies, which focus respectively on growth and wood physical traits (Chapter 2), and resistance to spruce budworm (SBW, Choristoneura fumiferana Clemens) (Chapter 3). We first analyzed wood and growth traits using a permissive threshold of statistical significance (P &lt; 0.05, without correction for multiple tests) to maximize the discovery of genes carrying significant SNPs. For each of the traits studied, between 229 and 292 such genes were identified, which were then subjected to further analyses based on their functional annotations and their tissue expression. The genes associated with wood traits were over-represented in a co-expression group that is preferential to secondary xylem. A network analysis of this group of genes allowed identifying certain transcription factors of MYB and NAC types with a high level of connectivity, including PgNAC-7 which was the most connected hub gene, suggesting a role in the regulation of wood physical traits. The study of SBW defence traits was based on three methods of association analysis considering one locus at a time, several loci, or several traits simultaneously. The analyses identified 33 genes with various metabolic functions, which explained a major proportion of the observed phenotypic variation. Weakly positive phenotypic correlations were also observed between growth and resistance traits against SBW, indicating the absence of a metabolic trade-off between these traits. Our results indicate that the strategies proposed in this thesis could be applied to other quantitative traits to develop a better understanding of the genetic architecture of complex traits in forest trees

Changes in the bacterial community colonizing extracted and non-extracted tannin-rich plants in the rumen of dromedary camels.

Leguminous trees and saltbushes provide potential alternatives to conventional feeds to overcome feed deficiency in arid and semi-arid countries. However, these plants are rich in antinutritional factors that have adverse effects on rumen microbiota and the host- animal. Some rumen microbiota detoxifies plants' secondary metabolites; thus, understanding plant-microbe interaction in the rumen could improve the plants' utilization. This study investigated the bacterial colonization and degradation of non-extracted and extracted tanniniferous plants: Atriplex halimus, Acacia saligna, and Leucaena leucocephala, in the rumen of three fistulated camels at 6 and 12 hours. The results showed that these plants have high nutritional value and tannins contents. The rumen degradation and microbial diversity of plant-attached bacteria varied according to plant type and phenols' extraction. Atriplex and leucaena showed higher microbial diversity at 6 and 12h, respectively. Bacteroidetes and Firmicutes were the main bacterial phyla, and the main genera were Prevotella, RC9_gut_group, Butyrivibrio that overrepresented in non-extracted plants (P<0.05). Fibrobacteres and Anaerovibrio showed sensitivity to plant toxins and Ruminococcus attached to plants with lower tannins. Several bacterial genera in the camel rumen have the potential to resist antinutritional factors in fodder plants, which could be used to improve the performance of grazing animals

Comparative analysis of the metabolically active microbial communities in the rumen of dromedary camels under different feeding systems using total rRNA sequencing

Breakdown of plant biomass in rumen depends on interactions between bacteria, archaea, fungi, and protozoa; however, the majority of studies of the microbiome of ruminants, including the few studies of the rumen of camels, only studied one of these microbial groups. In this study, we applied total rRNA sequencing to identify active microbial communities in 22 solid and liquid rumen samples from 11 camels. These camels were reared at three stations that use different feeding systems: clover, hay and wheat straw (G1), fresh clover (G2), and wheat straw (G3). Bacteria dominated the libraries of sequence reads generated from all rumen samples, followed by protozoa, archaea, and fungi respectively. Firmicutes, Thermoplasmatales, Diplodinium, and Neocallimastix dominated bacterial, archaeal, protozoal and fungal communities, respectively in all samples. Libraries generated from camels reared at facility G2, where they were fed fresh clover, showed the highest alpha diversity. Principal co-ordinate analysis and linear discriminate analysis showed clusters associated with facility/feed and the relative abundance of microbes varied between liquid and solid fractions. This provides preliminary evidence that bacteria dominate the microbial communities of the camel rumen and these communities differ significantly between populations of domesticated camels

Association genetics of acetophenone defence against spruce budworm in mature white spruce

Abstract Background Outbreaks of spruce budworm (SBW, Choristoneura fumiferana Clem.) cause major recurrent damage in boreal conifers such as white spruce ( Picea glauca [Moench] Voss) and large losses of forest biomass in North America. Although defensive phenolic compounds have recently been linked to chemical resistance against SBW, their genetic basis remains poorly understood in forest trees, especially in conifers. Here, we used diverse association genetics approaches to discover genes and their variants that may control the accumulation of acetophenones, and dissect the genetic architecture of these defence compounds against SBW in white spruce mature trees. Results Out of 4747 single nucleotide polymorphisms (SNPs) from 2312 genes genotyped in a population of 211 unrelated individuals, genetic association analyses identified 35 SNPs in 33 different genes that were significantly associated with the defence traits by using single-locus, multi-locus and multi-trait approaches. The multi-locus approach was particularly effective at detecting SNP–trait associations that explained a large fraction of the phenotypic variance (from 20 to 43%). Significant genes were regulatory including the NAC transcription factor, or they were involved in carbohydrate metabolism, falling into the binding, catalytic or transporter activity functional classes. Most of them were highly expressed in foliage. Weak positive phenotypic correlations were observed between defence and growth traits, indicating little or no evidence of defence-growth trade-offs. Conclusions This study provides new insights on the genetic architecture of tree defence traits, contributing to our understanding of the physiology of resistance mechanisms to biotic factors and providing a basis for the genetic improvement of the constitutive defence of white spruce against SBW

Rumen bacterial community profile and fermentation in Barki sheep fed olive cake and date palm byproducts

Rumen bacteria make the greatest contribution to rumen fermentation that enables the host animal to utilize the ingested feeds. Agro-industrial byproducts (AIP) such as olive cake (OC) and date palm byproducts (discarded dates (DD), and date palm fronds (DPF)) represent a practical solution to the deficiency in common feed resources. In this study, thirty-six growing Barki lambs were divided into three groups to evaluate the effect of untraditional diets including the AIP on the growth performance. Subsequently, nine adult Barki rams were used to evaluate the effect of experimental diets on rumen fermentation and rumen bacteria. Three rations were used: common concentrate mixture (S1), common untraditional concentrate mixture including OC and DD (S2), and the same concentrate mixture in S2 supplemented with roughage as DPF enriched with 15% molasses (S3). The animals in S2 group showed higher dry matter intake (DMI) and lower relative growth rate (RGR) as compared to the animals in S1 group. However, the animals in S3 group were the lowest in DMI but achieved RGR by about 87.6% of that in the S1 group. Rumen pH, acetic and butyric acids were more prevalent in animals of S3 group and rumen ammonia (NH3-N), total volatile fatty acids (TVFA), propionic acid were higher in S1. Rumen enzymes activities were higher in S1 group followed by S3 and S2. The bacterial population was more prevalent in S1 and microbial diversity was higher in the S3 group. Principal coordinate analysis revealed clusters associated with diet type and the relative abundance of bacteria varied between sheep groups. The bacterial community was dominated by phylum Bacteroidetes and Firmicutes; whereas, Prevotella, Ruminococcus, and Butyrivibrio were the dominant genera. Results indicate that diet S3 supplemented by OC, DD, and DPF could replace the conventional feed mixture