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Caffeine is the most consumed alkaloid stimulant in the world. It is synthesized through the activity of three known N-methyltransferase proteins. Here we are reporting on the 422-Mb chromosome-level assembly of the Coffea humblotiana genome, a wild and endangered, naturally caffeine-free, species from the Comoro archipelago. We predicted 32,874 genes and anchored 88.7% of the sequence onto the 11 chromosomes. Comparative analyses with the African Robusta coffee genome (C. canephora) revealed an extensive genome conservation, despite an estimated 11 million years of divergence and a broad diversity of genome sizes within the Coffea genus. In this genome, the absence of caffeine is likely due to the absence of the caffeine synthase gene which converts theobromine into caffeine through an illegitimate recombination mechanism. These findings pave the way for further characterization of caffeine-free species in the Coffea genus and will guide research towards naturally-decaffeinated coffee drinks for consumers. [ABSTRACT FROM AUTHOR]

Coffea arabica (the Arabica coffee) is an allotetraploid species originating from a recent hybridization between two diploid species: C. canephora and C. eugenioides. Transposable elements can drive structural and functional variation during the process of hybridization and allopolyploid formation in plants. To learn more about the evolution of the C. arabica genome, we characterized and studied a new Copia LTR-Retrotransposon (LTR-RT) family in diploid and allotetraploid Coffea genomes called Divo. It is a complete and relatively compact LTR-RT element (~5 kb), carrying typical Gag and Pol Copia type domains. Reverse Trancriptase (RT) domain-based phylogeny demonstrated that Divo is a new and well-supported family in the Bianca lineage, but strictly restricted to dicotyledonous species. In C. canephora, Divo is expressed and showed a genomic distribution along gene rich and gene poor regions. The copy number, the molecular estimation of insertion time and the analysis at orthologous locations of insertions in diploid and allotetraploid coffee genomes suggest that Divo underwent a different and recent transposition activity in C. arabica and C. canephora when compared to C. eugenioides. The analysis of this novel LTR-RT family represents an important step toward uncovering the genome structure and evolution of C. arabica allotetraploid genome. [ABSTRACT FROM AUTHOR]

The Coffea genus, 124 described species, has a natural distribution spreading from inter-tropical Africa, to Western Indian Ocean Islands, India, Asia and up to Australasia. Two cultivated species, C. arabica and C. canephora, are intensively studied while, the breeding potential and the genome composition of all the wild species remained poorly uncharacterized. Here, we report the characterization and comparison of the highly repeated transposable elements content of 11 Coffea species representatives of the natural biogeographic distribution. A total of 994 Mb from 454 reads were produced with a genome coverage ranging between 3.2 and 15.7 %. The analyses showed that highly repeated transposable elements, mainly LTR retrotransposons (LTR-RT), represent between 32 and 53 % of Coffea genomes depending on their biogeographic location and genome size. Species from West and Central Africa (Eucoffea) contained the highest LTR-RT content but with no strong variation relative to their genome size. At the opposite, for the insular species (Mascarocoffea), a strong variation of LTR-RT was observed suggesting differential dynamics of these elements in this group. Two LTR-RT lineages, SIRE and Del were clearly differentially accumulated between African and insular species, suggesting these lineages were associated to the genome divergence of Coffea species in Africa. Altogether, the information obtained in this study improves our knowledge and brings new data on the composition, the evolution and the divergence of wild Coffea genomes. [ABSTRACT FROM AUTHOR]

The completion of the genome assembly for the economically important coffee plant Coffea canephora (Rubiaceae) has allowed the use of bioinformatic tools to identify and characterize a diverse array of transposable elements (TEs), which can be used in evolutionary studies of the genus. An overview of the copy number and location within the C. canephora genome of four TEs is presented. These are tested for their use as molecular markers to unravel the evolutionary history of the Millotii Complex, a group of six wild coffee ( Coffea) species native to Madagascar. Two TEs from the Gypsy superfamily successfully recovered some species boundaries and geographic structure among samples, whereas a TE from the Copia superfamily did not. Notably, species occurring in evergreen moist forests of eastern and southeastern Madagascar were divergent with respect to species in other habitats and regions. Our results suggest that the peak of transpositional activity of the Gypsy and Copia TEs occurred, respectively, before and after the speciation events of the tested Madagascan species. We conclude that the utilization of active TEs has considerable potential to unravel the evolutionary history and delimitation of closely related Coffea species. However, the selection of TE needs to be experimentally tested, since each element has its own evolutionary history. Different TEs with similar copy number in a given species can render different dendrograms; thus copy number is not a good selection criterion to attain phylogenetic resolution. [ABSTRACT FROM AUTHOR]

Retrotransposons are the main component of plant genomes. Recent studies have revealed the complexity of their evolutionary dynamics. Here, we have identified Copia25 in Coffea canephora, a new plant retrotransposon belonging to the Ty1- Copia superfamily. In the Coffea genomes analyzed, Copia25 is present in relatively low copy numbers and transcribed. Similarity sequence searches and PCR analyses show that this retrotransposon with LTRs (Long Terminal Repeats) is widely distributed among the Rubiaceae family and that it is also present in other distantly related species belonging to Asterids, Rosids and monocots. A particular situation is the high sequence identity found between the Copia25 sequences of Musa, a monocot, and Ixora, a dicot species (Rubiaceae). Our results reveal the complexity of the evolutionary dynamics of the ancient element Copia25 in angiosperm, involving several processes including sequence conservation, rapid turnover, stochastic losses and horizontal transfer. [ABSTRACT FROM AUTHOR]

Coffea canephora is subject to enormous competitive challenges from other crops, especially for farmer sustainability and consumer requirements. Coffee breeding programs have to focus on specific traits linked to these two key targets, such as quality character, largely depending on the bean's biochemical composition and field yield. Two segregating populations A and B, from crosses between a hybrid (Congolese × Guinean) FRT58 parental clone and a Congolese FRT51 genotype and between two Congolese parents FRT67 and FRT51, respectively, were used to characterize the quantitative trait loci (QTL) involved in agronomic and biochemical traits. A consensus genetic map was established using 249 SSRs covering 1,201 cM. Three QTL detection models per population with MapQTL (model I) and MCQTL (model II) followed by a connected population approach with MCQTL (model III) were compared based on their efficiency, precision for QTL detection, and their genetic effect assessment (additive, dominance, and parental-favorable allele). The analysis detected a total of 143 QTLs, 60 of which were shared between the three models; 28 found with two models; and two, 13, and 40 specific from models I, II, and III, respectively. The last model III based on connected populations is much more efficient in detecting QTLs with low variance explained and led to the genetic characterization of favorable allele. Thanks to this comparison of three QTL detection models on our quantitative genetic study, we will give a new insight for coffee breeding programs dedicated to managing complex agronomic or qualitative traits. [ABSTRACT FROM AUTHOR]

Phenylalanine ammonia lyase (PAL) is the first entry enzyme of the phenylpropanoid pathway producing phenolics, widespread constituents of plant foods and beverages, including chlorogenic acids, polyphenols found at remarkably high levels in the coffee bean and long recognized as powerful antioxidants. To date, whereas PAL is generally encoded by a small gene family, only one gene has been characterized in Coffea canephora ( CcPAL1), an economically important species of cultivated coffee. In this study, a molecular- and bioinformatic-based search for CcPAL1 paralogues resulted successfully in identifying two additional genes, CcPAL2 and CcPAL3, presenting similar genomic structures and encoding proteins with close sequences. Genetic mapping helped position each gene in three different coffee linkage groups, CcPAL2 in particular, located in a coffee genome linkage group (F) which is syntenic to a region of Tomato Chromosome 9 containing a PAL gene. These results, combined with a phylogenetic study, strongly suggest that CcPAL2 may be the ancestral gene of C. canephora. A quantitative gene expression analysis was also conducted in coffee tissues, showing that all genes are transcriptionally active, but they present distinct expression levels and patterns. We discovered that CcPAL2 transcripts appeared predominantly in flower, fruit pericarp and vegetative/lignifying tissues like roots and branches, whereas CcPAL1 and CcPAL3 were highly expressed in immature fruit. This is the first comprehensive study dedicated to PAL gene family characterization in coffee, allowing us to advance functional studies which are indispensable to learning to decipher what role this family plays in channeling the metabolism of coffee phenylpropanoids. [ABSTRACT FROM AUTHOR]

Tomato ( Solanum lycopersicum) and coffee ( Coffea canephora) belong to the sister families Solanaceae and Rubiaceae, respectively. We report herein the mapping of a common set of 257 Conserved Ortholog Set II genes in the genomes of both species. The mapped markers are well distributed across both genomes allowing the first syntenic comparison between species from these two families. The majority (75%) of the synteny blocks are short (<4 cM); however, some extend up to 50 cM. In an effort to further characterize the synteny between these two genomes, we took advantage of the available sequence for the tomato genome to show that tomato chromosome 7 is syntenic to half of the two coffee linkage groups E and F with the putative break point in tomato localized to the boundary of the heterochromatin and euchromatin on the long arm. In addition to the new insight on genome conservation and evolution between the plant families Solanaceae and Rubiaceae, the comparative maps presented herein provide a translational tool by which coffee researchers may take benefit of DNA sequence and genetic information from tomato and vice versa. It is thus expected that these comparative genome information will help to facilitate and expedite genetic and genomic research in coffee. [ABSTRACT FROM AUTHOR]

The purpose of the study was to evaluate the possible genetic effect on vegetative propagation of Coffea canephora. Diversity for somatic embryogenesis (SE) ability was observed not only among two groups of C. canephora Pierre (Congolese and Guinean), but also within these different genetic groups. The results therefore showed that, under given experimental conditions, SE ability is depending on genotype. Furthermore the detection of quantitative trait loci (QTLs) controlling the SE and cutting abilities of C. canephora was performed on a large number of clones including accessions from a core collection, three parental clones and their segregating progenies. On the one hand we detected eight QTLs determining SE. Six positive QTLs for SE ability, whatever the criteria used to quantify this ability, were localized on one single chromosome region of the consensus genetic map. Two negative QTLs for SE ability (frequency of micro calli without somatic embryo) were detected on another linkage group. Deep analysis of the six QTLs detected for SE ability came to the conclusion that they can be assimilated to one single QTL explaining 8.6–12.2% of the observed variation. On the other hand, two QTLs for average length of roots and length of the longest sprouts of cuttings were detected in two linkage groups. These QTLs detected for cutting ability are explaining 12–27% of the observed variation. These observations led to conclude that SE and cutting abilities of C. canephora Pierre appeared to be genetic dependent but through independent mechanisms. [ABSTRACT FROM AUTHOR]

An EST database has been generated for coffee based on sequences from approximately 47,000 cDNA clones derived from five different stages/tissues, with a special focus on developing seeds. When computationally assembled, these sequences correspond to 13,175 unigenes, which were analyzed with respect to functional annotation, expression profile and evolution. Compared with Arabidopsis, the coffee unigenes encode a higher proportion of proteins related to protein modification/turnover and metabolism—an observation that may explain the high diversity of metabolites found in coffee and related species. Several gene families were found to be either expanded or unique to coffee when compared with Arabidopsis. A high proportion of these families encode proteins assigned to functions related to disease resistance. Such families may have expanded and evolved rapidly under the intense pathogen pressure experienced by a tropical, perennial species like coffee. Finally, the coffee gene repertoire was compared with that of Arabidopsis and Solanaceous species (e.g. tomato). Unlike Arabidopsis, tomato has a nearly perfect gene-for-gene match with coffee. These results are consistent with the facts that coffee and tomato have a similar genome size, chromosome karyotype (tomato, n=12; coffee n=11) and chromosome architecture. Moreover, both belong to the Asterid I clade of dicot plant families. Thus, the biology of coffee (family Rubiacaeae) and tomato (family Solanaceae) may be united into one common network of shared discoveries, resources and information. [ABSTRACT FROM AUTHOR]

Two related segregating populations of Theobroma cacao L. were analysed for their resistance to Phytophthora palmivora. The first F1 population was obtained by crossing two susceptible cacao clones of Catongo (a highly homozygous genotype) and Pound 12(a highly heterozygous genotype) and the second population was obtained by backcrossing a single F1 tree with Catongo. The genetic maps obtained for each population were compared. The F1 map includes 162 loci and the backcross has 140 loci. The two maps, F1 and BC1, exhibit high co-linear loci organization covering respectively, 772 and 944 cM. Phytophthora resistance was assessed by measuring the size increase of a lesion at five (DL5)and ten days (DL10) after pod inoculation. Six different QTL were detected in the F1 and BC1 populations. One QTL was found in both populations, and appeared to be a major component of disease resistance, and explaining nearly 48% of the phenotypic variance in the F1 population. The absence of some yield QTL detection in the BC1 in comparison with the F1 population is due to the lack of transmission of the favouring alleles for these QTL from the single F1 tree used for the backcross. The phenotypic variance explained by the action of the quantitative trait alleles indicated that genetic factors of both major and minor effects were involved in the control of the character studied. QTL conferring increased resistance to Phytophthorawere identified in both susceptible parents, suggesting the presence of transgressive traits and the possibility of selection in cacao. Pleiotropic and epistatic effects for the QTL were also detected. Finally, the use of marker assisted selection (MAS) in cacao breeding programs is discussed. [ABSTRACT FROM AUTHOR]

The aim of this study was to determine the genetic components controlling yield in an F1 cacao cross between Catongo and Pound 12 clones. Genetic maps were constructed for the two parents using molecular markers which detected 158 polymorphic loci covering 772 cM for the heterozygous genotype Pound 12 and only 4 loci representing 16.9 cM of a linkage group which indicated a high level of homozygosity of Catongo. Yield was recorded twice a month during 15 years on 55 individuals from this segregating population. Ten yield QTL were detected on eight linkage groups. Some of these QTL were frequently detected over 15 years of production, while others were specific for a given year. Total yield genetic variance, on a yearly basis, ranged from 0 to 56%. Two major QTL (E and I) each explained approximately 20% of the total variance of the average yield over 15 years. The analysis of potential cacao yield components, such as pod index and trunk diameter, suggested that some regions of the genome exert effects on more than one trait, providing a possible genetic explanation for the correlations detected between some of title traits studied. Data showed that correlation between successive annual yield decreased when the lag between corresponding seasons increased. When separated by more than 10 years, annual yields were no longer correlated. The utilisation of molecular markers alone or in combination with phenotypic selection showed an advantage in the early selection of the best cacao producer trees. Further use of molecular markers in breeding programs is discussed with a view to reducing the generation time of a selection procedure. [ABSTRACT FROM AUTHOR]