Abstract: Polyploid plants often produce new phenotypes, exceeding the range of variability existing in the diploid gene pool. Several hundred citrus allotetraploid hybrids have been created by somatic hybridization. These genotypes are interesting models to study the immediate effects of allopolyploidization on the regulation of gene expression. Here, we report genome-wide gene expression analysis in fruit pulp of a Citrus interspecific somatic allotetraploid between C. reticulata cv 'Willowleaf mandarin'+C. limon cv 'Eureka lemon', using a Citrus 20K cDNA microarray. Around 4% transcriptome divergence was observed between the two parental species, and 212 and 160 genes were more highly expressed in C. reticulata and C. limon, respectively. Differential expression of certain genes was confirmed by quantitative real-time RT-PCR. A global downregulation of the allotetraploid hybrid transcriptome was observed, as compared with a theoretical mid parent, for the genes displaying interspecific expression divergence between C. reticulata and C. limon. The genes underexpressed in mandarin, as compared with lemon, were also systematically repressed in the allotetraploid. When genes were overexpressed in C. reticulata compared with C. limon, the distribution of allotetraploid gene expression was far more balanced. Cluster analysis on the basis of gene expression clearly indicated the hybrid was much closer to C. reticulata than to C. limon. These results suggest there is a global dominance of the mandarin transcriptome, in consistence with our previous studies on aromatic compounds and proteomics. Interspecific differentiation of gene expression and non-additive gene regulation involved various biological pathways and different cellular components.
Abstract: The main components of citrus fruit quality (organic acids, sugars, and aromatic compounds) were studied in fruits of a somatic hybrid allotetraploid between Willow leaf mandarin (Citrus deliciosa Ten.) + Eureka lemon [Citrus limon (L.) Burm.] and the two diploid parents. The somatic hybrid (WLM + EUR) combined both nuclear genomes of the parents, with chloroplasts and mitochondria of mandarin. Variations in sugar and acid content were studied in fruit pulp during the maturing period, and the chemical composition of peel oils was investigated by capillary gas chromatography (GC), GC/mass spectrometry (MS), and (13)C NMR. The somatic hybrid was close to the lemon parent in the synthesis of organic acids and close to the mandarin parent in fructose content, while sucrose and glucose contents were between the two parents. The aromatic compounds of WLM + EUR were close to mandarin with a non-negligible effect of lemon, which inhibits the methyl N-methylanthranilate, a mandarin-specific compound. Our results lead us to conclude that biosynthesis of compounds involved in citrus fruit quality is not inherited in an additive way in the allotetraploid hybrid. We observed mandarin dominance for fructose and most of the aromatic compounds, lemon dominance for organic acid and methyl N-methylanthranilate, and codominance for sucrose and glucose.
Abstract: 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 beta-carotene and beta-cryptoxanthin were observed in Citrus limon, while beta-cryptoxanthin was a major component of C. reticulata, which also displayed high levels of phytoene, phytofluene, beta-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.
Abstract: A dinucleotide-enriched genomic library was obtained from mandarin orange (Citrus reticulata Blanco). A subset of 101 positive clones was sequenced and primers were designed. The loci were screened for levels of variation using 26-29 wild mandarin oranges collected in Vietnam. Forty-three loci were polymorphic with the number of alleles ranging from two to 18. The observed heterozygosity (H-O) and expected heterozygosity (H-E) were from 0.03 to 0.96 and from 0.03 to 0.92, respectively.
Abstract: Sugar, organic acid, and carotenoid are the most important indicators of fruit taste and nutritional and organoleptic quality. These components were studied on fruit pulp of the cybrid between Willow leaf mandarin ( Citrus deliciosa Ten.) and Eureka lemon [ Citrus limon (L.) Burm.] and the two parents. The cybrid possessed nuclear and chloroplast genomes of Eureka lemon plus mitochondria from Willow leaf mandarin. The impact of new mitochondria on fruit quality was studied during the mature period. Levels of organic acids were slightly higher in the cybrid fruit pulp than in Eureka lemon. No significant difference in sugar and carotenoid content was observed between the cybrid and the lemon. Results confirm that the main genetic information for the biosynthesis of sugars, organic acids, and carotenoids is contained in the nucleus. In Citrus, cybridization can be used as a strategy to breed specific traits associated with mitochondrial genomes, such as male sterility, without affecting the main organoleptic and nutritional qualities.
Abstract: This study focused on haploid induction in mandarin through in situ gynogenesis by pollination with irradiated pollen of 'Meyer' lemon. Pollination was carried out for three genotypes of mandarin with four levels of gamma-ray-irradiated pollen (150, 300, 600, and 900 Gy). The resulting seeds were characterised by a small size. Embryos were rescued in vitro and the ploidy level of the plantlets was determined by flow cytometry analysis. Haploid, diploid, triploid plantlets were obtained. The haploid parthenogenetic origin was confirmed using microsatellite marker analysis and chromosome count. Diploid and triploid plants were the result of crosses between mandarin and lemon. The induction of gynogenetic haploids of 'Fortune' (Citrus clementina Hort ex Tan. x Citrus tangerina Hort ex Tan.) and 'Ellendale' (Citrus reticulata Blanco x Citrus sinensis L. Osb) is reported here for the first time.
