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Genome of a citrus rootstock and global DNA demethylation caused by heterografting.

ABSTRACT: Grafting is an ancient technique used for plant propagation and improvement in horticultural crops for at least 1,500 years. Citrus plants, with a seed-to-seed cycle of 5-15 years, are among the fruit crops that were probably domesticated by grafting. Poncirus trifoliata, a widely used citrus rootstock, can promote early flowering, strengthen stress tolerance, and improve fruit quality via scion-rootstock interactions. Here, we report its genome assembly using PacBio sequencing. We obtained a final genome of 303 Mb with a contig N50 size of 1.17 Mb and annotated 25,680 protein-coding genes. DNA methylome and transcriptome analyses indicated that the strong adaptability of P. trifoliata is likely attributable to its special epigenetic modification and expression pattern of resistance-related genes. Heterografting by using sweet orange as scion and P. trifoliata as rootstock and autografting using sweet orange as both scion and rootstock were performed to investigate the genetic effects of the rootstock. Single-base methylome analysis indicated that P. trifoliata as a rootstock caused DNA demethylation and a reduction in 24-nt small RNAs (sRNAs) in scions compared to the level observed with autografting, implying the involvement of sRNA-mediated graft-transmissible epigenetic modifications in citrus grafting. Taken together, the assembled genome for the citrus rootstock and the analysis of graft-induced epigenetic modifications provide global insights into the genetic effects of rootstock-scion interactions and grafting biology.

SUBMITTER: Huang Y

PROVIDER: S-EPMC8012640 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:BACKGROUND: Citrus represents a substantial income for farmers in the Mediterranean Basin. However, the Mediterranean citrus industry faces increasing biotic and abiotic constraints. Therefore the breeding and selection of new rootstocks are now of the utmost importance. In Tunisia, in addition to sour orange, the most widespread traditional rootstock of the Mediterranean area, other citrus rootstocks and well adapted to local environmental conditions, are traditionally used and should be important genetic resources for breeding. To characterize the diversity of Tunisian citrus rootstocks, two hundred and one local accessions belonging to four facultative apomictic species (C. aurantium, sour orange; C. sinensis, orange; C. limon, lemon; and C. aurantifolia, lime) were collected and genotyped using 20 nuclear SSR markers and four indel mitochondrial markers. Multi-locus genotypes (MLGs) were compared to references from French and Spanish collections. RESULTS: The differentiation of the four varietal groups was well-marked. The groups displayed a relatively high allelic diversity, primarily due to very high heterozygosity. Sixteen distinct MLGs were identified. Ten of these were noted in sour oranges. However, the majority of the analysed sour orange accessions corresponded with only two MLGs, differentiated by a single allele, likely due to a mutation. The most frequent MLG is shared with the reference sour oranges. No polymorphism was found within the sweet orange group. Two MLGs, differentiated by a single locus, were noted in lemon. The predominant MLG was shared with the reference lemons. Limes were represented by three genotypes. Two corresponded to the 'Mexican lime' and 'limonette de Marrakech' references. The MLG of 'Chiiri' lime was unique. CONCLUSIONS: The Tunisian citrus rootstock genetic diversity is predominantly due to high heterozygosity and differentiation between the four varietal groups. The phenotypic diversity within the varietal groups has resulted from multiple introductions, somatic mutations and rare sexual recombination events. Finally, this diversity study enabled the identification of a core sample of accessions for further physiological and agronomical evaluations. These core accessions will be integrated into citrus rootstock breeding programs for the Mediterranean Basin.

Project description:Iron is an essential micronutrient for plants, and plants have evolved adaptive mechanisms to improve iron acquisition from soils. Grafting on iron deficiency-tolerant rootstock is an effective strategy to prevent iron deficiency-chlorosis in fruit-tree crops. To determine the mechanisms underlying iron uptake in iron deficiency, two iron deficiency-tolerant citrus rootstocks, Zhique (ZQ) and Xiangcheng (XC), as well as iron deficiency-sensitive rootstock trifoliate orange (TO) seedlings were studied. Plants were grown in hydroponics system for 100 days, having 50 ?M iron (control) and 0 ?M iron (iron deficiency) nutrient solution. Under iron deficiency, more obvious visual symptoms of iron chlorosis were observed in the leaves of TO, whereas slight symptoms were observed in ZQ and XC. This was further supported by the lower chlorophyll concentration in the leaves of TO than in leaves of ZQ and XC. Ferrous iron showed no differences among the three citrus rootstock roots, whereas ferrous iron was significantly higher in leaves of ZQ and XC than TO. The specific iron absorption rate and leaf iron proportion were significantly higher in ZQ and XC than in TO, suggesting the iron deficiency tolerance can be explained by increased iron uptake in roots of ZQ and XC, allowed by subsequent translocation to shoots. In transcriptome analysis, 29, 298, and 500 differentially expressed genes (DEGs) in response to iron deficiency were identified in ZQ, XC, and TO, respectively (Fold change ? 2 and Probability ? 0.8 were used as thresholds to identify DEGs). A Gene Ontology analysis suggested that several genotype-specific biological processes are involved in response to iron deficiency. Genes associated with cell wall biosynthesis, ethylene and abscisic acid signal transduction pathways were involved in iron deficiency responses in citrus rootstocks. The results of this study provide a basis for future analyses of the physiological and molecular mechanisms of the tolerance of different citrus rootstocks to iron deficiency.

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Genome of a citrus rootstock and global DNA demethylation caused by heterografting.

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