OPENRosaceae is the most important fruit-producing clade, and its key commercially relevant genera (Fragaria, Rosa, Rubus and Prunus) show broadly diverse growth habits, fruit types and compact diploid genomes. Peach, a diploid Prunus species, is one of the best genetically characterized deciduous trees. Here we describe the high-quality genome sequence of peach obtained from a completely homozygous genotype. We obtained a complete chromosome-scale assembly using Sanger whole-genome shotgun methods. We predicted 27,852 protein-coding genes, as well as noncoding RNAs. We investigated the path of peach domestication through whole-genome resequencing of 14 Prunus accessions. The analyses suggest major genetic bottlenecks that have substantially shaped peach genome diversity. Furthermore, comparative analyses showed that peach has not undergone recent whole-genome duplication, and even though the ancestral triplicated blocks in peach are fragmentary compared to those in grape, all seven paleosets of paralogs from the putative paleoancestor are detectable.
We characterized the response of root hair density to phosphorus (P) availability in Arabidopsis thaliana. Arabidopsis plants were grown aseptically in growth media with varied phosphorus concentrations, ranging from 1 mmol m−3 to 2000 mmol m−3 phosphorus. Root hair density (number of root hairs per mm of root length) was analysed starting at 7 d of growth. Root hair density was highly regulated by phosphorus availability, increasing significantly in roots exposed to low‐phosphorus availability. The initial root hairs produced by the radicle were not sensitive to phosphorus availability, but began to respond after 9 d of growth. Root hair density was about five times greater in low phosphorus (1 mmol m−3) than in high phosphorus (1000 mmol m−3) media. Root hair density decreased logarithmically in response to increasing phosphorus concentrations within that range. Root hair density also increased in response to deficiencies of several other nutrients, but not as strongly as to low phosphorus. Indoleacetic acid (IAA), the auxin transport inhibitor 2‐(p‐chlorophenoxy)‐2‐methylpropionic acid (CMPA), the ethylene precursor 1‐aminocyclopropane‐1‐carboxylic acid (ACC), and the ethylene synthesis inhibitor amino‐oxyacetic acid (AOA) all increased root hair density under high phosphorus but had very little effect under low phosphorus. Low phosphorus significantly changed root anatomy, causing a 9% increase in root diameter, a 31% decrease in the cross‐sectional area of individual trichoblasts, a 40% decrease in the cross‐sectional area of individual atrichoblasts, and 45% more cortical cells in cross‐section. The larger number of cortical cells and smaller epidermal cell size in low phosphorus roots increased the number of trichoblast files from eight to 12. Two‐thirds of increased root hair density in low phosphorus roots was caused by increased likelihood of trichoblasts to form hairs, and 33% of the increase was accounted for by changes in low phosphorus root anatomy resulting in an increased number of trichoblast files. These results show that phosphorus availability can fundamentally alter root anatomy, leading to changes in root hair density, which are presumably important for phosphorus acquisition.
Buds are specialized structures that protect fragile meristematic regions during dormancy and are part of the mechanism that plants use to survive unfavorable environmental conditions such as low temperature or dessication stress. The evergrowing (evg) mutant of peach [Prunus persica (L.) Batsch] does not form terminal vegetative buds in response to dormancy-inducing conditions such as short days and low temperatures, and the terminal meristems maintain constant growth (leaf addition and internode elongation). We genetically mapped the evg trait and identified the corresponding genomic region in a wild-type genome. We sequenced and annotated the 132-kb region. Nineteen genes were predicted to be in the sequenced region. Ten of the predicted genes were demonstrated to be expressed in the wild-type germplasm but six of these were not expressed in mutant tissues. These six genes are a cluster of MIKC-type MADS-box transcription factors similar to genes from Ipomoea batatas and Solanum tuberosum MADS-box, which also regulate meristem growth in vegetative tissues. A 41,746-bp deletion is present in this region of the mutant genome which results in the loss of all or part of four of the six MADS-box genes. The six MADS-box genes that are not expressed in the mutant are candidates for the regulation of growth cessation and terminal bud formation in peach in response to dormancy-inducing conditions and have been named dormancy-associated MADS-box (DAM) genes.
Summary• Chilling requirement, together with heat requirement, determines the bloom date, which has an impact on the climatic distribution of the genotypes of tree species. The molecular basis of floral bud chilling requirement is poorly understood, despite its importance to the adaptation and production of fruit trees. In addition, the genetic nature of heat requirement and the genetic interrelationships among chilling requirement, heat requirement and bloom date remain unclear.• A peach (Prunus persica) F 2 population of 378 genotypes developed from two genotypes with contrasting chilling requirements was used for linkage map construction and quantitative trait loci (QTL) mapping. The floral bud chilling and heat requirements of each genotype were evaluated over 2 yr and the bloom date was scored over 4 yr.• Twenty QTLs with additive effects were identified for three traits, including one major QTL for chilling requirement and two major QTLs for bloom date. The majority of QTLs colocalized with QTLs for other trait(s). In particular, one genomic region of 2 cM, pleiotropic for the three traits, overlapped with the sequenced peach EVG region.• This first report on the QTL mapping of floral bud chilling requirement will facilitate marker-assisted breeding for low chilling requirement cultivars and the map-based cloning of genes controlling chilling requirement. The extensive colocalization of QTLs suggests that there may be one unified temperature sensing and action system regulating chilling requirement, heat requirement and bloom date together
Mapping and sequencing of the non-dormant evg mutant in peach [Prunus persica (L.) Batsch] identified six tandem-arrayed DAM (dormancy-associated MADS-box) genes as candidates for regulating growth cessation and terminal bud formation. To narrow the list of candidate genes, an attempt was made to associate bud phenology with the seasonal and environmental patterns of expression of the candidates in wild-type trees. The expression of the six peach DAM genes at the EVG locus of peach was characterized throughout an annual growing cycle in the field, and under controlled conditions in response to a long day–short day photoperiod transition. DAM1, 2, 4, 5, and 6 were responsive to a reduction in photoperiod in controlled conditions and the direction of response correlated with the seasonal timing of expression in field-grown trees. DAM3 did not respond to photoperiod and may be regulated by chilling temperatures. The DAM genes in peach appear to have at least four distinct patterns of expression. DAM1, 2, and 4 are temporally associated with seasonal elongation cessation and bud formation and are the most likely candidates for control of the evg phenotype.
We previously identified a cluster of d ormancy-a ssociated M ADS-box transcription factors (DAM genes) in peach [Prunus persica (L.) Batsch] as potential candidates for control of the non-dormant phenotype observed in the evg mutant. Of these genes, DAM3, DAM5 and DAM6 were winter expressed, suggesting a role for these genes during endodormancy. We used peach cultivars with contrasting chilling requirements (CR) for bud break to observe the expression of DAM3, DAM5 and DAM6 in response to chilling accumulation in the field and controlled environments. Vegetative terminal and floral buds were sampled weekly from field grown 'Contender' (1050 h CR), 'Rubyprince' (850 h CR) and 'Springprince' (650 h CR) peach cultivars through winter 2008-2009. Flower and vegetative terminal bud break potential was evaluated at each sampling by forcing cuttings in a growth-permissive environment. We also measured vegetative terminal bud break and DAM gene expression in potted 'Contender' and 'Peen-To' (450 h CR) trees under controlled-environment cold exposure. DAM3, DAM5 and DAM6 are all suppressed by exposure to chilling temperatures in the field and in controlled conditions. Expression of DAM5 and DAM6 are higher in high chill cultivars prior to chilling accumulation and their expression level reaches a minimum in each cultivar coincident with acquisition of bud break competence. Expression levels of DAM5 and DAM6 in vegetative tips in controlled environment conditions were negatively correlated with the time required for bud break in forcing conditions. The expression patterns of DAM5 and DAM6 are consistent with a role as quantitative repressors of bud break.
Low-cost, high throughput genotyping methods are crucial to marker discovery and marker-assisted breeding efforts, but have not been available for many ‘specialty crops’ such as fruit and nut trees. Here we apply the Genotyping-By-Sequencing (GBS) method developed for cereals to the discovery of single nucleotide polymorphisms (SNPs) in a peach F2 mapping population. Peach is a genetic and genomic model within the Rosaceae and will provide a template for the use of this method with other members of this family. Our F2 mapping population of 57 genotypes segregates for bloom time (BD) and chilling requirement (CR) and we have extensively phenotyped this population. The population derives from a selfed F1 progeny of a cross between ‘Hakuho’ (high CR) and ‘UFGold’ (low CR). We were able to successfully employ GBS and the TASSEL GBS pipeline without modification of the original methodology using the ApeKI restriction enzyme and multiplexing at an equivalent of 96 samples per Illumina HiSeq 2000 lane. We obtained hundreds of SNP markers which were then used to construct a genetic linkage map and identify quantitative trait loci (QTL) for BD and CR.
Evergrowing (EVG) peach is one of only two described mutants affecting winter dormancy in woody perennial species. EVG peach does not set terminal buds, cease new leaf growth, nor enter into a dormant resting phase in response to winter conditions. The EVG mutation segregates in F2 progeny as a single recessive nuclear gene. A local molecular genetic linkage map around EVG was previously developed using amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) markers, and a bacterial artificial chromosome (BAC) contig that contains the EVG mutation was assembled. A MADS box coding open reading frame (ORF) was found in a BAC of this contig and used as a probe. The probe detected a polymorphism between the wild-type and mutant genomes, and the polymorphism is indicative of a deletion in EVG peach. The EVG gene region contained six potential MADS-box transcription factor sequences, and the deletion in EVG affected at least four of these. The deletion was bracketed using RFLP analysis, which showed that it is contained within a segment of the genome no greater than 180 kb.
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