Genotyping-by-sequencing of pear (Pyrus spp.) accessions unravels novel patterns of genetic diversity and selection footprints

Understanding of genetic diversity and marker-trait relationships in pears(Pyrus spp.)forms an important part of gene conservation and cultivar breeding.Accessions of Asian and European pear species,and interspecific hybrids were planted in a common garden experiment.Genotyping-by-sequencing(GBS)was used to genotype 214 accessions,which were also phenotyped for fruit quality traits.A combination of selection scans and association analyses were used to identify signatures of selection.Patterns of genetic diversity,population structure and introgression were also investigated.About 15000 high-quality SNP markers were identified from the GBS data,of which 25%and 11%harboured private alleles for European and Asian species,respectively.Bayesian clustering analysis suggested negligible gene flow,resulting in highly significant population differentiation(Fst=0.45)between Asian and European pears.Interspecific hybrids displayed an average of 55%and 45%introgression from their Asian and European ancestors,respectively.Phenotypic(firmness,acidity,shape and so on)variation between accessions was significantly associated with genetic differentiation.Allele frequencies at large-effect SNP loci were significantly different between genetic groups,suggesting footprints of directional selection.Selection scan analyses identified over 20 outlier SNP loci with substantial statistical support,likely to be subject to directional selection or closely linked to loci under selection.

Differential regulation of triterpene biosynthesis induced by an early failure in cuticle formation in apple

Waxy apple cuticles predominantly accumulate ursane-type triterpenes,but the profile shifts with the induction of skin russeting towards lupane-type triterpenes.We previously characterised several key enzymes in the ursane-type and lupane-type triterpene pathways,but this switch in triterpene metabolism associated with loss of cuticle integrity is not fully understood.To analyse the relationship between triterpene biosynthesis and russeting,we used microscopy,RNA-sequencing and metabolite profiling during apple fruit development.We compared the skin of three genetically-close clones of‘Golden Delicious’(with waxy,partially russeted and fully russeted skin).We identified a unique molecular profile for the russet clone,including low transcript abundance of multiple cuticle-specific metabolic pathways in the early stages of fruit development.Using correlation analyses between gene transcription and metabolite concentration we found MYB transcription factors strongly associated with lupane-type triterpene biosynthesis.We showed how their transcription changed with the onset of cuticle cracking followed by russeting and that one factor,MYB66,was able to bind the promoter of the oxidosqualene cyclase OSC5,to drive the production of lupeol derivatives.These results provide insights into the breakdown of cuticle integrity leading to russet and how this drives MYB-regulated changes to triterpene biosynthesis.

Multiple quantitative trait loci contribute to resistance to bacterial canker incited by Pseudomonas syringae pv.actinidiae in kiwifruit(Actinidia chinensis)

Pseudomonas syringae pv.actinidiae(Psa)biovar 3,a virulent,canker-inducing pathogen is an economic threat to the kiwifruit(Actinidia spp.)industry worldwide.The commercially grown diploid(2×)A.chinensis var.chinensis is more susceptible to Psa than tetraploid and hexaploid kiwifruit.However information on the genetic loci modulating Psa resistance in kiwifruit is not available.Here we report mapping of quantitative trait loci(QTLs)regulating resistance to Psa in a diploid kiwifruit population,derived from a cross between an elite Psa-susceptible‘Hort16A’and a resistant male breeding parent P1.Using high-density genetic maps and intensive phenotyping,we identified a single QTL for Psa resistance on Linkage Group(LG)27 of‘Hort16A’revealing 16–19%phenotypic variance and candidate alleles for susceptibility and resistance at this loci.In addition,six minor QTLs were identified in P1 on distinct LGs,exerting 4–9%variance.Resistance in the F1 population is improved by additive effects from‘Hort16A’and P1 QTLs providing evidence that divergent genetic pathways interact to combat the virulent Psa strain.Two different bioassays further identified new QTLs for tissue-specific responses to Psa.The genetic marker at LG27 QTL was further verified for association with Psa resistance in diploid Actinidia chinensis populations.Transcriptome analysis of Psa-resistant and susceptible genotypes in field revealed hallmarks of basal defense and provided candidate RNA-biomarkers for screening for Psa resistance in greenhouse conditions.