Trends in Fruit Quality Improvement From 15 Years of Selection in the Apple Breeding Program of Washington State University

Washington State University's apple breeding program (WABP) was initiated in 1994 to select new apple cultivars with improved eating quality, appearance, and storability that are suitable for production in the main growing regions of the state. Fruit quality is phenotyped using various instrumental measures, such as penetrometers (texture), titrator (acidity), and refractometer (soluble solids concentration; SSC), as well as sensory assessment. The selection regime of WABP occurs in three sequential phases: phase one (P1)—single, unreplicated seedlings at one site, phase two (P2)—replicated selections at three geographically diverse sites, and phase three (P3)—highly replicated elite selections at one to two grower sites. Most of the data collection of WABP occurs in P2. Knowledge of trends/changes associated with advancing selections is essential for understanding the selection criteria and progress of WABP throughout the changing compositions of advancing and culling selections. For each post-harvest trait, P2 data from harvest years 2005 to 2019 were split across sites, and between selections and reference cultivars (e.g., Cripps Pink, Gala, and Honeycrisp). Means of instrumental crispness (Cn) and inner cortex firmness for the advancing selections increased gradually over this period and were significantly higher than those for cultivars. Means of outer cortex firmness measurements were stable for selections but significantly higher than those for cultivars. The average fruit acidity of selections increased marginally over this period and was higher than that of the cultivars. Meanwhile, the average fruit SSCs of selections and cultivars were statistically indistinguishable. These 15-year trends indicate that WABP has been selecting apples with improved eating quality and storability through increased crispness and inner cortex firmness, respectively.

Phenotypic divergence between the cultivated apple (Malus domestica) and its primary wild progenitor (Malus sieversii)

An understanding of the relationship between the cultivated apple (Malus domestica) and its primary wild progenitor species (M. sieversii) not only provides an understanding of how apples have been improved in the past, but may be useful for apple improvement in the future. We measured 10 phenotypes in over 1000 unique apple accessions belonging to M. domestica and M. sieversii from Canada’s Apple Biodiversity Collection. Using principal components analysis (PCA), we determined that M. domestica and M. sieversii differ significantly in phenotypic space and are nearly completely distinguishable as two separate groups. We found that M. domestica had a shorter juvenile phase than M. sieversii and that cultivated trees produced flowers and ripe fruit later than their wild progenitors. Cultivated apples were also 3.6 times heavier, 43% less acidic, and had 68% less phenolic content than wild apples. Using historical records, we found that apple breeding over the past 200 years has resulted in a trend towards apples that have higher soluble solids, are less bitter, and soften less during storage. Our results quantify the significant changes in phenotype that have taken place since apple domestication, and provide evidence that apple breeding has led to continued phenotypic divergence of the cultivated apple from its wild progenitor species.

Fire blight QTL analysis in a multi-family apple population identifies a reduced-susceptibility allele in ‘Honeycrisp’

Breeding apple cultivars with resistance offers a potential solution to fire blight, a damaging bacterial disease caused by Erwinia amylovora. Most resistance alleles at quantitative trait loci (QTLs) were previously characterized in diverse Malus germplasm with poor fruit quality, which reduces breeding utility. This study utilized a pedigree-based QTL analysis approach to elucidate the genetic basis of resistance/susceptibility to fire blight from multiple genetic sources in germplasm relevant to U.S. apple breeding programs. Twenty-seven important breeding parents (IBPs) were represented by 314 offspring from 32 full-sib families, with ‘Honeycrisp’ being the most highly represented IBP. Analyzing resistance/susceptibility data from a two-year replicated field inoculation study and previously curated genome-wide single nucleotide polymorphism data, QTLs were consistently mapped on chromosomes (Chrs.) 6, 7, and 15. These QTLs together explained ~28% of phenotypic variation. The Chr. 6 and Chr. 15 QTLs colocalized with previously reported QTLs, while the Chr. 7 QTL is possibly novel. ‘Honeycrisp’ inherited a rare reduced-susceptibility allele at the Chr. 6 QTL from its grandparent ‘Frostbite’. The highly resistant IBP ‘Enterprise’ had at least one putative reduced-susceptibility allele at all three QTLs. In general, lower susceptibility was observed for individuals with higher numbers of reduced-susceptibility alleles across QTLs. This study highlighted QTL mapping and allele characterization of resistance/susceptibility to fire blight in complex pedigree-connected apple breeding germplasm. Knowledge gained will enable more informed parental selection and development of trait-predictive DNA tests for pyramiding favorable alleles and selection of superior apple cultivars with resistance to fire blight.

Self-incompatibility characterization in segregating populations of apple trees with DNA markers for S-alleles

The objective of this study was to characterize the parents and respective populations of apple trees regarding S-alleles to confirm their genealogy and to evaluate the efficiency of the molecular markers used. Sixteen specific sets of primers were used for identification of apple S-alleles by PCR. Two segregating populations of the Epagri Apple Breeding Program resulting from crosses between ‘Fred Hough’ × ‘Monalisa’ and ‘M-11/00’ × ‘M-13/91’ were evaluated. The expected segregations are 1:1:1:1 for full compatibility and 1:1 for semi-compatibility, which can be confirmed by the X2 test. The ‘Fred Hough’ (S5S19) × ‘Monalisa’ (S2S10) cross proved to be fully compatible; and two triploids were identified among the hybrids as well. The ‘M-11/00’ (S3S19) × ‘M-13/91’ (S3S5) cross was characterized as semi-compatible based on DNA markers, and the segregation of the S-alleles in the hybrids was 1:1, as expected. The segregation of the DNA markers occurred together with their respective S-alleles: S2, S3, S5, S10, and S19. Thus, characterization of the S-alleles not only allowed identification of compatibility between parents but also identified contaminations in segregating populations.

DNA MARKING WHEN CREATING APPLE VARIETIES WITH STABLE SCAB RESISTANCE

The use of chemical means of protection against Apple scab is associated with high material costs and causes great harm to the environment. Growing of apple varieties with stable resistance to scab (the pathogen Venturia inaequalis) will significantly limit the use of pesticides. On the territory of the Russian Federation, the Rvi6 gene is considered immune, which determines resistance to five scab races, including the most aggressive fifth. The country has scabimmune apple varieties created by scientists from VNIISPK, NCFSCHWWG, FRC named after I. V. Michurin , VSTISP and other scientific institutions. In recent years, the Rvi6 gene has been overcome by scab in many European countries, and the Rvi5 gene, which is immune to four scab races, has been overcome in Russia and Germany. When creating high-yielding apple varieties of a new generation, with good fruit flavor quality, long-term and stable resistance to scab, in addition to the rvi6 resistance gene, the most promising sources are the Rvi5, Rvi11, Rvi12, Rvi14 and Rvi15 genes. The Rvi2, Rvi4, Rvi6, Rvi7 and Rvi9 genes in the apple breeding process are best used in extended pyramids of genetic resistance to scab. This will allow you to combine several scab resistance genes that control the immune system in one apple genotype. The article describes the characteristics of DNA markers, the nucleotide sequence of primers, the size of target fragments of the PCR product, including the size of the dominant allele product for detecting Rvi genes of the Venturia inaequalis pathogen that are promising for apple breeding in varieties and hybrid material. Amplification programs were selected to identify resistance genes to various races of apple scab.

Eleven-year Correlation of Physical Fruit Texture Traits between Computerized Penetrometers and Sensory Assessment in an Apple Breeding Program

Fruit texture is a major target of apple (Malus domestica) breeding programs due to its influence on consumer preference. This multitrait feature is typically rated using sensory assessment, which is subjective and prone to biases. Instrumental measurements have predominantly targeted firmness of the outer region of fruit cortex using industry standard Magness–Taylor-type penetrometers, while other metrics remain largely unused. Additionally, there have been limited reports on correlating sensory attributes with instrumental metrics on many diverse apple selections. This report is the first to correlate multiyear historical fruit texture information of instrumental metrics and sensory assessment in an apple breeding program. Through 11 years of routine fruit quality evaluation at the Washington State University apple breeding program, physical textural data of 84,552 fruit acquired from computerized penetrometers were correlated with sensory assessment. Correlations among various instrumental metrics are high (0.63 ≤ r ≤ 1.00; P < 0.0001). In correlating instrumental outputs with sensory data, there is a significant correlation (r = 0.43; P < 0.0001) between the instrumental crispness value and sensory crispness. Additionally, instrumental hardness traits are significantly correlated (0.61 ≤ r ≤ 0.69; P < 0.0001) with sensory hardness. Outputs from two versions of computerized penetrometers were tested and shown to have no statistical differences. Overall, this report demonstrates potential use of instrumental metrics as firmness and crispness estimates for selecting apples of diverse backgrounds in a breeding program. However, in testing a large number and diversity of fruit, experimenters should perform data curation and account for lower limits/thresholds of the instrument.

Evidence of apple blotch resistance in wild apple germplasm (Malus spp.) accessions

Diplocarpon coronariae causes apple blotch, which results in significant economic losses. Resistance to apple blotch in commercial apple cultivars has not been reported in recent literature. The Malus germplasm collection from the Fruit Genebank of the Julius Kühn-Institut that is maintained in Dresden, Germany consists of 516 accessions of about 49 Malus species. This apple gene pool was evaluated for resistance to Diplocarpon coronariae. A four-year field survey was subsequently followed by artificial inoculation of field-selected accessions. Fifteen accessions that represent nine different species and hybrids were confirmed to be highly resistant following a repeated inoculation assay in the laboratory. The results from this investigation is of high interest to breeders for future apple breeding programmes and investigations of resistance to Diplocarpon coronariae.