Gene expression of DAM5 and DAM6 is suppressed by chilling temperatures and inversely correlated with bud break rate.

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.

Identification and mapping of resistance gene analogs (RGAs) in Prunus: a resistance map for Prunus.

The genetically anchored physical map of peach is a valuable tool for identifying loci controlling economically important traits in Prunus. Breeding for disease resistance is a key component of most breeding programs. The identification of loci for pathogen resistance in peach provides information about resistance loci, the organization of resistance genes throughout the genome, and permits comparison of resistance regions among other genomes in the Rosaceae. This information will facilitate the breeding of resistant species of Prunus. A candidate gene approach was implemented for locating resistance loci in the genome of peach. Candidate genes representing NBS-LRR, kinase, transmembrane domain classes, as well as, pathogen response (PR) proteins and resistance-associated transcription factors were hybridized to a peach BAC library and mapped by using the peach physical map database and the Genome Database for Rosaceae (GDR). A resistance map for Prunus was generated and currently contains 42 map locations for putative resistance regions distributed among 7 of the 8 linkage groups.

A deletion affecting several gene candidates is present in the Evergrowing peach mutant.

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.

Simple sequence repeat (SSR) analysis for assessment of genetic variability in apricot germplasm.

Thirty SSR primer combinations, developed from peach SSR-enriched genomic libraries and BAC libraries of peach [ Prunus persica (L.) Batsch.], were tested for cross amplification with 74 apricot ( Prunus armeniaca L.) germplasm accessions. Twelve primer pairs amplified 14 polymorphic SSR loci useful for discriminating most apricot cultivars, as well as for investigating patterns of variation in apricot germplasm. Levels of polymorphism were higher than the levels described using other codominant marker systems (i.e., isozymes, RFLP markers). Overall, 107 alleles were identified, and all but 11 accessions were unambiguously discriminated. Genetic differentiation of native germplasm into traditional ecogeographical groups was low, with a high level of genetic identity (> 0.75) between the groups. However, neighbor joining cluster analysis of marker distances between cultivars reflected the complex history of apricot domestication, producing groupings not evidently based on the geographical origin of the cultivars. Distant positioning of Chinese cultivars on UPGMA and neighbor joining dendrograms supports the authors' consideration of Chinese apricots as subspecies, Prunus armeniaca var. ansu Maxim., rather than a separate species.

Construction of a BAC library and its application to the identification of simple sequence repeats in peach [ Prunus persica (L.) Batsch].

The Rosaceae contains many economically important crop species, but their genomes are not well characterized, and comparative genetic mapping lags well behind that of other families. To facilitate genome comparisons and gene discovery in the Rosaceae, we have begun the development of genomic resources for peach as the model genome for this family. First, we developed a simplified, cost-effective method for constructing BAC libraries, particularly appropriate for plant species of relatively minor economic importance. Second, we used the library to investigate the abundance and local distribution of simple sequence repeats (SSRs) in peach. Our results indicate that microsatellite loci are locally much more highly abundant than previously estimated, and BAC sequencing results suggest that microsatellite repeats are not randomly distributed within gene-containing regions of the peach genome. This makes it relatively easy to identify SSRs in peach by hybridization to BAC clones, and even by random sequencing of BAC clones, not known a priori to contain SSRs.

Genetic mapping of the evergrowing gene in peach [Prunus persica (L.) Batsch].

In temperate locations, terminal apices on evergrowing (also called evergreen) peach trees keep growing in winter until killed by low temperatures, while the lateral buds go into dormancy. A recessive allele of a single gene (evergrowing or evg) controls this trait in peach. The amplified fragment length polymorphism (AFLP) technique and bulked segregant analysis were applied to construct a local genetic linkage map for the evg gene from the cross Empress op op dwarf x Evergrowing (P.I. 442380). This map, comprising nine AFLP markers and the evg locus, covers a total genetic distance of 79.3 cM. Four dominant AFLP markers (EAT/MCAC, ETT/MCCA2, EAT/MCTA, and ETT/MACC) were linked to the evg locus at distances of 1, 5.3, 6.7, and 11.7 cM, respectively. EAT/MCAC and EAT/MCTA were converted into polymorphic sequence-tagged sites. Microsatellite markers in the evg region were developed from peach bacterial artificial chromosome (BAC) clones that hybridized to the AFLP marker fragments. Using three microsatellite anchor markers (pchgms12, pchgms17, and pchgms19), the local genetic linkage map was integrated into one minor linkage group of a previously constructed peach rootstock genetic linkage map. Three AFLP markers from the rootstock genetic linkage map were found linked to the evg locus.

Reproduction and Development of Meloidogyne incognita and M. javanica on Guardian Peach Rootstock.

Guardian peach rootstock was evaluated for susceptibility to Meloidogyne incognita race 3 (Georgia-peach isolate) and M. javanica in the greenhouse. Both commercial Guardian seed sources produced plants that were poor hosts of M. incognita and M. javanica. Reproduction as measured by number of egg masses and eggs per plant, eggs per egg mass, and eggs per gram of root were a better measure of host resistance than number of root galls per plant. Penetration, development, and reproduction of M. incognita in Guardian (resistant) and Lovell (susceptible) peach were also studied in the greenhouse. Differences in susceptibility were not attributed to differential penetration by the infectivestage juveniles (J2) or the number of root galls per plant. Results indicated that M. incognita J2 penetrated Guardian roots and formed galls, but that the majority of the nematodes failed to mature and reproduce.

Impact of Meloidogyne incognita on the Incidence of Peach Tree Short Life in the Presence of Criconemella xenoplax.

The relationship between Cricenemella xenoplax alone and in combination with Meloidogyne incognitaon the incidence of peach tree short life disease was studied in field microplots during 1989-96. The presence of M. incognita suppressed the population density of C. xenoplax on Lovell peach. Tree trunk diameter was significantly reduced in the presence of both nematode species prior to 1993. Soil pH was lowest in the co-infection treatment as compared with the uninoculated control on three of the four sampling dates. In 1994, 80% of the trees growing in soil infested with C. xenoplax alone developed typical disease symptoms and died. The remaining tree died in 1995. No trees died in the M. incognita alone, C. xenoplax + M. incognita, or uninoculated control treatments. Parasitism by C. xenoplax, but not by M. incognita, made Lovell peach trees more susceptible to the disease. These findings were confirmed in an orchard site naturally infested with both C. xenoplax and M. incognita where Redhaven trees budded to Lovell rootstock exhibited a reduction of 1.6 years in average tree life for every centimeter increase in trunk diameter.