The USDA/ARS collection of Malus is held by the Plant Genetic Resources Unit in Geneva, N.Y. The collection comprises ≈2500 accessions, most of which must be maintained as clones in the field to provide propagating material for distribution to the user community. Field maintenance of replicated accessions places the collection at risk from weather extremes, pests, diseases, etc. and is extremely costly. Cryopreservation of dormant buds in a base, or backup, collection could reduce risks and decrease maintenance costs. Since 1988, we have developed and implemented protocols to cryopreserve dormant apple buds at the National Seed Storage Laboratory, Fort Collins, Colo. More than 500 accessions have been placed in cryogenic storage. Buds have been successfully recovered by grafting from >70% of the first 250 accessions cryopreserved. These results, and those from ongoing recovery tests, indicate cryopreservation may be a safe, cost-effective approach to back up collections of tree fruit germplasm. It also may be used to enhance management of the active collections of Malus, Vitis, and Prunus at Geneva.
Philip L. Forsline, Leigh E. Towill, John Waddell, and Loren Wiesner
Stan C. Hokanson, Phil L. Forsline, James R. McFerson, Warren F. Lamboy, Herb S. Aldwinckle, and Aimak D. Djangaliev
Malus sieversii, the main progenitor of domesticated apple, is native to areas in Central Asia. To better represent Malus wild germplasm in the USDA–ARS germplasm collections, maintained in Geneva, N.Y., a cooperative project was initiated with the Republic if Kazakhstan to collect and assess that country's wild populations of M. sieversii and to develop more secure in situ reserves to complement ex situ holdings in the United States and Kazakhstan. To date, four exploration trips to the region have included participants from the United States, Kazakhstan, Canada, New Zealand, and South Africa. Four Kazkh scientists have toured USDA–ARS sites, exchanged information, and collected germplasm in the United States greenhouse screens of 1600 have revealed potentially new sources of resistance to apple scab, cedar apple rust, and fire blight. An isozyme analysis of maternal half-sib families from four regions suggests the populations of M. sieversii collected represent a single panmictic population, with over 85% of total genetic variation due to differences among families. The most recent collections in 1995 were directed towards more ecologically diverse regions, including a site (Tarbagatai) at the most northern limit for M. sieversii equivalent to northern Minnesota in the United States. Some trees in this region produced fruit nearly 70 mm in diameter with excellent aroma, firmness, and color. This germplasm is being systematically characterized for horticultural traits, pest and disease resistance, and molecular markers.
Roberto Hauagge and James N. Cummins
Dormancy patterns throughout the season were studied in more than 90 apple (Malus ×domestica Borkh.) cultivars and related Malus spp. The seasonal apple bud dormancy pattern resembles a normal curve: it starts to intensify soon after bud formation and reaches maximum intensity by the time of leaf fall/senescence. Genotypes were grouped into three general classes based on maximum dormancy intensity. Maximum intensity of bud dormancy measured in cold winters is inversely related to adaptation to the subtropics. Low-chilling requirement (CR) cultivars have a shallow depth of dormancy with very little alteration throughout the year. High-CR cultivars have intense bud dormancy, the first stage of which can be induced by growing these cultivars at temperatures above 20C. Genotypes differed in their rates of dormancy dissipation. The efficiency of chilling unit (CU) accumulation to break dormancy was negatively correlated with CR, which indicates the importance of factors other than CU accumulation in terminating bud dormancy in low-CR cultivars. The inherent length of bud dormancy plays a major role in determining the time of budbreak in the spring. Deviations may be related to the genotypic efficiency in which chilling modifies dormancy and possibly the basal temperatures to which buds respond. Chill unit requirement and heat unit requirement are dependent factors. Heat requirement comparisons may be meaningless if the dormancy intensities of the genotypes are not taken into consideration.
M.T. Momol, W.F. Lamboy, P.L. Forsline, and H.S. Aldwinckle
Malus sieversii is one of the primary progenitors of the cultivated apple. Since 1989, several collecting trips have been made to central Asia by personnel of the USDA and Cornell Univ. to collect seeds of wild Malus sieversii from many diverse ecosystems. In 1992, an ex situ plot in Geneva, N.Y., was established with trees grown from seed that was collected in three different habitats in Kazakstan, Tajikistan, and Uzbekistan in 1989. In 1995, trees grown from seed that was collected in five additional habitats in Kazakstan and Kyrgyzstan in 1993 were added to the ex situ plot. In the summers of 1995 and 1996, tips of vigorously growing shoots of 1135 seedlings from 79 different populations were inoculated by hypodermic syringe with 5 × 108 cfu/ml of Erwinia amylovora strain Ea273. Seedlings from the 1989 collection were in the fourth and fifth field-growing seasons, with some beginning to bear fruit. Seedlings from the 1993 collection were in first and second field-growing seasons. Results from both seasons indicated that individuals within each of the 79 populations of M. sieversii are resistant to fire blight (defined as ≤20% shoot length infected). Resistance differed among populations, with some populations having no resistant individuals and others having >80% of the seedlings resistant. The range of resistance is quite similar to that seen among apple cultivars from North America and Europe. In another test, some accessions from 1989 collection had sufficient bloom for inoculation in 1995 and 1996. At full bloom, blossoms on these trees were inoculated with the E. amylovora suspensions (5 × 107 cfu/ml) using a backpack sprayer. These also gave diverse resistant reactions.
Roberto Hauagge and James N. Cummins
The chilling requirements (CR) to break bud dormancy in a broad range of apple cultivars (Malus ×domestica Borkh.) and related Malus spp. were assessed by periodic sampling and forcing of field-grown shoots as a function of chill unit (CU) accumulation and/or by the total growing degree hours (GDH) accumulated from leaf fall until the time of budbreak under a simulated subtropical winter. The mean number of CU required to break dormancy of field overwintered shoots varied between 218 ± 113 for `Anna' and 1516 ± 113 for `Wright #1'. However, most genotypes had CR between 800 and 1200 CU. Much wider variation for the length of bud dormancy was observed in plants growing under simulated subtropic winter conditions. Genotypes that had shown the lowest CR values under Geneva, N.Y., winters generally had the highest year-to-year variation in CR estimates. Cultivar bud CR values obtained under cold winters are related to field-observed CR estimates in a subtropical environment, but absolute values may differ markedly. Furthermore, several genotypes that show reasonable adaptation to the subtropics have similar or higher CR than apple cultivars with standard CR under Geneva conditions. In addition, enough CU accumulated under the simulated subtropic winters to break dormancy of standard apple cultivars. However, complete dormancy removal was observed only in cultivars well-adapted to a subtropical environment. This result indicates that in addition to CU accumulation, there are important interactions among cultivars and environmental factors that are responsible for terminating bud dormancy. Several cultivars and wild species have shown resistance to delayed foliation. Among the species, M. brevipes, M. rockii, M. spectabilis, and M. turesii are more tolerant than M. baccata and its hybrids, which are recognized for their adaptation to the subtropic environment.
James J. Luby, Peter A. Alspach, Vincent G.M. Bus, and Nnadozie C. Oraguzie
Incidence and severity of fire blight [Erwinia amylovora (Burr.) Winslow, Broadhurst, Buchanan, Krumwiede, Rogers, and Smith] following field infection were recorded using families resulting primarily from open-pollination of Malus sylvestris (L.) Mill. var.domestica (Borkh.) Mansf. cultivars and a few other Malus Mill. sp. The families were structured as three sublines, planted in three successive years (1992 to 1994), of a diverse population of apple germplasm established at HortResearch, Hawkes Bay, New Zealand. The incidence of fire blight varied among the sublines with the oldest planting exhibiting more fire blight. Flowering trees were more likely to be infected than nonflowering trees, in terms of both incidence and severity. Furthermore, the level of fire blight was related to flowering date, with later flowering trees having higher levels. Thus, family means and narrow-sense heritability estimates were computed after first adjusting the fire blight score for flowering date by fitting a linear model. Provenance of origin of the maternal parent explained little variation except that M. sieversii Lebed. families were more resistant than M. sylvestris var. domestica families in one subline. Family means computed using all trees, and those from only flowering trees were highly correlated. Families from open-pollination of M. honanensis Rehder and M. xhartwiggii Koehne females were among the more susceptible. Those from several European M. sylvestris var. domestica cultivars as well as from M. baccata (L.) Borkh. and M. toringoides (Rehder) Hughes females were among the more resistant families. Narrow-sense heritability estimates ranged from 0.05 to 0.85 depending on the subline, with most estimates between 0.12 and 0.36. They were higher in the two older sublines that consisted primarily of open-pollinated families from M. sylvestris var. domestica, and lower in the younger subline that consisted primarily of M. sieversii, due to lower incidence and severity in the latter subline. Breeders who consider potential complications of juvenility, tree size, and flowering date in relation to infection periods should be able to exploit field epidemics to perform effective selection.
Fenton E. Larsen and Stewart S. Higgins
Tree size, cumulative yield, yield efficiency and anchorage of 6 micropropagated (MP) apple (Malus domestica Borkh.) cultivars were determined in 1991 after 5 years of production, as compared with trees on seedling (sdlg) or M 7a roots. Trees were planted in 1984, with crops harvested from 1987 through 1991. Trees were generally smallest (trunk cross-sectional area) on M 7a and were largest with 4 cultivars (`Delicious', `Jonathan', `Rome', `Spartan') when micropropagated. `Golden Delicious' (GD) was largest on sdlg. Cumulative yield was affected by a scion × rootstock interaction, with few trends in scion or rootstock effects. Mean cumulative yield was 84 kg tree-1, 71 and 58 for M 7a, MP and sdlg, respectively. Yield efficiency was also affected by a scion × rootstock interaction. In 1991, mean yield efficiency was 0.5 kg cm-2 for sdlg and MP trees, but was 1.05 for M 7a. Efficiency on M 7a was superior to other rootstocks with all scions except `GD', while sdlg and MP trees were statistically similar with all scions. All trees leaned in response to prevailing westerly winds, with trees on sdlg tending to be more upright than MP or M 7a trees.
Wesley T. Watson*, David N. Appel, Michael A. Arnold, Charles M. Kenerley, and James L. Starr
Phymatotrichopsis omnivora (Duggar) Hennebert (syn. Phymatotrichum omnivorum Duggar) is a recalcitrant soilborne pathogen that causes serious root rot problems on numerous plant species in the southwestern United States and northern Mexico. Apple trees [Malus sylvestris (L.) Mill. var. domestica (Borkh.) Mansf. (syn. M. domestica Borkh. non Poir.)] are highly susceptible to P. omnivora with most tree death occurring in the summer months. Studies were conducted from 1996 to 1999 to examine when and at what rate infection and colonization of roots of apple trees by P. omnivora actually occurs. In three-year-old trees growing in orchard soils in 45-gallon containers (171,457 cm3) and inoculated with sclerotia in August 1997, infection occurred in the nursery after 12 weeks. For trees inoculated with sclerotia in February 1998, infection occurred within 15 weeks. After 18 weeks, 100% of trees were infected after inoculation in August and 80% of trees were infected after the February inoculation. This information is vital to understanding the epidemiology of Phymatotrichum root rot in apple orchards.
Nobuko Sugimoto and Randy Beaudry
The objective of the experiment was to determine developmental changes in major aroma profiles in `Jonagold' apple (Malus ×domestica Borkh.) and analyze climacteric fruit characteristics. Changes in internal ethylene production, respiration, skin color, texture, and aroma concentration were measured during maturation and ripening of `Jonagold' apple fruit. Patterns for skin color, starch, and internal ethylene content were typical for the variety. Volatile compounds and CO2 increased after a rapid increase in ethylene production. Total ester emission peak coincided with fruit softening. Hexyl acetate, 2-methylbutyl acetate, butyl acetate, and hexyl 2-methylbutanoate were found to be the major volatile compounds detected by GC/MS. Long chain esters, such as hexyl acetate and butyl acetate, contributed during the early stages of ripening and short chain esters such as n-propyl acetate and butyl propanoate increased later. Esters are formed by combining alcohol moiety with CoA derivative of fatty acid moiety by the action of alcohol acyl transferase (AAT). The alcohols butanol, 2-methylbutanol, propanol, and hexanol increased at an earlier developmental stage than the esters for which they acted as substrates.
Warren F. Lamboy, Jing Yu, Phil L. Forsline, and Norman F. Weeden
One of the primary progenitors of the cultivated apple (Malus ×domestica) is M. sieversii, a species native to the forested regions of Central Asia. Despite the horticultural importance of M. sieversii, little is known about its genetic variation. In this study, isozyme diversity at 18 loci was determined for 259 open-pollinated offspring belonging to 31 different maternal half-sib families collected from 14 different populations in 4 regions of central Asia. Genetic diversity statistics were computed from the resulting allele and phenotype frequencies. Cluster analysis of half-sib families showed that there was some grouping based on geographic region, but 16 of the half-sib families were most closely related to half-sib families from other regions. AMOVA, the analysis of molecular variance, indicated that most of the enzyme variability (85%) was attributable to differences among half-sib families within populations, none could be assigned to populations within regions, and 15% was due to differences among regions. In addition, no alleles were found that were both fixed in a region and unique to that region. These results suggest that plants belonging to M. sieversii effectively form a single panmictic population. Thus, a thorough sampling of a few large populations will efficiently capture most of the genetic diversity present in wild M. sieversii.