Natural variation in fruit abscission-related traits in apple (Malus)

Abscission or retention of ripening fruit is a major component of seed dispersal strategies and also has important implications for horticultural production. Abscission-related traits have generally not been targeted in breeding efforts, and their genetic bases remain mostly unknown. We evaluated 144 Malus accessions representing wild species, domestic cultivars, and hybrids for abscission-related traits. We found that seasonal timing of fruit abscission in wild species and hybrids showed a broad distribution similar to that seen for domestic cultivars, and that internal ethylene concentration at the time of abscission varied by over three orders of magnitude. Wild species, domestic cultivars, and hybrids all included representatives that showed abscission of fruit prior to substantial production of ethylene, as well as accessions that retained fruit for a significant period of time following ethylene production. For all accessions that retained fruit, fruit removal resulted in abscission of the pedicel, and exogenous ethylene promoted abscission, suggesting that the abscission zone was functional. Our results suggest important roles for mechanisms independent of fruit ethylene production in abscission.     

RFLP analysis of genetic variation in species of section Arachis, genus Arachis (Leguminosae)

Four A-genome species of the genus Arachis (A. cardenasii, A. correntina, A. duranensis, A. kempff-mercadoi), three B genomes species (A. batizocoi, A. ipaënsis and A. magna),the AABB allotetraploid A. hypogaea (cultivated peanut) and introgression lines resulting from a cross between A. hypogaea and A. cardenasii were analyzed by RFLP. The A genome species (cytologically characterized by the presence of a small chromosome pair ‘A’) were closely similar to each other and shared a large number of restriction fragments. In contrast, the B genome species differed more from one another and shared few fragments. The results of this study indicate that the absence of the small chromosome pair is not a good criterion for grouping species of section Arachis as B genome species, since their genome might be quite distinct from the B genome of A. hypogaea.The lowest genetic variation was detected within accessions of A. duranensis (17 accessions), followed by A. batizocoi (4 accessions) and A. cardenasii (9 plants of accession GKP 10017).The high level of genetic variation found in A. cardenasii might indicate that not all accessions of wild species of Arachis are autogamous, as reported for A. hypogaea.     

Evidence of gene introgression in apple using RAPD markers

Summary A genomic remnant of Malus floribunda clone 821 introgressed into the cultivated apple M. x domestica Borkh. was identified using randomly amplified polymorphic DNA (RAPD) markers obtained by the polymerase chain reaction (PCR). Using a set of 59 oligonucleotide decamer primers, polymorphic DNA markers were identified among three pooled DNA samples. Based on the presence or absence of bands among bulked apple scab-resistant selections and cultivars, bulked scab-susceptible cultivars, and a M. floribunda clone 821 sample, one primer, A 15, identified amplified fragments in the scab-resistant bulked sample that was also unique to the M. floribunda clone 821. The unique band from M. floribunda clone 821 was amplified in four out of 17 scab-resistant selections/cultivars. This RAPD, designated OA15₉₀₀, identifies an introgressed fragment that has as yet no known function.     

Dwarfing and lethal genes in apple progenies

Summary Three dwarf types are described, early dwarf, determined by two recessive genes d ₁ and d ₃, in the double homozygous state, and an additional recessive gene d ₄ and crinkle dwarf and sturdy dwarf each also determined by a recessive gene.The gene l, for pale green lethal was found to be closely linked to the gene V _f , for scab resistance.There was evidence of a regrowth promoting gene, G, in some seedlings homozygous for d ₁ and d ₃.     

DNA fingerprinting of apple (Malus spp.) rootstocks using Simple Sequence Repeats

Identification of apple clonal rootstocks based on visual morphological characteristics is often difficult because of the genetic similarity of many rootstocks and the restriction of observable characteristics at the nursery stage. In this study, a total of 66 rootstock clones representing a sub‐sample of a collection of apple rootstocks from different countries maintained at NIFTS, Morioka, Japan, was screened with seven simple sequence repeat (SSR) markers to establish genetic identities, estimate genetic diversity, verify parentage and examine genetic relationships. All seven SSR loci produced one or two amplified fragments and made it possible to differentiate among the rootstocks, except between virus‐free subclones and their original parents as well as between ‘Aotea 40’ and ‘Aotea 106’. The parentage of all but one rootstock clone was confirmed. The placement of individual rootstock clones into subgroups in a phenogram generated from un‐weighted pair group average cluster analysis was generally based on parentage or genetic origin. Based upon the ability of the SSR data to differentiate these apple rootstocks and the high level of gene diversity, it is proposed that all commercial apple rootstocks be fingerprinted to provide a mechanism to verify the identity of individual clones.     

Inheritance and selection of some fruit characters of apple. II. The relation between leaf and fruit pH as a basis for preselection

Summary The leaf pH of two year old apple seedlings was found to be sufficiently related to the fruit pH, measured when the seedlings were six to seven years old, to serve as a basis for pre-selection. Discarding all seedlings with an above average leaf pH (40% of the total) eliminated the majority (74%) of seedling with low-acid cum sweet fruits. The method was not effective for reducing the proportion of seedlings with high-acid cum sour fruits.     

DNA-RAPDs detect genetic variation and paternity in Malus

Summary DNA amplification fingerprinting using arbitrary primer(s) was applied to the identification of Malus species. Highly variable DNA fragment patterns were clearly detected by polyacrylamide gel electrophoresis of the amplified extension products, although three sports of Delicious exhibited the same fingerprint as the original cultivar. The fingerprinting of two triploid apple cultivars suggested that the parent contributing the 2n gamete was the female. We applied this fingerprinting to paternity analysis of an apple cultivar of which the pollen parent was unknown. By using 5 arbitrary primers and RFLP analysis of the amplified products, one cultivar was singled out for paternity among six putative candidates.     

Genetic variation in a Norwegian germplasm collection of white clover (Trifolium repens L.)

Genetic variation and genotype × environment interactions for important agronomic characters were estimated for eleven Norwegian populations of white clover (Trifolium repens L.), originating from between 58^°52′N and 69^°30′N latitude, and from altitudes of 10 to 450 m a.s.l. The populations were evaluated in monoculture clonal field plots for two years at two locations in Norway. The medium-leaved Danish cultivar Milkanova was also included in the experiment as a standard market cultivar. Considerable variation between populations was observed for winter survival, spring growth, morphological characteristics, dry matter yield, general performance, earliness and seed yield. Winter survival, measured as plant cover after two years, was significantly different at the two locations. All the populations had sufficient winter survival at the southern, continental highland site (61^°07′N).At the northern coastal location (69^°30′N),where the winter climate is unstable with physical stresses such as frost, ice-encasement and water-logging frequently occurring, only the northernmost, coastal populations showed sufficient winter survival. Highly significant interactions were found between populations and locations, and between populations and years for all characters measured at both locations and in both years, respectively. Interaction effects between populations, locations and years were significant only for leaflet length. The largest leaf lamina's were observed in the most southern populations. However, genotypes possessing relatively long leaflets were also found in the northernmost populations, although at low frequencies. In general, the populations originating from the northernmost locations had lower biomass production, and were smaller-leaved, more prostrate and exhibited shorter internodes and stolons compared to the populations from South and Middle Norway. These populations also initiated inflorescences earliest, and were among the lowest seed yielding populations. Otherwise, no clear clinal variation in growth characteristics was observed which could be related to the latitude or altitude of origin of the populations. This revised version was published online in July 2006 with corrections to the Cover Date.     

Microsatellite mapping of complementary genes for purple grain colour in bread wheat (Triticum aestivum) L.

Complementary genes for purple grain colour Pp1, Pp2, Pp3 (now designated Pp1, Pp3b, Pp3a, respectively) were mapped using crosses between purple-grained hexaploid wheats ‘Purple Feed’ – Pp1Pp1/Pp2Pp2 (Pp1Pp1/Pp3bPp3b), ‘Purple’ – Pp1Pp1/Pp3Pp3 (Pp1Pp1/Pp3aPp3a) with non-purple-grained cultivars ‘Novosibirskaya 67’ (‘N67’) and ‘Saratovskaya 29’ (‘S29’). The genes Pp2 (Pp3b) and Pp3 (Pp3a) were inherited as monofactorial dominant when purple-grained wheats were crossed to ‘N67’. Both were mapped in the centromeric region of the chromosome 2A. Therefore, they were suggested being different alleles at the same locus and designated Pp3a and Pp3b. In the crosses between purple-grained wheats and ‘S29’ a segregation ratio of 9 (purple) to 7 (non purple) was obtained suggesting a complementary interaction of two dominant genes, Pp1 and Pp3. To map Pp1 as a single gene, the influence of the other Pp gene was taken into consideration by determining the Pp3 genotype of the F₂ plants. The gene was mapped on chromosome 7BL, about 24 cM distal to the centromere. The Pp1gene was shown to be non allelic to the Rc-1 (red coleoptile) and Pc (purple culm) genes, contrary to what was previously suggested. The colouration caused by the Pp genes has no effect on pre-harvest sprouting.     

Intravarietal polyploidy in the apple (Malus pumila Mill.) cultivar Hazratbali

Summary A triploid cytotype of cv. Hazratbali (Malus pumila Mill.), was found growing in orchards of diploid trees. Morphological comparisons between the two cytotypes reveal that the triploid is more vigorous, and bears more attractive fruit. The meiotic behaviour of the two cytotypes has been described. The impact of polyploidy on quality characteristics of apple at the intravarietal level is analysed. The possible mode of origin of the triploid cytotype is discussed.     

Inheritance and selection of some fruit characters of apple. I. Inheritance of low and high acidity

Summary Fruit acidity as such appears to be gouverned by a single major gene, low acidity being a recessive character. The additional genetic variation is explainable by the additive gene action model. This mode of inheritance causes the pH distribution of seedlings in progenies to shift with a higher mid-parent pH towards more low-acid cum sweet (pH 3.8) and fewer high-acid cum sour (pH 2.9) seedlings or vice versa with a lower midparent pH. As a result, in crosses between heterozygotes (Mama) the total proportion of undesirable seedlings -low plus high-acid ones-, remains similar, irrespective of the mid-parent pH. This proportion constituted about one third of the seedlings. A same proportion, but of mainly sour plants, was found in crosses between relatively acid homozygotes (Mama) or between these and similar heterozygotes. It was less than one quarter if the heterozygous partner was medium-acid and a few percentages only when both the homozygous and heterozygous partner were medium-acid. Choosing parents in the absence of information, chances are that the very large majority of the medium-acid types are heterozygotes, that among the relatively acid ones one in two is homozygous and that the odds to find homozygotes among the sour types are probably best. Attention was drawn to the importance of a high sugar content — which is quantitatively inherited — in parents with a relatively low pH to increase the chances of selecting seedlings with a good sugar/acid ratio.     

The ‘European Apple Genome Mapping Project’-developing a strategy for mapping genes coding for agronomic characters in tree species

Summary A recently initiated collaborative project involving apple breeders in seven European countries is described. The objective is to improve the European apple crop by molecular-aided breeding to increase efficency and reduce the time-scale in breeding for resistance, tree habit and fruit quality. The strategy adopted provides a model for similar studies in fruit, forest and other woody species. The project is based on progenies from a small number of crosses involving many important agronomic genes. Replication of these reference progenies by vegetative propagation will enable studies to be carried out simultaneously in each country. By developing a range of molecular markers, including isozymes, RFLPs and sequence-tagged DNA probes, an integrated molecular map is being constructed for use in a wide range of breeding and genetic studies. Construction of a database recording many mapped molecular markers will enable efficient exploitation of data in future genetic, breeding and physiological studies of apple. Aspects of the adopted strategy, techniques and management are discussed in the context of mapping genes in perennial crop genomes.     

Molecular marker analysis of genetic variation characterizing a grasspea (Lathyrus sativus) collection from central Italy

Levels of genetic similarity characterizing 20 grasspea (Lathyrus sativus L.) populations collected in central Italy (17 populations in the Marche region and three populations in the Abruzzo region) were analysed with amplified fragment length polymorphism (AFLP) molecular markers. Two main clusters were found: one included large‐seeded populations from farms that were not market‐oriented (named Household populations) and the second, small‐seeded populations, cultivated in market‐oriented farms (named Commercial populations). Relationships among populations collected in different regions were found, although one population of the Abruzzo region was placed between the two main clusters, suggesting a possible further genetic differentiation within this grasspea germplasm collection. Principal component analysis based on AFLP marker frequency was effective in identifying polymorphic markers showing high discriminating ability between clusters H and C. In particular, seven markers showing high positive and three markers with low negative PC1 scores showed an almost cluster‐specific distribution. These results will be useful for enhancing Italian grasspea germplasm use in plant‐breeding programmes and for extending grasspea cultivation within the sustainable agricultural systems of central Italy.     

Screening for resistance to Nectria galligena Bres. in cut shoots of apple

Summary To find a fast and reliable test to assess resistance to Nectria galligena in apple, different methods of inoculation were compared using macroconidia of N. galligena and one-year-old cut shoots from mature trees of Cox's Orange Pippin, IVT 69078-19, James Grieve and Jonathan.With the best inoculation method 11 genotypes were screened for resistance. Elstar, Golden Delicious, Jonathan and Lombart's Calville were highly resistant and the level of resistance of Ingrid marie, Gloster, Melrose, IVT 69078-19, Cox's Orange Pippin, James Grieve and Idared decreased in this order.The best inoculation method proved to be simple, giving results within four to nine weeks after inoculation.     

Pollination with gamma-irradiated pollen and development of fruits,seeds and parthenogenetic plants in apple

Summary Four apple (Malus X domestica) genotypes, Erovan, Golden Delicious, R1-49 and X6677, were pollinated with a marked pollen irradiated by -rays at doses ranging from 125 to 1000 Gy. Pollination with such irradiated pollen affected fruit set, seed number and seed contents, and induced the formation of parthenocarpic fruits and the development of parthenogenetic embryos. The immature embryos extracted from seeds. 2 and 3 months after pollination, were cultured in vitro and germinated after 2 months of cold treatment (3°C). Haploid plants were obtained in all 4 genotypes, after pollination with pollen irradiated at doses from 200 to 500 Gy. The optimum conditions for induction of apple haploids, by irradiated pollen approach, have been established.     

Geographic distribution of variation in quantitative traits in a world lentil collection

Summary In a world lentil collection the distribution of variation amongst accessions from 13 major lentil-producing countries was examined on the basis of nine quantitative morphological characters by discriminant analysis and canonical analysis. Stepwise discriminant analysis revealed major differences between accessions from different countries. Three major regional groups were apparent: 1) a levantine group (Egypt, Jordan, Lebanon and Syria, 2) a more northern group composed of Greece, Iran, Turkey, and USSR, and 3) accessions from India and Ethiopia with strikingly similar quantitative morphological characters. Misclassifications of individuals within groups were frequent. Characters useful in discriminating between accessions from different countries were in descending order of importance: time to maturity, lowest pod height and 100-seed weight. The regional grouping indicates the importance of local adaptation through clusters of associated characters with phenological adaptation to the ecological environment as the major evolutionary force in the species.     

Pollen grain germination inside the anthers of two chasmogamous angiosperms: Almond (Prunus amygdalus L. Batsch) and apple (Malus pumila Mill.)

Summary Pollen grain germination inside anthers has so far been known in only eleven chasmogamous angiospermous species. The discovery of this phenomenon in some varieties of Prunus amygdalus (almond) and Malus pumila (apple) is therefore significant. The anomaly appears to be genetically controlled, the gene expression occurring under specific environmental conditions.     

The effects of apple rootstock genotypes on mineral content of scion leaves

Summary The relative effects of year to year variation and rootstock genotypes on the major content of leaves of four apple scion cultivars were studied. For nitrogen and phosphorus the main source of variation was due to years. For potassium, magnesium and calcium there were significant genetic effects. The results are discussed in relation to breeding for improved efficiency in mineral uptake in apple rootstocks. Particular reference is made to uptake of minerals associated with physiological disorders of stored apples.