New sources of major gene resistance in Lactuca to Bremia lactucae

Summary A total of 1789 accessions of several lettuce collections was screened to find new major gene resistance to the downy mildew fungus Bremia lactucae Regel. The accessions belonged to the species Lactuca sativa (N=1288), L. serriola (N=399), L. saligna (N=52) and L. virosa (N=50). A total of 20 races of B. lactucae were used, 14 of which were NL-races, isolated from cultivated lettuce in the Netherlands. The other six races were isolated from wild L. serriola in Czechoslovakia. The accessions were initially screened with two races: NL1 and NL3. Accessions with resistance to one or both of these races were tested with the other races. Phenotypes with new resistance were found in accessions of all four Lactuca species. Of L. sativa, four accessions were found with resistance phenotypes that could not be explained by combinations of known major genes. Many accessions of L. serriola had resistance phenotypes that indicated the presence of unknown resistance genes. All interactions between accessions of L. saligna and races of B. lactucae were incompatible in leaf disc tests, except for four accessions, which showed some sporulation with race NL6. Several accessions of L. virosa were resistant to all races used. Other accessions of L. virosa gave a race-specific interaction with B. lactucae.     

Inheritance of resistance to Zucchini yellow mosaic virus in the interspecific cross Cucurbita maxima × C. ecuadorensis

Summary More than 200 accessions of three wildLactuca species were screened in the laboratory for race-specific resistance toBremia lactucae. Only a fewLactuca entries showed resistance both as seedlings and in a leaf disc test. Accessions ofL. serriola andL. sativa had a low degree of resistance. The best entries were also compared under field conditions. TheL. saligna entries were totally free from disease in the field test. High resistance was also recorded in a fewL. serriola entries and in lettuce cultivars such as Saffier and Mariska.     

Resistance in Lactuca spp. to Microdochium panattoniana (lettuce anthracnose)

Summary Four hundred and forty-nine lines of Lactuca spp. were screened for resistance to an isolate of Microdochium panattoniana. Eighty-seven lines were resistant. Forty-four lines were then screened for resistance to four isolates; twelve groups of lines were identified on the basis of their pattern of resistance. A differential set of lines was used to analyse nine further isolates; this revealed five races of M. panattoniana. Salad Bowl was the only cultivar found to be resistant to three races. A line of L. saligna, UC83US1, was the only line found to be resistant to all the isolates tested.     

Resistance of lettuce (Lactuca) to the leaf aphid Nasonovia ribis nigri. 1. Transfer of resistance from L. virosa to L. sativa by interspecific crosses and selection of resistant breeding lines

Summary Interspecific crosses were carried out between the three Lactuca species L. virosa, L. serriola and L. sativa to transfer resistance to the leaf aphid Nasonovia ribis nigri from L. virosa to the cultivated lettuce. L. sativa, L. serriola was used as an intermediate parent between the other two species. Many irregularities were observed in the interspecific hybrids, ranging from premature dying of F₁ plants resulting from a kind of bastard necrosis till complete male and female sterility and deviating microsporogenesis. Using in vitro culture and after several backcrosses male and female fertile plants were obtained with a L. sativa habit and with resistance to the leaf aphid.     

Identification of sources of resistance in lettuce to the lettuce root aphid, Pemphigus bursarius

In a series of glasshouse and laboratory experiments, lettuce varieties and wild Lactuca species were screened for resistance to the lettuce root aphid, Pemphigus bursarius (Ron.). Several new sources of resistance to P. bursarius development and survival were identified. Extremely high levels of resistance were found in the wild species L. saligna, L. perennis and L. virosa. Resistance was also identified in the lettuce variety Grand Rapids. This resistance is controlled by one or two genes. At least one of these genes is not allelic to the existing Lra gene. Grand Rapids therefore represents a new source of resistance to P. bursarius. This revised version was published online in August 2006 with corrections to the Cover Date.     

The identification and characteristics of field resistance to lettuce downy mildew (Bremia lactucae Regel)

Summary Eighty-one accessions of three Lactuca species which showed no recognisable race specific resistance to Bremia lactucae when tested in the laboratory as seedlings, exhibited different degrees of susceptibility when exposed to natural field infection. As a group, crisp genotypes had less mildew and a slower rate of disease development than other types of lettuce. Wild forms of Lactuca sativa and Lactuca serriola were particularly susceptible. In a further trial, the low field susceptibility of three lettuce cultivars (Iceberg, Batavia blonde de Paris and Grand Rapids) was confirmed. Disease development on cv. Iceberg was compared to that on the highly susceptible cv. Hilde in experiments where the two cultivars were grown either in close proximity or in isolation. The absolute level of attack on cv. Iceberg depended upon disease pressure and differences between the two types only became apparent approximately 8 wk after sowing.     

Giemsa N-banding pattern in cabbage and Chinese kale

Summary In cabbage (Brassica oleracea var. capitata) and Chinese kale (B. oleracea alboglabra) four types of N-bands can be distinguished: pericentromeric, telomeric (terminal), intercalary and satellite bands. Typical NOR bands were not observed. The pericentromeric bands appear at the pericentric regions, possibly even at the centromeres of all chromosomes. Telomeric bands are observed on the short arms of chromosomes 1,5 and 6 in cabbage and chromosomes 1 and 5 in Chinese kale. Intercalary bands stained weakly in the long arms of chromosome 3 in cabbage and chromosome 2 in Chinese kale. Satellite bands cover the entire satellites in both Brassica species. The N-banding pattern is very similar in appearance to the C-banding pattern in both species and much more convenient to apply.     

Sexual and somatic hybridization in the genus Lactuca

Summary Various genes for disease resistance identified in wild Lactuca are difficult, even impossible to exploit in lettuce breeding, due to sexual incompatibility between L. sativa and wild Lactuca sp. We adapted two cellular biology techniques to overcome these interspecific barriers: in vitro embryo rescue and protoplast fusion. In vitro rescue of immature embryos was used successfully for sexual hybridization between L. sativa and L. virosa. Vigorous hybrid plants were produced between L. sativa and seven accessions of L. virosa. Protoplast fusion permitted the regeneration of somatic hybrids between L. sativa and either L. tatarica or L. perennis. Hybrids between L. sativa and L. tatarica were backcrossed to L. sativa.     

The location and characteristics of novel sources of resistance to Bremia Lactucae Regel (downy mildew) in wild Lactuca L. Species

Summary Twenty-one accessions of 3 wild Lactuca species which could be hybridised with L. sativa, the cultivated lettuce, were inoculated at different stages of plant development with 3 multivirulent isolates of Bremia lactucae. Nineteen sources of resistance to B. lactucae, not attributable to the previously recognised resistance factors 1–11 were identified. Two lines of L. serriola showed similar resistance patterns as lines carrying R11. The resistance of some accession was incomplete particularly at the seedling stage and this phenomenon may be race specific.Tests on segregating F₂ populations of crosses between 2 different L. serriola accessions and L. sativa cultivars showed that the resistance in one line (LSE/18) appears to be inherited as a single dominant gene, which is sometimes incomplete in expression and allelic to either Dm6 or R7. The segregation patterns for resistance in PI 281876 did not give readily interpretable ratios.To assess the frequency of occurrence in B. lactucae populations of virulence factors to overcome this novel resistance, 11 of the novel sources of resistance were inoculated with numerous collections of the pathogen from the UK, Czechoslovakia and elsewhere and found to show a high level of resistance.     

Resistance of wild emmer wheat to stem rust: Ecological,pathological and allozyme associations

Summary Seedling resistance to wheat stem rust was determined in populations of wild emmer wheat, Triticum dicoccoides, and characterized by means of ecological factors and allozyme genotypes. Reactions to wheat stem rust were studied in 102 single plant accessions of T. dicoccoides from ten populations by inoculation with Puccinia graminis tritici race 14, isolate GSR-739. Six populations displayed different degrees of response polymorphism with reactions ranging from high resistance to complete susceptibility, whereas four populations contained only susceptible plants. In some of the accession, unexplained intrasib variation in resistance and intraplant variation of infection-types were found. Resistance to stem rust was negatively correlated with two ecological factors, altitude and number of Sharav (hot-dry) days which are unfavorable to disease development. Variation in stem rust response was shown to exist in ecogeographic regions where climatic variables enhanced the development of the fungus, conceivably maintained by natural selection. Likewise, allozyme genotypes, single or in multiple loci combinations, appeared to be associated with resistance or susceptibility to rust. Such association need to be verified by genetic studies in order to become established as useful markers.     

Partial resistance in lettuce to downy mildew (Bremia lactucae). 1. Search for partially resistant genotypes and the influence of certain plant characters and environments on the resistance level

Summary In three bicyclic inoculation experiments in the field 756 genotypes from the IVT Lactuca gene bank were screened for differences in partial resistance against Bremia lactucae. Significant differences between genotypes were found. No linkage was found between the presence or absence of R-genes and partial resistance nor was there a clear relationship between plant characters such as size, erectness and lettuce type and resistance level. Comparison of test results for genotypes which were tested twice and in different years and/or environments revealed a rather good relationship. So no clear genotype × environment interactions for resistance level were found. Results imply that the bicyclic inoculation experiments are useful to determine levels of partial resistance.     

Differences in resistance of wild Lactuca species to natural infection of lettuce powdery mildew (Erysiphe cichoracearum)

Summary The natural infection of Erysiphe cichoracearum on 29 accessions of five wild Lactuca species (L. serriola, L. saligna. L. virosa, L. aculeata and L. dentata) and on one hybrid of L. serriola x L. sativa has been investigated for two and three years, respectively. No infection was observed on L. serriola (PI 255665), L. saligna (LSA/92/1 and LSA/92/2), L. virosa (LVIR/26 and LVIR/57/1) and L. dentata (PI 234204). The level of susceptibility of the control cultivars of L. sativa was found to be moderate (Fila) to fairly high (Hilde).     

An electrophoretic study of esterase and peroxidase isozymes in Brassicoraphanus

Summary A comparative electrophoretic study of esterase and peroxidase isozymes from the leaves of Brassicoraphanus and its parental species (Brassica japonica and Raphanus sativus) was carried out by means of the polyacrylamide gel isoelectrofocusing technique. The isozyme bands of Brassicoraphanus could be regarded as a summation of parent-derived bands, some of which were missing, in addition to some new bands. The qualitative and quantitative variation of isozyme patterns among individual plants was found within each strain of Brassicoraphanus as well as each parental species. The range of the enzymatic variation of a certain strain seemed to reflect the genetic homogeneity of the strain in question. Every strain of Brassicoraphanus was less variable in esterase patterns than the parental species, but in peroxidase patterns the variations of Brassicoraphanus were intermediate between those of both parents. Some strains of Brassicoraphanus were uniform in isozyme patterns, whereas others were variable. The difference in enzymatic variation among strains was perhaps due to the difference in the source of the strains and the selection which they received.     

Genome size and Giemsa C-banded karyotype of tetraploid Bromus ciliatus L.

Summary Tetraploid Bromus ciliatus L. is a North American bromegrass that has been placed in the Pnigma section of Bromus. The objective of this study was to characterize the genome of tetraploid B. ciliatus by cytogenetic methods and compare it to the genomes of other species included in the section Pnigma. All the plants of the accession (USDA PI 232214) selected for chromosome counting were tetraploids (2n = 28). The mean 2C nuclear DNA content for tetraploid B. ciliatus was 19.13± 0.07 pg as determined by flow cytometry which is significantly greater than the tetraploid DNA content of B. inermis Leyss. (11.74± 0.16 pg). C-banding procedures were used to identify individual mitotic chromosomes and to develop a karyotype for B. ciliatus. The genome of the tetraploid B. ciliatus consisted of 16 median chromosomes, eight submedian chromosomes, and four chromosomes with satellites which included one pair with a large satellite and one pair with a small satellite. The general pattern of the distribution of constitutive heterochromatin in B. ciliatus was quite different than the other bromegrasses that have been analyzed to date. Except for two pairs of chromosomes, all chromosomes in tetraploid B. ciliatus had telomeric bands on one or both arms. Some of the chromosomes with telomeric bands had centromeric bands that were located at one or both sides of the centromere and intercalary bands which were generally absent in the other bromegrass species. It was possible to identify all chromosomes of tetraploid B. ciliatus and to match the pairs of homologous chromosomes by using chromosome lengths, arm length ratios and C-banding patterns. The results of this study indicate that tetraploid B. ciliatus has different genomes than the European species evaluated to date in the section Pnigma.     

Identification of Vigna spp. through specific seed storage polypeptides

Summary Seed globulins of ten Vigna species were separated by means of SDS electrophoresis. Both inter and intraspecific variation were observed.Groups of specific bands which do not vary in the species being present in all the accessions of the species were identified. These species-specific bands, allow the identification of the 10 Vigna spp. analyzed.     

PCR-RFLP analysis of the whole chloroplast DNA from three cultivated species of cotton (Gossypium L.)

Variations of the chloroplast DNA (cpDNA) from three of the four cultivated species of cotton (Malvaceae); Gossypium barbadense L., Gossypium hirsutum L., Gossypium arboreum L. and its synonym Gossypium nanking Meyen., were analyzed. Using specific set of primers, the whole circular cpDNAs from the four test species were amplified. These were subsequently digested with the use of seven restriction enzymes. The amplified fragments of the whole cpDNAs of the diploid cultivated cotton G. arboreum and its synonym G. nanking did not show any differences. However, the allotetraploid cultivated cottons G. barbadense and G. hirsutum, showed some fragment length differences directly visible after amplification and two types of restriction fragment length polymorphism (RFLP), the first appeared as slightly lengthened bands and the other as gain or loss of a restriction site. The results also showed that the chloroplast genomes of the allotetraploid cultivated cottons are highly similar to the diploid cultivated cottons tested in terms of length and digestion patterns. The detected amplified length differences, RFLPs and the restriction sites can be considered as species specific markers for the allotetraploid cultivated cottons, which could be a useful tool for future studies of the cpDNA of the genus Gossypium L.     

Application of DNA fingerprints for identification and genetic analysis of Carica papaya and other Carica species

Summary Various DNA fingerprint probes were applied to Carica papaya and other Carica species for both identification and genetic analysis.Each of the Carica papaya cultivars is characterized by a specific DNA fingerprints pattern. Various Carica species also have specific patterns which distinguish them from one another. Band sharing levels were used to estimate the relatedness between the various Carica species.Genetic analysis of 11 progeny from a cross between the Carica papaya cultivars 17/82 and 112 suggests that application of DNA fingerprinting to Carica papaya breeding, could make the process more efficient. Genetic analysis of the DNA fingerprint bands revealed no linkage or allelic relationship among the bands analyzed, indicating that these loci are not clustered in the Carica genome.     

Genetic diversity of wheat wild relatives in the Near East detected by AFLP

In order to reveal the molecular genetic diversity of wheat wild relatives, an AFLP analysis was conducted with 16 accessions of five Triticum andAegilops species originating from the Near East. Variation within population was studied with at least seven individuals per accession. Four primer combinations were used for selective amplification. Based on the scored bands, we estimated percentage of polymorphic bands, 1 – proportion of shared bands (1-psb) and nucleotide diversity (π). Of the five species used in this study, Ae. speltoides had the highest level of `within population' variation. This species had also the highest value of the variation among populations. As for Triticum species, the level of variation within population was low in diploid species (T. urartu and T. boeoticum),whereas two tetraploid species (T. dicoccoides and T. araraticum) had relatively high levels of variation within population. While the two diploid Triticum indicated a clear interspecific divergence, the two tetraploid wild wheats were not clearly divergent in this study. The variance portioning analysis indicated that the variation detected for diploid Triticum species was mainly composed of `between species' variation, on the other hand that for tetraploid Triticum was mostly composed of `within population' variation. In conclusion, AFLP analysis reveals molecular variation in all accessions used in this study, suggesting a potential genetic diversity of the wheat wild relatives in natural populations. These results have implications for the design of strategies to maintain genetic diversity within genebank collections. This revised version was published online in August 2006 with corrections to the Cover Date.     

Assessment of genetic relationships in Mentha species

A set of 60 random primers was used to analyse 11accessions from six taxa of Mentha developed byCIMAP. These accessions were maintained in the nationalgene bank for medicinal and aromatic plants at CIMAP.A total of 630 bands could be detected as amplifiedproducts upon PCR amplification, out of which 589 werepolymorphic (93.5%). Further analysis of these RAPDprofiles for band similarity indices clearlydifferentiated five of the Mentha arvensis L.accessions from the rest. Among two accessions of Mentha spicata L. CIMAP/C33 could bedistinguished from CIMAP/C32. Mentha × gracilis Sole cv. cardiaca showed a muchhigher similarity with Mentha spicata L. as wellas Mentha arvensis L. which amongst themselvesshowed rather a greater distance indicating that Mentha × gracilis Sole cv. cardiaca might have evolved as a natural hybridbetween M arvensis L. and M. spicataL. In terms of uniqueness of amplified bands fordeveloping RAPD markers, it was observed that at taxalevel 298 bands were unique to one of the six taxa,singly amounting to 47.3% of total amplifiedfragments. Primers MAP 10 and 17 produced polymorphismonly in case of M. spicata L. and Menthaspicata L. cv. viridis while MAP 08 producedpolymorphic bands in all 4 other species than thesetwo. Similarly unique patterns were observeddifferentiating all six species and could be used asRAPD markers for differentiating Mentha species.     

Species affinities between Cajanus cajan and some Atylosia species based on esterase isoenzymes

Summary Esterase isozymes were studied in seed extracts of Cajanus cajan and six Atylosia species by polyacrylamide gel electrophoresis and isoelectrofocusing. The isozyme patterns were stable and accession specific. Within the accessions of the Atylosia species, A. albicans and A. scarabaeoides showed three common bands indicating that they are more closely related to each other than to the other species. Of the accessions of Atylosia only A. cajanifolia shares the esterase isozyme of C. cajan and hence seems to be the closest wild relative of C. cajan.