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.     

Application of the TRAP technique to lettuce (<Emphasis Type="Italic">Lactuca sativa</Emphasis> L<Emphasis Type="Italic">.</Emphasis>) genotyping

Summary To demonstrate the applicability of the target region amplification polymorphism (TRAP) marker technique to lettuce genotyping, we fingerprinted 53 lettuce (Lactuca sativa L.) cultivars and six wild accessions (three from each of the two wild species, L. saligna L. and L. serriola L.). Seven hundred and sixty-nine fragments from 50 to 900 bp in length were amplified in 10 PCR reactions using 10 fixed primers in combination with four fluorescent labeled arbitrary primers. Three hundred and eighty-eight of these fragments were polymorphic among the 59 Lactuca entries and 107 fragments were polymorphic among the 53 lettuce cultivars and the six wild accessions; 251 fragments were present only in the wild species. These markers not only discriminated all cultivars, but also revealed the evolutionary relationship among the three species: L. sativa, the cultivated species, is more closely related to L. serriola than to L. saligna. Cluster analysis grouped the cultivars by horticultural types with a few exceptions. These results are consistent with previous findings using RFLP, AFLP, and SAMPL markers. The TRAP markers revealed significant differences in genetic variability among horticultural types, measured by the average genetic similarity among the cultivars of the same type. Within the sample set, the leaf type and butterhead types possessed relatively high genetic variability, the iceberg types had moderate variability and the romaine types had the lowest variability. The genetic behavior of TRAP markers was assessed with a mapping population of 45 recombinant inbred lines (RILs) derived from an interspecific cross between L. serriola and L. sativa. Almost all the markers segregated in the expected 1:1 Mendelian ratio and are being incorporated into the existing lettuce linkage maps. Our results indicate that the TRAP markers can provide a powerful technique for fingerprinting lettuce cultivars. The U.S. Government's right to retain a non-exclusive, royalty-free license in and to any copyright is acknowledged.     

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.     

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 of Sources of Field Resistance to Bremia lactucae in Wild Lactuca Species

Thirty-one accessions of four Lactuca species (L. serriola, L. saligna, L. aculeata, L. squarrosa) and one L. serriola×L. sativa hybrid were screened for field resistance to Bremia lactucae. In the L. serriola group, significant differences in the level of field resistance were distinguished. The accessions PI 281876 and PI 253467 were free of infection. Moreover, all L. saligna accessions tested were totally free from disease during three-year-investigation.     

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.     

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).     

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 to a new biotype of the lettuce aphid Nasonovia ribisnigri in Lactuca virosa accession IVT280

Host plant resistance is an effective protection strategy to control aphids in many crops. However, the evolution of insensitive aphid biotypes necessitates the search for new resistance sources. Wild relatives of crop plants can be important sources for resistance genes to be introgressed into new cultivars. Nasonovia ribisnigri (Mosely) (Homoptera: Aphididae) is an important pest of cultivated lettuce, Lactuca sativa. Since 1982, resistance introduced into lettuce cultivars has relied on the Nr-gene, originating from a wild relative, Lactuca virosa. In 2007 first reports appeared that Nr-based resistance had become ineffective against certain populations of N. ribisnigri. The objective of this study was to establish if the original donor of the Nr-gene L. virosa accession, IVT 280, is resistant against recently emerged virulent (Nr:1) N. ribisnigri biotypes. To this end we investigated feeding and penetration behaviour of virulent and avirulent (Nr:0) aphids on the resistant L. virosa, IVT 280, and two susceptible L. virosa accessions, using the electrical penetration graph method. Additionally, aphid performance was analysed in terms of survival, development time and reproduction on these accessions. L. virosa accession IVT 280 was resistant against all populations of N. ribisnigri tested. The ingestion of phloem was strongly reduced on the resistant accession compared to the susceptible L. virosa accessions. Additionally, none of the aphids survived on the resistant accession that, therefore, constitutes a good source of resistance in lettuce against both biotypes of N. ribisnigri.     

The response to saprol (systemic fungicide) in lettuce species and cultivars and its inheritance

Summary The response of Lactuca sativa, L. serriola and L. saligna to saprol was studied. Spraying the whole plant or part of a leaf caused wilting of susceptible plants. A similar effect was observed when leaves were soaked in a solution containing saprol.Inheritance studies with L. sativa and L. serriola showed that resistance to the wilting reaction caused by saprol is controlled by a single recessive gene.Contribution no. 198-E, 1978 series.     

Sources and inheritance of resistance to stemphylium leaf spot of lettuce

Summary None of the tested cultivars of lettuce was found resistant to Stemphylium leaf spot, a common disease in Israel. Within a Lactuca saligna population collected in wild lettuce in Israel, resistance was traced. Interspecific crosses of L. saligna x L. sativa were made and the mode of inheritance of resistance to this disease was studied. Resistance is apparently controlled by two genes: one dominant (Sm₁) and one recessive (sm₂).Contribution No. 1176-E 1984 series, from the ARO.     

DNA polymorphisms in Oryza sativa L. and Lactuca sativa L. amplified by arbitrary primed PCR

Summary Thirty-five rice (Oryza sativa L.) varieties, including 18 japonica, 5 javanica and 12 indica subspecies and 12 lettuce (Lactuca sativa L.) varieties were identified taxonomically, using PCR with originally designed 21 RAPD (Random Amplified Polymorphic DNA) primers and 8 sequence-specific primers, used for amplifying four specific DNA fragments. Use of these primers revealed polymorphisms among varieties in rice and lettuce and facilitates DNA fingerprinting. Dendrograms of both species based on polymorphisms were constructed and genetical relationships were established. In rice, half the number of amplified bands were polymorphic and almost all varieties differentiated. However, differentiation of minor genetic alterations among somaclonal variants or mutants and their mother varieties was not feasible. In L. sativa, 47% of the amplified fragments were polymorphic and all 12 varieties were differentiated. Some of the PCR fragments were variety or type specific, which could be used for indicators for type-selection. The dendrogram obtained showed differentiated clusters of crisphead, leaf and butterhead type, findings in good accord with the classification based on the genetic background.     

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.     

Inheritance of insensitivity of lettuce to a surplus of exchangeable manganese in steam-sterilized soils

Summary After steam sterilizing of certain glasshouse soils a surplus of exchangeable manganese (SEM) occurs. Great differences exist between lettuce genotypes for sensitivity to SEM.Research on the inheritance of insensitivity was carried out by intererossing five lettuce genotypes: Lactuca sativa cys. Neptune (sensitive). Plenos. Troppo and Celtuce (all insensitive) and an accession of L. scrriola (insensitive). From analysing various populations (as F₁'s. F₂'s and B₁'s) it appeared that different numbers of genes for insensitivity were present in the parents varying from one gene in Plenos and Troppo, two genes in L. serriola to possibly four genes in Celtuce. Genes in the three first mentioned parents were linked in repulsion phase and a chromosome map for these genes was made.     

Molecular diversity and genome-wide linkage disequilibrium patterns in a worldwide collection of <Emphasis Type="Italic">Oryza sativa</Emphasis> and its wild relatives

Genetic diversity analysis within a species is vital for understanding evolutionary processes at the population and genomic levels. We report a detailed study of molecular diversity, polymorphism and linkage disequilibrium in three groups of rice (Oryza) germplasm accessions based on 176 SSR markers. The first group included 65 rice (O. sativa L.) accessions introduced from seven countries, including five regions of China. The second group included 58 US rice varieties released in the past 25 years. The third group consisted of 54 accessions of rice wild relatives represented by ten different species. The number of alleles per SSR marker ranged from 4 to 32 with a mean of 16 alleles and the polymorphism information content values ranged from 0.43 to 0.91 with a mean of 0.70. The variation in SSR alleles was a significant contribution to the genetic discrimination of the 177 accessions within the three Oryza groups. Analysis of molecular variance identified deviation from Hardy–Weinberg equilibrium. Principal coordinates analysis clearly separated the accessions into their respective three groups. Neighbor-joining phylogenetic cluster reflects the ordination of each accession. Linkage disequilibrium (D′) averaged 0.75 in wild Oryza spp., and about 0.5 in both US and international O. sativa accessions. Our results showed that LD among adjacent loci in both O. sativa and Oryza spp. accessions is strong enough to be detecting marker-trait association via genome-wide scans.     

Screening wild Lycopersicon species for resistance to powdery mildew (Oidium lycoperiscum)

Since the late 1980s powdery mildew, designated Oidium lycopersicum, frequently invaded the tomato crop in Western Europe. All commercial cultivars are susceptible. To screen for resistance in wild species a reliable and efficient disease test was developed. Young plants with two to three true leaves are inoculated at high relative humidity by spraying with a freshly prepared suspension of 2×10⁴ conidia, ml^–1. Symptoms are periodically evaluated according to a scale based on the percentage of leaf area with mycelium.One hundred and twenty seven accessions, representing eight wild Lycopersicon species, were screened for resistance to O. lycopersicum. A large variation in resistance was found between species. L. hirsutum was the most resistant species; L. pennellii was moderately resistant; species of the subgeneric group of L. esculentum and of the peruvianum-complex were all susceptible. L. parviflorum was classified separately due to a large variation between accessions. Except for this species, a low variation was found between accessions within species. High levels of resistance were observed in four accessions of L. hirsutum, in one of L. parviflorum and in one of L. peruvianum. This resistance is characterized by a very low disease incidence and a strongly restricted mycelium growth and lack of sporulation.     

High salt tolerance potential in Lycopersicon species during germination

Summary The potential to improve seed germination responses to salinity was evaluated for 13 accessions representing six wild Lycopersicon species and 20 accessions of L. esculentum. Germination response times increased in all accessions at 100 mM NaCl. Analysis indicated that one accession of L. peruvianum (PI126435) germinated faster under high salinity than all other accessions and was closely followed by L. pennellii (LA716). The fastest germinating L. esculentum accession, PI174263, ranked third. Additional wild ecotypes exhibiting rapid germination at 100 mM NaCl were identified among L. pimpinellifolium and L. peruvianum.     

Response of Lettuce Cultivars Carrying the Resistance Gene Dm11 to Isolates of Bremia lactucae from Lactuca serriola

Seven Bremia lactucae isolates from Lactuca serriola were tested for pathogenicity on 41 Lactuca sativa cultivars carrying a race specific resistance gene Dm11 which originated from L. serriola. Some of these cultivars also carried other Dm genes. Sporulation was observed in 34 % of the 287 cultivar/isolate combinations, but it was limited in most cases. A compatible response was recorded in 7.3% of cultivar/isolate. Cultivars with Dm 11 alone, or in combination with Dm 2, exhibited a higher decree of susceptibility then those also carrying Dm 3. There were differences in specific virulence among the isolates under study. Nearly all the isolates studied were virulent on newly described resistance genes Dm 15, Dm 16, and Dm 7 +? which originate from L. serriola.