Characterization and inheritance of partial resistance to downy mildew, Peronospora parasitica, in breeding material of broccoli, Brassica oleracea convar. botrytis var. italica

Twenty doubled haploid broccoli lines from breeding material were evaluated for resistance to Peronospora parasitica at the seedling stage. All lines supported sporulation of the pathogen but to varying extents and intensities. Partial resistance of the more resistant lines,‘br8’and ‘br9′, reduced conidia production on cotyledons by 50-70% compared with the most susceptible lines. Inoculation of the two most resistant lines with 13 isolates of different geographical origins revealed that ‘br9′, showed a rather uniform level of resistance to all isolates while ‘br8’showed some isolate specificity Partial resistance was evaluated in six of the broccoli lines in a half diallel set of crosses. Disease assessment of seedlings showed that additive genetic effects explained 45.8% and 31.8% of the total variation of sporulation score and conidia production, respectively This suggests that recurrent selection for partial resistance to P. parasitica in early generation in breds or in populations of broccoli will be efficient to obtain cotyledon resistance.     

A single dominant gene for downy mildew resistance in broccoli

Downy mildew, incited by Peronospora parasitica (Pers.: Fr.) Fr., is a destructive disease of broccoli (Brassica oleraceaL., Italica Group). Resistant cultivars represent a desirable control method to provide a practical, environmentally benign, and long-term means of limiting damage from this disease. Doubled-haploid (DH) lines developed by us exhibit a high level of downy mildew resistance at the cotyledon stage. To determine the mode of inheritance for this resistance, a resistant DH line was crossed to a susceptible DH line to make an F₁, from which F₂ and backcross (BC) populations were developed. All populations were evaluated for response to artificial inoculation with P. parasitica at the cotyledon stage. All F₁ plants (including reciprocals) were as resistant as the resistant parent, indicating no maternal effect for this trait. F₂ populations segregated approximately 3resistant to 1 susceptible, BC populations using the resistant parent as the recurrent parent contained all resistant plants, and the BC to the susceptible parent segregated 1 resistant to 1 susceptible. These results indicate that resistance is controlled by a single dominant gene. This gene should be easily incorporated into F₁ hybrids and used commercially to prevent downy mildew at the cotyledon stage. This revised version was published online in August 2006 with corrections to the Cover Date.     

Variation in virulence and resistance in the bean-bean rust pathosystem in Ecuador

Twenty two Uromyces appendiculatus isolates were tested on 20 differential and 25 Ecuadorian Phaseolus vulgaris cultivars in the seedling stage. Based on the infection types 20 races could be discerned. The Ecuadorian cultivars differed greatly in their reaction to the isolates, from resistant to only one isolate (`Red Small Garden') to resistant to all isolates (’G2333‘). The isolates showed a wide range of virulence to the Ecuadorian cultivars, from virulent to only two cultivars (isolate 13) to virulent to 21 cultivars (isolates 5 and 23). Seven cultivars with a basically susceptible infection type appeared to differ greatly in quantitative resistance when tested in three consecutive crop cycles. The disease severities in percentage leaf area affected averaged over the three cycles ranged between 83.9% for ‘Red Small Garden’ and 13.1% for ‘INIAP-414’. Race-specific resistance does not seem an advisable breeding strategy, but the quantitative resistance offers a good alternative.     

Evidence for the production of unreduced gametes by tetraploid Rosa hybrida L.

The objective of this study was to determine the mode of inheritance of field resistance to downy mildew (Peronospora parasitica (Pers. ex Fr.) Fr.) in broccoli (Brassica oleracea var.italica) at the adult plant stage. The F₁, F₂ and F₃ progeny of resistant and susceptible plants of broccoli were tested in the field under natural infection, in central Portugal, from August to December in two successive years. The plants were evaluated for resistance to downy mildew at maturity using a five-class scale of increasing susceptibility to the disease, which took into account the number of infected leaves and the size of the sporulating lesions. The F₁ was completely resistant, the F₂ segregated a clear 3 resistant: 1susceptible and the F₃ confirmed the F₂ segregation, which suggests a dominant character controlled by a single locus. This resistance has good potencial for direct use in commercial broccoli breeding or for transfer to other Brassica vegetables. This revised version was published online in August 2006 with corrections to the Cover Date.     

The use of wheat/goatgrass introgression lines for the detection of gene(s) determining resistance to septoria tritici blotch (Mycosphaerella graminicola)

At the IPK Gatersleben a series of 85 bread wheat (T. aestivum)/goatgrass (Aegilops tauschii) introgression lines was developed recently. Based on the knowledge that chromosome 7D of this particular Ae. tauschii is a donor of resistance to septoria tritici blotch (Mycosphaerella graminicola), a sub-set of thirteen chromosome 7D introgression lines was investigated along with the susceptible recipient variety ‘Chinese Spring’ (CS) and the resistant donor line ‘CS (Syn 7D)’. The material was inoculated with two Argentinian isolates of the pathogen (IPO 92067 and IPO 93014) at both the seedlings (two leaf) and adult (tillering) stages at two locations over 2 years (2003, 2004). The resistance was effective against both isolates and at both developmental stages, and the resistance locus maps to the centromeric region of chromosome arm 7DS. On the basis of its relationship with the microsatellite marker Xgwm44, it is likely that the gene involved is Stb5. Stb5 is therefore apparently effective against M. graminicola isolates originating from both Europe and South America.     

Morphological and molecular characterization of melon accessions resistant to Fusarium wilts

Fusarium wilt incited by Fusarium oxysporum f. sp. melonis (F.o.m) is one of the most widespread and devastating melon diseases. While resistance to physiological races 0, 1, and 2 is relatively frequent in different botanical varieties, sources of resistance to race 1,2 are restricted to a few Far-Eastern accessions. In this work, the results of a screening for resistance to F.o.m. race 1,2 among 32 accessions are presented. Three Japanese accessions (‘Kogane Nashi Makuwa’, ‘C-211’, and ‘C-40’) showed the highest resistance levels, but useful levels of resistance were also detected in one Russian ‘C-160’ and two Spanish (‘C-300’ and ‘Mollerusa-7’) accessions. These resistant materials, together with other accessions previously described as resistant to F.o.m. races 0, 1, and/or 2 have been morphologically and molecularly characterized. Based on cluster analysis, these accessions have been grouped according to the botanical subspecies they belong to. Assessment of genetic diversity indicated that the resistant accessions to races 0, 1 and 2, are scattered along the established clusters. On the other hand, high levels of resistance to the race 1,2 could be found only among accessions belonging to Cucumis melo subsp. agrestis, nevertheless, a certain degree of resistance to this race could also be found within some accessions belonging to subsp. melo. As far as we know, this is the first report of resistance to F.o.m race 1,2 found out from the Far-Eastern melon material. Based on fruits characteristics, it appears that several inodurus and cantalupensis accessions could be exploited in breeding programs as resistance sources to F.o.m races 0, 1 and/or 2 for the improvement of these melon types. The accessions with the highest levels of resistance to the race 1,2 appeared to be very distant both molecularly and morphologically from the commercial types. Nevertheless ‘C-160’, ‘C-300’, and ‘Mollerusa-7’ classified as var. inodorus are morphologically very similar to the Spanish commercial types and might be used as resistant sources in breeding these melon types.     

New inheritance studies related to Potato Virus Y (PVY) resistance in Capsicum annuum L. ‘Serrano Criollo de Morelos-334’

Summary C. annuum L. ‘Serrano Criollo de Morelos-334’ (SCM-334) is a well-known source of resistance against pepper diseases, including Potato Virus Y (PVY). In this work, inheritance studies involving ‘SCM-334’ and related to PVY resistance were described. For this purpose, ‘SCM-334’ derived lines were used, and plant material from the cross between ‘SCM-334’ and ‘Yolo Wonder’ (YW) was employed as well. Five PVY isolates were used for the inoculations. The first resistance described is a monogenic and recessive gene, controlling a specific-isolate resistance, and provisionally named as pvr1 ⁴. The second one is related to the inheritance of systemic necrotic symptoms after PVY inoculation. In this case a codominant gene is proposed and expressed only when the Pvr4 gene is not present.     

Isolation of wheat mutants with increased resistance to powdery mildew from small induced variant populations

Small variant wheat populations created by induced mutagenesis (n = 69) or adventitious regeneration (n = 66) were intensively screened for an altered response (compared to the parent variety ‘Guardian’) to the causal pathogen of powdery mildew in wheat, Blumeria graminis f. sp. tritici. Intensive field screening following natural infection of replicated plots of wheat lines over two years revealed a total of 13 mutants exhibiting significantly greater resistance than ‘Guardian’: eight from induced mutagenesis (11.6%) of the M2 population and five from adventitious regeneration (7.6%). Complete resistance was identified in two lines, (one (M66) developed following induced mutagenesis, and the other (SC240) by adventitious regeneration). The complete resistance in the induced mutant was stable over two generations and was associated with a high frequency of leaf flecking, and consequently a low grain yield. Resistance in SC240 proved to be unstable; SC240 exhibited complete resistance to powdery mildew in the SC₂ and SC₃ generations, but only 20% of the SC₄ plants were completely resistant, while the remainder were indistinguishable in mildew response to ‘Guardian’. The mildew response of all the SC₅ generation of SC240 was not significantly different from ‘Guardian’. Yield analysis of the thirteen mutants with increased resistance in the presence of powdery mildew indicated that eleven exhibitedgrain yields at least as high as that of ‘Guardian’, while the mutant M19 exhibited a yield significantly higher than that of ‘Guardian’. This revised version was published online in July 2006 with corrections to the Cover Date.     

Molecular mapping of quantitative trait loci determining resistance to septoria tritici blotch caused by Mycosphaerella graminicola in wheat

A set of 65 recombinant inbred lines of the ‘International Triticeae Mapping Initiative’ mapping population (‘W7984’בOpata 85’) was analysed for resistance to septoria tritici blotch at the seedling and adult plant stages. The mapping population was inoculated with two Argentinean isolates (IPO 92067 and IPO 93014). At the seedling stage, three loci were discovered on the short arms of chromosomes 1D, 2D and 6B. All three loci were detected with both isolates. At the adult plant stage, two isolate-specific QTL were found. The loci specific for isolates IPO 92067 and IPO 93014 were mapped on the long arms of chromosomes 3D and 7B, respectively. This revised version was published online in August 2006 with corrections to the Cover Date.     

Screening of Portuguese cole landraces (Brassica oleracea L.) with Peronospora parasitica and Plasmodiophora brassicae

Summary Forty-four landraces of Portuguese coles (Brassica oleracea L.), representing different morphotypes, were screened at the cotyledon stage for downy mildew resistance, caused by Peronospora parasitica. Two isolates of the pathogen, PHW 630 and PHW 828, were used in the experiment. The mean disease interaction phenotype varied among the landraces. Overall, the Portuguese coles have shown resistance to both isolates, particularly to isolate PHW 828. Some landraces were identified with differential reaction to the two P. parasitica isolates. Some of the Portuguese coles are, therefore, good sources of resistance genes to P. parasitica, and could be used in breeding programs or in the development of a downy mildew host differential set. Among the morphotypes tested, Couve Algarvia (ISA 207), Couve Galega (ISA 243 and 187) and Couve Glória de Portugal (ISA 84) have presented the highest resistance interaction. For the development of host differential sets, the landraces ISA 2 (Penca de Mirandela), ISA 35 (Couve de Valhascos) and ISA 600 (Penca Espanhola) could be further selected for the differentiation of P. parasitica isolates. They are resistant to PHW 828 and susceptible to PHW 643.The same morphotypes and landraces were also screened at the seedling stage for resistance to clubroot, caused by Plasmodiophora brassicae Woron.. Although some variation for resistance could be noticed in some the landraces tested, mainly Galega kale types, none of them presented a sufficient level of resistance as to be directly be useful in breeding for resistance to race 6 of P. brassicae.     

Comparison of the resistance and susceptibility to downy mildew [<Emphasis Type="Italic">Hyaloperonospora parasitica</Emphasis>, constant. (Pers. Ex Fr)] of nine <Emphasis Type="Italic">Brassica olearacea</Emphasis> accessions in laboratory,seedbed, and field screens

The relationship between resistance in seedlings, young and adult plants is studied for the pathosystem Brassica oleracea–Hyaloperonospora parasitica. Genotypes identified in the laboratory as resistant or susceptible or exhibiting a differential reaction to a selection of H. parasitica isolates were tested in 1997–1999 in seedbeds and fields under natural infestation. Isolates tested in the laboratory were grouped in five pathotypes, of which four were presented by isolates from Brittany, France. Genotypes susceptible to all pathotypes in the laboratory were also susceptible in the seedbed and field tests, while genotypes expressing a differential response to pathotypes were either resistant or susceptible. Accessions Everest, DEGC, ESPG and RS1105 exhibiting resistance to all pathotypes except I, were resistant in all environments and remained resistant in 2000–2002. Pathotype I was not prevailing in the field and results support the hypothesis that accessions resistant under laboratory conditions will be resistant under field conditions, provided the same pathotypes are present under the laboratory and field conditions.     

Identification of pathogenic races ofPhytophthora fragariae Hickman in Germany

Summary The fungusPhytophthora fragariae Hickman is the causal agent of red stele disease in strawberry (Fragaria × ananassa), this being a major disease in many areas with cool and moist conditions. Success of resistance breeding can be nullified by the appearence of specific races of the fungus capable of overcoming the introduced resistance. In some countries (USA, UK, Japan and Canada) races were identified by using a differential set of strawberry cultivars. The absence of an international standard differential set and the use of different test methods make a comparison of the identified races difficult or even lead to contradictory results for one variety/race combination. The aim of this study was to obtain information about the spectrum of pathogenic races in Germany as a basic contribution to research on resistance breeding against the fungus. The susceptibility of different strawberry cultivars to German isolates ofP. fragariae was evaluated. The inoculation was done by modifying a method described by Milholland et al. (1989). Rootedin vitro plants, four weeks after transferring them to the soil, were used for the investigation. Inoculation was done with a zoospore suspension of defined concentration. Up to now three German isolates, G-1, G-2, and G-3, can be separated by their ability to infect and produce oospores in the roots of the strawberry cultivars ‘Senga Sengana’, either ‘Saladin’, ‘Redgauntlet’ or ‘Climax’ andFragaria chiloensis clone ‘YaquinaB’. These results confirm the existence of pathogenic races ofP. fragariae in Germany and should be taken into consideration for resistance breeding against the fungus.     

Effect of incubation temperature regimes and culture medium on broccoli microspore culture embryogenesis

Conditions for reliable induction of embryogenesis from isolated microspores were studied in ten genotypes of broccoli. Embryo yields were significantly increased in almost all of the broccoli genotypes by the incubation at 32.5 ^°C for 1 day, than when the standard incubation at 30 ^°C for 2 days was used. Treatments of 48 hours at 32.5 ^°C produced less than optimal results suggesting that broccoli microspores are more sensitive to high temperatures than those of B. napus. The use of the ¹/₂ NLN-13 medium yielded greater number of embryos than the standard NLN-13. The magnitude of the response to the redution of the concentration of major salts by half in the NLN medium varied with the different genotypes. High embryogenic broccoli cultivars, such as ‘Shogun’, ‘SDB9’, and ‘Green Valiant’, presented a better response to the reduction of the concentration of major salts by half in NLN-13. Reduction never produced a detrimental effect on embryo yield and seems not to have any effect in the subsequent development of embryos in plants. This revised version was published online in July 2006 with corrections to the Cover Date.     

Screening methods of apple trees for resistance toValsa canker

Summary Valsa canker is the most serious wood rot disease of apple in Japan. Dormant, excised twig assay in vitro was used to determine resistance to Valsa canker. ‘Fuji’, ‘Starking Delicious’, ‘M.26 EMLA’, and ‘MM.106’ were considered to be highly susceptible, while Malus sieboldii showed moderate resistance. Several cultivars belonging toM. prunifolia, M. baccata, andM. × domestica showed comparative resistance. The phloridzin content in the bark ofMalus species was studied in relation toValsa canker resistance. Phloridzin content ofM. sieboldii was lower than inM. × domestica. Comparatively resistant cultivars had similar or higher phloridzin content compared to susceptible cultivars. No clear correlations were found between resistance and phloridzin contents in the bark ofMalus species.     

New source of resistance to mosaic virus transmission by Aphis gossypii in melon

The melon accession ‘TGR-1551’ shows a clear and total resistance to the transmission of several isolates of CMV and ZYMV when A. gossypii was used as the vector. The genetic analysis of the progenies obtained crossing this accession with the susceptible Spanish cultivar ‘Bolade Oro’ showed that the resistance to mosaic virus transmission is conferred by the presence of a dominant gene. There are also evidences that this dominant gene is different from ‘Vat’ gene. This revised version was published online in July 2006 with corrections to the Cover Date.     

Resistance to <Emphasis Type="Italic">Meloidogyne incognita</Emphasis> race 1 in the lettuce cultivars Grand Rapids and Salinas-88

The objective of this work was to check the possible allelism between two sources of resistance to the root-knot nematode Meloidogyne incognita race 1 in lettuce (‘Grand Rapids’ and ‘Salinas-88’). The experiments were carried out in greenhouses, in expanded 128-cell polystyrene trays filled with commercial substrate. Lettuce cultivars ‘Salinas 88’ and ‘Grand Rapids’ were tested along with the populations F₁ (‘Grand Rapids’ × ‘Salinas-88’), F₂ (‘Grand Rapids’ × ‘Salinas-88’), F₃ (‘Grand Rapids’ × ‘Salinas-88’), and with F₄ families derived from the latter population. Seedlings were inoculated 15 days after sowing with a nematode egg suspension equivalent to 30 eggs ml⁻¹ of substrate. Plants were evaluated for apparent gall incidence, gall scores, egg mass scores and extracted egg numbers 45 days after the inoculation date. There was evidence that two different genes are involved in control of resistance to M. incognita race 1 in lettuce cultivars Grand Rapids and Salinas-88. Lines with higher levels of nematode resistance than either Grand Rapids or Salinas-88 could be selected in the F4 generation of the cross between these resistant parental lines.     

Segregation distortion of Brassica carinata derived black rot resistance in Brassica oleracea

Three segregating F₂ populations were developed by self-pollinating 3 black rot resistant F₁ plants, derived from across between black rot resistant parent line 11B-1-12 and the susceptible cauliflower cultivar ‘Snow Ball’. Plants were wound inoculated using 4 isolates ofXanthomonas campestris pv. campestris (Xcc) race 4, and disease severity ratings of F₂ plants from the three populations were scored. A total of 860 arbitrary oligonucleotide primers were used to amplify DNA from black rot resistant and susceptible F₂ plants and bulks. Eight RAPD markers amplified fragments associated with completely disease free plants following black rot inoculation,which segregated in frequencies far lower than expected. Segregation of markers with black rot resistance indicates that a single, dominant major gene controls black rot resistance in these plants. Stability of this black rot resistance gene in populations derived from 11B-1-12 may complicate introgression into B. oleracea genotypes for hybrid production. This revised version was published online in July 2006 with corrections to the Cover Date.     

Zingiberene-mediated resistance to the South American tomato pinworm derived from Lycopersicon hirsutum var. hirsutum

The Lycopersicon hirsutum var. hirsutum accession PI 127826 is recognized as a good source of resistance to arthropod pests due to the action of the allelochemical zimgiberene, a sesquiterpene present in its glandular trichomes. Five genotypes were selected from the F₂ generation of the interspecific cross Lycopersicon esculentum ‘TOM-556’ × Lycopersicon hirsutum var. hirsutum ‘PI 127826’, based on their low levels (BPX-368-clone#56) or high levels(BPX-368-clone#92, BPX-368-clone#105,BPX-368-clone#179, BPX-368-clone#250) of zingiberene. The five F₂ genotypes were tested for resistance to the South American tomato pinworm Tuta absolutaalong with accession L. esculentum ‘TOM-556’ (pinworm susceptible), and the accessions L. hirsutum var. hirsutum ‘PI 127826’ and L. pennellii ‘LA716’ (resistant). The F₂ clones selected for high foliar zingiberene levels showed lower scores for leaflet lesion type(LLT), percent leaflets attacked (PLA) and overall plant damage (OPD) than the low zingiberene genotypes. The results indicated that zingiberene mediates resistance to the South American pinworm, based on feeding and on ovipositing deterrence, in populations derived from the interspecific cross between Lycopersicon. esculentum and Lycopersicon hirsutum var. hirsutum. Indirect selection for high foliar zingiberene content is suggested as an efficient technique for breeding tomatoes for resistance to the South American tomato pinworm. This revised version was published online in July 2006 with corrections to the Cover Date.     

Detecting and estimating segregation distortion and linkage between glufosinate tolerance and blackleg resistance in Brassica napus L.

Summary The segregation and linkage between glufosinate (transgenes ‘Rf3’ and ‘T177’) and blackleg resistance genes in canola (Brassica napus L.) were assessed using F₁ microspore-derived doubled haploid (DH) populations from four crosses including reciprocals, two involving the transgene ‘Rf3’ and the other two involving the transgene ‘T177’. To relax the assumption of no segregation distortion required for the conventional analysis of segregation and linkage, we employed Bailey's analysis that allows detecting segregation distortion at linked loci. The significant departures from the 1:1 segregation were detected in the crosses involving the transgene ‘T177’ but not in the crosses involving the transgene ‘Rf3’. The apparent deficit of the herbicide tolerant DH lines in the crosses with the transgene ‘T177’ is likely due to differential selection against the gametes carrying ‘T177’ during microspore culture. The linkage was strong between blackleg resistance and the transgene ‘Rf3’ but weak or absent between blackleg resistance and the transgene ‘T177’, suggesting that the two transgenes are probably inserted into distant regions of the genome. The observed linkage offers an opportunity to develop new canola cultivars with both glufosinate tolerance conferred by transgene ‘Rf3’ and blackleg resistance.     

Yield, yield components and plant architecture in the F3 generation of common bean (Phaseolus vulgaris L.) derived from a cross between the determinate cultivar ‘Prelude’ and an indeterminate landrace

Growth trials were conducted outdoors in the UK to determine the yield, yield components and plant architectural differences between determinate and indeterminate Phaseolus vulgaris bean plants. F3 lines derived from crosses between ‘Prelude’, a determinate cultivar and ‘V8’, an indeterminate landrace, were grown together with the parents and ‘Carioca’, an indeterminate landrace from Brazil. Data were recorded on flowering date, number of nodes on main stem at flowering, plant height at maturity, number of pods/plant, number of seeds/pod, number of seeds/plant, 100-seed weight, seed mass/plant, percentages of diseased and healthy seeds/plant. Determinate F3 lines had significantly lower (P < 0.05) seed mass/plant, fewer pods/plant, fewer seeds/pod, fewer seeds/plant, lower harvest index, shorter stems, earlier date of flowering, fewer nodes at flowering and fewer healthy seeds/plant than indeterminate F3 lines. However, determinate genotypes had significantly larger (P < 0.05) pods to accommodate their larger seeds compared with indeterminate lines. A similar situation was found when the yield and yield components of ‘V8’, ‘Prelude’ and ‘Carioca’ were evaluated; the highest seed mass/plant was given by ‘V8’ and the lowest by ‘Prelude’. Furthermore, some indeterminate genotypes with Type IV growth habit yielded significantly higher (P < 0.05) than the high-yielding parent, ‘V8’, indicating a positive influence on seed yield by crossing different genotypes.