The exploitation of genetic resources of Brassica oleracea in breeding for resistance to clubroot (Plasmodiophora brassicae)

Summary About 1000 Brassica oleracea accessions were evaluated in glasshouse tests for response to Plasmodiophora brassicae (clubroot). Resistance was confirmed in some north and west European kales and cabbage. A new source of resistance in cabbage, from Eire, is reported. Most other accessions were highly susceptible but lower levels of susceptibility were observed in open pollinated Brussels sprouts and forms of south European cabbage, cauliflower and broccoli. Modern breeding (as in the production of hybrid cultivars) appears to have resulted in increased susceptibility in several crop types. The implications of these results for the exploitation of germplasm are discussed.     

The exploration of genetic resources of Portuguese cabbage and kale for resistance to several Brassica diseases

Summary Twenty three accessions of nine Portuguese cabbage and kale land races from different geographic origins were tested at the seedling stage for resistance to several important brassica diseases. Resistance to downy mildew (Peronospora parasitica), expressed as necrosis of the cotyledon mesophyll, was found in all the accessions. Type A resistance to cabbage yellows (Fusarium oxysporum f. sp. conglutinans race 1) was present in most of the landraces. Resistance to clubroot (Plasmodiophora brassicae race 6) was found in one accession of the Portuguese tree kale. High resistance to blackleg (Leptosphaeria maculans) and white rust (Albuco candida) was not detected, although several accessions showed 20 to 30% of plants with intermediate expression of resistance. All Portuguese cole accessions were susceptible to blackrot (Xanthomonas campestris pv. campestris).     

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.     

Genetic analysis of resistance to clubroot (Plasmodiophora brassicae Woron) in two Brassica oleracea groups (ssp. acephala and ssp. botrytis) through diallel analysis

Clubroot, caused by Plasmodiophora brassicae Woron, is a major disease of cruciferous crops and few sources of resistance have been detected and genetically studied in the Brassica oleracea species. In this study, two diallel mating systems using resistant kale lines and susceptible cauliflower lines were performed. Resistance was assessed from a disease index evaluated on young plants artificially inoculated with local isolates of the parasite. Both general and specific combining ability effects (GCA and SCA) and partly reciprocal effects were demonstrated. Resistance inheritance was, however, characterized by a predominance of additive genetic effects (GCA). Three groups of parent lines could be separated; kale lines with very high resistance, kale lines with intermediate resistance and susceptible cauliflower lines. In the two kale groups, two genetically different resistance types were suggested and various recurrent selection procedures are proposed following the diallel results.     

Association of RFLP markers with trait loci affecting clubroot resistance and morphological characters in Brassica oleracea L.

Summary Resistance to Plasmodiophora brassicae Wor. race 7, the causal agent of the disease clubroot, was examined in an F₂ population of a cross between a clubroot resistant broccoli (Brassica oleracea var. italica) and a susceptible cauliflower (B. oleracea var. botrytis). A genetic linkage map was constructed in the same population based on the segregation of 58 dispersed restriction fragment length polymorphism (RFLP) markers. Associations between the inheritance of RFLP marker genotypes and segregation for disease resistance, morphological and maturity characteristics were examined. For each triat examined, several chromosomal regions marked by RFLP probes appeared to contain trait loci, suggesting that each trait was under polygenic control. RFLP marker linkage to a major factor imparting dominance for clubroot resistance from the broccoli parent was observed in this population. Additionally, RFLP marker linkage to an independently segregating factor contributing clubroot resistance from the cauliflower parent was observed, indicating that it should be possible to use RFLP markers to facilitate selection of transgressive segregants having the combined resistance from both parental sources. In some instances, RFLP markers from the same or closely linked chromosomal regions were associated with both clubroot resistance and morphological traits. Analysis of RFLP marker genotypes at linked loci should facilitate the selection of desired disease resistant morphotypes.     

Variation in the expression of specificity in two maize diseases

Summary Ten inbred lines from the open-pollinated maize variety Jarvis were selected from 51 randomly collected lines to represent a wide range of susceptibility to one isolate each of Bipolaris maydis or Colletotrichum graminicola. Ten isolates of each pathogen were selected for a range of virulence on a maize line with average resistance. Resistance and virulence ratings were based on lengths of lesions that developed on leaves of greenhouse-grown seedlings inoculated with 5 l droplets of suspensions of known spore concentrations. For each disease the ten maize lines were inoculated in all possible combinations with the ten pathogen isolates. The experiment was run six times with each pathogen. Analysis of variance for individual trials indicated a significant interaction between maize lines and B. maydis isolates in all six trials and between maize lines and C. graminicola isolates in four of six trials. For both diseases. the combined analysis over all six trials revealed no significant interaction. Apparently the expression of specificity in these host-pathogen interactions is variable.     

Resistance in two Lycopersicon species to an Arkansas isolate of tomato spotted wilt virus

Summary Tomato spotted wilt virus (TSWV) was obtained from infected tomatoes in commercial fields in Arkansas in 1985. A greenhouse screening procedure for identifying tomatoes resistant to TSWV was established using an enzyme-linked immunosorbent assay (ELISA) to detect infected plants. Symptom expression was variable and symptom expression was not reliable for identifying infected plants. Germplasm evaluated for resistance to one typical Arkansas isolate (85–9) of TSWV included: twenty cultivars and breeding lines of Lycopersicon esculentum Mill, 52 accessions of L. pimpinellifolium (Jusl.) Mill and 8 accessions of L. peruvianum (L.) Mill. All cultivated accessions and breeding lines evaluated were susceptible. Some individual plants in several accessions of L. pimpinellifolium were resistant and nearly all plants of the L. peruvianum accessions that were evaluated were resistant to isolate 85–9.Dept. of Plant Pathology     

The transfer of characters from Brassica campestris L. to Brassica napus L.: Production of clubroot-resistant oil-seed rape (B. napus ssp oleifera)

Summary Four oil-seed rape lines were crossed with a clubroot resistant Brassica campestris line from the European Clubroot Differential sct. The allotriploid hybrids were backcrossed to the rape lines to introgress clubroot resistance into oil-seed rape. Using a combination of screening for disease resistance and chromosome number, a high proportion of 38-chromosome, clubroot resistant selections were obtained.     

Inheritance of resistance against Phytophthora infestans in Lycopersicon pimpenellifolium L3707

Summary Lycopersicon pimpenellifolium L3707, resistant to the late blight oomycete Phytophthora infestans was crossed with the susceptible Lycopersicon pimpenellifolium 14377 or the susceptible Lycopersicon esculentum ZH. Progeny F1 and F2 generations were scored at the 5-leaf stage for resistance against 175 field and recombinant isolates of the pathogen. F1 plants exhibited various levels of moderate resistance and F2 plants segregated 3:6:7 resistant/moderately resistant/susceptible. The data support the hypothesis that race-non-specific resistance in L3707 is controlled by two independent genes: a partially-dominant gene and a dominant epistatic gene.     

Contribution of host-pathogen interactions to the expression of the blackleg disease of spring rape (Brassica napus L.) caused by Leptosphaeria maculans (Desm.) Ces. et de not

Variation in the responses of selected genotypes of spring rape to infection by a wide range of isolates of Leptosphaeria maculans was analyzed in this study.Differences in severity of disease damage to seedlings between host and pathogen genotypes were highly significant. Interactions between host and pathogen were also highly significant. The generally resistant cultivar Wesreo was susceptible to a number of isolates whereas the normally susceptible cultivars Zollern-gold and Ceska were resistant to some isolates. Overall, however, Wesreo was resistant to a far greater number of isolates than were the susceptible genotypes.Significant differences in severity of disease damage were also observed between groups of isolates from different sites. All host genotypes were resistant to a substantially greater proportion of isolates from certain sites than from others. Interactions were again detected in that the ranking of sites on the basis of the proportion of isolates causing severe damage varied between host genotypes.The significance of host-pathogen interactions in the B. napus-L. maculans relationship is discussed with special reference to current views on the nature of horizontal resistance.     

Resistance to Brevicoryne brassicae in horticultural brassicas

Summary A wide range of cultivated brassica accessions including broccoli, Brussels sprouts, Chinese cabbage, cauliflower, collard, kale and swede material was tested against the cabbage aphid, Brevicoryne brassicae, at HRI, Wellesbourne in the field and laboratory in both 1992 and 1993. In the field, B. brassicae attack was assessed as the proportion of infested plants and the numbers of aphid colonies present. In the laboratory, aphid performance was measured in terms of the pre-reproductive and reproductive period, population increase, and insect mortality. Interpretation of the data was facilitated by plotting sorted accession means against normal order statistics. This statistical approach indicated the spread of variation amongst the accessions and permitted identification of extremes. Partial levels of antixenosis resistance were discovered in red brassicas. Glossy accessions of cabbage and cauliflower possessed antixenosis and antibiosis resistance that lasted throughout the season of crop growth in the field. Other accessions were shown to withstand aphid attack and therefore possessed tolerance. The laboratory studies provided information on mechanisms of antibiosis resistance. The potential value of the different sources of resistance is discussed.     

Brassica coenospecies: a rich reservoir for genetic resistance to leaf spot caused by Alternaria brassicae

Development of leaf spot resistant mustard cultivars is a relevant objective in view of heavy crop losses caused by this pathogen. Thirty-eight species belonging to 9 genera, including cultivated and wild allies, of the genus Brassica were evaluated under epiphytotic conditions for two years. Inoculations were done on whole plants (in vivo) and on detached leaves (in vitro). Data on incubation period, number of lesions per leaf, lesion size and leaf area covered by lesions were recorded. Species which never produced disease symptoms throughout the growing period in pots and until 72 hours after inoculation in detached leaf assays during both years were treated as resistant, while those that produced symptoms were classified as moderately resistant, susceptible or highly susceptible depending upon incubation period, size of lesions and leaf area covered by disease symptoms. Eight species (Brassica desnottesii, Camelina sativa, Coincya pseuderucastrum, Diplotaxis berthautii, D. catholica, D. cretacea, D. erucoides, and Erucastrum gallicum) were found completely resistant, whereas others were classified as moderately resistant (12), susceptible (11) or highly susceptible (9). Since resistance is unavailable within the cultivated species, these 8 resistant wild species could be used as donor parents for introgressing resistance to leaf spot disease in Indian mustard. This revised version was published online in August 2006 with corrections to the Cover Date.     

Genetic variation in Zantedeschia spp. (Araceae) for resistance to soft rot caused by Erwinia carotovora subsp. carotovora

Bacterial soft rot caused by Erwiniacarotovora subsp. carotovora is amajor disease in Zantedeschia spp.,particularly in cultivars from the sectionAestivae. The disease can partiallybe controlled by cultivation measures, so acombination with resistant cultivars couldeffectively protect the crop. However,resistant commercial Aestivaecultivars are not available yet. By meansof a recently developed non-destructiveresistance test, variation inaggressiveness was observed among fiveisolates of Erwinia carotovora subsp.carotovora without interactionsbetween the isolates and three Zantedeschia accessions. Within elevenaccessions of Z. aethiopica,variation was observed from almost completeto moderate resistance, while theZ. odorata accession was susceptible.All 21 Aestivae cultivars weresusceptible. Within the Aestivaespecies, Z. elliotiana and Z.pentlandii were also susceptible, butwithin twelve accessions of Z.albomaculata, as well as in six accessionsof Z. rehmannii, variation was foundfrom susceptible to moderately resistant.Hence, new sources of resistance wereidentified that show good potentials forresistance breeding.     

Prehaustorial resistance to the wheat leaf rust fungus, Puccinia triticina, in Triticum monococcum (s.s.)

Diploid wheat, Triticum monococcum s.l., is a host for the wheat leaf rust fungus, Puccinia triticina. Some accessions have been reported to show a high degree of prehaustorial resistance. This is non-hypersensitivity resistance, which acts before the formation of haustoria by the pathogen. To assess the frequency of prehaustorial resistance 598 accessions of diploid wheat were inoculated with the wheat leaf rust isolate Felix. Most T. monococcum s.s. accessions (84%) were resistant whereas all T. urartu and all but three T. boeoticumaccessions were susceptible. Histological components analysis revealed that a high percentage of prehaustorial resistance to P. triticina was found in only three T. monococcum accessions. No haustoria were observed in such infection units confirming the prehaustorial nature of the resistance. Prehaustorial abortion of certain infection units in an accession always coincided with posthaustorial abortion of the other infection units. This revised version was published online in July 2006 with corrections to the Cover Date.     

Diversity among Finnish net blotch isolates and resistance in barley

Summary Seedlings of a differential barley (Hordeum vulgare L.) series (21 genotypes) and of six check genotypes were used in the greenhouse to assess variation in virulence among 20 single-spore isolates of the net blotch pathogen. Pyrenophora teres Drechs. f.teres Smedeg., collected from various sites in Finland. The experiment was run twice and symptom expression was recorded on the first three leaves. Analysis of second leaf symptom scores from Run 1 indicated differences in virulence between isolates, all of which were pathogenic, and differential resistance among the barleys. The virulence of P. teres isolates appeared to be conditioned by the host barley from which the isolate derived; the average virulence of isolates collected from a susceptible host was greater than that of isolates collected from a less susceptible host. Results from Run 2 were similar regarding resistance within the barleys, but variation in virulence among the P. teres isolates was not consistent with that from Run 1. CI 9819 caries duplicate genes for resistance to P. teres (Rpt1b and Rpt1c), and CI 7548 possesses Rpt3d. Both genotypes were highly resistant to all isolates. The Rpt1a gene of Tifang (CI4407) conferred resistance to all isolates in Run 2, but only to about half of the isolates in Run 1. The checks, including two of the symptomatically most resistant Nordic barley genotypes, were universally susceptible during these stringent tests. No selective pressure has been placed on Finnish isolates of P. teres through previous deployment of major resistance genes, and it is speculated that any variation in virulence among isolates is likely to be due to a combination of evolutionary forces including, natural selection, random genetic drift and gene flow.     

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.     

Screening and identification of resistance to bacterial soft rot in Brassica rapa

Mist-chamber, field, and detached leaf inoculation procedures identified plants resistant to bacterial soft rot [Erwinia carotovora subsp. carotovora, (Ecc)] in Brassica rapa and related species. The mist-chamber seedling inoculation provided the best correlation of mean disease severity ratings with the field plant inoculation (r = 0.67^**) and was used to identify resistant materials. The optimum mist-chamber incubation conditions to distinguish the resistance of accessions were 23 ^°C and 100% RH and were used for primary screening. A total of 752 accessions of B. rapa and related Cruciferae were screened. In general, accessions of B. oleracea were more resistant than accessions of B. rapa. Within B. rapa, subspecies pekinensis and chinensis were more susceptible than other subspecies. No completely resistant material was found. In species of B. rapa, only 7% of accessions showed some degree of resistance with plant-to-plant variation within the accessions. G30444, G30449, and AVRDC2837 were identified as the most resistant materials in B. rapa by both mist-chamber and field inoculations. The resistance was correlated between an USA isolate (Geneva-1) and two Chinese isolates (RL4-1 and RL-19) of Ecc. This revised version was published online in July 2006 with corrections to the Cover Date.     

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.     

Evaluation of F2 and F3 plants introgressed with QTLs for clubroot resistance in cabbage developed by using SCAR markers

Clubroot disease caused by Plasmodiophora brassicae is one of the major diseases of Brassica crops, often devastating to the cultivation of cruciferous crops in temperate regions. In a previous study (Moriguchi et al. 1999) identified three major quantitative trait loci (QTLs) for clubroot resistance, each in a separate linkage group, in a population derived from a cross between a clubroot‐susceptible inbred cabbage line, Y2A and a resistant inbred kale line, K269. In this study, the original random amplified polymorphic DNA (RAPD) and restriction fragment length polymorphism (RFLP) markers were converted into sequence‐characterized amplified region (SCAR) markers to facilitate large‐scale marker‐assisted screening of clubroot resistance in cabbage breeding. Of 15 RAPD markers closely linked to the three QTLs, nine SCARs were developed as dominant markers after cloning and sequencing. In addition, two RAPD markers were converted into co‐dominant cleaved amplified polymorphic sequence (CAPS) markers, and one RFLP marker out of three tested was converted to a dominant SCAR marker. The effect of selection for resistance by the improved markers was evaluated in progeny plants in the F2 and F3. A total of 138 F2 plants were genotyped with nine SCARs and 121 well‐distributed makers consisting of 98 RAPD, 19 RFLP, two isozymes, and two morphological markers in order to estimate the level of resistance and the proportion of undesirable alleles from the kale in non‐target areas in each of the F2 populations. An F2 plant, YK118, had kale alleles at QTL1, QTL3 and QTL9. Three F2 plants, namely, YK107, YK25 and YK51 had kale alleles at only QTL1, QTL3 and QTL9, respectively. These F2 plants were selected for their low proportion of alleles derived from kale in non‐target regions. YK118, like the resistant kale parent, expressed very high resistance to three field isolates of Plasmodiophora brassicae, whereas the mean disease index in the F2 and F3 plants carrying only single QTLs was intermediate. The QTLs showed no differential response to the isolates. These plants with improved resistance will be useful as parental inbred lines for F1 hybrids.     

Inheritance of resistance to spotted stem borer, Chilo partellus, in sorghum, Sorghum bicolor

The spotted stem borer, Chilo partellus, is one of the most important pests of sorghum, and host plant resistance is an important component for the management of this pest. Most of the sorghum hybrids currently under cultivation are based on cytoplasmic male-sterility (CMS). In order to develop a strategy for resistance to stem borer, we studied the traits associated with resistance, and their nature of gene action in F₁ hybrids derived from resistant, moderately resistant, and susceptible CMS and restorer lines. The hybrids based on stem borer-resistant, moderately resistant, or susceptible CMS and restorer lines were equally resistant or susceptible as the parents for leaf feeding [Damage rating (DR) 5.8 to 6.6 vs. 5.9 to 6.6], and had significant and decreasing trend in deadheart formation (resistant CMS × resistant restorer lines < moderately resistant CMS × moderately resistant restorer lines < susceptible CMS × susceptible restorer lines), respectively. Proportional contributions of restorer lines were greater than those of the CMS lines for leaf feeding, deadhearts, recovery and overall resistance, stalk length, nodes per plant, stem borer holes per plant, and peduncle tunneling. The general (GCA) and specific combining ability (SCA) estimates suggested that leaf feeding score, number of nodes, overall resistance score, panicle initiation, recovery score, and stalk length (dominance type of gene action) have been found to be associated with resistance to spotted stem borer, governed by additive type of gene action, their correlation and direct effects in the same direction, and explained 65.3% of the variation in deadhearts, and thus could be used as marker traits to select and breed for resistance to C. partellus in sorghum. The parents having significant SCA effects for two or more resistance traits for either or more parents have also been discussed for their use in the stem borer resistance breeding.