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).     

Introgression of clubroot resistance into an elite pak choi inbred line through marker‐assisted introgression breeding

Clubroot is a soilborne disease that severely infects cruciferous species. Pak choi (Brassica rapa subsp. chinensis) is an economically important cruciferous crop cultivated throughout the world. However, no clubroot‐resistant germplasms have been identified in pak choi to date. To improve disease resistance, we used marker‐assisted selection (MAS) to introgress the clubroot resistance (CR) trait from the ‘CCR13685’ Chinese cabbage (B. rapa subsp. pekinensis) inbred line into an elite pak choi inbred line, ‘GHQ11021’. Genetic analysis of F₂ and BC₁ progeny showed that CR of ‘CCR13685’ was controlled by a single dominant gene. We designed nine candidate sequence‐characterized amplified region markers, K‐1 to K‐9, based on two molecular markers linked to the CR gene. We found that K‐3 co‐segregated with CR and an inoculation test confirmed that K‐3 could be used for MAS. Two introgression lines, BC₃‐1‐4 and BC₃‐2‐18, were developed using K‐3 for foreground selection. These lines displayed the same phenotypic properties as ‘GHQ11021’, but were highly resistant to clubroot, indicating that the CR gene of ‘CCR13685’ had been successfully introduced into pak choi.     

Expression of resistance to downy mildew at cotyledon and adult plant stages in Brassica oleracea L.

This research compared the expression of resistance to downy mildew at cotyledon and adult plant stages in seven Brassica oleracea genotypes against two P. parasitica isolates of different virulence. Seven day old seedlings were dual inoculated under controlled environment by depositing two 10 μl droplets of a spore suspension of a different isolate on each cotyledon and the interaction phenotype (IP) evaluated 7 days later using a 0–5 scale of increasing susceptibility. The seedlings were transplanted to 16 cm pots and grown in the greenhouse for 110 days (15 to 28 leaves). Adult plants were tested using a single leaf inoculation method that allowed the same plant to be simultaneously inoculated with the two P. parasitica isolates. Leaves were scored 10 days after inoculation using a 0–5 scale of increasing susceptibility. The inoculation of the same plant with the two isolates produced different combinations of cotyledon (CT) and adult-plant (AP) interaction-phenotype according to the genotype. CrGC 3.1 was susceptible at CT and AP stages, ‘Algarvia’ resistant at CT and AP stages, and broccoli ‘A’ susceptible at CT and resistant at AP stage against the two isolates. ‘Murciana’ and broccoli ‘B’ were differential at CT and AP stages. Savoy and Shetland cabbages were differential at CT stage and resistant to both isolates as adult plants. Cotyledon resistance could not be used to predict adult-plant resistance since the two types of resistance were very poorly correlated. Resistance can be race specific either at cotyledon or at adult-plant stage. This revised version was published online in July 2006 with corrections to the Cover Date.     

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.     

Evaluation of French Brassica oleracea landraces for resistance to Plasmodiophora brassicae

A total of 240 kale, 38 cabbage and 126 winter cauliflower French landraces from the B. oleracea genepool of INRA were assessed for resistance to clubroot caused by Plasmodiophora brassicaeWoron. Two French isolates of the pathogen (K and SJ) were used in the experiments under controlled conditions. The reaction of the 126 cauliflower accessions to naturally occurring clubroot was also evaluated in field trials. Kales exhibited considerable variation for expression of disease resistance and high levels of resistance were found in several accessions. In this group, single resistant plants were observed in most of the morphological types and from quite different geographical origins. Cabbage accessions were moderately to highly susceptible to both isolates. All cauliflower populations proved to be highly susceptible to K isolate and moderately susceptible to SJ isolate. In field trials, cauliflowers were also severely infected. Two lines selected from a resistant kale population were highly resistant against a large range of pathotypes of the pathogen. These lines presented a sufficient level of resistance to be directly useful in the breeding program in order to develop cauliflower and broccoli hybrids resistant to clubroot. This revised version was published online in July 2006 with corrections to the Cover Date.     

Resistance to the cabbage root fly, <Emphasis Type="Italic">Delia radicum</Emphasis> (Diptera,Anthomyiidae), of turnip varieties (<Emphasis Type="Italic">Brassica rapa</Emphasis> subsp. <Emphasis Type="Italic">rapa</Emphasis>)

The cabbage root fly Delia radicum L. (Diptera: Anthomyiidae) is one of the major pests of many Brassica crops in the temperate areas of Europe and North America. At present, turnip (B. rapa ssp. rapa L.) varieties resistant to the pest does not exist. With the aim to fill this gap, a no-choice tolerance test of 56 accessions among turnips, turnip tops and turnip greens was performed under controlled conditions by introducing D. radicum eggs. Plant survival, leaf and root conditions, pupae number and weight significantly varied among plant accessions. Ten putatively resistant and ten susceptible accessions (control group) were selected from this first screening, transplanted in the field and exposed to natural infestation to detect antibiosis and antixenosis mechanisms. Both in the laboratory and in the field, pupae number significantly varied within accessions and between resistant and susceptible group, although pupal weight did not, indicating the absence of antibiosis effect on this early stage. In the field, the number of galleries was significantly lower in the resistant group in comparison with the control. Resistant accessions had smaller size, and a smaller, white and mostly buried root. Within the resistant and susceptible accessions, larger plants harboured more pupae, however purple roots were those most preferred, and the hosted pupae weighed most. Three accessions from the resistant group (MBGBR0178, MBGBR0570 and MBGBR0371) stand out for resistance to D. radicum possibly through antixenosis mechanisms.     

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.     

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.     

Regeneration of Chinese cabbage transgenic plants expressing antibacterial peptide gene and cowpea trypsin inhibitor gene

Summary An efficient transformation system for Chinese cabbage cotyledon explants was developed using Agrobacterium tumefaciens strains LBA4404 harbouring the plasmid pMOG 411 and the plasmid pBinΩSCK respectively. Various factors affecting the transformation efficiency and subsequent regeneration were identified. The age of seedlings, growth conditions and status of Agrobacterium suspension, preculture of explants, cocultivation time, ratio between Agrobacterium and explants, acetosyringone and concentration of kanamycin had a significant influence on transformation frequency and plant regeneration. The presences of the antibacterial peptide gene and the cowpea trypsin inhibitor gene in those selected shoots in kanamycin medium were confirmed by PCR, Southern blotting and Northern blotting, The frequency of regeneration from cotyledons was analyzed in eight parental lines of Chinese cabbage and five lines were suitable for obtaining transformed plants, among which Qingmaye C and Qingmaye D were more responsive than other lines.     

Resistance to stripe rust in Triticum turgidum,T. tauschii and their synthetic hexaploids

Resistance to stripe rust (caused by Puccinia striiformis Westend.) of 34 Triticum turgidum L. var.durum, 278 T. tauschii, and 267 synthetic hexaploid wheats (T. turgidum x T. tauschii) was evaluated at the seedling stage in the greenhouse and at the adult-plant stage at two field locations. Mexican pathotype 14E14 was used in all studies. Seedling resistance, expressed as low infection type, was present in all three species. One hundred and twenty-eight (46%) accessions of T. tauschii, 8 (23%) of T. turgidum and 31 (12%) of synthetic hexaploid wheats were highly resistant as seedlings. In the field tests, resistance was evaluated by estimating area under disease progress curve (AUDPC). Synthetic hexaploid wheats showed a wide range of variability for disease responses in both greenhouse and field tests, indicating the presence of a number of genes for resistance. In general, genotypes with seedling resistance were also found to be resistant as adult plants. Genotypes, which were susceptible or intermediate as seedlings but resistant as adult plants, were present in both T. turgidum and the synthetic hexaploids. Resistances from either T. turgidum or T. tauschii or both were identified in the synthetic hexaploids in this study. These new sources of resistance could be incorporated into cultivated hexaploid wheats to increase the existing gene pool of resistance to stripe rust.     

Screening for Fusarium resistance in seedling populations of Asiatic hybrid lily

A test to select Fusarium resistant seedlings of the Asiatic hybrid lily is described. Young seedlings of 28 populations, obtained from an incomplete diallel between eight parents with different levels of Fusarium resistance, were tested for resistance. Significant differences in Fusarium resistance between and within populations were detected. The average percentage of selected seedlings ranged from 34% in resistant × resistant crosses to 2% in susceptible × susceptible crosses. Although resistant descendants were obtained in susceptible × susceptible crosses, using at least one resistant parent produced higher percentages of resistant seedlings. The resistance level of the parents correlated highly with the general combining ability for Fusarium resistance based on the seedling test. For eight populations, seedlings selected for Fusarium resistance and non-tested (control) seedlings of the same cross were compared, after propagation, in a clonal test. Variation between and within populations, found at seedling level, was confirmed at clonal level. A positive selection response was found for all eight populations. In the seedling test, approximately 18% of the seedlings were selected as resistant of which 15% (2.7% of seedlings tested) appeared to be susceptible escapes. Comparison between selection at seedling level and at clonal level indicated that approximately 25% of the seedlings tested were missed (rejected resistant plants) in the seedling test. The practical use of a seedling test for Fusarium resistance in lily breeding programs is discussed.     

Histological and inheritance studies of partial resistance in the Brassica napus–Albugo candida host-pathogen interaction

Traditional and doubled haploid (DH) genotypes of oilseed Brassica spp. resistant, partially resistant, moderately susceptible, and susceptible to Albugo candida were compared for phenotypic development of host‐pathogen interaction and histology of host‐pathogen interaction. The partially resistant genotype showed pinhead‐size pustules, mainly on the upper surface of cotyledonary leaves. Relatively less mycelium was observed in the partially resistant genotype compared with the susceptible genotype. In resistant B. napus genotypes, there was neither pustule development nor any mycelial growth. In the moderately susceptible genotype, the pustules were similar to those in the partially resistant genotype in being of pinhead‐size and occasionally coalescing. However, ample mycelial growth in the mesophyll tissue in the moderately susceptible genotype was similar to that in the susceptible control B. rapa cv. ‘Torch’. The susceptible genotype B. rapa cv. ‘Torch’ also showed large coalescing pustules. In the non‐host B. juncea cv. ‘Commercial Brown’, no pustules were formed although some mycelial growth was observed beneath the epidermal cell layer and in the mesophyll cell layer of the cotyledonary leaf tissue. For inheritance studies, two partially resistant B. napus genotypes were crossed with a resistant B. napus genotype. Various generations viz., F1, F1(reciprocal), F₂, and DHs produced from the crosses were inoculated with a zoospore suspension of race 7v of A. candida. The partially resistant phenotype appeared to be controlled by a single recessive gene designated as wpr with variable expression. The simple inheritance of partial resistance has implications for disease resistance breeding against white rust, as this type of resistance can be easily incorporated into elite breeding lines through conventional and DH breeding methods.     

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.     

Genetic and histological characterization of downy mildew resistance at the cotyledon stage in <Emphasis Type="Italic">Raphanus sativus</Emphasis> L.

Downy mildew (DM; Hyaloperonospora brassicae) is one of the most serious diseases in radish and responsible for high production losses. The objectives of this study were (1) to identify new sources of DM resistance at the cotyledon stage; (2) to determine the inheritance of the resistance; and (3) to clarify the histological and cellular response involved in DM infection. We screened 200 radish accessions for DM resistance at the cotyledon stage. Radish accessions Rd001, Rd004, Rd048 and Rd150 showed a complete resistance response (hypersensitive reaction), and accession Rd193 exhibited a partial resistance with confined sporulation. The highly susceptible accession Rd197 was then crossed with all resistant genotypes for a genetic analysis. F₂ segregation analysis suggested that cotyledon resistance in Rd001, Rd004, Rd048 and Rd150 is conferred by a single dominant gene while resistance in Rd193 is polygenic. Genetic data suggests Rd193 resistance might be conferred by two dominant genes with complementary action, althought further experiments are needed to confirm this hypothesis. To conduct a histological study using light and fluorescence microscopy, we collected cotyledons from three genotypes with either differing levels of resistance (Rd004 and Rd193) and high susceptibility (Rd197) at ten different time points, from 3 h to 7 days post inoculation. Tissue necrosis was responsible for the inhibition and reduction of H. brassicae growth in Rd004 and Rd193 within 24 h of inoculation. For susceptible accession Rd197, the first evidence of necrotic cells appeared 48 h post inoculation. Callose deposition associated with intercellular H. brassicae growth occurred 3 h post inoculation for all genotypes and was the lowest in resistant accession Rd004. Cotyledon resistance to DM identified in radish is of high commercial interest and has the potential to be used in breeding programs.     

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.     

New sources of resistance of cowpea (Vigna unguiculata) toStriga gesnerioides,a parasitic angiosperm

Summary Thirty-seven accessions of cowpea and yard-long bean were assessed for resistance toStriga gesnerioides. Cowpea plants were grown using anin vitro method, then inoculated with young seedlings ofS. gesnerioides produced from seed from three West African countries. Resistance was assessed by comparing the number and size ofS. gesnerioides tubercles on these accessions with those on a known susceptible cowpea, cv. Blackeye. Two cowpea landraces, APL-1 and 87-2, were completely resistant toS. gesnerioides from Burkina Faso, Mali and Cameroon and partially resistant toS. gesnerioides from Niger. Complete resistance was expressed either as a hypersensitive response of infected root tissues or as a severely retarded development of successful infections. All other accessions, including three samples of yard-long bean were susceptible toS. gesnerioides. The original 87-2 plants segregated for resistance and susceptibility. However, uniformly resistant progeny were obtained by producing seed from vegetatively propagated clones of single resistant 87-2 plants. Resistance of APL-1 and 87-2 toS. gesnerioides was confirmed in pot and field trials. Neither of these cowpeas were resistant toAlectra vogelii. Varieties APL-1 and 87-2 provide additional sources of resistance to most races ofS. gesnerioides, including a newly discovered virulent race from Benin.Abbreviations ICRISAT International Crops Research Institute for the Semi-Arid Tropics - IITA International Institute of Tropical Agriculture - SAFGRAD Semi-Arid Food Grain Research and Development     

Inheritance of Russian wheat aphid resistance from tetraploid wheat accessions during transfer to hexaploid wheat

Identification of new sources of resistance to Russian wheat aphid (RWA) (Diuraphis noxia (Kurdjumov) in wheat (Triticum aestivum L.) has become very important with the identification of several new biotypes since 2003. Our objective was to characterize inheritance and expression of resistance to RWA biotype 2 from three tetraploid wheat landraces (Triticum turgidum L. subsp. dicoccon) during transfer to hexaploid wheat. Resistant tetraploid accessions PI 624903, PI 624904, and PI 624908 were crossed to the susceptible hexaploid cultivars ‘Len’ and ‘Coteau’. Resistant F1 progeny were advanced to the F2:3 by self-pollination and to the BC1F2 and BC2F1 by backcrossing. Leaf rolling and chlorosis were recorded in standard seedling screening tests on F1 and F2:3 individuals while the F2, BC1F1, BC1F2, and BC2F1 were scored as resistant or susceptible. Segregation in the BC1F1 and BC2F1 fit a 1:1 resistant:susceptible ratio, indicative of control by a single dominant gene. Segregation for resistance in the F2 did not fit 3:1, 13:3, or 15:1 ratios for any of the resistant accessions. Expression of resistance in homogeneous resistant F2:3 lines was greater than susceptible checks, similar to the resistant tetraploid accessions, and less than a line carrying the Dn7 resistance gene. Resistance derived from these tetraploid accessions will be useful to broaden the base of RWA resistance available for use in wheat breeding.     

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.     

Chromosomal location of resistance to Septoria tritici in seedlings of a synthetic hexaploid wheat, Triticum spelta and two cultivars of Triticum aestivum

Chromosomal location of resistance to two virulent Argentinean isolatesof Septoria tritici was studied in two wheat (Triticum aestivumL.) cultivars (Cappelle-Desprez & Cheyenne), a synthetic hexaploid(Synthetic 6x) and Triticum spelta in seedlings. Substitution lines of these(resistant or moderately resistant) genotypes into (susceptible) ChineseSpring were selected from a previous screening. For Synthetic 6x,resistance was clearly located in chromosome 7D. Chinese Spring with the7D chromosome substituted by Synthetic 6x showed almost completeresistance, similar to the level of Synthetic 6x. For the substitutions withCappelle-Desprez, Cheyenne, and T.spelta there were no lines with abehaviour similar to the resistant parent. However, some substitutions weremore resistant than the susceptible parent suggesting that severalchromosomes could be involved in the resistance of these genotypes toSeptoria leaf blotch.     

低温对大白菜幼苗生长及生理指标的影响

比较不同基因型大白菜对低温冷害的响应,为黑龙江大白菜种植及耐低温品种选育提供依据。白菜幼苗生长至3叶1心时,转入人工气候箱中进行低温处理3天,对6个白菜品种进行生物产量及生理性状测定与分析。结果表明：低温胁迫条件下,6 个供试品种生物产量及叶绿素含量均呈下降趋势。除‘新锉四号’外,其余5 个大白菜品种的可溶性蛋白含量在低温处理下均高于常温对照。除‘抗病91-12’外,其余5 个品种的过氧化物酶活性在低温处理下均高于常温对照。6 个大白菜品种耐冷性能力依次为：‘新北京三号’＞‘农学昌五小根’＞‘牡丹江二号’＞‘义和九号’＞‘抗病91-12’＞‘新锉四号’。因此,‘新北京三号’可作为耐冷性品种在黑龙江地区进行种植。