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.     

Inheritance of downy mildew resistance at cotyledon and adult-plant stages in ‘Couve Algarvia’ (Brassica oleracea var. tronchuda)

Parasitic nematodes damage white clover (Trifolium repens) roots, negatively impacting forage yield and persistence. No single gene resistance to nematodes has been identified in white clover. Trifolium semipilosum (2n = 2x = 16) genotypes exhibiting either complete resistance or susceptibility to infection by the clover root-knot nematode (CRKN), Meloidogyne trifoliophila, were identified. F₁ progeny (n = 92) of a pair-cross between ‘TsR’, a plant heterozygous for the resistance phenotype and ‘TsS’, a plant homozygous for the susceptible phenotype, were challenged with infective CRKN juveniles and evaluated subsequently for root galling. Segregation analysis indicated the resistance phenotype may be conferred by a single dominant allele at a locus (designated TRKR, Trifolium Root-knot Resistance) subject to segregation distortion. TsR, TsS, and bulked resistant and bulked susceptible F₁ progeny (n = 12/bulk) were screened using T. repens microsatellite (SSR) markers. Three SSRs revealed polymorphism in TsR and the resistant bulk, of which prs090 and prs247 map to loci on T. repens linkage group D2. Progeny were genotyped with these three SSRs and 23 additional SSRs from T. repens groups D1 and D2. Linkage analysis in both TsR and TsS demonstrated macro-synteny between T. repens group D homoeologues and the T. semipilosum linkage group (designated D_Ts) containing the TRKR locus. Significant segregation distortion was detected in TsR, and recombination in the central region of the T. semipilosum linkage group was suppressed relative to T. repens in both parents. These data demonstrate the opportunities and challenges for comparative mapping in Trifolium and characterisation of loci conferring resistance to plant-parasitic nematodes.     

Linkage in Spinacia oleracea L. of two race-specific genes for resistance to downy mildew Peronospora farinosa f. sp. spinaciaeByford

Summary The genetics of resistance to races 1 and 2 of downy mildew Peronospora farinosa f. sp. spinaciae in the spinach variety Nores was investigated in crosses and backcrosses with the susceptible variety Eerste Oogst. Resistance appears to be governed by two closely linked genes in coupling phase with a recombination percentage of 4.6.     

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.     

Inheritance of downy mildew resistance in Indian cauliflower (group III)

Summary Inheritance of downy mildew (Peronospora parasitica) resistance in three resistant x susceptible crosses, one susceptible x susceptible and one resistant x resistant cross were studied in Indian cauliflower (Group III) over the two years (1990 and 1991). No significant difference was observed between the years for various estimates and hence pooled data are presented. Downy mildew resistance in crosses cc×HR 5-4 and 3-5-1-1×244 (R×S) is governed by single dominant gene PPA3 but in cross cc×244 (R×S), recessive epistasis was observed. The resistance level was not improved in both the cc×3-5-1-1 (R×R) and 244×267-6-9 (S×S) crosses. Exploitation of downy mildew resistance from cc and 3-5-1-1 in F₁ hybrid is explained in detail.     

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.     

Transfer of powdery mildew resistance from Brassica carinata to Brassica oleracea through embryo rescue

Interspecific hybrid plants and backcross 1 (BC₁) progeny were produced through sexual crosses and embryo rescue between Brassica carinata accession PI 360883 and B. oleracea cvs Titleist’and‘Cecile’to transfer resistance to powdery mildew to B. oleracea. Four interspecific hybrids were obtained through application of embryo rescue from crosses with B. carinata as the maternal parent, and their interspecific nature confirmed through plant morphology and random amplified polymorphic DNA (RAPD) analysis. Twenty‐one BC₁ plants were obtained through sexual crosses and embryo rescue although embryo rescue was not necessary to produce first backcross generation plants between interspecific hybrids and B. oleracea. All interspecific hybrids and eight of the BC₁ plants were resistant to powdery mildew.     

Development of black rot resistant interspecific hybrids between Brassica oleracea L. cultivars and Brassica accession A 19182, using embryo rescue

Black rot is a bacterial disease of Brassica oleracea caused by Xanthomonas campestris pv. campestris. Resistance to the major black rot races 1 or 4 has been identified in related Brassica species including B. carinata and B. napus. In this study, two B. juncea accessions (A 19182 and A 19183) that are resistant to races 1 and 4 of Xcc were used as maternal and paternal parents to generate interspecific hybrids with B. oleracea cultivars. Interspecific hybrids were recovered using the embryo rescue technique and confirmed through inheritance of paternal molecular markers. Twenty-six interspecific hybrid plants were obtained between A 19182 and B. oleracea cultivars, but no interspecific hybrids were obtained using A 19183. Although interspecific hybrid plants were male sterile, they were used successfully as maternal parents to generate backcross plants using embryo rescue. All hybrid and BC₁ plants were resistant to black rot races 1 and 4.     

Selection for partial resistance to downy mildew in peas by means of greenhouse tests

Summary A partially downy mildew resistant pea line was back-crossed to a susceptible cultivar with more pods per node and lower seed weight. Breeding lines with different degrees of infection in a greenhouse test were selected. These lines and the two parental lines were investigated in field trials and tested in the greenhouse for four generations. Significant genetic variation among lines was found for infection of seedlings in greenhouse tests, and infection of pods, pod set and seed weight in field-trials. Infection of seedlings in the greenhouse was correlated with infection of pods in the field. In greenhouse tests, the non-genetic variance component was large in comparison with the genetic component and a significant genotype trial interaction was found. Significant repeatability was obtained for downy mildew on seedlings and pods, number of pods per node and seed weight. An unfavourable correlation between susceptibility to downy mildew and number of pods per node was found. No single breeding line showed the ideal combination of good resistance, high number of pods per node and small seeds. However, one line showing better resistance than the susceptible parent, with smaller seeds and more pods per node than the resistant parent was found. The susceptible parent also carries some resistance factor that is not present in the resistant parent.     

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.     

RAPD markers linked to resistance to downy mildew disease in soybean

To determine and utilize RAPD markers linked to resistance to downymildew incited by Peronospora manshurica in soybean, a resistantcultivar `AGS129' was crossed to a susceptible cultivar `Nakhon Sawan 1'(NS1). F₂ and BC₁ populations were advanced from the F₁ and evaluatedfor resistance to the disease. ²-test demonstrated that the resistancewas controlled by a single dominant gene (Rpmx). Near-isogenic lines(NILs) and bulked segregant analysis (BSA) were used to identify RAPDmarkers linked to the gene. Six DNA bulks namely F₅(R), F₅(S),BC₆F₃(R), BC₆F₃(S), F₂(R) and F₂(S) were set up by pooling equalamount of DNA from 8 randomly selected plants of each disease responsetype. A total of 180 random sequence decamer oligonucleotide primerswere used for RAPD analysis. Primer OPH-02 (5 TCGGACGTGA 3 andOPP-10 (5 TCCCGCCTAC 3) generated OPH-02₁₂₅₀ and OPP-10₈₃₁fragments in donor parent and resistant bulks, but not in the recurrentparent and susceptible ones. Co-segregation analysis using 102 segregatingF₂ progenies confirmed that both markers were linked to the Rpmxgene controlling downy mildew disease resistance with a genetic distance of4.9 cm and 23.1 cm, respectively. Marker OPH-02₁₂₅₀ was presentin 13 of 16 resistant soybean cultivars and absent in susceptible cultivars,thus confirming a potential for MAS outside the mapping population.     

自花授粉诱导的甘蓝功能基因BoSPI的克隆与表达分析

自交不亲和性是甘蓝在长期进化过程中形成的防止自交衰败、促进杂交优势的一种复杂而完善的遗传机制。克隆自交不亲和性相关基因对甘蓝自交不亲和性的深入研究和利用有重要意义。本研究通过挖掘0~60 min自花和异花授粉的甘蓝柱头转录组数据, 筛选到一个受自花授粉诱导上调表达的基因, 命名为BoSPI。BoSPI开放阅读框534 bp, 编码177个氨基酸, 理论等电点为4.21, 不包含信号肽和跨膜区, 含有4个保守的EF-hand结构域。BoSPI基因起始密码子上游2000 bp的核苷酸序列中含有真菌诱导响应、代谢调节以及器官形成等应答元件。BoSPI基因在大肠杆菌中可诱导表达为17 kD的蛋白。BoSPI在柱头中表达量最高, 在花瓣、萼片、叶片、雄蕊表达量较低。BoSPI蛋白被定位在细胞膜和细胞质。自花授粉30 min后对BoSPI基因的诱导表达显著增强。表明BoSPI参与了柱头响应自花花粉刺激的分子过程, 可能是实现甘蓝自交不亲和性相关的某种新功能基因。     

Differential response and genes for resistance to Peronospora parasitica (downy mildew) in Brassica juncea (mustard)

The response of a wide range of Brassica juncea accessions to 14 isolates of Peronospora parasitica, 12 from India (IP00A, IP02, IP03, IP04, IP04A, IP05, IP05B, IP33 and IP33A were derived from B. juncea; IP09, IP14 and IP13A from B. rapa) and two from B. napus in the UK (R1 and P003), was screened. Sixteen differential host response groups to these isolates (classified as groups A‐P) were identified. Groups‘A’and‘B’expressed the widest resistance profiles to these isolates. Group‘A’was susceptible to isolates IP05 and IP05B, moderately resistant to isolate IP33 and resistant to all other isolates. Group‘B’was susceptible to isolates IP03, IP04 and IP04A, and resistant to the other isolates. Putative homozygous lines resistant to all 14 isolates were selected from the F₄ progeny of crosses involving lines RESBJ‐200 from group‘A’(selection from cv. Kranti) and RESBJ‐190 from group‘B’(selection from cv. Krishna). Both selections were selfed and tested for uniformity of reactions to all isolates for three generations. The resistance of RESBJ‐200 to isolates IP00A, IP04A and IP33A seems to be conditioned by single dominant genes. The resistance of RESBJ‐190 to isolates IP00A, IP05B and IP33A was also conditioned by single dominant genes. The gene for resistance to IP00A and IP33A in RESBJ‐200 seems to be independent of the genes for resistance to the same isolates in RESBJ‐190. The new genes for differential resistance to P. parasitica will be of value in future studies of the genetics of the host‐pathogen interaction and for breeding for disease resistance.     

The transfer of triazine resistance from Brassica napus L. to B. oleracea L. III. First backcross to parental species

Summary Atrazine resistant Brassica napus × B. oleracea F₁ hybrids were backcrossed to both parental species. The backcrosses to B. napus produced seeds in both directions but results were much better when the F₁ hybrid was the pollen parent. Backcrosses to B. oleracea failed completely but BC₁s were rescued by embryo culture both from a tetraploid hybrid (2n = 4x = 37; A₁C₁CC) and sesquidiploid hybrids (2n = 3x = 8; A₁C₁C). Progeny of crosses between the tetraploid hybrid and B. oleracea had between 25 and 28 chromosomes. That of crosses between the sesquidiploid hybrid and B. oleracea had between 21 and 27. A few plants that had chromosome counts outside the expected range may have originated from either diploid parthenogenesis, unreduced gametes or spontaneous chromosome doubling during in vitro culture. Pollen stainability of the BC₁s ranged from 0% to 91.5%. All the BC₁s to B. oleracea were resistant to atrazine.     

SCAR and CAPS markers flanking the Brassica oleracea L. Pp523 downy mildew resistance locus demarcate a genomic region syntenic to the top arm end of Arabidopsis thaliana L. chromosome 1

We recently mapped the Pp523 locus that includes a single, dominant gene conferring resistance to downy mildew expressed in adult plants to a 75.1 cm long linkage group on a genetic linkage map of Brassica oleracea L. More recently, we identified a new AFLP marker 2.8 cm downstream from the resistance gene. The five DNA markers within an 8.5 cm region encompassing the Pp523 gene were cloned and sequenced. Three of these markers were transformed into SCARs (sequence characterised amplified regions), however, two among them were monomorphic and were analysed as CAPS (cleaved amplified polymorphic sequence) markers among the mapping population. Searched against genomic databases, the five B. oleracea DNA-marker sequences matched Arabidopsis thaliana L. gene sequences that delimit a conserved syntenic region in the top arm end of chromosome 1 of this last species. Considering the close genetic relatedness between both species, the information on this specific genomic region in A. thaliana is particularly useful for the construction of a fine-scale map of the corresponding genomic region in B. oleracea. The identified SCAR and CAPS markers can be used for marker assisted selection (MAS) in breeding programs aimed at the introgression of the Pp523 resistance locus, allowing the reliable indirect identification of plants harbouring the resistance gene with a margin of error of approximately six in ten-thousand selected plants.     

Genetic analysis of adult plant resistance to powdery mildew in pea (Pisum sativum L.)

Summary An analysis of adult plant resistance of powdery mildew in 15 F₁, F₂ and F₃ populations of pea derived from crossing 15 diverse and susceptible lines with one resistant line revealed that resistance to powdery mildew is controlled by duplicate recessive genes. The genes were designated as er₁ and er₂.Disease reaction showed independent segregation with three known markers in the resistant parent, namely, af (afila, chromosome 1), st (stipule reduced, chromosome 3) and tl (clavicula, chromosome 7).Contribution form the Department of Genetics and Plant Breeding Banaras Hindu University, Varanasi-221005, India.     

Transfer of resistance to race 2 of Plasmodiophora Brassicae from Brassica napus to cabbage (B. oleracea var. Capitata). I. Interspecific hybridization between B. napus and B. oleracea var. Capitata

Summary Interspecific hybridization between Brassica napus L. (2n=38, a₁a₁c₁c₁) and B. oleracea var. capitata L. (2x- and 4x-cabbage; 2n=2x=18, cc and 2n=4x=36, cccc) was carried out for the purpose of transferring clubroot disease resistance from the amphidiploid species to cabbage. Nineteen hybrids with three different chromosome levels (2n=28, a₁c₁c; 2n=37, a₁c₁cc and 2n=55, a₁c₁cccc) were obtained. The F₁ plants were mostly intermediate between the two parents but as the number of c genomes in the hybrids increased, the more closely the hybrids resembled the cabbage parent. All F₁ hybrids were resistant when tested against race 2 of Plasmodiophora brassicae wor. The complete dominance of resistance over susceptibility suggested that the gene(s) controlling resistance to this particular race of the clubroot pathogen is probably located on a chromosome of the a genome in Brassica.Contribution No. J654.     

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.