Creating cold resistant strawberry via interploidy hybridization between octoploid and dodecaploid

Strawberry cultivars showed limited cold resistance in the Northeast of China, while we obtained a synthetic dodecaploid strawberry hybrid ‘YH15-10’ (2n = 12x = 84) which showed sufficient cold resistance in this area. The reciprocal crosses between F. × ananassa cv. ‘Allstar’ (2n = 8x = 56) and ‘YH15-10’ (2n = 12x = 84) were carried out to select cold resistant strawberry in this study. The 134 seedlings were obtained from the cross of Allstar × YH15-10, while failed in its reciprocal cross. The 30 randomly selected seedlings were examined in terms of morphological characters, chromosome numbers and cold resistance. Most morphological characters were widely separated among F1 progeny with a high broad-sense heritability, which showed that these variations mainly resulted from genetic effect. Some hybrids exhibited heterosis, especially in growth vigor and runner production. Among the 30 tested hybrids, 28 decaploids (2n = 10x = 70), one octoploid (2n = 8x = 56) and one enneaploid (2n = 9x = 63) were observed. The 63.3% hybrids demonstrated higher cold resistance than that of ‘Allstar’ at P < 0.05. These high polyploidy strawberries have potential values in commercial production and modern cultivar improvement.     

Resistance screening of breeding lines and commercial tomato cultivars for <Emphasis Type="Italic">Meloidogyne incognita</Emphasis> and <Emphasis Type="Italic">M. javanica</Emphasis> populations (Nematoda) from Ethiopia

Soil and root samples were collected from major tomato growing areas of Ethiopia during the 2012/2013 growing season to identify root-knot nematode problems. DNA-based and isozyme techniques revealed that Meloidogyne incognita and M. javanica were the predominant Meloidogyne species across the sampled areas. The aggressiveness of different populations of these species was assessed on tomato cultivars Marmande and Moneymaker. The two most aggressive populations of each species were selected and further tested on 33 tomato genotypes. The resistance screening and mechanism of resistance was performed after inoculation with 100 freshly hatched (<24 h) second-stage juveniles (J2). Eight weeks after inoculation the number of egg masses produced on each cultivar was assessed. For the resistance mechanism study, J2 penetration and their subsequent development inside the tomato roots were examined at 1, 2, 4 and 6 weeks after inoculation. On both cultivars Marmande and Moneymaker all M. incognita and M. javanica populations formed a high number of egg masses indicating highly aggressive behaviour. Populations from ‘Jittu’ and ‘Babile’ for M. incognita and ‘Jittu’ and ‘Koka’ for M. javanica were selected as most aggressive. None of the 33 tomato genotypes were immune for these M. incognita and M. javanica populations. However, several tomato genotypes were found to have a significant effect on the number of egg masses produced indicating possible resistance. For M. javanica populations there were more plants from cultivars or breeding lines on which no egg masses were found compared to M. incognita populations. The lowest number of egg masses for both populations of M. incognita was produced on cultivars Bridget40, Galilea, and Irma while for M. javanica it was on Assila, Eden, Galilea, Tisey, CLN-2366A, CLN-2366B and CLN-2366C. Tomato genotypes, time (weeks after inoculation) and their interaction were significant sources of variation for J2 penetration and their subsequent development inside the tomato roots. Differential penetration was found in breeding lines such as CLN-2366A, CLN-2366B and CLN-2366C, but many of the selected tomato genotypes resistance for the tested M. incognita and M. javanica populations were expressed by delayed nematode development. Therefore, developing a simple screening technique to be used by local farmers or extension workers is crucial to facilitate selection of a suitable cultivar.     

Obtainment of an intergeneric hybrid between <Emphasis Type="Italic">Forsythia</Emphasis> and <Emphasis Type="Italic">Abeliophyllum</Emphasis>

Forsythia suspensa and F. ‘Courtaneur’ were used as female parents to cross with Abeliophyllum distichum in 2011 and an intergeneric hybrid of F. suspensa × A. distichum was obtained, though with very low seed set. The morphological characteristics, flower fragrance and volatile organic compounds of flowers were analysed. The intergeneric hybrid had intermediate morphological characteristics of both parents and flower fragrance and was confirmed as a true intergeneric hybrid by amplified fragment length polymorphism (AFLP) markers. Compared with its mother parent (F. suspensa), flowers of the intergeneric hybrid are pale yellow with delicate fragrance. Volatile organic compounds of flowers were retrieved by purge-and-trap techniques, and determined by gas chromatography and mass spectrometry (GC–MS). The main volatile organic components of F. suspensa were isoprenoids, while the main volatile organic components of A. distichum and the hybrid of F. suspensa × A. distichum were aliphatics. To determine the time and the site of intergeneric hybridizing barriers occured, the pollen tubes’ behavior after pollination was observed under fluorescence microscopy. It was found that significant pre-fertilization incompatibility existed in intergeneric crossing combinations [F. ‘Courtaneur’ (Pin) × A. distichum (Thrum) and F. suspensa (Pin) × A. distichum (Thrum)], and only a few pollen tubes of A. distichum penetrated into the ovaries of Forsythia. In our research, an intergeneric hybrid between Forsythia and Abeliophyllum was obtained for the first time, which will provide a solid foundation for expanding the flower color range of Forsythia and breeding fragrant-flowered cultivars.     

Hybrids of <Emphasis Type="Italic">Passiflora</Emphasis>: <Emphasis Type="Italic">P. gardneri</Emphasis> versus <Emphasis Type="Italic">P. gibertii,</Emphasis> confirmation of paternity,morphological and cytogenetic characterization

Two new varieties of interspecific hybrids of Passiflora have been developed from the cross between P. gardneri versus P. gibertii, both registered under the Passiflora Society International. Twelve putative hybrids were analyzed. Hybridization was confirmed using RAPD and SSR markers. Primer UBC11 (5′-CCGGCCTTAC-3′) generated informative bands. Primer SSR Pe75 has amplified species-specific fragments and a heterozygote status was observed with two parent bands 300 and 350 bp. The molecular markers generated have been analyzed for the presence or absence of specific informative bands. Based on the morphological characterization, we have identified two hybrid varieties: P. ‘Gabriela’ and P. ‘Bella’. P. ‘Gabriela’ produced flowers in bluish tones, bluish petals on the adaxial and abaxial faces, light blue sepals on the adaxial and light green on the abaxial faces, corona with the base of filaments in intense lilac color and white apex. P. ‘Bella’ produced flowers in lilac tones, intense lilac petals on the adaxial and abaxial faces, dark lilac sepals with whitish edges on the adaxial and light green on the abaxial faces, corona with the base of filaments in intense lilac color and white apex. The cytogenetic analysis verified that the hybrids have the same chromosomal number as the parents (2n = 18); the formation of bivalents between the homeologous chromosomes (n = 9) was observad, leading to regular meiosis, which allows the sexual reproduction and use of these hybrids in breeding programs.     

Adaptation to iron deficiency and high pH in evergreen azaleas (<Emphasis Type="Italic">Rhododendron</Emphasis> spp.): potential resources for breeding

The growth of evergreen azaleas (Rhododendron spp.) can be altered by iron (Fe) chlorosis when plants are cultivated in a neutral-alkaline substrate. In this study, morphological and physiological responses to alkalinity and Fe deficiency were evaluated in five diploid Japanese azaleas to assess their potential as resources for breeding. R. obtusum ‘Kirin’, R. indicum ‘Shinsen’, R. × pulchrum ‘Sen-e-oomurasaki’, R. indicum ‘Osakazuki’, and R. ripense were pot cultivated in a peat-based substrate for 10 weeks, in acid and alkaline growing media with both adequate and inadequate Fe nutrition. Plant performance was generally affected by high pH of the substrate, while Fe deficiency by itself influenced few of the evaluated parameters, possibly due to the complex adaptive response mechanisms of these slow growing ornamental shrubs. According to the biochemical and physiological variations recorded on a long period of cultivation, R. indicum ‘Osakazuki’ reported the best performance. This azalea could be a valuable resource for breeders.     

Development of high-density interspecific genetic maps for the identification of QTLs conferring resistance to <Emphasis Type="Italic">Valsa ceratosperma</Emphasis> in apple

Valsa canker (Valsa ceratosperma (Tode ex Fr.) Maire) is one of the most destructive fungal diseases of apple, especially in Eastern Asia. In this study, the first high density genetic linkage map of Malus asiatica × Malus domestica was constructed by 640 simple sequence repeats (SSRs) and 490 single nucleotide polymorphisms (SNPs), which spanned 1497.5 cM with an average marker interval of 1.33 cM per marker. Quantitative trait loci (QTLs) for apple’s resistance to V. ceratosperma isolates 03-8 and xc56 were identified using the linkage map. Lesion lengths were used as the phenotypic data for the QTL analysis, which were measured on 1-year-old shoots inoculated with conidia of the two isolates. One QTL for resistance to isolate 03-8 was mapped on LG 16, and one QTL for resistance to isolate xc56 was detected on LG 9. Our research not only promoted the further understanding of the genetic basis of apple’s resistance to Valsa canker but also provided two molecular markers that might be used in future marker-assisted selection for resistance in apple breeding programs.     

Induced expression of selected plant defence related genes in pot azalea, <Emphasis Type="Italic">Rhododendron simsii</Emphasis> hybrid

A set of putative marker genes to study plant defense responses against Polyphagotarsonemus latus, a key pest in the production of Rhododendron simsii hybrids, was selected and validated. Genes belonged to the biosynthetic pathway of phytohormones jasmonic acid (JA) (RsLOX, RsAOS, RsAOC, RsOPR3 and RsJMT) and salicylic acid (SA) (RsPAL and RsICS). Furthermore, RsPPO, a putative marker gene for oxidative stress response was successfully cloned from R. simsii. A CTAB-based extraction protocol was optimized to assure excellent RNA quality for subsequent RT-qPCR analysis. The RT-qPCR protocol was extensively tested and RsRG7 and RsRG14 were selected as reference genes from a geNorm pilot study. Validation of the marker genes was done after application with elicitors [methyl jasmonate (MeJA), coronatine, β-aminobutyric acid and acibenzolar-Smethyl] or wounding. Both 100 μM MeJA and 0.1 μM coronatine had a significant effect on the expression of all marker genes. Foliar application of MeJA on the shoots resulted in a significantly earlier response when compared to root application and subsequent sampling of the shoots. Expression patterns after MeJA treatment were generally the same in six R. simsii genotypes: ‘Nordlicht’, ‘Elien’, ‘Aiko Pink’, ‘Michelle Marie’, ‘Mevrouw Gerard Kint’ and ‘Sachsenstern’. Wounding resulted in the same expression patterns as MeJA treatment except for RsJMT. None of the genotypes showed a significant induction of the latter gene 6 h upon wounding. Findings of these experiments indicated that the tolerant genotype ‘Elien’ has low basal expression levels of RsPPO. This might be the first step towards the breeding of mite-tolerant genotypes.     

Fine mapping of <Emphasis Type="Italic">Ren3</Emphasis> reveals two loci mediating hypersensitive response against <Emphasis Type="Italic">Erysiphe necator</Emphasis> in grapevine

Grapevine (Vitis vinifera L.) is economically very important for the production of wine, table grapes and raisins. However, grapevine is threatened by a brought range of pathogens. A destructive disease worldwide is powdery mildew caused by the ascomycete Erysiphe necator. In the grapevine cultivar `Regent’ a resistance locus against E. necator, Ren3, was previously reported. It spans an interval of approximately seven Mb on chromosome 15. We attempted to delimit this interval to facilitate its further molecular analysis. New simple sequence repeat markers targeted to the Ren3 region were designed. They were applied for fine mapping in the cross populations of ‘Regent’ × ‘Lemberger’ and ‘Regent’ × ‘Cabernet Sauvignon’ that segregate for E. necator resistance. Complementarily we scored E. necator infection levels of ‘Regent’ × ‘Lemberger’ progeny at different time points over the course of the vegetation period in 2015 and 2016. Subsequent QTL analysis revealed a maximum LOD value that shifted during the season from marker GF15-10 located at 2.2 Mb to marker GF15-53 located at 3.5 Mb and to marker ScORA7* located at 9.4 Mb on chromosome 15 (positions according to the grapevine reference genome of PN40024). To investigate the Ren3-encoded resistance mechanism we performed detached leaf infection assays for microscopic studies. These revealed that Ren3 carrying individuals react with a hypersensitive response. Results of detached leaf assays on recombinants in the Ren3 locus indicate that not only one, but two distinct genetic regions on chromosome 15 mediate hypersensitive response against E. necator.     

Mapping of new resistance (<Emphasis Type="Italic">Vr2</Emphasis>, <Emphasis Type="Italic">Rm1</Emphasis>) and ornamental (<Emphasis Type="Italic">Di2</Emphasis>, <Emphasis Type="Italic">pl</Emphasis>) Mendelian trait loci in peach

Peach powdery mildew is one of the major diseases of the peach. Various sources of resistance to PPM have thus been identified, including the single dominant locus Vr2 carried by the peach rootstock ‘Pamirskij 5’. To map Vr2, a linkage map based on microsatellite markers was constructed from the F₂ progeny (WP²) derived from the cross ‘Weeping Flower Peach’ × ‘Pamirskij 5’. Self-pollinations of the parents were also performed. Under greenhouse conditions, all progenies were scored after artificial inoculations in two classes of reactions to PPM (resistant/susceptible). In addition to Vr2, WP² segregated for three other traits from ‘Weeping Flower Peach’: Rm1 for green peach aphid resistance, Di2 for double-flower and pl for weeping-growth habit. With their genomic locations unknown or underdocumented, all were phenotyped as Mendelian characters and mapped: Vr2 mapped at the top of LG8, at 3.3 cM, close to the CPSCT018 marker; Rm1 mapped at the bottom of LG1, at a position of 116.5 cM, cosegregating with the UDAp-467 marker and in the same region as Rm2 from ‘Rubira’^®; Di2 mapped at 28.8 cM on LG6, close to the MA027a marker; and pl mapped at 44.1 cM on LG3 between the MA039a and SSRLG3_16m46 markers. Furthermore, this study revealed, for the first time, a pseudo-linkage between two traits of the peach: Vr2 and the Gr locus, which controls the red/green color of foliage. The present work therefore constitutes a significant preliminary step for implementing marker-assisted selection for the four major traits targeted in this study.     

Characterization of the resistance to <Emphasis Type="Italic">Phytophthora infestans</Emphasis> in local potato cultivars in Bolivia

This experiment was carried out to investigate whether and how much field resistance to late blight, caused by Phytophthora infestans, is present in the local cultivated potato germplasm. In total 36 entries were compared in a field experiment in an area highly conducive to late blight development. Of the 36 cultivars 32 were local cultivars belonging to five Solanum species, S. tuberosum (1 accession), S. andigena (18), S. juzepczukii (2), S. stenotomum (9) and S. ajanhuiri (2). The other four cultivars were derived from breeding programmes, one being the Dutch cultivar Alpha used as a highly susceptible control. The 36 cultivars were planted according to a simple 6 × 6 lattice design with three replicates. Each replicate was divided in six incomplete blocks each with six cultivars. The disease severity was assessed weekly during 9 weeks starting 48 days after planting. The area under the disease progress curve (AUDPC) was used as a measure of the field resistance. Nine isolates from surrounding potato fields were tested for their virulence to the resistance genes R1–R11 using 22 differential cultivars. The components of the field resistance of 19 of these cultivars were compared in the greenhouse using a local isolate with virulence to all known R-genes, except to R9. The nine isolates represented seven races with a race complexity varying from 7 to 10 virulence factors. All isolates carried virulence against R1, R2, R3, R7, R10 and R11, while virulence against R9 was absent. The AUDPC among the 32 local cultivars ranged from very large, significantly larger than that of ‘Alpha’ to very small. The AUDPC from S. stenotomum accessions ranged from very large to intermediate, those from S. andigena accessions from large to very small. Especially among the S. andigena accessions interesting levels of field resistance were found. Four components of field resistance were assessed, latency period (LP), lesion size (LS), lesion growth rate (LGR) and relative sporulation area (RSA). All four showed a considerable variation among the cultivars. The LP ranged from 3½ to 6 days. The LS ranged from 225 mm² to 20 mm². The LGR varied about six-fold, the RSA more than 10-fold. The components tended to vary in association with one another. LP and LGR were well associated with each other and had a significant correlation with the AUDPC.     

Effective selection of soybean cultivars to wildfire disease pathogen Pseudomonas amygdali pv. tabaci

Wildfire, caused by Pseudomonas amygdali pv. tabaci is one of the most destructive bacterial diseases and was recognized as a disease of soybean in 1943. Wildfire has been seen a steady increase in the incidence and prevalence on some cultivars of soybean in Korea by climatic changes but there is little information on effective control measures for wildfire or soybean varieties showing complete resistance to the disease. In this study, the efficient and reliable screening method to evaluate soybean genotype for resistance to P. amygdali pv. tabaci in field had been developed. In order to determine the host resistance of the soybean cultivar against P. amygdali pv. tabaci, development of symptom by infiltration inoculation was evaluated. Significant differences between susceptible plants and resistant plants were observed through these assays. Based on these results, ‘Shinpaldal2’, ‘Daepung’ are resistant to wildfire compared to ‘Hwangeum’, ‘Taekwang’. The optimum temperature of this pathogen was between 20-25°C and when the pathogen was in the optimum temperature, the responses of susceptible or resistant cultivar were dramatically different. Prior to initiation of resistance breeding of soybean wildfire, it is imperative to set uniform resistance screening techniques. The obtained results can be effectively used to enhance the selection of wildfire resistance as well as directly applied in resistant soybean development. Resistant lines identified through this assay could be directly used in soybean breeding programs for wildfire resistance.     

Transformation of <Emphasis Type="Italic">Campanula</Emphasis> by wild type <Emphasis Type="Italic">Agrobacterium rhizogenes</Emphasis>

Compact growth is an important quality criterion in horticulture. Many Campanula species and cultivars exhibit elongated growth which is suppressed by chemical retardation and cultural practice during production to accommodate to the consumer’s desire. The production of compact plants via transformation with wild type Agrobacterium rhizogenes is an approach with great potential to produce plants that are non-GMO. Efficient transformation and regeneration procedures vary widely among both plant genera and species. Here we present a transformation protocol for Campanula. Hairy roots were produced on 26–90% of the petioles that were used for transformation of C. portenschlagiana (Cp), a C. takesimana × C. punctata hybrid (Chybr) and C. glomerata (Cg). Isolated hairy roots grew autonomously and vigorously without added hormones. The Cg hairy roots produced chlorophyll and generated plantlets in response to treatments with cytokinin (42 µM 2iP) and auxin (0.67 µM NAA). In contrast, regeneration attempts of transformed Cp and Chybr roots lead neither to the production of chlorophyll nor to the regeneration of shoots. Agropine A. rhizogenes strains integrate split T-DNA in T_L- and T_R-DNA fragments into the plant genome. In this study, regenerated plants of Cg did not contain T_R-DNA, indicating that a selective pressure against this T-DNA fragment may exist in Campanula.     

Pollen characteristics and stigma receptivity for <Emphasis Type="Italic">Anemone coronaria</Emphasis> L.

To obtain crossing products, an efficient pollination and subsequent fertilization is essential. This efficient pollination is achieved by pollen germination and tube growth. Here, these pollen characteristics of 2 genetically differentiating cultivars of Anemone coronaria L. were investigated in vitro. For the essential components boron, calcium and an osmoticant, only calcium showed to be crucial for pollen germination. Boron concentrations influenced the pollen tube length with a concentration of 100 mg l^?1 H₃BO₃ resulting in the longest pollen tubes. For the osmoticant sucrose, a concentration of 100 g l^?1 caused a significant positive effect on both pollen germination and pollen tube length for the 2 cultivars. The cultivars reacted similarly with respect to the investigated compounds. Next to this, the pollen development was correlated with 8 different flower stages in A. coronaria. As a final point, the germination of an optimized pollen germination medium was compared with in vivo pollen germination in cross-pollinations within the same cultivar (identified by aniline blue staining). For ‘Mistral Wine’, pollen germination percentage was lower in vitro than in vivo, while ‘Wicabri Blue’ pollen showed no significant difference in germination rates in vivo and in vitro. To achieve fertilization a following requisite is that the stigma is receptive. To study this, the most receptive female flower stage of the 8 different flower stages for A. coronaria was characterized by aniline blue staining. A. coronaria clearly showed protogyny.     

Genetic mapping of resistance to <Emphasis Type="Italic">Puccinia hordei</Emphasis> in three barley doubled-haploid populations

The success of breeding for barley leaf rust (BLR) resistance relies on regular discovery, characterization and mapping of new resistance sources. Greenhouse and field studies revealed that the barley cultivars Baronesse, Patty and RAH1995 carry good levels of adult plant resistance (APR) to BLR. Doubled haploid populations [(Baronesse/Stirling (B/S), Patty/Tallon (P/T) and RAH1995/Baudin (R/B)] were investigated in this study to understand inheritance and map resistance to BLR. The seedlings of two populations (B/S and R/B) segregated for leaf rust response that conformed to a single gene ratio (\({\text{X}}_{1:1}^{2}\) = 0.12, P > 0.7 for B/S and \({\text{X}}_{1:1}^{2}\) = 0.34, P > 0.5 for R/B) whereas seedlings of third population (P/T) segregated for two-gene ratio (\({\text{X}}_{1:1}^{2}\) = 0.17, P > 0.6) when tested in greenhouse. It was concluded that the single gene in Baudin and one of the two genes in Tallon is likely Rph12, whereas gene responsible for seedling resistance in Stirling is Rph9.am (allele of Rph12). The second seedling gene in Tallon is uncharacterized. In the field, APR was noted in lines that were susceptible as seedlings. A range of disease responses (CI 5–90) was observed in all three populations. Marker trait association analysis detected three QTLs each in populations B/S (QRph.sun-2H.1, QRph.sun-5H.1 and QRph.sun-6H.1) and R/B (QRph.sun-1H, QRph.sun-2H.2, QRph.sun-3H and QRph.sun-6H.2), and four QTLs in population P/T (QRph.sun-6H.2, QRph.sun-1H.2, QRph.sun-5H.2 and QRph.sun-7H) that significantly contributed to low leaf rust disease coefficients. High frequency of QRph. sun-5H.1, QRph. sun-6H.1, QRph. sun-1H.1, QRph. sun-2H.2, QRph. sun-6H.2, QRph. sun-7H (based on presence of the marker, closely associated to the respective QTLs) was observed in international commercial barley germplasm and hence providing an opportunity for rapid integration into breeding programmes. The identified candidate markers closely linked to these QTLs will assist in selecting and assembling new APR gene combinations; expectantly this will help in achieving good levels of durable resistance for controlling BLR.     

Inheritance of type IV glandular trichome density and its association with whitefly resistance from <Emphasis Type="Italic">Solanum galapagense</Emphasis> accession LA1401

Tomato is affected by a large number of arthropod pests, among which the whitefly (Bemisia tabaci) is considered to be one of the most destructive. Several accessions of the wild species of Solanum galapagense, including accession LA1401, are considered resistant to whitefly (B. tabaci). This resistance has been associated with the presence of type IV glandular trichomes on the leaf surface. Our research aimed to study the inheritance of type IV glandular trichome density and its association with resistance to whitefly (B. tabaci biotype B) in populations derived from the interspecific cross Solanum lycopersicum × S. galapagense ‘LA1401.’ High estimates for both broad-sense and narrow-sense heritabilities of type IV glandular trichome densities suggest that inheritance of this trait is not complex. Whitefly resistance was associated with high density of type IV glandular trichomes. F₂ (S. galapagense × S. lycopersicum) population plants selected for the highest densities of type IV glandular trichomes showed similar levels of resistance to those found in the donor of resistance LA1401.     

Resistance to <Emphasis Type="Italic">Cucumber green mottle mosaic virus</Emphasis> in <Emphasis Type="Italic">Cucumis sativus</Emphasis>

Cucumber green mottle mosaic virus (CGMMV) is a severe threat for cucumber production worldwide. At present, there are no cultivars available in the market which show an effective resistance or tolerance to CGMMV infection, only wild Cucumis species were reported as resistant. Germplasm accessions of Cucumis sativus, as well as C. anguria and C. metuliferus, were mechanically infected with the European and Asian strains of CGMMV and screened for resistance, by scoring symptom severity, and conventional RT-PCR. The viral loads of both CGMMV strains were determined in a selected number of genotypes using quantitative RT-PCR. Severe symptoms were found following inoculation in C. metuliferus and in 44 C. sativus accessions, including C. sativus var. hardwickii. Ten C. sativus accessions, including C. sativus var. sikkimensis, showed intermediate symptoms and only 2 C. sativus accessions showed mild symptoms. C. anguria was resistant to both strains of CGMMV because no symptoms were expressed and the virus was not detected in systemic leaves. High amounts of virus were found in plants showing severe symptoms, whereas low viral amounts found in those with mild symptoms. In addition, the viral amounts detected in plants which showed intermediate symptoms at 23 and 33 dpi, were significantly higher in plants inoculated with the Asian CGMMV strain than those with the European strain. This difference was statistically significant. Also, the amounts of virus detected over time in plants did not change significantly. Finally, the two newly identified partially resistant C. sativus accessions may well be candidates for breeding programs and reduce the losses produced by CGMMV with resistant commercial cultivars.     

Interspecific hybridization in <Emphasis Type="Italic">Sarcococca</Emphasis> supported by analysis of ploidy level,genome size and genetic relationships

Knowledge of ploidy level differences, genome size and genetic relationships between species facilitates interspecific hybridization in ornamentals. For Sarcococca (Buxaceae) only limited (cyto)genetic information is available. The aim of this study was to determine the genome size and chromosome number and to unravel the genetic relationships of a breeder’s collection using AFLP marker analysis. Based on these results, interspecific crosses were made and the efficiency and hybrid status was verified. Two groups of diploid plants (2n = 2x = 24) were observed, with either a genome size of 4.11–4.20 or 7.25–9.63 pg/2C. All the tetraploid genotypes (2n = 4x = 48) had genome sizes ranging from 7.91 to 8.18 pg/2C. In crosses between parents with equal ploidy level and genome size a higher crossing efficiency (on average 58% of the hybridizations resulting in fruits) and more true hybrids (on average 96% of the offspring) were obtained compared to crosses between plants with different genome size and ploidy level (on average 23% fruits and 24% hybrids, respectively). In none of the cross combinations, the ploidy level or genome size was found to be a complete hybridization barrier, although unilateral incongruity was found in some cross combinations. Distant genetic relationships did not hamper the hybridization within Sarcococca genotypes. Our findings will contribute to a more efficient breeding program and a faster achievement of hybrids with an added value.     

Development of late-bolting F<Subscript>1</Subscript> hybrids of Chinese cabbage (<Emphasis Type="Italic">Brassica rapa</Emphasis> L.) allowing early spring cultivation without heating

We developed new F₁ hybrids of Chinese cabbage (Brassica rapa L. ssp. pekinensis) that allow cultivation earlier in spring without heating by introducing extremely late-bolting alleles at two homologs of the flowering repressor Flowering Locus C (BrFLC2 and BrFLC3) from non-heading ‘Leafy Green Parental Line No. 2’. These new F₁ hybrids were produced by the following four steps. First, the extremely late bolting selected lines were developed. These selected lines headed in spring after overwintering cultivation, whereas the conventional F₁ cultivars flowered. Secondly, an investigation of the three plantings showed that our F₁ hybrids formed heads when seeds were sown from mid-February to early March, whereas the conventional F₁ cultivar did not form heads because of premature bolting. Thirdly, we identified some F₁ hybrids with extremely late bolting during early spring cultivation in an investigation of many F₁ hybrids. Finally, based on an investigation across four cold regions for 2 years, we compared the commercialization rate, defined as the proportion of plants greater than 2000 g in weight and with a flowering stalk less than 10 cm long. Then we identified a F₁ of MS02 × 12-04 which had a high commercialization rate on average (92%), whereas the rates of three conventional F₁ cultivars were only 0–2%. In the near future, these F₁ hybrids will be valuable late-bolting cultivars despite climate change, permitting stable cultivation and harvest over wide regions.     

Determination of responses of different bean cultivars against races of <Emphasis Type="Italic">Pseudomonas syringae</Emphasis> pv <Emphasis Type="Italic">phaseolicola</Emphasis>, causal agent of halo blight of bean

Use of resistant plant varieties combined with other disease management practices is regarded as the most practical approach to control of seed-borne bacterial disease agents. In this study, responses of different bean cultivars to nine different races of Pseudomonas syringae pv phaseolicola, the causal agent of bacterial halo blight of common bean (Phaseolus vulgaris L.), were determined. During compatible interaction in susceptible cultivars, virulent bacterial races caused water soaked lesion at sites of inoculation. Similar lesions developed in moderately resistant cultivars but symptoms were later associated with more tissue browning around the sites of inoculation. In contrast, the resistant response, produced the characteristic hypersensitive reaction (HR), was characterized as a small discrete browning and tissue collapse at site of inoculation. No local cultivars showed complete resistance to all races tested. Bean cultivars Sehirali-90 and Göynük-98 were found to be resistant or moderately resistant to five different bacterial races. Bean cultivar, Karaca?ehir-90, on the other hand, was found to be resistant or moderately resistant to six different bacterial races. Analysis of bacterial growth and the accumulation of isoflavonoid bean phytoalexin, phaseollin in planta were carried out for tissues expressing compatible and incompatible interactions to enable a link to be made between reaction phenotypes and restriction of bacterial growth and phytoalexin accumulations. Development of the HR was clearly associated with the restricted multiplication of bacteria during incompatible interactions. A time-course accumulation analysis on pods treated with different races of bacterial agent showed that a strong correlation was observed between the timing and extent of cell death and accumulation of phaseollin, being rapid and extensive in incompatible interactions compared to compatible interaction.