Inheritance of resistance to early blight disease in a diploid potato population

Early blight disease, caused by Alternaria solani Sorauer, is a serious disease of potato foliage and tubers that occurs in most potato‐growing regions world‐wide. Developing new potato cultivars with resistance to early blight may reduce losses in the field and in storage, and lessen the need for fungicide applications. A total of 280 clones, derived from 72 maternal half‐sib families from a diploid random‐mated hybrid population of Solarium phureja×Solarium stenotomum were examined for resistance to early blight. The clones that were evaluated in a replicated field trial for 2 years in Pennsylvania, USA, had similar early blight intensity both years. Significant differences were found among families, within families and for the interaction of years × within families. Broad‐sense heritability for resistance, measured as area under the disease progress curve (AUDPC), was estimated as 0.73, with a 95% confidence interval of 0.65‐0.78, and narrow‐sense heritability was estimated as 0.61 ± 0.29 (P = 0.05). The correlation of AUDPC for early blight between years was 0.57 (P < 0.0001). These results suggest that this diploid population is worthy of use in breeding for early blight resistance.     

Transmission of Verticillium wilt resistance to tetraploid potato via unilateral sexual polyploidization

Summary Verticillium wilt is a serious disease of potato and is caused by the soil-borne fungi Verticillium dahliae and V. albo-atrum. No major cultivar is resistant to this disease. Two diploid interspecific potato clones, C287 and C545, were previously identified with consistently high levels of Verticillium wilt resistance and are thought to have the same genotype for the loci that confer resistance to V. dahliae stem colonization. The purpose of this study was to determine whether resistance to V. dahliae stem colonization could be transferred to the tetraploid level in potato via unilateral sexual polyploidization (USP). Progenies in eight families obtained by crossing C287 and C545 to two tetraploid breeding clones, S438 and S440, and the cultivar ‘Atlantic’ were planted in a V. dahliae-infested field and a field without a significant V. dahliae infestation. Resistance was evaluated relative to C545 and C287. There were differences among families for stem colonization and yield. No differences between the mean stem colonization of C545 and C287 progeny were detected. Family differences due to the tetraploid parents indicate that they contributed resistance to the progeny. Stem colonization data from this experiment were consistent with the proposed complementary two-gene model for Verticillium wilt resistance in the diploid parents. Unilateral sexual polyploidization is an effective method for transferring V. dahliae stem colonization resistance to the tetraploid level.     

Variation for resistance to head blight caused by Fusarium graminearum in wild emmer (Triticum dicoccoides) originating from Israel

Head blight of wheat (FHB, scab) caused by Fusarium spp. has been associated with yield and quality losses in many wheat-growing regions. In tetraploid wheat sources of resistance are scarce. In the search for novel sources of resistance, 151 Triticum dicoccoides genotypes, originating from 16 habitats in Israel and one habitat in Turkey together with several control genotypes, were evaluated for reaction to fungal spread (Type II resistance) in replicated greenhouse experiments. Significant genetic diversity was found among the tested genotypes, the broad sense heritability for Type II FHB resistance was 0.71. Most of the tetraploid accessions were highly susceptible, only a few showed moderate resistance. Among the eight T. dicoccoides lines with the lowest relative infection rates, five originated from the Mt. Gerizim population, and three from the Mt. Hermon population. None of the T. dicoccoides lines reached the level of resistance present in the common wheat cultivar Sumai3. This revised version was published online in July 2006 with corrections to the Cover Date.     

Identification of somatic hybrids of dihaploid Solanum tuberosum lines and S. brevidens by species specific RAPD patterns and assessment of disease resistance of the hybrids

Summary Symmetric somatic hybrids were produced by electrofusion of protoplasts of two dihaploid tuber-bearing potato (Solanum tuberosum L.) lines and Solanum brevidens Phil., a diploid non-tuber-bearing wild potato species. A total of 985 plants was obtained. Verification of nuclear hybridity of putative hybrids was based on additive RAPD patterns, general morphological characteristics and chromosome counts. 53 (90%) calli regenerated into plants which were identified as somatic hybrids. Most of the hybrids were aneuploids at the tetraploid (4×) or hexaploid (6×) level. The 20 hybrids tested expressed a high level of resistance to potato virus Y (PVY ^N ) characteristic of the S. brevidens parent. Resistance to late blight (Phytophthora infestans (Mont.) de Bary) varied between hybrids, but was on average better than that of the fusion parents. Resistance of hybrids to bacterial stem rot (Erwinia carotovora subsp. atroseptica (van Hall) Dye) was not superior to that of commercial potato cultivars.     

Barriers to interspecific hybridization in Ulmus americana

Summary Crossability barriers between the tetraploid Ulmus americana and five other diploid elm species were investigated using both fluorescence and scanning electron microscopy. Both pollen germination and pollen tube development were inhibited in all interspecific crosses. Pollen tube behavior on the stigma of U. americana was unchanged when pollen of a colchicine-induced U. pumila tetraploid was substituted for diploid U. pumila pollen.Self-incompatibility was observed in one of two American elms studied and manifested itself via reduced pollen germination and inhibited pollen tube growth on the stigma surface.The results indicate that crossing barrier(s) between U. americana and other elm species occur at the stigma surface and operate independently of ploidy level. This conclusion is discussed in relation to current breeding efforts to transfer Dutch elm disease resistance into the American elm via interspecific hybridization with Eurasian elms.     

Development of a promising interspecific hybrid in black pepper (Piper nigrum L.) for Phytophthora foot rot resistance

For the first time in the history of black pepper cultivation, a partly fertile interspecific hybrid having partial resistance to the dreaded disease Phytophthora foot rot was developed through hybridizing Piper nigrum with the wild species Piper colubrinnum. Hybridity of interspecific progenies was established through morphology, anatomy, cytology, and molecular studies. The hybrid, whose chromosome number is 2n = 39, is a triploid hybrid between a tetraploid and diploid species. The hybrid designated as Culture P5PC-1 exhibited distinct anatomical and morphological feature with a large number of long spikes with reduced setting percentage. The RAPD primers OPE 07 and OPG 08 were identified as hybrid specific molecular markers. Functional evaluation revealed partial introgression of genes—responsible for Phytophthora foot rot resistance—into the hybrid. This hybrid is considered as a successful breakthrough for introgression of resistance to the cultivated species Piper nigrum from the wild species Piper colubrinnum.     

Immunity to Clavibacter michiganensis subsp. sepedonicus: Screening of exotic Solanum species

Summary Accessions from exotic Solanum species, including diploid and tetraploid species, were screened for immunity to Clavibacter michiganensis subsp. sepedonicus, the causal agent of potato ring rot. The diploid species included S. infundibuliforme, S. lesteri, S. megistacrolobum, S. tuberosum Group Phureja, S. polyadenium, S. pinnatisectum, S. raphanifolium, S. sparsipilum, S. sanctae-rosae, S. tuberosum Group Stenotomum, S. toralapanum, and S. verrucosum. The tetraploid species included S. tuberosum Group Andigena, S. acaule, S. fendleri, S. hjertingii, S. oplocense, S. polytrichon, and S. stoloniferum. Apparent immunity was initially found in several diploid species, but was not present during subsequent retesting. Immunity was found in nine accessions of tetraploid S. acaule. These accessions maintained their immunity during testing over an eight-month period. S. acaule appears to be a good source of immunity for introgression studies.     

Genetic diversity of wheat wild relatives in the Near East detected by AFLP

In order to reveal the molecular genetic diversity of wheat wild relatives, an AFLP analysis was conducted with 16 accessions of five Triticum andAegilops species originating from the Near East. Variation within population was studied with at least seven individuals per accession. Four primer combinations were used for selective amplification. Based on the scored bands, we estimated percentage of polymorphic bands, 1 – proportion of shared bands (1-psb) and nucleotide diversity (π). Of the five species used in this study, Ae. speltoides had the highest level of `within population' variation. This species had also the highest value of the variation among populations. As for Triticum species, the level of variation within population was low in diploid species (T. urartu and T. boeoticum),whereas two tetraploid species (T. dicoccoides and T. araraticum) had relatively high levels of variation within population. While the two diploid Triticum indicated a clear interspecific divergence, the two tetraploid wild wheats were not clearly divergent in this study. The variance portioning analysis indicated that the variation detected for diploid Triticum species was mainly composed of `between species' variation, on the other hand that for tetraploid Triticum was mostly composed of `within population' variation. In conclusion, AFLP analysis reveals molecular variation in all accessions used in this study, suggesting a potential genetic diversity of the wheat wild relatives in natural populations. These results have implications for the design of strategies to maintain genetic diversity within genebank collections. This revised version was published online in August 2006 with corrections to the Cover Date.     

Diallel analyses reveal the genetic control of resistance to ascochyta blight in diverse chickpea and wild Cicer species

Ascochyta blight is a major fungal disease affecting chickpea production worldwide. The genetics of ascochyta blight resistance was studied in five 5 × 5 half-diallel cross sets involving seven genotypes of chickpea (ICC 3996, Almaz, Lasseter, Kaniva, 24B-Isoline, IG 9337 and Kimberley Large), three accessions of Cicer reticulatum (ILWC 118, ILWC 139 and ILWC 184) and one accession of C. echinospermum (ILWC 181) under field conditions. Both F₁ and F₂ generations were used in the diallel analysis. The disease was rated in the field using a 1–9 scale. Almaz, ICC 3996 and ILWC 118 were the most resistant (rated 3–4) and all other genotypes were susceptible (rated 6–9) to ascochyta blight. Estimates of genetic parameters, following Hayman’s method, showed significant additive and dominant gene actions. The analysis also revealed the involvement of both major and minor genes. Susceptibility was dominant over resistance to ascochyta blight. The recessive alleles were concentrated in the two resistant chickpea parents ICC 3996 and Almaz, and one C. reticulatum genotype ILWC 118. The wild Cicer accessions may have different major or minor resistant genes compared to the cultivated chickpea. High narrow-sense heritability (ranging from 82% to 86% for F₁ generations, and 43% to 63% for F₂ generations) indicates that additive gene effects were more important than non-additive gene effects in the inheritance of the trait and greater genetic gain can be achieved in the breeding of resistant chickpea cultivars by using carefully selected parental genotypes.     

Polyploidy and sexual polyploidization in the genus Vaccinium

Genus Vaccinium,consisting of blueberries, cranberries, lingonberries and many related wild species, includes diploid, tetraploid, and hexaploid species. Most evidence indicates that the tetraploid species are autotetraploids, with non-preferential bivalent chromosome pairing. Although homoploid interspecific crosses usually produce numerous fertile hybrids if the parents are from the same section of the genus, inter sectional crosses at the diploid level normally produce no seedlings, weak seedlings, or seedlings that are very low in fertility. There is a strong but not complete triploid block within Vaccinium. Even with insections, tetraploid × diploid (and the reciprocal) crosses normally give only a few tetraploid and a few triploid hybrids. Hexaploid × diploid crosses within sections are very hard to make, and the few hybrids that have been obtained are pentaploid. The frequency of 2n gametes varies,both among genotypes within species and among species. Vaccinium pollen is normally shed in tetrads, and the frequency of large pollen grains shed in dyads can be used to estimate 2n gamete frequency. Cultivated blueberries occur at both the tetraploid and the hexaploid levels, and there are important genetic resources in the diploids. Unreduced gamete production has permitted transfer of genetic material from the diploid to the tetraploid level and from the diploids and tetraploids to the hexaploid level via triploid hybrids. Intersectional crosses can occasionally produce tetraploid Vaccinium hybrids that appear to behave as amphidiploids and have medium to high fertility. CommercialVaccinium cultivars are normally propagated by cuttings. Intersectional hybridization, chromosome doubling, and asexual propagation could permit the production of novel hybrid combinations with value as ornamentals or in fruit production. This revised version was published online in July 2006 with corrections to the Cover Date.     

Heritability of resistance to foliar late blight in a diploid hybrid potato population of Solanum phureja×Solanum stenotomum

The emergence of new races of Phytophthora infestans has necessitated the search for additional sources of potato germplasm with resistance to late blight. This study examined 281 clones, derived from 72 families of a diploid random-mated hybrid population of Solanum phureja×Solanum stenotomum. The clones were evaluated in a replicated field trial for 2 years with the control cultivar ‘Atlantic’ in Pennsylvania, USA. The P. infestans US-8 A2 mating type culture was used to inoculate spreader rows of susceptible S. tuberosum cv.‘Russet Burbank’. Percent defoliation caused by the late blight fungus was estimated visually in each plot three times near the end of the growing season. Area under the disease progress curve (AUDPC) was estimated and showed that late blight was more severe in 1997 than in 1996. Genetic differences among clones and significant, but small, clone–environment interaction were detected for AUDPC. Broad-sense and narrow-sense heritability estimates, over years, were 0.79 ± 0.05 (P = 0.05) and 0.78 ± 0.29, respectively. Seventy-five percent of the diploid clones had a significantly lower mean AUDPC than Atlantic. These results support the idea that this diploid population is worthy of use in breeding for late blight resistance in tetraploid potato cultivars.     

The production and behaviour of hexaploid hybrids between Hordeum vulgare and H. Bulbosum

Summary To produce hexaploid (or other polyploid) hybrids, diploid or tetraploid Hordeum vulgare was crossed with hexaploid or octoploid H. bulbosum, and perennial triploid hybrids between the two species were treated with colchicine. The crosses did not yield viable plants: seedset was low, the seed aborted and embryo culture was unsuccessful. The colchicine treatments geve rise to plants in which hexaploid chromosome numbers were observed. At the hexaploid level chromosomal instability occurred, resulting in chromosome elimination.The colchicine-treated triploid hybrids showed in the first years after the treatment better fertility after open flowering than untreated plants, but the level of fertility remained very low. The offspring consisted of haploid, diploid and approximately triploid plants like H. vulgare, tetraploid and approximately tetraploid plants like H. bulbosum, and plants with hybrid morphology and unstable chromosome number, which were highly sterile. Thus the crossing barrier between H. vulgare and H. bulbosum could not be broken down at higher ploidy level.     

Polyploid and maternal effects on Rhizoctonia root rot resistance in sugarbeet

Summary Three sugarbeet breeding lines partially resistant to the root-rotting fungus, Rhizoctonia solani, were converted to the tetraploid condition without selection. These three diploid and tetraploid lines were crossed with three diploid male-sterile lines to produce equivalent diploid and triploid hybrids. The triploid hybrids were significantly more resistant to Rhizoctonia than were the diploid hybrids. However, the tetraploid resistant limes were no different than their diploid equivalent lines. Reciprocal crosses provided no evidence of maternal effect on resistance. Cytoplasm that included the male-sterility factor had no influence on resistance. Triploid hybrids, where the resistant parent is tetraploid, should be advantageous in the breeding of rhizoctonia-resistant hybrid varieties.Joint contribution of the Agricultural Research Service, U.S. Department of Agriculture, the Colorado State University Experiment Station, and the Beet Sugar Development Foundation. Published with the approval of the Director of the Colorado State University Experiment Station as Scientific Paper Series No. 2072.     

The breeding of the cultivated potato species solanum x juzepczukii and S. x curtilobum II. The resynthesis of S. x juzepczukii and S. x curtilobum

Summary The possibility of combining anew the genomes of wild and cultivated progenitors of triploid S. x juzepczukii and pentaploid S. x curtilobum by following the known evolutionary pathway of these species was investigated.The resynthesis of S. x juzepczukii was easy, and a wide range of synthetic forms was bred. Among these were forms with higher frost resistance (-5°C) than has been found in natural S. x juzepczukii. The total tuber glycoalkaloid content of several synthetic hybrids was lower than or as low as that of natural clones. Most synthetic hybrids were more vigorous than natural S. x juzepczukii and produced about the same types of tubers as are found in the natural range of variation. The best diploid parents were found in the species S. goniocalyx.The attempt to resynthesize pentaploid S. x curtilobum has not been successful but tetraploid plants were obtained in the process. An explanation for the occurrence of tetraploids resulting from triploid x tetraploid and/or diploid crosses is offered.The newly bred tetraploids contain at least one genome from S. acaule (possibly two) and hybridize easily with ssp. andigena. They thus provide a means for the transfer of S. acaule germ plasm into the tetraploid cultivated gene pool which would profit from the frost resistance of S. acaule.     

The resistance of Hordeum bulbosum and its hybrids with H. vulgare to common fungal pathogens

Summary Two tetraploid and two diploid clones of Hordeum bulbosum were screened for resistance to five isolates of powdery mildew which are virulent on cultivated barley. All were resistant and this resistance was also expressed in hybrids with H. vulgare. The tetraploid genotypes were also resistant to isolates of yellow rust and brown rust. These results show that H. bulbosum contains useful genes for resistance to these diseases and that there is a potential to transfer these into cultivated barley.     

Analysis of nucleolar organizer regions by silver staining in two Lolium species

Summary The nucleolar organizer activity of diploid and tetraploid Lolium perenne L. and diploid Lolium multiflorum Lam. varieties was analyzed by the silver staining procedure. Both species show six or twelve Ag-NORs (silver stained nucleolar organizer regions) at the diploid and tetraploid levels, respectively. A positive correlation between the size of secondary constrictions and Ag-NORs was observed.     

Genetic linkage of QTLs for late blight resistance and foliage maturity type in six related potato progenies

A set of test crosses of diploid potatoes was used to identify QTLs for foliage resistance against Phytophthora infestans and QTLs for foliage maturity type, and to assess their genetic relationship. The most important locus for both traits was found on chromosome 5 near marker GP21: the allele of marker GP21 that is associated with resistance to late blight is also associated with late foliage maturity. An additional QTL with a small effect on foliage maturity type was identified on chromosome 3, and additional QTLs for late blight resistance were found on chromosomes 3 and 10. Another QTL was detected on chromosome 7 when resistance was adjusted for the effect of foliage maturity type. The additional QTLs for resistance against P. infestans on chromosomes 3 and 10 seem to be independent of foliage maturity type and are not affected by epistatic effects of the locus on chromosome 5. The effects of the additional QTLs for resistance are small, but early maturing genotypes that necessarily have the allele for susceptibility for late blight on chromosome 5 may benefit from the resistance that is provided by these QTLs on chromosomes 3 and 10.     

Ploidy Manipulation and Introgression of Resistance to Alternaria Helianthi from Wild Hexaploid Helianthus Species to Cultivated Sunflower (H. annuus L.) Aided by Anther Culture

The present investigation discusses the scope for transferring of resistance to leaf spot disease incited by Alternaria helianthi from two hexaploid wild species (H. tuberosus and H. resinosus) to diploid cultivated sunflower. Interspecific hybrids produced between sunflower and these two hexaploid species were partially fertile with tetraploid chromosome status. Backcrosses of these interspecific hybrids with cultivated sunflower resulted in the formation of sterile triploid plants. To overcome the problem of sterility and facilitate backcrosses with cultivated sunflower, anther culture of the tetraploid interspecific hybrids was carried out to bring down their chromosome number to diploid status. Anthers from both interspecific hybrids were cultured on basal Murashige and Skoog media supplemented with varying concentrations of organics and the growth regulators benzyladenine and naphthaleneacetic acid. Anthers of interspecific hybrids involving H. resinosus responded well and regenerated through an embryogenic route at a frequency of 98.7%. But in interspecific hybrids with H. tuberosus, anthers formed callus and subsequently regenerated shoots through an organogenic pathway. DNA ploidy analysis of anther culture plants of interspecific hybrids derived from H. tuberosus crosses was carried out to identify plants with desired diploid status. In vitro screening of parents, interspecific hybrids and anther culture plantlets against A. helianthi showed resistance in 68.5% of the anther culture plants of interspecific hybrids from H. tuberosus and in 24.3% of the plants derived from interspecific hybrids involving H. resinosus.     

Discovery of 2n gametes in tetraploid oat Avena vaviloviana

Summary Sexual polyploidization via the action of 2n gametes (gametes with the sporophytic chromosome number) has been identified as the most important evolutionary mode of polyploidization among plant genera. This study was conducted to determine whether 2n gametes are present in the tetraploid level of the genus Avena (2n=4×=28) Twenty tetraploid Avena lines, representing four species and one interspecific hybrid, were screened for pollen grain size in order to differentiate between n and 2n pollen. Avena vaviloviana (Malz.) Mordv. line PI 412767 was observed to contain large pollen grains at a 1.0% frequency. Cytogenetic analyses of pollen mother cells of PI 412767 revealed cells with double the normal chromosome number (i.e., 56 chromosomes at metaphase I and anaphase I). The mode of chromosome doubling was found to be failure of cell wall formation during the last mitotic division that preceded meiosis. The resulting binucleate cells underwent normal meiotic divisions and formed pollen grains with 28 chromosomes. Based on the formation and function of 2n gametes, three models involving diploid and tetraploid oat lines are proposed to describe possible evolutionary pathways for hexaploid oats. If stable synthetic hexaploid oat lines could be developed by utilizing 2n gametes from diploid and tetraploid oat species through bilateral sexual polyploidization, the resulting hexaploids could be used in breeding programs for transferring genes from diploids and tetraploids to cultivated hexaploids.     

Transmission of bacterial wilt resistance by First Division Restitution (FDR) 2n pollen via 4x×2x crosses in potatoes

Summary Resistance to bacterial wilt (Pseudomonas solanacearum) found in tuber-bearing Solanum species was transferred into a diploid potato breeding population. Simultaneous selections were made for agronomic characters, production of first division restitution (FDR) 2n pollen, and resistance. Diploid resistant genotypes were identified via inoculation with a virulent isolate (CIP-204) of race 3 of Pseudomonas solanacearum. These resistant diploid genotypes were crossed to susceptible tetraploid potatoes. An investigation was made to assess whether resistant diploid genotypes transmit resistance to bacterial wilt, which is a quantitatively inherited trait, to tetraploid potatoes via FDR 2n pollen. Tetraploid seedlings from 4x×2x crosses were inoculated with the same isolate CIP-204, and the percentage of surviving seedlings was scored. Some 4x×2x families from resistant diploid genotypes demonstrated a high level of survival rate. The transmission of bacterial wilt resistance was achieved by the use of FDR 2n pollen. It was speculated that a female x male interaction effect on the survival rate of the evaluated progeny may exist. Selecting proper 4x and 2x parents would be important for obtaining a higher frequency of transmission of resistance to bacterial wilt in the progeny.