Discrepancy between spindle anomalies and 2N pollen production insolanum interspecific hybrids

The cytological basis of 2n pollen production was studied in a diploidSolanum progeny coming from a cross between aS. tuberosum S. chacoense hybrid which does not produce2n pollen (T710) and a S.phurejar S. tuberosum hybrid (W5295.7) which does. Both parental clones showed anomalous orientation of spindles at Metaphase II of microsporogenesis, but only clone W5295.7 produced dyads. All plants of the progeny T710 × W5295.7 showed anomalous spindles at Metaphase II, but only 60% of them produced 2n pollen, thus evidencing a discrepancy between spindle anomalies and dyad/2n pollen production. This may suggest that both parallel and fused spindles are a necessary but insufficient condition for 2n pollen production, and that some other mechanisms expected to be more highly correlated with them may be involved in 2n pollen production in this population.     

Quantification of Field Resistance to Verticillium dahliae in Eight Russet-Skinned Potato Cultivars Using Real-Time PCR

Changes in potato production over the past 10 to 20 years, have resulted in increased emphasis being placed on breeding for resistance to Verticillium wilt, caused by Verticillium dahliae Kleb. While many russet-skinned cultivars recently have been released with reported resistance to Verticillium wilt, information is lacking on the level of pathogen colonization, and therefore, the level of true genetic resistance is not known. Eight russet-skinned cultivars were grown in field trials with low and high levels of V. dahliae in the soil, and evaluated for wilt, stem colonization, yield, and tuber vascular discoloration. A recently developed QPCR assay was validated, with strong relationships to culture plating assays over three stem sampling dates. Additionally, stem colonization levels, as determined by QPCR, were related to wilt and tuber vascular discoloration. However, total yield did not exhibit a strong relationship to any other parameter evaluated in this study. Results from these studies indicate that varying levels of true resistance are present in the russet-skinned cultivars evaluated, and that the QPCR assay can be reliable in rapidly evaluating resistance to V. dahliae under field conditions. Based on pathogen quantification using stem colonization derived from traditional plating assays and QPCR, the resistance level of several cultivars is more clearly defined and discussed.     

Germplasm Release: Three Tetraploid Potato Clones with Resistance to Common Scab

Common scab caused by the soil-borne bacterium Streptomyces scabies is a serious disease for the potato industry. We have identified a strong source of resistance in the diploid wild relative Solanum chacoense. This resistance has been introgressed into tetraploid cultivated potato via unilateral sexual polyploidization. This paper describes three hybrid clones (M8, M16, M17) for use by potato breeding programs to enhance resistance to common scab. They were created by crossing a diploid clone (50% S. tuberosum, 50% S. chacoense) to tetraploid cultivars, producing hybrids with 75% cultivated and 25% wild germplasm. The clones are male and female fertile, and are adapted to temperate zone production environments.     

Disease resistance inSolanum phureja and diploid and tetraploidS. tuberosum × S. phureja hybrids

Summary DiploidSolanum phureja, diploid and tetraploidS. tuberosum × S. phureja hybrid, and tetraploidS. tuberosum backcross clones were assessed for resistance to commercially important diseases. There was a reduction in the expression ofS. phureja characteristics with increasing dosage ofS. tuberosum genetic material. The generally high resistance of theS. phureja group to common scab, potato leafroll virus and potato virus Y decreased, while resistance to gangrene, foliage and tuber blight tended to increase. Resistance genes were probably well dispersed throughout theS. phureja genotype so that many were lost on hybridisation, butS. phureja could be a useful source of scab and virus resistance in the production ofS. tuberosum cultivars.     

Use of RAPD markers in potato genetics: Segregations in diploid and tetraploid families

The objective of the present study was to evaluate random amplified polymorphic DNA (RAPD) as a source of markers for use in investigations in potato genetics. Segregation of 18 loci in diploidSolanum goniocalyx xS. phureja and 12 loci in tetraploidS. tuberosum ssp.tuberosum ×S. tuberosum ssp.andigena families fitted Mendelian and tetrasomic ratios, respectively. Eight loci in the diploid progeny were arranged in three linkage groups. Segregations of these markers fitting expected ratios indicate that they can be effectively used in potato genetics, breeding and evolution.     

Evaluation of potato clones for severity of Verticillium wilt, yield and specific gravity in Maine

In 1994 and 1995, the effect of Verticillium wilt, caused byVerticillium dahliae andV. albo-atrum, on tuber yields, number and weight of U.S. No. 1 and B size tubers, and specific gravity was studied in northern Maine, an area with a short growing season. Seven clones (four resistant and three susceptible) were evaluated in a split-plot design with three replications. Clones were the whole-plot factor, and seed pieces in sub-plots were either uninoculated or inoculated with 50 ml of 4 × 10⁴ cfu/mlVerticillium spp. at planting. Individual plants were scored for Verticillium wilt symptoms before harvest on a 1= <3% wilt to 10= >97% wilt. Differences among clones for wilting and specific gravity were significant. The inoculation treatment had no effect on any of the tuber traits measured. However, there were significant clone x inoculation interactions for most tuber traits. Reductions in yield, weight and number of U.S. No. 1 potatoes, and specific gravity were greater in the Verticillium wilt susceptible clones than in the resistant clones. These results suggest that breeding clones with resistance toVerticillium spp. will reduce yield losses, while maintaining tuber size and specific gravity under disease pressure.     

Effect of Soil Temperature,Injection Depth,and Metam Sodium Rate on the Management of Verticillium Wilt of Potato

Verticillium wilt, caused by Verticillium dahliae Kleb., is a primary component of the early dying complex of potato (Solanum tuberosum L.) in the United States. Although genetic resistance to V. dahliae exists and has been incorporated into several potato cultivars, the commercial potato industry is still dominated by cultivars susceptible to the pathogen. As a result, soil fumigation with metam sodium remains an important means by which Verticillium wilt is controlled, despite its expense and potentially negative environmental impact. Recent restrictions on metam sodium use by the Environmental Protection Agency directed at reducing exposure to vapor emissions have increased the need to improve shank injection of the soil fumigant. In studies reported here, the application of metam sodium reduced the severity of Verticillium wilt, however, soil temperature at the time of injection, metam sodium injection depth, and application rate had little overall effect. In 2011, temperature at the time of metam sodium injection did not result in significant differences in any parameter evaluated. However, in 2012, soil populations of V. dahliae, wilt severity and host colonization were significantly reduced when metam sodium was applied at 4 °C compared to 13 or 15 °C. No significant differences were observed between a single or two metam sodium injection depths in any parameter evaluated across the 2 years the study was conducted. While all rates of metam sodium significantly reduced soil populations of V. dahliae compared to the non-treated control, significant differences across rates were rarely observed. Improved control of Verticillium wilt and increased yield can be achieved as a result of these studies. The effective control of Verticillium wilt can be obtained by using metam sodium at a comparatively low rate of 373 l/ha, particularly when applied at a relatively cold soil temperature of 4 °C using a single injection depth of 25 cm. The potential impact of these application modifications of metam sodium in reducing emissions and non-target exposure is discussed.     

Genetic mapping of a dwarfing gene found inSolanum phureja clone 1.22

Dwarf plants were obtained in an F₂ population of a cross betweenSolanum chacoense (clone chc 525-3) andS. phureja (clone 1.22). Segregation analyses in F₂ and backcross populations suggested that the dwarfism was controlled by a single recessive gene transmitted fromS. phureja clone 1.22. The dwarf plants responded to treatment with gibberellic acid, which recovered normal growth. We named this gene‘ga ₂’. Linkage analysis of the genega ₂ with RFLP and RAPD markers indicated that the genega ₂ was located on the most or near distal end of chromosome 7.     

Field assessment of commercial cotton cultivars for Verticillium wilt resistance and yield

Verticillium wilt is responsible for substantial yield losses in cotton. The use of cultivars with partial resistance to Verticillium wilt is a preferred management strategy. Cotton breeders utilize rating scales based on severity of foliar symptom expression and/or defoliation when making cultivar selections. The objective of this research was to determine the yield relationship with cultivars grouped by both incidence of Verticillium wilt during the boll filling stage, and late season defoliation in fields with a history of Verticillium wilt. Small plot, replicated cultivar trials were conducted over an 11 year period in west Texas. Cultivars were divided into four classes (A–D) based on normalized wilt incidence (NW) and four classes based on normalized defoliation ratings (ND). Classes were determined through a mixed model analysis of cultivars, with T-test comparisons between a partially resistant check (Fibermax 2484B2F) and a susceptible check (Deltapine 0912B2RF). The A and C classes did not differ (P = 0.05) from the partially resistant and susceptible checks, respectively. Classes were used in a mixed model analysis with lint yield. In a model with only NW classes, lint yield decreased significantly as NW increased with least square mean values of 1421, 1385, 1284 and 1204 kg/ha, for classes A, B, C and D, respectively. In a model with both NW and ND classes, the A/A class combination of NW and ND had a significantly (P ≤ 0.05) higher lint yield (1776 kg/ha) than any other combination. NW/ND class combinations of A/B, B/A, B/B, and C/A had intermediate yields, and ND classes of C or D had the lowest yields. Cultivars with both low wilt and defoliation incidences should result in higher yields in fields infested with Verticillium dahliae. Furthermore, these two rating criteria could be used to standardize the process for developing resistant cultivars.     

Pollen-pistil incompatibility in a diploid hybrid potato population with cultivated and wild germplasm

The basis of low seed set was investigated in a diploid hybrid population with germplasm from the cultivated speciesSolanum tuberosum spp.tuberosum andS. tuberosum ssp.phureja and the wild speciesS. chacoense. Controlled crosses were performed following an incomplete diallel mating design which included the hybrid population and the parental species. Pollen grain germination and pollen tube growth were observed with fluorescence microscopy in 174 intrapopulation and interspecific combinations of genotypes. Fifty percent of the combinations of genotypes within the hybrid population and 46% of those between this population and the parental species were incompatible; overall, 71% of the interspecific combinations of genotypes were compatible when plants of the hybrid population were used as females, vs. 5% when used as males. Although the site of reaction could vary in a given intrapopulation or interspecific cross with the combinations of genotypes, pollen tube inhibition occurred in the first third of the style in 42% of them. Since the gene pool of the hybrid population had been widened with germplasm from other geographic areas and the genotypes ofS. tuberosum ssp.tuberosum, S. chacoense andS. tuberosum ssp.phureja used in this study were not the ones involved in the original crosses, it is concluded that the S-locus is not controlling the incompatibility reaction but rather that a cross-incompatibility system, possibly governed by more than one locus, is acting.     

Resistance to the potato nematode Heterodera rostochiensis Woll. in clones derived from Solanum vernei

Previous research had shown that the diploidS. vernei clone EBS181 contained at least two genes for resistance to the golden nematode. Resistant tetraploids obtained by backcrossing to tuberosum clones, with selection against Race A, possessed only one gene. Crosses between these tetraploids produced evidence that at least two genes exist within this set of tetraploids. Crosses between tetraploids with resistance from vernei EBS181 and tetraploids with resistance from andigena CPC1673 produced results that can be explained by one of the genes from EBS181 segregating at the same locus as the one from CPC1673.     

Agronomic performance of potato germplasm selected for high resistance to verticillium wilt

A three-year study was conducted from 1991 through 1993 to evaluate the field performance of potato clones that had been selected for resistance to wilt symptom development and stem colonization byVerticillium dahliae. The total yield, size distribution, and specific-gravity of these highly resistant clones were compared with standard cultivars and two parent clones with high Verticillium resistance and high yield, A66107-51 and A68113-4. Two groups of Verticillium resistant germplasm were selected in 1991. One group was the highly resistant progeny from a cross between A66107-51 and A68113-4. The second group consisted of hybrids between wild species accessions with high Verticillium resistance and cultivated diploid and tetraploid germplasm. Twenty-one out of 125 progeny clones from the A66107-51 × A68113-4 cross were highly resistant to Verticillium infection and were tested in yield trials for two years. Another eight selections had high Verticillium resistance but such low yield of seed tubers that they were tested only one year for yield. Eleven of the original 29 highly resistant selections were significantly lower in total yield than Russet Burbank. Only one clone was significantly higher in yield than RB, and none outyielded either of the resistant parents. Reddale and Century Russet had moderate to high Verticillium resistance, respectively. Both outyielded Russet Burbank and were comparable with the high yielding resistant parents. Of 15 interspecific hybrids tested in 1991, only four had sufficient yield to produce seed for yield trials and good wilt resistance after three successive seasons of evaluation. We concluded from this study that selecting at early stages of variety development strictly for Verticillium resistance is likely to be inefficient. Rather, selection should be for yield and other agronomic criteria in Verticillium infested fields.     

Transmission of Scab Resistance to Tetraploid Potato Via Unilateral Sexual Polyploidization

Resistance to common scab continues to be a high priority trait for potato breeders. We have identified a source of resistance in the diploid wild potato relative Solanum chacoense and have introgressed it into cultivated potato by crossing it to a dihaploid. A clone generated by crossing two full-sib hybrids is highly resistant and produces both 2n pollen and 2n eggs. This clone, named 4–48, is homozygous for a major QTL for scab resistance derived from S. chacoense. Clone 4–48 was reciprocally crossed to three round white cultivars Megachip, Pike, and White Pearl. Common scab resistance was transferred to tetraploid offspring, with transmission through the male parent generally more effective than through the female parent. The majority of progeny (70%) exhibited stability for resistance to common scab across environments. This germplasm will be useful for breeding new resistant cultivars.     

Assessment of PCR-based simple sequence repeats to fingerprint North American potato cultivars

Seven primer pairs flanking di- and tri-nucleotide repeat sequences, identified from previously sequenced regions of the potato genome, were examined for their potential use in DNA-fingerprinting of thirty-nineSolanum tuberosum subsp. tuberosum cultivars (released between 1861 and 1988) and one diploidS. phureja breeding line. Of the simple sequence repeats (SSRs), the primers for six SSRs amplified DNA sequences within the potato genome between cultivars for a total of 14 bands. The polymerase chain reaction (PCR)based amplification products generated from each primer pair consisted of 1 to 2 bands per cultivar but band variation among cultivars demonstrated up to 4 bands per SSR. A similarity matrix generated from five SSRs was able to distinguish 24 of the 40 cultivars. However, when the potato cultivars were grouped by tuber type (round white-skinned, long white-skinned, russetskinned, red-skinned, and yellow flesh) only five pairs of cultivars remained indistinguishable: Atlantic/Katahdin, Belchip/Wauseon, Red LaSoda/Bliss Triumph, Red Pontiac/Norland, and Burbank/Spunta. Although SSRs did not generate unique fingerprints for all of the North American genotypes examined, the potential to discriminate most cultivars should increase as additional SSRs are identified in potato.     

Bacterial wilt resistance in the progenies of the fusion hybrids between haploid of potato andSolanum commersonii

Bacterial wilt of potatoes, which used to be a widespread disease in tropics and subtropics, has become a threat to potato production in temperate region. The diploid species Solanum commersonii has several desirable characteristics including cold tolerance and resistance to several diseases. Selected somatic hybrids between S. tuberosum dihaploid and bacterial-wilt-resistant S. commersonii clones were cross pollinated with S. tuberosum cultivars for further selection of bacterial wilt resistance. The chromosome numbers of the fusion parents were confirmed as 24, and the three fusion hybrids crossed were all tetraploids. The chromosome number of 11 backcross 1 progenies (BC1) was 48 and that of the other six was close to the tetraploid number. Backcross 2 progenies (BC2) were obtained from only three of the 44 BC1 clones crossed. The S. commersonii parent clone, LZ3.2, was the most resistant to bacterial wilt among wild species clones tested. The first sexual progenies segregated for resistance, with one clone highly susceptible and four clones highly resistant. Three highly resistant BC1 clones, CT02-4, CT08-4, and CT10b-4, were backcrossed to cultivars. Two cross combinations produced mostly susceptible BC2 progenies; however seven clones were resistant or highly resistant for both race 1 and race 3. The highly resistant three clones, CT204-3, CT206-9 and CT206-10, were selected for the further testing as cultivars or breeding materials.     

Field evaluation of mutant and hybrid lines of mint for resistance to Verticillium wilt and yield

Mint lines that were previously identified with low wilt-severity values in the greenhouse were evaluated for resistance to Verticillium wilt and yield in the field in the presence of naturally occurring inoculum and in plots artificially infested with the soilborne fungus. Severity of Verticillium wilt varied significantly among mint lines and cultivars in the inoculated and non-inoculated sub-plots in two field trials. Verticillium wilt was significantly less severe for mutant lines 87M0109-1, 84M0107-7, and M90-11 than for Black Mitcham in 2002 and 2003. Verticillium wilt was significantly less severe for mutant line M83-14 and for Murray Mitcham than for Black Mitcham in 2003 and 2004. At the completion of the first trial, fresh hay yields of both peppermint hybrids 87M0109-1 and 84M0107-7 and peppermint mutant M90-11 were significantly higher than the standards Black Mitcham and Redefined Murray Mitcham. Oil yield in the first trial of standard Black Mitcham declined in years 3 and 4, while the decline in oil yield was less dramatic with the other four peppermint lines. In the second trial, hay yields of both B90-9 and M83-14 were comparable to, but not significantly different from Black Mitcham and Redefined Murray Mitcham. Line 92(B-37 × M0110)-1 yielded significantly less oil than all peppermint lines including both standards Black Mitcham and Redefined Murray Mitcham in six of the seven cuttings in the second trial. Verticillium rating of the lines and cultivars inoculated in trials from a previous greenhouse study were highly correlated with the Verticillium wilt rating of the lines and cultivars the second and third years when inoculated in both field nurseries in this study. Verticillium wilt resistance and yield can be improved in commercial mint cultivars and form a basis for managing the disease.     

Early detection of Verticillium wilt resistance in a potato breeding program

Nine potato clones/cultivars selected for varying degrees of Verticillium wilt (VW) resistance based on degree of vascular colonization, were intercrossed and outcrossed to 13 uncharacterized clones. Based on growth characteristics of 3,535 seedling transplants and an assay quantifyingV. dahliae in potato vascular tissue, 404 clones were selected for further tests. The average number ofVerticillium dahliae colony forming units (CFU) expressed as Log₁₀(CFU + l)/0.1 ml of plant sap for the 404 clones was 2.3, and 1.3 and 3.7 for resistant Reddale and susceptible Kennebec checks, respectively. Thirty-five clones (8.66%) had CFU values equal to or less than Reddale suggesting that VW resistance is a readily transmitted trait.     

Glycoalkaloid content in tubers of hybrid and backcross populations from aSolanum tuberosumx s. chacoense cross

The glycoalkaloid contents of tubers were measured in hybrid and back-cross populations derived from a cross ofSolarium chacoense Bitter (PI 472810, 2n=2x=24) withSolanum tuberosum L. (2n=4x=48). In tubers sampled from near the median size in each population and parent, the mean total glycoalkaloid (TGA) contents of theS. chacoense andS. tuberosum parent genotypes, and the F₂ and F₄ hybrid genotypes were 245, 4, 39, and 35 mg/ 100 g fresh weight (fw), respectively. Backcrossing the F₂ genotypes toS. tuberosum reduced the mean TGA content to 15 mg/100 g fw. A second backcross further reduced the content to 9 mg/100 g fw. However, some genotypes in both backcross populations (25% and 10%) had TGA contents above the generally accepted upper limit of 20 mg/100 g fw. Tuber size was significantly smaller in theS. chacoense parent and the hybrid populations compared with the size in theS. tuberosum parent and backcross populations. In a second sampling of tubers from each population, intentionally selected to equalize tuber size among populations, the mean tuber size for populations was reduced by 79%. The mean population TGA content increased by 44%. However, the differences and rankings among parent, hybrid, and backcross populations were the same for both tuber samplings.     

Study of gene effects for cotton yield and Verticillium wilt tolerance in cotton plant (Gossypium hirsutum L.)

Verticillium wilt (VW), caused by Verticillium dahliae Kleb., is a destructive disease of cotton (Gossypium hirsutum L.). The use of resistant cultivars has long been considered the most practical and effective mean of control. The aim of this work was to study the quantitative genetic basis of Verticillium wilt resistance in Upland cotton by using five genotypes and their possible crosses without reciprocals selecting simultaneously for resistance and desirable agronomic characteristics. Five cotton cultivars and 10 F1s from half-diallel crosses were analyzed for VW resistance. The seed cotton yield, the number of bolls/ plant, and boll weight were measured and Verticillium wilt index (VWI) was estimated during two crop seasons in two different sites each year always on plots with naturally infested soil. Genetic components of variance were analyzed using the Hayman model. Analysis of variance for all characters showed significant differences between genotypes, without genotype-site interaction in most cases. Both, additive genetic variance component (D) and dominance genetic variance components (H1 and H2) were presented in all characters, except for VWI. D was the most important component for boll weight and VWI. Boll weight was the most correlated character with seed cotton yield and VWI. Broad sense heritability was high for boll weight and VWI, moderate for seed cotton yield and low for bolls per plant. Narrow sense heritability was moderate for boll weight, and high for VWI.     

Segregation of leptines and other glycoalkaloids inSolanum tuberosum (4x) ×S. chacoense (4x) crosses

A clone, 8380-1, selected fromSolanum chacoense (PI 458310) for its high foliage content of the leptine glycoalkaloids, a factor in resistance to Colorado potato beetle, was doubled in chromosome number from 2n=2x=24 to 2n=4x=48. Three 4x clones were crossed with sixS. tuberosum (4x) clones. Foliage glycoalkaloid contents were measured for 452 F1 hybrids from 15 crosses. The 4xchacoense parental clones were not different in respect to glycoalkaloid contents and were similar to the original 2x clone. All F1 hybrids synthesized foliage leptines ranging from 9 to 369 mg/100 g fresh weight (fw) with a mean content of 113 mg/100 g fw. The proportion of leptines in the total glycoalkaloid content ranged from 1% to 62% with a mean of 25%. The 4xchacoense parent mean leptine content was 1482 mg/ 100 g fw which was 90% of the total glycoalkaloid content. Tubers from 136 hybrids and the threechacoense parental clones were tested for glycoalkaloid contents. The tuber solanine + chaconine contents of the 136 hybrids ranged from 30 to 180 mg/100 g fw with a mean of 79. The mean tuber content of the threechacoense parental clones was 157 mg/100 g fw. Leptines were not found in any of the tubers.