Comparison of different pollen viability assays to evaluate pollen fertility of potato dihaploids

Summary The main obstacle in breeding potato (Solanum tuberosum L.) dihaploids is the severe limitation of male fertility. To determine pollen viability assays that correlate well to fertility in crosses, results of five different pollen viability assays were compared by correlation analysis with fruit and seed set characters in test crosses, and to pollen tube growth in situ (PL-test). The methods used were: staining the pollen cells with carmino acetic acid (CAA-test); in vitro pollen germination (PG-test); and detection of pollen staining rates after incubation with fluoresceine diacetate (FDA-test), 2-3-5-triphenyle tetrazolium chloride (TTC-test), and 5-bromo-4-chloro-3-indolyle--galactoside (X-Gal-test).The results of test crosses and pollen tube growth in situ correlated with the results of all other assays with the following ranking, from highest to lowest: enzyme activity assays (X-Gal-test, FDA-test, TTC-test), in vitro pollen germination (PG-test), and pollen staining by CAA. The newly developed X-Gal-test for monitoring -galactosidase activity showed the least variation of all assays investigated. Thus, this highly reproducible simple procedure is recommended for male fertility screening.Abbreviations B/F Berries obtained per 100 flowers - CAA Carmino acetic acid - FDA Fluoresceine diacetate - PG Pollen germination rate in vitro - PL Pollen tube growth in situ - S/B Seeds per berry - S/F Seeds per pollinated flower - TTC 2-3-5-triphenyle tetrazolium chloride - X-Gal 5-bromo-4-chloro-3-indolyle--galactoside     

Genetics of self-compatibility in dihaploids of Solanum tuberosum L. 5. The S-genotype of tetraploid potato cultivar gineke

Summary Investigations of the genetics of self-compatibility and self-incompatibility in dihaploids and diploid derivatives from cv. Gineke revealed the presence of S ₁, S₂ and S ₃ at the S-locus of Gineke and in addition an S ₁-allele on a translocation. By means of a complete tester set involving the S-alleles S ₁, S₂ and S ₃ (all from Gineke) and S ₄ (from Black 4495) it was demonstrated that some Gineke dihaploids were compatible with all six testers. This indicated a fourth S-allele in Gineke, which differs from those in the tester series and was therefore assigned S ₅. Additional evidence was obtained from an analysis of F₁'s from crosses of two S ₅-bearing dihaploids and one of the testers. So the S-genotype of cv. Gineke was identified as S ₁S₂S₃S₅/S₁, the second S ₁ being the S-allele on a translocated fragment.     

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.     

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.     

Meiotic restitution mechanisms and 2n pollen formation in a Solanum tuberosum dihaploid and in dihaploid × wild species hybrids*

A Solatium tuberosum dihaploid that proved to be a 2n pollen producer, exhibited two types of meiotic abnormality involved in 2n pollen formation: premature cytokinesis at prophase II and parallel/tripolar spindles in meiosis II. For this reason such a dihaploid was preferentially used as a seed parent in crosses with the diploid species S. tarijense, S. sanctae-rosae, and S. phureja. Diploid interspecific hybrids were evaluated for male fertility, for 2n pollen production and stability over three years and for meiotic nuclear restitution mechanisms. Pollen stainability ranged from 6 to 85% and 2n pollen from 5 to 35% among hybrids. The meiotic mechanism generating 2n pollen was that of spindle abnormality in meiosis II and consequent dyad/triad formation. High correlation was found between fused/parallel spindles and dyads.     

Transmission of resistance to common scab from the diploid to the tetraploid level via 4x-2x crosses in potatoes

Summary Diploid potato clones selected for their reaction to common scab and their ability to produce 2n male gametes were used in a series of crosses to a susceptible tetraploid female parent (cv. Shepody). In addition, two tetraploid clones were also selected for their reaction to common scab and crossed with Shepody as a female parent. Results indicated that resistance to common scab can be effectively transmitted from the diploid to the tetraploid level via 4x-2x crosses. Diploid parents producing 2n pollen via either first division or second division restitution can be used to transmit scab resistance. A relatively small proportion of resistant individuals could be recovered from susceptible x susceptible crosses in both 4x-2x and 4x-4x combinations. The data support a previously developed hypothesis that scab resistance is relatively simply inherited.     

Comparative performance of diploid and tetraploid progenies from 2x.2x crosses in potato

Summary Selection criteria for agronomic characters in a potato breeding program at the diploid (2x) level may differ from selection criteria used when selecting breeding lines at the tetraploid (4x) level. Differences between selection criteria are expected, (1) when expression of the characters is different at both ploidy levels and/or (2) when the effect of diploid breeding lines on agronomic characters of tetraploid progenies is different from the effect of tetraploid breeding lines. In this investigation sets of diploid and tetraploid progenies, each set derived from the same 2x.2x cross, were compared as to the expression of six agronomic characters. Diploid progenies had significantly lower yields (due to smaller tubers) and significantly higher under water weights than tetraploid progenies. Vine maturity and chip colour were similarly expressed at both ploidy levels. Correlations between yield and yield components, and between under water weight and chip colour were similar at both ploidy levels. The lower yields and higher under water weights found in diploids point to the need of different selection criteria for selecting diploid and tetraploid breeding lines.     

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.     

Selection of vigorous and fertile S-homo- and heterozygous tester clones from self-incompatible diploid potato, Solanum tuberosum L.

For the selection of diploid (2n = 2x = 24) potato (Solanum tuberosum) genotypes that are useful for the molecular and genetic analysis of the phenomenon of gametophytic self-incompatibility, three different types of basic populations were investigated. These populations were derived from three primary dihaploid clones, G609, G254 and B16, which possessed the S-allele combinations S1S2, S1S3 and S3S4 respectively. In order to select highly vigorous, profusely flowering, fertile and tuberising progenies, three types of populations, derived from the above mentioned diploid genotypes, were screened for performance and classified for the expression of self-incompatibility. Although the selection for well defined S-genotypes was sometimes complicated due to the occurrence of pseudo-compatibility and of a self-compatibilising factor, the use of a combination of criteria, viz., Iso Electric Focusing (IEF), pollen tube growth in the styles and the extent of berry and seed set made the selection of sufficient representatives of all six types of S-heterozygotes (S1S2, S1S3, S1S4, S2S3, S2S4 and S3S4) possible. After evaluating the strength of the self-incompatibility reaction in these heterozygotes, those with high expression were selfed, and intercrossed within their S-allele incompatibility group through the method of counterfeit pollination. In these progenies, well-performing S-homozygotes (S1S1; S2S2; S3S3; S4S4) for all four S-alleles with high expression of self-incompatibility were selected. As a result, all possible S-homo- and heterozygous genotypes with a predictable type of self-incompatibility are available and maintained both vegetatively and as botanical seed. The development of this material has paved the way for more critical analysis of molecular factors involved in self-incompatibility in diploid potato. This revised version was published online in July 2006 with corrections to the Cover Date.     

外来杂草刺萼龙葵及其风险评估

外来入侵生物已对世界各国的自然生态系统、农业生产及公众健康造成了严重危害。刺萼龙葵(Solanum rostratum)起源于北美洲,其繁殖和适应能力很强,是具有潜在危险的外来有害杂草。对刺萼龙葵的分布、生物生态学特性、经济和生态影响及其控制和管理措施等进行了系统综述。根据生物生态学特性、潜在危险及管理控制的难度等特点,对刺萼龙葵进行了风险评估,得出其风险值为86,属高度危险的检疫性有害生物。应当高度重视刺萼龙葵的入侵,积极采取措施控制其危害蔓延。