Assignment of Endosperm Balance Numbers to the tuber-bearing Solanums and their close non-tuber-bearing relatives

The Endosperm Balance Number (EBN) of over 80 species and subspecies of the tuber-bearing Solanums and their close non-tuber-bearing relatives representing 13 taxonomic series has been determined, with no species assigned to more than one EBN level. Among North American species, most diploids are 1EBN, most tetraploids are 2EBN and all hexaploids are 4EBN; however, among South American species most diploids are 2EBN, most tetraploids are 4EBN and again all hexaploids are 4EBN. Thus species may be isolated from others of the same ploidy level by EBN differences, e.g., 4×(2EBN) from 4×(4EBN), while other species differing in ploidy but having the same EBN may be intercrossed, e.g., 4×(2EBN) and 2×(2EBN). Chromosome doubling or 2n gametes can be used to make a lower EBN species compatible with a higher EBN species. These findings also explain the major crossing difficulties previously inherent in the use of North American species in potato improvement. They also have direct implications for potato improvement, barring the occurrence of other incompatibility barriers. Any 4×(4EBN) cultivar is endosperm compatible and thus will cross with 4×(4EBN) and 6×(4EBN) species. The 2×(2EBN) haploids of 4×(4EBN) cultivars likewise will hybridize with 2×(2EBN) and 4×(4EBN) species. All 2×(1EBN) species are crossable with 2×(2EBN) haploids through 2n gametes or chromosome doubling.Cooperative investigation of the U.S. Department of Agriculture, Agriculture Research Service, and the Wisconsin Experiment Station.     

Crossability relationships among wild potato species with different ploidies and Endosperm Balance Numbers (EBN)

Three wild potato species with different ploidies and Endosperm Balance Numbers (EBN) were crossed in a complete diallel design and the development of the embryo and endosperm as well as the type of seeds produced were analyzed. The compatible crosses – intraspecific intra-EBN and interspecific intra-EBN – produced more than 89% plump seeds, whereas in the incompatible crosses – intraspecific inter-EBN and interspecific inter-EBN – more than 85% of the seeds were not as well developed or were shrunken. The histological analysis revealed that inviable seeds had less developed or collapsed endosperms and thicker endotheliums than viable ones. A gradation of crossabilities was observed among species. The self-compatible species Solanum acaule had good performance as a female but not as a male parent. Among the self-incompatible species, Solanum gourlayi was the best male parent but had a poor performance as a female, whereas Solanum commersonii had an intermediate behavior. Differences in crossabilities among genotypes within species were also observed. These results can not be solely explained by the EBN hypothesis. It is, therefore, suggested that the EBN may be part of a more complex system of interspecific barriers acting at the pre- and post-zygotic levels. This revised version was published online in July 2006 with corrections to the Cover Date.     

A database for wild and cultivated potatoes

A database has been developed for the collections of wild and cultivated potatoes examined by the author in the world herbaria and in his own collections. A total of some 9,000 collections were examined, amounting to approximately 27,000 herbarium sheets when duplicates in different herbaria were taken into account. The information is arranged in twenty-one fields and the system used is DBase IV, version 1.1. 15.00 (including postage and packing) by applying to the author at the above address. Sterling cheques should be made payable to J.G. Hawkes.     

Crossability between tritordeum and triticale

Seed set over three years in crosses between three tritordeums used as female parents and four triticale lines, showed that there are significant differences in crossability attributable to both parents and that most of these differences are consistent over the three years. When used as the female parent tritordeum line HT67 had an average seed set of 29.62%, tritordeum line HT9 an average of 12.73%, and tritordeum line HT31 an average of only 6.58% averaged over the four triticales lines used as pollinators. These data show genotype effect that is highly significant (P < 0.001) both for tritordeum and triticale genotypes and highly significant (P < 0.001) female ×year, male × year and female × pollinator interactions. The behaviour of F₁ tritordeum hybrids when crossed with one of the triticale pollinators supports the conclusion that the parents' crossability behaviour is genetically controlled. Analysis of segregation ratio of F₂ hybrids plants from high and low crossability tritordeum genotypes crossed with the same triticale pollinator genotype is consistent with 9:3:3:1 ratio. This revised version was published online in August 2006 with corrections to the Cover Date.     

Expression of nonacclimated freezing tolerance and cold acclimation capacity in somatic hybrids between hardy wild Solanum species and cultivated potatoes

The expression of freezing tolerance was characterized in interspecific somatic hybrids between S. tuberosum (tbr) and two cold-hardy wild species, S. brevidens (brd) and S. commersonii (cmm). The nonacclimated freezing tolerance (NA) and acclimation capacity (ACC, increase in freezing tolerance in response to low nonfreezing temperature), two main genetic components of freezing tolerance, were evaluated separately. In contrast to cmm, which exhibited excellent NA and ACC, the freezing tolerance of brd was mainly due to ACC. However, the ACC of brd was only moderately expressed in the somatic hybrids. The NA of cmm was also suppressed in combination with tbr genomes. However, with acclimation, some of the tbr (+) cmm somatic hybrids achieved freezing tolerance comparable to pure hardy species such as brd used in this study. Analysis of chloroplast DNA type by RFLP markers revealed no significant difference in ACC between somatic hybrids carrying chloroplasts from either tbr or cmm. The reasons for the reduced expression of freezing tolerance from either the brd or cmm parent and the utilization of these somatic hybrids in breeding programs are discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Vertical resistance to late blight in wild potato species from Argentina

The possible presence of vertical resistance to late blight conferred by R genes in Argentine wild Solanum species, which presumably have not evolved under the pressure of the fungus, was investigated. Solanum microdontum, S. commersonii and S. chacoense clones were tested in the greenhouse and with detached leaves in the laboratory after inoculation with a complex race and a non-virulent race of Phytophthora infestans. Two progeny tests were carried out to assess the performance of contrasting parents in regards to their resistance. A varying frequency of incompatible clones was detected among the species, with S. microdontum having the higher proportion of clones yielding incompatible reactions both in the laboratory and in the greenhouse. S. chacoense and S. commersonii showed a small but still considerable frequency of incompatible clones. The progeny tests confirmed the presence of R genes in a S. commersonii cross and their absence in a S. chacoense cross. The unexpected presence of R genes, its evolutionary significance and the consequences on the use of these species in breeding is discussed. This revised version was published online in August 2006 with corrections to the Cover Date.     

Transmission of a Bacillus thuringiensis cry3Aa transgene from diploid to tetraploid potato using 4x-2x hybridization: effect of ploidy increase on transgene expression and implications for TPS hybrid production

The study examined the effect of ploidy elevation through unreduced gametes on transgene expression in potato. Tetraploid transgenic progenies were obtained from one tetraploid potato cultivar crossed with 2n pollen producing diploid clones harbouring an exogenous transgene (cry3Aa). Both single‐ and multiple‐insert diploid transgenic lines that were regenerated by Agrobacterium tumefaciens leaf disc inoculation were used in crosses. A DAS‐ELISA system and no‐choice feeding bioassay enabled characterization of the parental lines as either ‘high’ or low’ expressers of the Cry3Aa protein. High Cry3Aa expression was observed for both single‐insert transgenic diploids and their 4x‐2x progeny. On the contrary, 68% of 4x‐2x progeny derived from a multiple‐insert, diploid transgenic had significantly reduced Cry3Aa expression compared with the parent, with 32% demonstrating nearly complete silencing of the transgene. Multiple copies of a transgene, like homologous native genes, may be susceptible to transgene silencing following polyploidization. Therefore, incorporation of exogenous transgenes into a true potato seed (TPS) production system is feasible if a single‐insert diploid parent is used. Gene‐centromere mapping of the cry3Aa transgene demonstrated that a non‐transgenic refuge might be naturally created in a TPS hybrid system through genetic recombination.     

The Use of Potato Haploids to Put 2x Wild Species Germplasm into a Usable Form

An effective method of incorporating germplasm of wild 2x Solanum species into the cultivated potato is through hybridization with haploids of 4x S. tuberosum Group Tuberosum. This procedure provides for the introduction of desirable traits and genetic diversity from wild species into potato. Haploid and wild species genotypes can be evaluated for parental value based on the presence of good tuberization, desirable traits, fertility, and 2n gametes in their hybrid progeny. Haploid-wild species hybrids may be intercrossed and evaluated at the 2x level to take advantage of diploid genetic ratios. Selected 2x genotypes may then be brought to the 4x level through sexual polyploidization with 2n gametes.     

Selection of diploid tuberous Solanum hybrids for 2n-egg formation using 2x⋅4x-crosses

Diploid families of Tuberosum hybrids as well as Tuberosum-wild species F₁ hybrids were generated to select 2n-egg producing genotypes of different genetic backgrounds. Plants were selected if they produced more than four seeds per berry after pollination with tetraploid males (2x⋅ot4x-crosses). From the nine families of Tuberosum hybrids that were derived from one or two 2n-egg producing parents, 12 out of 82 (15%) 2n-egg producing plants were selected. From the 32 families of Tuberosum-wild species F₁ hybrids, 21 of 274 (8%) 2n-egg producing plants were selected. The level of 2n-egg formation was estimated in 25 selected diploid hybrids and five control clones (three high, one moderate and one low 2n-egg producers) on the basis of seed set following 2x⋅4x-crosses using 13 tetraploid males in four crossing years. Besides the effect of the diploid 2n-egg producing female on the seed set following 2x⋅4x-crosses (P = 0.0001), there was a significant effect of the tetraploid male (P = 0.0001), whereas the effect of the crossing year (P = 0.0688) was less significant. On the basis of differences in the seed set following 2x⋅4x-crosses as compared to the control clones for low, moderate and high level of 2n-egg formation, the level of 2n-egg formation in two of the newly selected hybrids was classified as very high, four were high, sixteen were moderate and three were low. The significance of the selected hybrids for the genetic analysis of various restitution mechanisms of 2n-egg formation is discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

六倍体裸燕麦与普通小麦属间可交配性研究

以4个不同的六倍体裸燕麦品种和普通小麦进行正反杂交,确定裸燕麦和普通小麦的可交配性。调查杂种结实率并诱导杂种愈伤组织。结果表明：六倍体裸燕麦与普通小麦的可交配性和愈伤组织诱导率因品种和正反交方式的不同而不同。在以六倍体裸燕麦为母本的正交试验中,‘白燕2号’母本结实率最高,为12.98%。在以六倍体裸燕麦为父本的反交试验中,‘白燕5号’为父本时结实率最高,为6.25%。相同品种间正反交结实率也有差异。4个裸燕麦品种与春小麦的杂种胚均有较高的愈伤组织诱导率。此研究对燕麦的优异基因资源的利用和裸燕麦、小麦种质创新具有重要的理论和现实意义。     

Introgression of resistance to Alternaria leaf spot from wild species into susceptible cultivated safflower

Alternaria leaf spot (ALS) caused by Alternaria carthami Chowdhury can cause yield loss up to 90% in safflower (Carthamus tinctorius L.) under severe conditions. Even though a definite source of ALS resistance is not available in cultivated species, a few of the wild species, viz. C. palaestinus and C. lantaus, are known to be tolerant to ALS. Therefore, an attempt was made to introgress Alternaria resistance from these species into cultivated species. F₁-F₈ generations of crosses (PI537632 × C. palaestinus), (C. palaestinus × PI537632), (“Nira” × C. palaestinus) and ([MS 6(O) × C. lanatus] × C. palaestinus) were screened against ALS. ALS infection (%) was recorded in field and quantified using Windias Leaf Image Analysis system. Detached leaf technique was used in laboratory to confirm resistance in interspecific selections. Six resistant and 29 moderately resistant interspecific lines resembling mostly cultivated species were developed. Inheritance of ALS resistance indicated involvement of multiple minor alleles having small effects on resistance. The identified resistant lines could provide potential source of resistance to ALS for safflower breeding programmes.     

Pollen-style compatibility relations in natural populations of the wild diploid potato species Solanum spegazzinii Bitt.

Solanum spegazzinii Bitt., a wild diploid potato species endemic to Argentina, possesses resistance to adverse biotic and abiotic agents. Northwestern Argentinean populations grouped according to their morphological variability (G1 = localities East of the Calchaquí Valley, G2 = Calchaquí Valley and G3 = locality Belén) presented problems in seed set following controlled crosses. Thus, to evaluate if the morphological variability previously reported was sustained on a genetic basis, three introductions per group and five plants per introduction were used in controlled crosses (genotypic combinations). These were classified according to the number of seeds produced per fruit. Pollen-style compatibility relations were studied in those crosses that did not set seeds or that produced less than 30 seeds each. Within groups, G1 produced seeds normally, whereas hybridization barriers were found in 71.4% and 50.0% of the combinations within G2 and G3, respectively; on the other hand, from 46.7% to 73.3% of the combinations among groups exhibited similar barriers. Variability in the expression of the sites of pollen tube inhibition in the style was observed both within and among groups. Speculations are made on the possible hybrid origin of the populations. This revised version was published online in July 2006 with corrections to the Cover Date.     

Evaluation of FDR diploid and tetraploid parents in potato under two different day-length environments

Five tetraploid clones of potato were crossed to each of eight diploid first division restitution (FDR) 2n pollen producers and four tetraploid males using a line × tester mating design. A total of 59 families were obtained and evaluated under long days at Rhinelander (USA). A subset of 49 families were grown under short days at four Peruvian locations. Tuber yield (kg per plant) of 4x-2x families was similar to, or greater than, that of 4x-4x families in short day environments. The diploid clones had breeding values greater than or equal to those of the tetraploid clones. Only families derived from FDR 2n pollinators combined high tuber yield with stability and high dry matter content. Reciprocal recurrent selection would be the best breeding scheme, in which the diploids are testers of the tetraploids and vice-versa. Intra-population improvement could be achieved through phenotypic recurrent selection. The best materials from both breeding pools could then be crossed to produce tetraploid hybrids through unilateral sexual polyploidization (4x-2x crosses).     

Characterization of resistance to Globodera rostochiensis pathotype Ro1 in cultivated and wild potato species accessions from the Vavilov Institute of Plant Industry

Potato cyst nematodes (PCN) collected in six localities in the Leningrad region of North West Russia were identified as Globodera rostochiensis pathotype Ro1 and were used for subsequent resistance tests. Seventy‐nine accessions of cultivated and closely related wild potato species from the VIR collection in Russia were screened on resistance to G. rostochiensis pathotype Ro1 and on the presence of molecular markers for H1 and Gro1‐4 resistance genes. No associations were detected between the resistance level of diploid and tetraploid Andean and tetraploid Chilean potato landraces (indigenous cultivars) and their related wild species and their geographical distribution or presence of PCR‐based markers that are associated with the H1 and Gro1‐4 genes. At the same time, all susceptible genotypes lacked such markers. New sources of resistance were found and could be used in breeding.     

Introgression of the low browning trait from the wild Mexican species Solanum hjertingii into cultivated potato (S. tuberosum L.)

Internal discoloration of tubers resulting from impact damage (blackspot bruise) is a serious quality problem in potato production and utilization, reducing profits to growers and increasing costs for processors. Resistance to blackspot bruise has been identified in the wild species Solanum hjertingii and is therefore a potential germplasm resource for genetic resistance to this problem. A bridging cross between S. hjertingii and a cultivated diploid clone was used to produce a triploid hybrid population that exhibited very low tuber browning potential, indicating a dominant pattern of inheritance for this trait. The triploid progeny were subjected to in vitrochromosome doubling and the resulting hexaploid clones were screened for browning potential. A hexaploid clone selected for low browning was reciprocally crossed with cultivated S. tuberosum cultivars exhibiting high susceptibility to blackspot bruise. Tubers obtained from the seed progeny of these 4x-6x crosses (hereafter referred to as the BC₁ populations) were evaluated for browning potential and polyphenol oxidase (PPO) activity. Tubers from the BC₁ populations displayed a very low potential for melanin production, while PPO activity was quite variable. The low Pearson correlation coefficient (r² = 0.45), between browning potential and PPO activity suggests that the mechanism of blackspot bruise resistance derived from S. hjertingii cannot be explained simply as a reduction in the initial PPO activity. The expression of substantial resistance to browning and dominant expression pattern in these BC₁ progeny indicate that utilizing genetic elements derived from S. hjertingii provides a robust approach for developing blackspot bruise resistant potato varieties. This revised version was published online in August 2006 with corrections to the Cover Date.     

Phylogenetic differentiation of cultivated rice,XXIII. Potentiality of wild progenitors to evolve the Indica and Japonica types of rice cultivars

Summary Studies of intermediate wild-cultivated plants suggested that the Indica-Japonica differentiation of cultivars of Oryza sativa L. has taken place with the domestication of wild plants. The wild progenitor, which is considered to be the Asian form of O. perennis Moench, shows no trace of differentiation into two such types as the Indica and Japonica. To elucidate the potentiality of the wild progenitor to evolve the two types when domesticated, the selfed progenies from an Indica × wild and a japonica × wild cross were examined with regard to three characters known to be useful for distinguishing between the two types. The data showed that Japonica-like plants were derived from the Indica × wild cross and Indica-like plants from the Japonica × wild cross. The Indica-Japonica differentiation was considered to have resulted from selection of incipient domesticates having such potentiality under different climatic conditions; the dynamics was discussed.     

Evaluation of Potato Hybrids Obtained from Tetraploid-Diploid Crosses I. Parent-Offspring Relationships

A total of 45 tetraploid-diploid (4x—2x) hybrid progenies were obtained by crossing each of ten diploid parents as males to each of five tetraploid parents. A relatively low degree of determination (R²) of both 4x and 2x parents on the performance of their 4x—2x hybrid progenies was found for all six traits studied. This implies that test crossing and subsequent progeny analysis is necessary before such parents can be used in a breeding program. Since marketable yield was found to have a higher predictability than total yield, it is the preferred yield trait to be measured. Diploids which produce 2n gametes via FDR usually give rise to 4x progenies with a greater amount of heterosis than similar progenies derived from diploids which produce 2n gametes via SDR, although such heterosis is not necessarily expressed in terms of marketable yield.     

Interspecific hybrids between three eggplant (Solanum melongena L.) cultivars and two wild species (Solanum torvum Sw. and Solanum sisymbriifolium Lam.)

Three Greek eggplant cultivars, ‘Langada’, ‘Tsakoniki’ and ‘Emi’ (2n= 24), were crossed with two wild species (Solanum torvum Sw., 2n= 24 and Solanum sisymbriifolium Lam., 2n= 24). Ovules isolated 15-27 days after pollination were cultured in a modified MS medium at 24°C and a 16h photoperiod. Fifty days later, the ovules were dissected and the interspecific embryos were cultured in the same medium. Interspecific hybrids were achieved only from crosses between the eggplant cultivars and S. torvum. The hybridity of the putative interspecific F¹ hybrid (Solanum melongena×S. torvum) was confirmed by using morphological and biochemical (isozyme isocitrate dehydrogenase A, phosphoglucomutase A, phosphoglucose isomerase B, 6-phosphogluconate dehydrogenase A, 6-phosphogluconate dehydrogenase B) markers. The F¹ plants (‘Langada’×S. torvum) were selfpollinated and backcrossed to both parents. Fruits, however, were produced only when the F¹ hybrid was backcrossed as female with the eggplant cultivar ‘Langada’.     

Tolerance to low temperatures and tuber soft rot in hybrids between Solanum commersonii and Solanum tuberosum obtained through manipulation of ploidy and endosperm balance number (EBN)

The objectives of this study were to evaluate the tolerance to low temperatures and tuber soft rot in hybrids between Solanum commersonii and Solanum tuberosum. The experimental materials consisted of F₁ triploid, BC₁ pentaploid‐near pentaploid and BC₂ tetraploid–near tetraploid hybrids. The F₁ triploids had a freezing tolerance and acclimatization capacity closest to S. commersonii. This indicated that the endosperm barriers which prevent the introgression of 1EBN S. commersonii into 4EBN S. tuberosum had been overcome. Indeed, the triploids produced 2n eggs, thus giving a compatible maternal to paternal EBN ratio in the hybrid endosperm generated by the 3x(2EBN) × 4x(3EBN) crosses. The tolerance to low temperatures of BC₁ and BC₂ hybrids was lower than that of the F₁. However, a number of genotypes were identified which were able to withstand temperatures down to ‐5°C. Some BC₂ hybrids were also tested for their tolerance to tuber soft rot, and some resistant hybrids were detected. A number of them combined the capacity for cold acclimatization with tolerance to tuber soft rot. These hybrids have an EBN of 4; they are fertile and have been used in backcrosses with 4EBN S. tuberosum.     

Introduction of an amylose-free (amf) mutant into breeding of cultivated potato,Solanum tuberosum L.

Summary Using an amylose-free (amf)mutant of diploid potato (Solanum tuberosum), diploid and tetraploid clones with different genotypes at the amf-locus were produced. In order to make use of the diploid material in analytic breeding of amf-potatoes, clones were selected that produced a considerable frequencies of 2n-pollen and 2n-eggs. Successful attempts were made to select normal synaptic as well as desynaptic clones producing 2n-gametes. Based on the phenotype of starch in the microspores, tetraploid clones with nulliplex, simplex and duplex genotypes at the amf-locus were selected. Prospects of using this material in conventional as well as in analytic breeding of potato are discussed.