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.     

The nature of the Bolivian weed potato species Solanum sucrense Hawkes

Summary Two theories for the origin of the Bolivian weed potato species Solanum sucrense Hawkes have been tested. The first was that it had an allopolyploid origin as a hybrid of the cultivated tetraploid S. tuberosum ssp. andigena (Juz. et Buk.) Hawkes with the wild/weed tetraploid cytotype of S. oplocense Hawkes. The second hypothesis postulated that it was a cross of the weed diploid species S. sparsipilum (Bitt.) Juz. et Buk. with S. oplocense.Synthetic hybrids of S. tuberosum ssp. andigena x S. oplocense and of S. sparsipilum x S. oplocense have been compared morphologically with S. sucrense accessions. Their crossability has also been investigated.The crossability and morphological studies strongly suggested that the former hypothesis was the more likely. This was borne out by observations made on the 1974 University of Birmingham Expedition when segregating populations of S. sucrense and S. oplocense were collected growing with feral S. tuberosum ssp. andigena.The evidence from this study indicates that the S. tuberosum ssp. andigena x S. oplocense hybrid has formed the basis of the S. sucrense gene pool. It is suggested that this hybridogenic taxon be maintained under the name Solanum x sucrense.The clarification of the taxonomic relationships of these three species will be of interest to Globodera resistance breeders, in view of the fact that these taxa are resistant to various pathotypes of the golden nematode.     

Use of Vaccinium octoploids to facilitate 4x-6x gene transfers

The use of an 8x Vaccinium produced through doubling a tetraploid hybrid with colchicine was studied as a means of facilitating gene exchange between V. corymbosum L. and V. ashei Reade. Analysis included meiotic observations and crossability studies of the 8x plant, as well as attempting to develop 6x breeding lines. Meiotic analysis revealed the presence of micronuclei in Telphase II products and polyspory in sporads. In the crossability studies five 4x highbush plants were pollinated with 8x o2 4x pollen. Data gathered included: % fruit set, average weight/fruit, average number of seeds/fruit, average number of seedlings/pollination, and number of 6x seedlings. Octoploid pollinations were significantly lower than 4x pollinations in all parameters. One 6x plant was produced from the 4x-8x crosses but was found to be mitotically unstable, having somatic cells with chromosome numbers ranging from 48 to 168.     

Natural triploid hybrids between Solanum acauleBitter and S. infundibuliformePhilippi in the province of Jujuy,Argentina

Summary The occurrence of natural triploid hybrids between tetraploid Solanum acauleBitter subsp. acaule and diploid S. infundibuliformePhilippi both tuber-bearing Solanums, in the Puna region of Jujuy, is reported. The natural hybrids, which are morphologically intermediate between their putative parents and completely male sterile, have a somatic chromosome number of 36. The artificial hybrids, obtained by crossing both parental species, closely resemble the natural ones thereby confirming their parentage. It is proposed to designate the natural hybrids Solanum x viirsooi hybr. nov.     

Breeding for quantitative resistance to potato cyst nematode (Globodera pallida) in tetraploid potatoes using dihaploids and unreduced gametes

Summary Dihaploids were produced from tetraploids resistant to potato cyst nematode (Globodera pallida (Stone)). High levels of resistance were found in the dihaploids and three were used to produce tetraploid progenies by crossing them with susceptible tetraploid cultivars. One dihaploid, PDH505, produced more highly resistant offspring than the other two, PDHs 417 and 418. The latter gave progenies whose levels of resistance were similar to those obtained from susceptible dihaploids crossed with resistant tetraploids.The differences between the progenies of the resistant dihaploids were probably due to different modes of unreduced gamete formation (PDH505 producing gametes by first division restitution (FDR) and PDHs 417 and 418 by second division restitution (SDR)) although cytological studies would be necessary to confirm this. The methods by which dihaploids could be utilised in a tetraploid potato breeding programme are discussed in relation to the mode of unreduced gamete formation.     

Origin and variation of gliadin proteins associated with pasta quality in durum wheat

Summary Studies were made of the presence and frequency of occurrence of gliadin bands 42 and 45 in three samples of Aegilops sharonensis Eig and 59 samples of wild tetraploid wheat (Triticum dicoccoides Korn.) from natural distributions of these species in Israel.Two samples of Ae. sharonensis possessed a band in position 45 and one possessed no bands corresponding to either band 45 or band 42. In T. dicoccoides, band 45 was either present or not and 42 was always absent. In its grassy and intermediate growth habit forms, (believed to be more primitieve than the cercal forms) band 45 appeared to be more frequent than in the cereal form.The presence of band 45 in the Ae. sharonensis, and its relatively high frequency in T. dicoccoides, populations from Mt. Hermon (likely to be relatively free from introgression from cultivated tetraploid wheat) indicate the likelihood of a primary origin of the allele coding for band 45. The absence of band 42 from all Ae. sharonensis and T. dicoccoides populations in this study, indicates a more recent evolutionary origin of the allele coding for this band, possibly arising as a mutation during the domestication of tetraploid wheat.The results have implications for breeding programmes in tetraploid wheat.     

Natural hybridization between Solanum acaule Bitt. and S. megistacrolobum Bitt. in the province of jujuy,Argentina

Summary Evidence is provided that hybridization of the tetraploid (2n=4x=48), self-fertile tuber-bearing species Solanum acaule Bitt. with the diploid (2n=2x=24), self-incompatible, tuber-bearing S. megistacrolobum Bitt. takes place in several localities of the province of Jujuy in the high, cold plateaux of the Argentine Puna. The triploid hybrids (2n=3x=36) closely resemble S. acaule in growth habit, leaf morphology and floral structures and for this reason they can be easily overlooked for that species in the field. Experimental data show that S. acaule can be crossed with S. megistacrolobum though the crossability is rather low and variable according to the particular cross considered. The artificial hybrid obtained compares well with the natural hybrid in morphology and chromosome number. The hybrids, though almost completely male sterile, are successful colonizers of disturbed areas around farmers' dwellings, cattle enclosures and other areas where the soil is rich in organic matter.There is some evidence that the Tilcara material of S. acaule subsp. aemulans and the hybrids of S. acaule x S. megistacrolobum have some characters in common, which can be interpreted as having a similar origin.It is postulated that S. acaule subsp. aemulans, in Jujuy at any rate, is not a primitive form of S. acaule as thought by Hawkes and Hjerting, but rather a fertile hybrid derivative of S. acaule x S. megistacrolobum through the functioning of 2n gametes.We also provide evidence that S. bruecheri Correll should not be considered a hybrid of S. acaulle x S. megistacrolobum but a synonym for S. gourlayi Hawkes. The new name, S. x indunii Okada et Clausen, is proposed to designate this hybrid.     

Molecular markers linked to <Emphasis Type="Italic">Bn</Emphasis>;<Emphasis Type="Italic">rf</Emphasis>: a recessive epistatic inhibitor gene of recessive genic male sterility in <Emphasis Type="Italic">Brassica napus </Emphasis>L.

7–7365AB is a recessive genic male sterile (RGMS) two-type line, which can be applied in a three-line system with the interim-maintainer, 7–7365C. Fertility of this system is controlled by two duplicate dominant epistatic genes (Bn;Ms3 and Bn;Ms4) and one recessive epistatic inhibitor gene (Bn;rf). Therefore an individual with the genotype of Bn;ms3ms3ms4ms4Rf_ exhibits male sterility, whereas, plant with Bn;ms3ms3ms4ms4rfrf shows fertility because homozygosity at the Bn;rf locus (Bn;rfrf) can inhibit the expression of two recessive male sterile genes in homozygous Bn;ms3ms3ms4ms4 plant. A cross of 7–7365A (Bn;ms3ms3ms4ms4RfRf) and 7–7365C (Bn;ms3ms3ms4ms4rfrf) can generate a complete male sterile population served as a mother line with restorer in alternative strips for the multiplication of hybrid seeds. In the present study, molecular mapping of the Bn;Rf gene was performed in a BC₁ population from the cross between 7–7365A and 7–7365C. Bulked segregant analysis (BSA) and amplified fragment length polymorphism (AFLP) technique was used to identify molecular markers linked to the gene of interest. From a survey of 768 primer combinations, seven AFLP markers were identified. The closest marker, XM5, was co-segregated with the Bn;Rf locus and successfully converted into a sequence characterized amplified region (SCAR) marker, designated as XSC5. Two flanking markers, XM3 and XM2, were 0.6 cM and 2.6 cM away from the target gene, respectively. XM1 was subsequently mapped on linkage group N7 using a doubled-haploid (DH) mapping population derived from the cross Tapidor × Ningyou7, available at IMSORB, UK. To further confirm the location of the Bn;Rf gene, additional simple sequence repeat (SSR) markers in linkage group N7 from the reference maps were screened in the BC₁ population. Two SSR markers, CB10594 and BRMS018, showed polymorphisms in our mapping population. The molecular markers found in the present study will facilitate the selection of interim-maintainer.     

The genetics of seedling resistance to leaf rust,Puccinia hordeiOtth. in some spring barley cultivars

Summary The three Dutch isolates studied carried virulence genes against the resistance genes Pa, Pa-2, Pa-4 and Pa-5, substantiating the widespread occurrence of these virulences in Western Europe as reported by others. The cultivars Cebada Capa, La Estanzuela, Gondar and Dabat carry the same dominant to semi-dominant gene, which is also found in Forrajera Klein and H2212. It is proposed to designate this gene Pa-7. This gene segregated independently from the Pa-3 gene in Rika x (Baladi x Rika) substantiating the data of Johnson (1968), but disagreeing with those of Roane & Starling (1970). EP 75 seems to carry a single dominant or semi-dominant gene conferring an intermediate resistance to the isolates used. This gene, tentatively designated as Pa-z, is different from the Pa, Pa-2, Pa-4, Pa-5 and Pa-7 genes. The intermediate resistance of Monte Christo is probably based on gene(s) different from EP 75.     

Identification of tetraploid regenerants from cotyledons of diploid watermelon cultured in vitro

Summary Adventitious shoots were obtained from the diploid watermelon [Citrullus lanatus (Thunb.) Matsum. & Nakai] cultivars Dixielee, Jubilee II, Mickylee, Minilee, and Royal Sweet by culturing excised cotyledons on shoot regeneration medium for six weeks. Tetraploid and diploid regenerants were identified by counting the number of chloroplasts per guard cell pair from leaves of regenerated plants. Cross fertilization of putative tetraploids with diploid pollinators and the production of triploid seed confirmed the efficacy of this approach. The mean number of chloroplasts for tetraploid regenerants was 19.1 whereas diploids averaged 11.2. These values were similar to tetraploid and diploid plants from seed. Ovary diameter, petal, and anther diameter of male flowers, and leaf length by width ratio were also good indicators of plant ploidy. Progeny obtained from self-fertile tetraploids of Mickylee were crossed with various diploid pollinators to produce triploid hybrid seed. All triploid plants from tissue culture-derived tetraploids produced fruit comparable in quality to fruit produced by currently-available triploid hybrids, demonstrating that in vitro tetraploid induction can be used to produce high quality tetraploid plants for use in triploid hybrid seed production.     

Methods of overcoming interspecific barriers in Trifolium

Summary Barriers to interspecific hybridization in Trifolium were investigated by manipulation of mentor pollen treatments, ploidy levels, and compatibility and male sterility systems. Crosses involving the addition of mentor pollen produced fewer seeds and hybrids than crosses involving normal pollination. Lower seed set with mentor pollen was deduced to result from the use of less viable pollen, approximately half the pollen having been killed by alcohol. Pollinations at the diploid level resulted in more hybrids than at the tetraploid level, perhaps because genes for male sterility produced higher female sterility in the tetraploids. The self-compatible stock produced more seeds, mostly selfs, than the self-incompatible stock, but produced more hybrids only in one cross, T. pratense L. × T. diffusum Ehrh. The use of male-sterile female parents reduced selfing but produced fewer hybrids than male-fertile female parents. Techniques of this study were designed to affect prefertilization barriers, but the lack of effect may indicate that postfertilization barriers in Trifolium are of greater importance.Journal Article No. 98-3-208 of Kentucky Agricultural Experiment Station. Published with approval of the Director.     

The inheritance of thornlessness from tissue culture-derived ‘Thornless evergreen’ blackberry

Summary Thornless Evergreen (TE) blackberry (Rubus laciniatus Willd.) is a tetraploid thornless periclinal chimera which produces thorny adventitious root suckers and breeds as if it were genetically thorny. Using tissue culture procedure, several pure thornless (non-chimeral) plants were obtained from TE. These tissue cultured-derived plants (TE_tc) were grown to maturity, flowered, hybridised with various thorny and thornless cultivars, and selfed. When TE and TE_tc were used as female parents, a high degree (ca 94%) of the seedlings were apomictic. However, both thorny and thornless sexual offspring occurred in all crosses. The thornless: thorny segregation ratios suggest that the thornless gene(s) of TE_tc is (are) dominant to the thorny alleles.     

Comparative study on vegetative,reproductive and qualitative traits of seven diploid and tetraploid watermelon lines

Seven tetraploid watermelon lines developed by colchicine treatments were compared with their diploid counterpart for plant, flower, fruit, seed and qualitative characteristics. Tetraploid genotypes attained statistically higher vine thickness (8.04 mm), leaf area (298.9 cm²) and chlorophyll content (55.6) while internode length and chlorophyll fluorescence was similar to their corresponding diploid. Both pistillate and staminate flower organs (pedicel, anther, ovary, stigma, petals) were larger in tetraploid plants; however, the percent increase in flower components varied across the tetraploid lines. Fruit weight and total sugar content (^∘Brix) in both ploidy fruits was similar. Rind thickness in fruits varied significantly and averaged 12.7 and 17.2 mm in diploid and tetraploid fruits, respectively. Tetraploid genotypes showed sterility, yielded lower number of seed per fruit (37.9), and tetraploid seed was larger and thicker than diploid seed. Overall β-carotene (0.89), lycopene (1.16), fructose (5.43%) and glucose (2.38%) contents were higher in tetraploid than diploid fruits.     

Breaking the crossability barriers between disomic tetraploid Solanum acaule and tetrasomic tetraploid S. tuberosum

Summary A combination of compatible second pollinations and embryo rescue was applied for systematic production of true tetraploid hybrids from crosses between disomic tetraploid Solanum acaule and tetrasomic tetraploid potato, S. tuberosum. Several genotypes of tetraploid potatoes were pollinated with S. acaule, and the compatible second pollinations were made on the following day, with a genotype of S. phureja, IvP 35 to promote fruit development. Embryo rescue was carried out in 21 families, 14 to 27 days after the first pollination. A total of eight plants were obtained from the embryo rescue and their chromosome numbers were counted in the root tips. Three of the eight plants were identified as tetraploid, and five others as diploid. Morphology, isozyme banding patterns, and pollen stainability, as well as potato spindle tuber viroid (PSTVd) resistance, indicated the hybrid nature of the three plants. This is the first report of successful tetraploid hybrid production between disomic tetraploid S. acaule (4x) and tetrasomic tetraploid potatoes. Seed set from the crosses between one of hybrids and diploid potatoes indicated workable levels of both male and female fertility for introgression of valuable genes from S. acaule into the cultivated potato gene pool. The methodology used may be applied to other disomic tetraploid tuber-bearing Solanum species and with some modifications also to distantly related solanaceous species and genera.     

Seasonal changes in the incidence of apomixis of diploid,triploid, and tetraploid plants of Paspalum cromyorrhizon

Summary Changes in incidence of apomixis were determined at different intervals of the flowering period in a highly sexual diploid cytotype of Paspalum cromyorrhizon Trin., a natural tetraploid cytotype of the same species, and in the triploid and tetraploid intraspecific hybrids that were produced by pollinating the 2x cytotype with pollen of the tetraploid. Reproductive behaviour was determined from observations of embryo sacs in mature ovules. Gametophytic apomixis in this species in characterized by aposporous embryo sacs which differ from normal sacs by their number, orientation inside the ovule, their dimensions and shape, and the absence of antipodal cells.The potential for apomictic reproduction increased in relation to the ploidy level, suggesting a gene dosage effect in the incidence of apomixis. In the three ploidy levels, the highest incidence of apomixis was observed when the plants reached the maximum number of flowering heads. These findings suggest that the same environmental conditions that favour flowering should also be responsible for increasing the incidence of apomixis. An additional experiment under controlled conditions indicated that the light regime should be one of the factors that govern the amount of apomictic reproduction. Thus, polyploidy and apomixis should be of special significance in the dispersion and evolution of this grass species. The 4x facultative apomictic cytotype is far more frequent in nature than the highly sexual 2x. Moreover, most of the seeds produced in a flowering season should arise asexually. So, apomixis rapidly increases the number of plants bearing the successful maternal genotype, and sexual reproduction becomes important in adverse environmental conditions.     

Disomic alien chromosome substitution and addition in hexaploid oat

Summary True-breeding, leaf rust (Puccinia coronata Cda. f. sp. avenae Fraser and Led.) resistant oat lines with somatic chromosome numbers of 42 and 44, were recovered in the advanced generations of a pentaploid hybrid from the cross between C.I. 7232, a leaf rust resistant and dark lemma colored derived tetraploid and Clarion, a susceptible hexaploid oat. Microsporocyte observations and breeding behaviour of these lines and their F₁ hybrids with susceptible hexaploid oat cultivars indicated that the 42- and 44-chromosome lines are disomic substitutions (20+1A) and disomic additions (21+1A), respectively, of the same alien chromosome which carries genes for dark colored lemma and leaf rust resistance. The substitution and addition lines possess good field resistance to leaf rust. Plants of substitution lines were less vigorous, shorter, with poor straw and reduced fertility. Addition lines had reduced tillering ability, plant vigour, culm thickness, leaf width and fertility and were late maturing. Progeny tests for resistance indicated that substitution lines are more stable than addition lines. It is suggested that these lines may be of value in a program of radiation-induced gene transfer.     

Cytological and geographical characterizations of Hemarthria

Summary Chromosme numbers were determined for 56 Hemarthria altissima (Poir.) Stapf et C. E. Hubb. introduction, forty of these were diploid (2n=18) and 16 were tetraploid (2n=36). These results, combined with previous findings, show that all but three of the tetraploids in the USDA Hemarthria altissima collection originated north of 20 S latitude. All other introductions collected throughout southern Africa were diploid. Meiosis was regular in a hexaploid, 6 tetraploid and 32 diploid introductions but a low frequency of quadrivalents was found in the teraploids. H. uncinata R. Br. and H. uncinata var. spathacea (Domin) Vickery were found to have 2n=36 chromosomes H. compressa (L.f.) R. Br., introduced from Japan, had 2n=54 chromosomes. No major cytological abnormalities exist in these Hemarthria species. Significant variation exists among the ploidy levels for cold tolerance and in vitro organic matter digestion to provide the basis for a forage improvement program.Joint contribution from the Department of agronomy. Institute of food and Agric. Sciences (IFAS), Univ. of Florida. Gainesville, Fl and, Plant Genetics and Germplasm Institute, USDA, Beltsville, MD Published as IFAS Journal Series No. 2849.     

Resistance to the gall mite and American gooseberry mildew in black currants in relation to season of leafing out

Summary In black currant (Ribes nigrum) backcross progenies segregating for resistance to the gall mite, Cecidophyopsis ribis Westw. (gene Ce from Ribes grossularia L.), and to American gooseberry mildew, Sphaerotheca mors-uvae (Schw.) Berk. (gene Sph ₃from R. glutinosum), significant differences in season of leafing out occurred between the resistant and susceptible classes. The data are consistent with the hypothesis that two additive genes, Lf ₁and Lf ₂, controlled season of leafing out and that Ce and Sph ₃ were linked with Lf ₁.     

Crosses between Beta intermedia bunge and B. vulgaris L.

Summary A tetraploid annual male sterile form of Beta vulgaris L. (2n=4x=36) was crossed with the wild beet species Beta intermedia Bunge (2n=36). The resulting F₁-plants were male sterile annuals being two or three times back-crossed to diploid and tetraploid sugar and fodder beets in the next years. Apart from tetraploid material (36 chromosomes) hexaploid (54 chromosomes) and a number of aneuploid plants developed.The results obtained justify the conclusion that, at a tetraploid level the material mostly propagates apomictically after the F₁ generation. The presence of penta-, hexa-, septa-and even octaploid plants might be explained by assuming that no meiosis has taken place in the crossing partners. Triploid plants are sometimes found in the progeny of hexaploid material and may presumably be considered haploids. Moreover some pentaploid plants were found in the progeny of the open pollinated F₁ which after two generations of bagging are still pentaploids although they produce no pollen. This is another clear indication of apomictic reproduction.The tetraploid generation from the cross between the hexaploid material and diploid sugar beets probably contains the best prospects for breeding.     

The introduction into bread wheat of a major gene for resistance to powdery mildew from wild emmer wheat

Summary A new source of resistance to wheat powdery mildew caused by Erysiphe graminis has been transferred to hexaploid bread wheat, Triticum aestivum, from the wild tetraploid wheat, Triticum dicoccoides. The donor was crossed to bread wheat and the pentaploid progeny was then self-pollinated. Plants having a near stable hexaploid chromosome complement were selected in the F₃ progeny and topcrossing and backcrossing of these to a second wheat cultivar to improve the phenotype was undertaken. Monosomic analysis of early backcross lines showed the transferred gene to be located on chromosome 4A. The gene has been designated Pm16.