Variable Penetrance among Different Sources of the Male Fertility Restoration Allele of Onion

Hybrid onion (Allium cepa) seed is produced using cytoplasmic male sterility (CMS). For the most commonly used source of onion CMS, male fertile plants possess male sterile (S) cytoplasm and dominant allele(s) at one nuclear male fertility locus (Ms). Because male fertility restoration is not necessary for bulb production, it is desirable to purge dominant alleles at Ms from populations and breeding lines to facilitate the development of male sterile inbreds for hybrid production. In this research, we used molecular markers to establish the cytoplasms and genotypes at Ms in progenies from testcrosses of male sterile lines with plants from three populations [B2354, Ailsa Craig (AC), and Sapporo-Ki (Ski)] possessing the dominant Ms allele. We scored male fertility of testcross progenies by visual examination of flowers and acetocarmine staining of pollen. Different sources of the dominant Ms allele showed significantly different amounts of male fertility restoration and proportions of stainable pollen, complicating visual selection against the dominant Ms allele. For AC and Ski, molecular markers correctly predicted male sterility vs. male fertility of progenies in the greenhouse and field. However, for B2354, male fertility restoration was less clear and especially difficult to score under field conditions, consistent with reduced penetrance of male fertility restoration for this source of the dominant Ms allele. These results will be of interest to onion breeders selecting S-cytoplasmic male sterile lines for hybrid onion development.

Production and Verification of Hydrangea macrophylla × H. angustipetala Hybrids

The genetic diversity among H. macrophylla (Thunberg) Seringe taxa is limited as a result of the restricted native distribution and multiple breeding programs that used the same taxa and targeted similar breeding goals. This study assessed the compatibility of interspecific crosses between Hydrangea macrophylla and H. angustipetala Hayata as a source of genetic diversity. Two lacecap cultivars of H. macrophylla, ‘Lady in Red’ and Midnight Duchess® (‘HYMMAD II’), were compatible with H. angustipetala. Hybridity of progeny was confirmed by simple sequence repeat markers and morphological comparisons. Some hybrids had red- or purple-pigmented stems, which are characteristic of ‘Lady in Red’ or Midnight Duchess®, respectively. All hybrids had white lacecap inflorescences. Some of the hybrid flowers were fragrant. Winter leaf retention of the hybrids ranged from deciduous to semievergreen. Male fertility of progeny was evaluated by fluorescein diacetate staining of pollen. ‘Lady in Red’, Midnight Duchess®, and H. angustipetala had 62%, 58%, and 79% stainable pollen, respectively, whereas the ‘Lady in Red’ × H. angustipetala and Midnight Duchess® × H. angustipetala hybrids had means of 48% and 47% stainable pollen, respectively. Selected progeny were used to develop F2 and BC1 populations. The interspecific hybrids produced in this study were attractive, fertile plants that are being used in further breeding to develop new cultivars.

Analysis of Ploidy Level and Its Effects on Guard Cell Length, Pollen Diameter, and Fertility in Hydrangea macrophylla

Ploidy level was estimated in Hydrangea macrophylla (Thunb) Ser. using flow cytometry. For H. macrophylla ssp. macrophylla, 42 diploid and 19 triploid cultivars were identified. All 14 H. macrophylla ssp. serrata (Thunb.) Makino cultivars tested were diploids. Somatic chromosome counts confirmed the ploidy of three diploid (2n = 2x = 36) and three triploid (2n = 3x = 54) cultivars. Stomatal guard cell length and pollen diameter of H. macrophylla ssp. macrophylla diploid cultivars were smaller than those of triploid cultivars. However, because the range of measurements for the diploids overlapped that of the triploids, neither guard cell nor pollen measurements are recommended for determining ploidy of H. macrophylla cultivars. Fertility was estimated using pollen staining and controlled pollinations. Stainable pollen for triploid cultivars averaged 63% and ranged from 25% in ‘Masja’ to 85% in ‘Marechal Foch’. Viable seed was obtained when four triploid cultivars were used as pistillate or staminate parents in controlled pollinations to diploid H. macrophylla ssp. macrophylla cultivars. A bimodal distribution of pollen sizes, which is suggestive of unreduced gamete production, was observed in one cultivar; however, more detailed genetic and cytologic studies are needed to elucidate the mechanism behind triploid formation in H. macrophylla taxa.

Backcross Hybrids of Zinnia angustifolia and Z. violacea: Embryology, Morphology, and Fertility

True-breeding lines of Zinnia marylandica Spooner, Stimart and Boyle [allotetraploids of Z. angustifolia H.B.K. and Z. violacea Cav. (2n = 46)] were reciprocally backcrossed with diploid and autotetraploid forms of Z. angustifolia (2n =22 or 44) and Z. violacea (2n =24 or 48). In most cases, backcrosses were more successful with Z. angustifolia and Z. violacea as autotetraploids than as diploids. Seed-generated, backcross (BC1) families were obtained by crossing Z. marylandica (as female) with autotetraploid Z. angustifolia or autotetraploid Z. violacea. BC1 plants were phenotypically intermediate between the two parental lines for most morphological characters. Crosses between Z. marylandica and autotetraploid Z. angustifolia yielded BC1 plants with 33% stainable pollen, whereas crosses between Z. marylandica and autotetraploid Z. violacea yielded BC1 plants that produced malformed, poorly-stained pollen. No embryos were observed in capitula collected from field-grown BC1 plants. BC1 hybrids of Z. marylandica and autotetraploid Z. violacea produced larger capitula and more ray florets than Z. marylandica, and exhibited novel combinations of floral pigments not observed in Z. marylandica ray florets. BC1 hybrids of Z. marylandica and Z. violacea have commercial potential as seed-propagated, bedding plants.

Cytogenetics off interpopulation Cuphea lanceolata hybrids

Cuphea lanceolata Ait. (Lythraceae) is an annual diploid (x = 6) with medium-chain fatty acid rich seed oils. Wild C. lanceolata populations are classified as C. lanceolata f. silenoides or C. lanceolata f. lanceolata on the basis of flower pigment differences. Although these taxa are taxonomically close, their interfertility has not been demonstrated. We describe meiotic phenomena underlying the sterility of hybrids between C. lanceolata f. silenoides (LNS-43) and C. lanceolata f. lanceolata (LNC-78) populations. We assayed metaphase and anaphase I microsporocytes of the parent and hybrid populations. The hybrids were female and male sterile. The mean percentage of stainable pollen was 94.9% for the parents and 1.1% for the hybrids. Chromosomes paired and disjoined normally in the parents (LNS-43 and LNC-78) and abnormally in the hybrids (LNS-43 × LNC-78 and LNC-78 × LNS-43). Univalents, unequal chromosome distributions, and laggards were observed in the hybrids. The mean number of univalents per cell was 0.00 for the parents and 5.95 for the hybrids, the mean number of bivalents per cell was 6.00 for the parents and 1.51 for the hybrids, and the mean number of chiasmata per cell was 9.19 for the parents and 4.04 for the hybrids. The most frequently observed (75%) anaphase I chromosome distribution for the hybrids was 7:5:0 (pole–pole–laggards). The genome affinities of the hybrids were half those of the parents (a mean of 0.5 for the hybrids as opposed to 1.0 for the parents). Although C. lanceolata f. silenoides and C. lanceolata f. lanceolata freely hybridize, their progeny are sterile, and the genetic diversity of LNC-78, and perhaps of C. lanceolata f. lanceolata as a whole, cannot be accessed through hybrids with C. lanceolata f. silenoides or C. viscosissima.Key words: Lythraceae, Cuphea, medium-chain fatty acids, interspecific hybrids, meiosis.

Fertility Losses in Congruity Backcross (CBC), Intraspecific Phaseolus vulgaris Hybrids

Decreases in fertility are most common among interspecific, wide crosses of Phaseolus; intraspecific hybrids are less likely to exhibit sterility. Intraspecific CBC hybrid pedigrees were created to test for comparative fertility losses. Eight P. vulgaris cultivars from different centers of origin, polymorphic for seed proteins (15, 20, 50 kDa), were used to create 16 CBC populations: dry (`Cuarenteño', `Great Northern Harris', `Sulfur', `Swedish Brown') and snap beans (`Purple Pod Pole', `Romano Bush', `Royal Burgundy Bush', `White Half Runner'). Despite repeated attempts, two crosses failed to produce primary hybrids. Primary hybrids had decreased percent stainable pollen from the parents. Female sterility was more severe, necessitating the screening of the F1–F3 before producing the next CBC. Yield was significantly lower than midparent values for all F3 CBC pedigrees. In several cases, phaseolin was no longer the major seed protein. Other hybrid breakdown symptoms were similar to those found with wide crosses, indicative of incongruity between centers of origin.

Effects of Safflower Sterility Genes on the Inflorescence and Pollen Grains

Three interacting, unlinked, nuclear genes are reported to control the inheritance of male-female sterility in safflower (Carthamus tinctorius L.). In a cross between US-10 cultivar (genotype S(1)S(1)s(2)s(2)s(3)s(3)) and a geographically distant Indian line 57-147 (genotype s(1)s(1)S(2)S(2)S(3)S(3)), F-2 plants segregated 57 fertile:7 sterile. Sterile genotypes set no seed under natural conditions. This study reports the effects of sterility genes on the inflorescence and pollen grains. Floret elongation was limited in sterile plants at bloom. Lengths of florets in the sterile individuals showed a 17% and a 40% reduction one day before and on the day of anthesis, respectively. This resulted in a conspicuous pinched appearance of the capitula. Lack of seed development in sterile plants hindered the expansion of capitula and prevented the dried florets from lying against the imbricate bracts as in normal fertile plants. The percentage pollen stainability was 96.1 in US-10, 87.5 in 57-147, 91.3 in the F-1, 90.5 in fertile F-2, and 1.4 in sterile F-2 plants. However, none of the stainable pollen grains of the sterile F-2 plants was viable. The mean pollen diameter was 53.2 in US-10, 56.3 in 57-147, 53.9 in the F-1, 58.1 in fertile F-2, and 35.8 mu m in sterile F-2 plants. Sterile plants exhibited a greater variation in pollen diameter than the fertile plants. Light and scanning electron microscopy observations revealed the absence of normally sculptured and tricolpate pollen from the sterile plants. The results suggest that the sterility genes have their effects primarily on reproductive organs, but do not hinder vegetative development and growth of sterile plants before they bloom.

Genome analysis of Elytrigia caespitosa, Lophopyrum nodosum, Pseudoroegneria geniculata ssp. scythica, and Thinopyrum intermedium (Triticeae: Gramineae)

To elucidate the genome constitutions of the tetraploid (2n = 4x = 28) species Elytrigia caespitosa, Lophopyrum nodosum, and Pseudoroegneria geniculata ssp. scythica and the hexaploid (2n = 6x = 42) Thinopyrum intermedium, meiotic pairing was studied in these species as well as 10 hybrids. Karyotype analysis with aceto-orcein stained root-tip cells was performed for the four species and the hybrids of T. bessarabicum with E. caespitosa, P. geniculata ssp. scythica, and T. intermedium. Karyotype analysis by Giemsa C-banding was carried out with the three tetraploid species and the two triploid hybrids involving T. bessarabicum. The species behaved as strict allopolyploids. All hybrids were male sterile with few stainable pollen grains. It is concluded from the results that the three tetraploid species have the genome formula JeJeSS and T. intermedium has the formula JeJeJeJeSS. The chromosomes of the Je and S genomes in these species had C-banding patterns differing from each other and from those of the extant diploid species. Based on these findings, the four species investigated should be placed in the same genus or the same section of a genus. However, new combinations are not proposed at this time pending future taxonomic investigation of the genome constitution of Elytrigia repens (L.) Nevski.Key words: genome, hybrid, meiosis, karyotype, chromosome banding, speciation.

2n gametes in Solanum commersonii and cytological mechanisms of triplandroid formation in triploid hybrids of Solanum commersonii × Solanum gourlayi

Solanum commersonii Dun. (cmm) is a diploid wild species of potential value for potato breeding. Ploidy level manipulations are necessary to cross this species with the common potato. This could be achieved by the use of "bridge" species and sexual polyploidization in which 2n gametes are involved. Eight introductions of cmm were screened for production of (i) diplandroids, through cytological observations confirmed by controlled 4x Solanum acaule Bitt. (acl) × 2x cmm crosses, and (ii) diplogynoids, by controlled 2x cmm × 2x Solanum gourlayi Haw. (grl) crosses. Eleven plants produced 1–3% large size pollen and one plant produced more than 3% of this type of pollen. Twelve triploid and seven tetraploid hybrids were obtained when these plants were used in crosses with acl. On the other hand, 75 triploid hybrids were obtained from the second type of cross. The average pollen stainability in the diplogynoid triploid cmm × grl hybrids was 18.9%, with a range of 2–34.4%. Forty-four to 54.6% of meiocytes with parallel spindles in metaphase II and anaphase II were observed in microsporogenesis of these hybrids. For each clone, the frequency of triplandroids predicted according to the frequency of meiocytes with parallel, fused and tripolar spindles in anaphase II were 52.3, 36.6, and 45.2%, respectively. These values were highly superior to the frequency of triplandroids predicted according to the number of dyads and triads observed, 8.8, 0.7, and 3.9%, respectively. However, triplandroid frequencies predicted according to the frequencies of fused and tripolar spindles agree with the triplandroid frequencies observed. The frequencies of stainable pollen grains are not completely accounted for by the formation of fused and tripolar spindles. In view of the variability encountered in size, it is assumed that a high proportion of gametophytes with aneuploid chromosome numbers are also stainable. A hypothesis is put forward regarding the possible genetic control of triplandroid formation in the diplogynous triploid hybrids analyzed.Key words: 2n gametes, triploid hybrids, spindle orientation, Solanum commersonii Dun.