Allelism of Endosperm Balance Number (EBN) in Mexican tuber-bearing Solanum species

Summary Endosperm Balance Number (EBN) is a genetic, dose-dependent crossability system functioning in tuber-bearing Solanum species. Each species has been assigned 1EBN, 2EBN, or 4EBN. Species thus designated cross only within their EBN group. Doubling of chromosome number also doubles the EBN. The ploidy: EBN ratio is not consistent among Solanum species. Some diploids are 2EBN while others are 1EBN. Some tetraploids are 4EBN while others are 2EBN. Species from Mexico typically have EBNs that are one-half of their ploidy [e.g. 2x(1EBN), 4x(2EBN)]. Hybrids of Mexican species and a South American species, 2x(1EBN) S. Commersonii, and its 4x(2EBN) colchicine derivative were made and crossed to 1, 2, and 4EBN standard testers to determine the relationship of the genetic organization of EBN among and within these species. Diploid hybrids crossed only to 1EBN standard testers. Hybrids of 4x(2EBN) S. commersonii and 4x(2EBN) Mexican species crossed almost exclusively to 2EBN standard testers. Complex tetraploid hybrids involving S. commersonii, S. stenophyllidium (a Mexican diploid), and Mexican tetraploids of series Longipedicellata also crossed only to 2EBN testers. The apparent lack of recombination and segregation for EBN in these hybrids indicates that the genomes of the Mexican diploid and tetraploid species carry EBN in a way genetically similar to that of the South American species S. Commersonii.Cooperative investigation of the U.S. Department of Agriculture, Agricultural Research Service and the Wisconsin Experiment Station. Supported in part by the USDA/Cooperative States Research Service Competitive Grant No. 83-CRCR-1-1253     

The role of 2n gametes and endosperm balance number in the origin and evolution of polyploids in the tuber-bearing Solanums

Polyploidization has played a major role in the origin and evolution of polyploid species. In this article we outline the unique characteristics of 2n gametes and implications of their participation in the evolution of polyploid Solanum species. The genetic consequences of 2n gametes indicate that sexual polyploidization results in greater variability, fitness, and heterozygosity than does somatic doubling. Further, the mechanisms of 2n gamete formation and the frequency of 2n gamete-forming genes in present polyploids and their ancestral species provide additional evidence of their involvement. Equally important is the endosperm, via the endosperm balance number (EBN) incompatibility system, in complementing the role of 2n gametes. In fact, the EBN system acts as a screen for either 1n or 2n gametes, depending on the EBN and chromosome numbers of parental species. EBN in combination with 2n gametes maintains the ploidy integrity of diploid ancestral species, while providing the flexibility for either unilateral or bilateral sexual polyploidization.     

The occurrence and frequency of 2n pollen in three diploid solanums from Northwest Argentina

Summary A group of wild, tuber-bearing species from Northwest Argentina, belonging to the series Tuberosa, Solarium spegazzini Bitt. (spg, 2n=2x=24), S. gourlayi Hawkes (grl, 2n=2x=24 and 2n=4x=48) and S. oplocense Hawkes (opl, 2n=6x=72), and Cuneolata, S. infundibuliforme Phil (ifd, 2n=2x=24), is being used to investigate the mode of origin of polyploids in the genus Solanum. 2n gametes have been detected in the diploid species ifd and spg and in a diploid race of grl, using cytological and breeding approaches. Twenty-two introductions of spg, 8 of grl and 26 of ifd have been tested for 2n pollen; 59%, 63% and 54% of them, respectively, had at least one 2n pollen producing plant. These introductions comprised 238, 76 and 235 plant respectively, of which 20, 16, and 32 plant produced 5% or more 2n pollen. The mechanism of 2n pollen formation was determined in several plant of 2x spg, 2x grl and 2x ifd. All of them were found to form diplandroids via parallel spindles. This mechanism, which gives meiotic products genetically equivalent to first division restitution gametes, is under control of the Mendelian recessive ps. The results suggest that the allele ps is widely distributed in natural populations of the three diploids, and that its frequency is very high. These species are seen as valuable material for population genetic studies, and for the eventual incorporation into a breeding scheme involving sexual polyploidization via 2n gametes.     

The use and limits of AFLP data in the taxonomy of polyploid wild potato species in Solanum series Conicibaccata

Solanum sect. Petota (tuber-bearing wild and cultivated potatoes) are a group of approximately 190 wild species distributed throughout the Americas from the southwestern United States south to Argentina, Chile, and Uruguay. Solanum series Conicibaccata are a group of approximately 40 species within sect. Petota, distributed from central Mexico to central Bolivia, composed of diploids (2n = 2x = 24), tetraploids (2n = 4x = 48) and hexaploids (2n = 6x = 64); the polyploids are thought to be polysomic polyploids. This study initially was designed to address species boundaries of the four Mexican and Central American species of series Conicibaccata with AFLP data with the addition of first germplasm collections of one of these four species, Solanum woodsonii, as a follow-up to prior morphological, chloroplast DNA, and RAPD studies; and additional species of series Conicibaccata from South America. AFLP data from 12 primer combinations (1722 polymorphic bands) are unable to distinguish polyploid species long thought to be distinct. The data suggest a complex reticulate history of the tetraploids or the need for a broad downward reevaluation of the number of species in series Conicibaccata, a trend seen in other series of sect. Petota. Separately, through flow cytometry, we report the first ploidy level of S. woodsonii, as tetraploid (2n = 48). The U.S. Government’s right to retain a non-exclusive, royalty-free license in and to any copyright is acknowledged.     

Cytological basis of 2n pollen formation in a wide range of 2x, 4x, and 6x taxa from tuber-bearing Solanum species.

Cytological mechanisms of 2n pollen formation were investigated in microsporogenesis and sporad development in 2x, 4x, and 6x taxa of tuber-bearing Solanum species that were originally collected from Mexico, and Central and South America. Parallel spindles and tripoles were seen at anaphase II in the majority of genotypes observed. The occurrence of parallel spindles and tripoles were well correlated with the occurrence of dyads and triads at the sporad stage. Further, the frequency of 2n pollen predicted by frequencies of parallel spindles and tripoles and by frequencies of dyads and triads were also well correlated with observed 2n pollen frequencies. Premature cytokinesis at telophase I to prophase II was seen only in 2x and 4x species from series Conicibaccata. The frequency of premature cytokinesis corresponded well with the frequency of dyads leading to 2n pollen formation. It was concluded that the major mechanisms of 2n pollen formation in tuber-bearing Solanum species is parallel spindles.     

Use of Ipomoea trifida germ plasm for sweet potato improvement. 3. Development of 4x interspecific hybrids between Ipomoea batatas (L.) Lam. (2n=6x=90) and I. trifida (H.B.K) G. Don. (2n=2x=30) as storage-root initiators for wild species

Summary More than 28,000 pollinations were carried out between 5 Ipomoea batatas and 41 diploid I. trifida accessions of diverse origins to obtain 4x interspecific hybrids. From the resultant 730 seeds, 248 plants were finally obtained. Ploidy level determination of the progeny showed unexpected results: 52 individuals were hexaploid, 5 were pentaploid, 190 were tetraploid, as expected, and one was not determined. The existence of 5x and 6x progenies from 6x x 2x crosses not only confirmed the presence of 2n gametes but also their successful function in gene flow between ploidy levels and polyploidization within this genus. The progeny and their cultivated parents were planted in an observation field. The cultivated parents produced 0.49 kg/plant or less. Most 4x progenies did not produce storage roots or had very poor yields; nonetheless, and despite their cultivated parents' poor yields, 8 genotypes yielded between 0.81 and 1.50 kg/plant.A new scheme, using the 4x interspecific hybrids, is proposed for evaluating 2x and 4x wild accessions of the section Batatas to which the sweet potato belongs. Other possible uses of the 4x hybrids in breeding and genetics of the sweet potato are also discussed.     

Allelism of endosperm balance number (EBN) in Solanum acaule Bitt. and other wild potato species

The inheritance of endosperm balance number (EBN), a genetic, dose-dependent crossability system functioning in tuber-bearing Solanum (potato) species, was investigated for certain wild potato species having an EBN equal to one half of their ploidy. The EBN of Solanum acaule, a disomic 4(2EBN) South American species, was investigated by producing F₁ and F₂ hybrids with artificial 4x(2EBN) S. commersonii. This allowed assessment of recombination among the two genomes of disomic S. acaule and that of S. commersonii. When crossability of the hybrids with 1EBN, 2EBN and 4EBN standards was tested, no variation for EBN was detected. The apparent lack of recombination and segregation for EBN in these hybrids indicates that the genomes of S. acaule and S. commersonii carry EBN in a genetically-similar way. Combined with previous reports, these data indicate that the inheritance of EBN is similar in widely-separated taxa from South America and Mexico.     

Molecular evolution of 35S rDNA and taxonomic status of Lycopersicon within Solanum sect. Petota

To clarify the taxonomic status of tomatoes (“Lycopersicon”) and their relationship to the members of sect. Petota of genus Solanum L., organization of the rDNA external transcribed spacer (5′ ETS) was studied in 33 Solanum and “Lycopersicon” species. Phylogenetic reconstruction revealed that three major groups can be distinguished. Non-tuber-bearing species of ser. Etuberosa as well as tuber-bearing Central American diploids appeared as a paraphyletic group. The first of two well-defined clades embraced all tuber-bearing South American species and Central American polyploids. The other clade (named “tomato clade”) contains non-tuber-bearing species of ser. Juglandifolia and tomato species of ser. Neolycopersicon, which appears to be imbedded in sect. Petota. The new 5′ ETS variant D characterized by a cluster of downstream subrepeats is characteristic for the tomato clade. The variant D originated directly from the most ancestral variant A found in ser. Etuberosa and the Central American diploids, whereas variants B and C specific for the tuber-bearing South American species and Central American polyploids represent a parallel lineage of molecular evolution. The sequence analysis demonstrates the existence of an evolutionary trend of parallel multiplication of specific motifs in 5′ ETS in different groups of sect. Petota.     

Polyploidy‐associated genomic instability in Arabidopsis thaliana

Formation of polyploid organisms by fertilization of unreduced gametes in meiotic mutants is believed to be a common phenomenon in species evolution. However, not well understood is how species in nature generally exist as haploid and diploid organisms in a long evolutionary time while polyploidization must have repeatedly occurred via meiotic mutations. Here, we show that the ploidy increased for two consecutive generations due to unreduced but viable gametes in the Arabidopsis cyclin a1;2‐2 (also named tardy asynchronous meiosis‐2) mutant, but the resultant octaploid plants produced progeny of either the same or reduced ploidy via genomic reductions during meiosis and pollen mitosis. Ploidy reductions through sexual reproduction were also observed in independently generated artificial octaploid and hexaploid Arabidopsis plants. These results demonstrate that octaploid is likely the maximal ploidy produced through sexual reproduction in Arabidopsis. The polyploidy‐associated genomic instability may be a general phenomenon that constrains ploidy levels in species evolution. genesis 48:254–263, 2010. © 2010 Wiley‐Liss, Inc.     

Molecular evolution of rDNA external transcribed spacer and phylogeny of sect. Petota (genus Solanum)

The 5(') external transcribed spacer (ETS) region of ribosomal DNA of 30 species of Solanum sect. Petota and the European Solanum dulcamara were compared. Two structural elements can be distinguished in the ETS: (i). a variable region (VR), demonstrating significant structural rearrangements and (ii). a conservative region (CR), evolving mainly by base substitutions. In VR, a conservative element (CE) with similarity to the ETS of distantly related Nicotiana is present. The ancestral organization of ETS (variant A) was found for non-tuber-bearing species of ser. Etuberosa, tuber-bearing wild potatoes of Central American ser. Bulbocastana, Pinnatisecta, and Polyadenia and S. dulcamara. Duplication of CE took place in the ETS of species from ser. Commersoniana and Circaeifolia (variant B). South American diploids and Mexican polyploids from superser. Rotata also possess two CE, and additionally two duplications around CE1 are present in VR (variant C). Three major lineages could be distinguished: non-tuber-bearing species of ser. Etuberosa, tuber-bearing Central American diploids and all South American species radiated from a common ancestor at early stages of evolution, indicating a South American origin of the tuber-bearing species. Later, Central and South American diploids evolved further as independent lineages. South American species form a monophyletic group composed of series with both stellata and rotata flower morphology. Solanum commersonii represents a sister taxon for all rotata species, whereas ser. Circaeifolia diverged earlier. Two main groups, C1 and C2, may be distinguished for species possessing ETS variant C. C1 contains ser. Megistacroloba, Conicibaccata, Maglia, and Acaulia, whereas all diploids of ser. Tuberosa are combined into C2. A closer relationship of Solanum chacoense (ser. Yungasensa) to the C2 group was found. The origin of polyploid species Solanum maglia, Solanum acaule, Solanum tuberosum, Solanum iopetalum, and Solanum demissum is discussed.     

Unreduced gametes in diploid Medicago and their importance in alfalfa breeding

Summary In the genus Medicago, it is known that 2n gametes have been important in the evolution and breeding of cultivated alfalfa, which is a natural polysomic polyploid (2n=4x=32), however little is known on the frequency of male and female 2n gametes in diploid relatives of alfalfa. To obtain data on the frequency of 2n gametes, more than 12,000 2x–4x and 4x–2x crosses were made in 1982 at Madison (USA). Diploid parents in crosses were from four populations of M. coerulea, two of M. falcata and one diploid population of cultivated M. sativa which was derived by haploidy. The tetraploid seed parent in the crosses was a male-sterile M. sativa clone and vigorous tetraploid M. sativa plants were used as pollen parents. Each of 274 diploid plants was utilized both as male and as female. Of the 548 cross combinations, 266 crosses produced variable quantities of seeds which were sown in 1983 in a greenhouse at Perugia (Italy); the plants were subsequently space transplanted in the field in 1984. The identification of ploidy level of these genotypes was made on the basis of morphological characters, plant fertility, pollen stainability and chromosome counts.Of the 515 plants analyzed, the majority behaved as normal tetraploids indicating that many diploid plants produced 2n gametes. Diplogynous and diplandrous gamete production was not correlated with each other, which indicated a different genetic control of 2n sporogenesis in the 2 sexes. Only 4 F₁ triploid plants confirmed the presence of a very effective triploid block in alfalfa. In consequence, bilateral sexual polyploidization is a more likely alternative for the origin of tetraploid alfalfa than triploid bridges. The present study showed that it is possible to efficiently identify genotypes able to produce high frequencies of 2n gametes within natural populations of diploids Medicago that are useful in alfalfa breeding.Part of this study was conducted at the Agronomy Department, University of Wisconsin, Madison, Wis, USA, while one of us (F. Veronesi) was in receipt of financial assistance provided by the National Research Council of Italy; part was conducted at Centro di Studio per il Miglioramento Genetico delie Piante Foraggere, C.N.R., Perugia, Italy. The paper was presented at the Eucarpia Fodder Crops Section Meeting, Svalöv, Sweden, 16–19th September 1985     

Occurrence of 2n pollen and ps gene frequencies in cultivated groups and their related wild species in tuber-bearing Solanums

Summary The gene frequency for parallel spindles (ps) was estimated from the frequency of plants producing 2n pollen in three cultivated groups: 2x Phureja (phu), 2x Stenotomum (stn), and 4x Andigena (adg), as well as in four related wild taxa: 2x Solanum brevicaule (brc), 2x S. sparsipilum (spl), 4x S. gourlayi (grl) and 4x S. gourlayi-S. infundibuliforme hybrids (grl-ifd). Plants with more than 1% large pollen were considered as 2n pollen producers. Observations of meiosis in a sample of 2n pollen-producing plants indicated that parallel spindles is the mechanism of 2n pollen formation. The number of plants with 2n pollen among the total examined was 228 plants (15.5%) of 1,473 in 2x spl, 31 (26.7%) of 116 in 2x brc, 92 (17.4%) of 528 in 2x stn, 665 (22.1%) of 3,008 in 2x phu, 731 (51.4%) of 1,421 in 4x adg, 591 (41.2%) of 1,436 in 4x grl, and 36 (64.3%) out of 56 in 4x grl-ifd. The ps gene frequencies assuming Hardy-Weinberg equilibrium were: 0.393 for 2x spl, 0.462 for 2x brc, 0.417 for 2x stn, 0.470 for 2x phu, 0.847 for 4x adg, 0.801 for 4x grl, and 0.895 for 4x grl-ifd. Twenty-five adg clones were randomly selected from a large population and were crossed with 2x clone W5295.7, which produces 2n pollen by parallel spindles (ps). The 4x progenies from 4x×2x crosses were used to determine the genotypes at the ps locus by screening 10–20 plants in each family for 2n pollen. Based on chromosome segregation at the ps locus, 9, 14, 1, and 1 clones were nulliplex, simplex, simplex or duplex, and duplex, respectively. The frequency of the ps gene in the adg population was estimated to be 0.825 and 0.815 for chromosome and chromatid segregation, respectively. The high frequencies of 2n pollen and the ps gene in cultivated 2x and 4x groups, and in wild taxa closely related to them, provide evidence for sexual polyploidization in the tuber-bearing Solanums.Paper No. 3032 from the Laboratory of Genetics. Research supported by the College of Agriculture and Life Sciences; International Potato Center; USDA, SEA, CGRO 84-CRCR-1-1389; and Frito Lay, Inc.     

CHLOROPLAST DNA EVIDENCE FOR GENOME DIFFERENTIATION IN WILD POTATOES (SOLANUM SECT. PETOTA: SOLANACEAE)

Chloroplast DNA restriction site analysis has been used to test Hawkes's phylogenetic interpretations of the genomic data in Solanum sect. Petota. Hawkes hypothesized a diploid (2n = 24) origin of the tuber-bearing members of this group (subsection Potatoe) in Mexico and Central America (as a B genome) with later migrations and evolution to an A genome in South America, later followed by a return migration of the A genome to Mexico and Central America with A × B hybridizations and polyploidizations to produce ser. Longipedicellata (4x) and Demissa (6x). Our results provide partial support for this hypothesis by demonstrating the paraphyletic and primitive nature of the B genome species group, and the monophyletic and derived nature of all A genome and A × B genome species, including S. verrucosum, a hypothesized A genome progenitor of ser. Demissa. Thus, the Mexican and Central American polyploid species must have obtained their cytoplasm from the A genome. However, our results question the Stellata/Rotata hypothesis of Hawkes and the taxonomic placement of S. chomatophilum in ser. Conicibaccata.     

Pollen production and spontaneous polyploidization in diploid populations of Anthoxanthum alpinum

The most common form of polyploidization is that which occurs via the fertilization of unreduced 2 n gametes, i.e. gametes that possess the somatic chromosome number of the species. However, very few data are available concerning the frequency of spontaneous polyploidization in diploid plant populations. In this study we have quantified both the frequency of In pollen production and the frequency of polyploid seed production in diploid populations of the grass Anthoxanthum alpinum. More than 6000 seeds from four different populations collected during two years of study were screened for ploidy using a flow cytometer. In parallel, the frequency both of plants with large pollen (LP) and of LP production on these plants was quantified in two populations. No tetraploid seeds were detected, although in each population a few triploid seeds were regularly produced at a frequency of two triploid seeds per thousand. The frequency of LP producers was similar in the four populations, as was the frequency of LP production. The frequency of LP was ten times that of triploid seeds. These finding suggest that the 'triploid block' is not enough to prevent the regular occurrence of triploids in diploid populations of this species. The role played by the hypothesized 'triploid bridge' in the genesis of autotetraploids is discussed in the light of these results.     

Ploidy level manipulations in potato through sexual hybridisation

There is no better use of sexual reproduction in regard to breeding and genetic research than the ploidy level manipulations possible in the potato and its relatives. Unique reproductive characteristics of tuber‐bearing Solanum species make possible: the production of gametes with unreduced chromosome number; the presence of an endosperm dosage system that regulates success of interploidy/interspecific crosses; the possibility to easily extract maternal haploids following crosses with S. phureja. This paper reviews results obtained in scaling genomic multiples up and down in potato, and relates these manipulations to breeding strategies for the genetic improvement of the cultivated potato. Several ploidy series have been developed, ranging from the monoploid to the hexaploid level. Cultivated tetraploids were scaled down to the diploid and monoploid level by haploidy. Scaling upward was achieved by sexual polyploidisation via 2n gametes that resulted in triploid, tetraploid, pentaploid, and hexaploid genotypes with a broad genetic base. Altogether, the success of ploidy level manipulations constitutes further proof that sexual polyploidisation played an important role in the polyploid evolution of Solanum species, and supports the idea that gene flow can be relatively easily accomplished through interploid and bridge crosses.     

Origin of triploid Arachis pintoi (Leguminosae) by autopolyploidy evidenced by FISH and meiotic behaviour

Background and Aims

Polyploidy is a dominant feature of flowering-plant genomes, including those of many important crop species. Arachis is a largely diploid genus with just four polyploid species. Two of them are economically important: the cultivated peanut and A. glabrata, a tropical forage crop. Even though it is usually accepted that polyploids within papilionoid legumes have arisen via hybridization and further chromosome doubling, it has been recently suggested that peanut arose through bilateral sexual polyploidization. In this paper, the polyploid nature of the recent, spontaneously originated triploid cytotype of the tropical lucerne, A. pintoi, was analysed, and thereby the mechanism by which polyploids may arise in the genus.

Methods

Chromosome morphology of 2x and 3x A. pintoi was determined by the Feulgeńs technique and the rDNA sites were mapped by FISH. To investigate whether polyploidization occurred by means of unreduced gametes, a detailed analysis of the microsporogenesis and pollen grains was made.

Key Results

The 2x and 3x plants presented 9m + 1sm and a satellited chromosome type 2 in each haploid genome. Physical mapping revealed a cluster of 18S–26S rDNA, proximally located on chromosome 6, and two 5S rDNA loci on chromosomes 3 and 5. Diploid plants presented 10II in meiosis while trivalents were observed in all triploids, with a maximum of 10III by cell. Diploid A. pintoi produced normal tetrads, but also triads, dyads and monads. Two types of pollen grains were detected: (1) normal-sized with a prolate shape and (2) large ones with a tetrahedral morphology.

Conclusions

Karyotype and meiotic analysis demonstrate that the 3x clone of A. pintoi arose by autopolyploidy. The occurrence of unreduced gametes strongly supports unilateral sexual polyploidization as the most probable mechanism that could have led to the origin of the triploid cytotype. This mechanism of polyploidization would probably be one of the most important mechanisms involved in the origin of economically important species of Arachis, either by triploid bridge or bilateral sexual polyploidization.     

Evidence for the existence of 2n gametes in Lotus tenuis Wald. et Kit. (2n=2x=12): their relevance in evolution and breeding of Lotus corniculatus L. (2n=4x=24)

Summary Crosses between male sterile L. corniculatus (2n=4x=24) and L. tenuis (2n=2x=12) plants were performed in order to verify the presence of 2n gametes in L. tenuis. All but one of the plants from these crosses had 2n=4x=24 and the L. corniculatus phenotype; this plant had 2n=2x=12 and the L. tenuis phenotype. The plants also showed good quantity of pollen at tripping, good pollen fertility and good percentage of seed setting in the backcross to L. corniculatus. On the whole, both cytological and morphological observations, showing that all but one of the plants from L. corniculatus x L. tenuis were normal tetraploids, suggest the existence of diploandrous gametes in L. tenuis. On the other hand, haploid parthenogenesis probably gave origin to the dihaploid plant 2n=2x=12.     

Studies on the Asian Eupatorias

Analysis of 265 plants derived from 13 sites in the Rokko Mountains shows that the widely distributed eastern AsianEupatorium chinense var.simplicifolium consists of seven cytotypes. These polymorphic karyotypes comprise four levels of ploidy based on X=10 and partial deficiency, occurring either alone or in combination. The polyploid cytotypes exhibit anomalous meiosis and highly variable pollen stainability in contrast to the nearly-normal behaviour of the diploid. The polyploid cytotypes were revealed as being agamospermous. They are readily distinguishable from the diploid cytotypes by several morphological characters and by growth habits closely related to their respective niches. Two or more cytotypes occurred in all sites examined, the most common being the coexistence of triploid and tetraploid cytotypes. The frequencies of occurrence of the pentaploid plants and the cytotypes involving a deficient chromosome were relatively low. While the diploid cytotype is restricted to fragile gravitational slopes and rocky areas which are poor in species and lack tall competitors, the polyploids occur widely in the grasslands, the roadsides or the forest-margins, closely associated with tall grasses and forbs such asMiscanthus sinensis. Based on these data, theE. chinense var.simplicifolium complex is divided into two distinct groups: the diploid cytotype and the polyploid cytotypes.     

Ploidy doubling by in vitro culture of excised protocorms or protocorm-like bodies in <Emphasis Type="Italic">Phalaenopsis</Emphasis> species

Endopolyploidy was observed in the protocorms of diploid Phalaenopsis aphrodite subsp. formosana with ploidy doubling achieved by in vitro regeneration of excised protocorms, or protocorm-like bodies (PLBs). Thirty-four per cent of the PLBs regenerated from the first cycle of sectioned protocorms were found to be polyploids with ploidy doubled once or twice as determined by flow-cytometry. The frequency of ploidy doubling increased as the sectioning cycles increased and was highest in diploid followed by the triploid and tetraploid. Regeneration of the endopolyploid cells in the tissue of the protocorms or PLBs is proposed as the source of the development of ploidy doubled plantlets. The frequency of ploidy doubling was similar in seven other Phalaenopsis species, although the rate of increase within cycles was genotype specific. In two species, a comparison of five parameters between 5-month-old diploid and tetraploid potted plants showed only the stomata density differed significantly. The flowers of the tetraploid plant were larger and heavier than those of the diploids. This ploidy doubling method is a simple and effective means to produce large number of polyploid Phalaenopsis species plants as well as their hybrids. The method will be beneficial to orchid breeding programs especially for the interspecific hybridization between varieties having different chromosome sizes and ploidy levels.     

Occurrence and Inheritance of 2 n Pollen in Musa

Sexual polyploidization explains the formation of most polyploidcrop species and provides means for their genetic improvement.This paper reports the occurrence and inheritance of 2n pollenin diploid and polyploidMusa . Pollen samples of male fertileaccessions available in the field gene bank of the InternationalInstitute of Tropical Agriculture were microscopically examinedthroughout a calendar year. Based on pollen diameter 2n pollenproducers were identified. The presence of 2n pollen in diploidspecies suggests that unilateral sexual polyploidization (2nxn) could have been involved in the origin of triploidMusa species.Segregation data suggests that at least one dominant gene controls2n pollen production inMusa . Further introgression of desirablealleles from diploid species to polyploids could be achievedeither through unilateral or bilateral (2nx2n ) sexual polyploidizationinMusa . Musa spp.; Musa hybrids; 2n gametes; evolution; ploidy manipulations; sexual polyploidization