4x Progeny Research Articles

Haploidy in Tobacco Induced by PsASGR-BBML Transgenes via Parthenogenesis.

Background: Engineering apomixis in sexually reproducing plants has been long desired because of the potential to fix hybrid vigor. Validating the functionality of genes originated from apomictic species that contribute to apomixis upon transfer to sexually reproducing species is an important step. The PsASGR-BABYBOOM-like (PsASGR-BBML) gene from Pennisetum squamulatum confers parthenogenesis in this apomict, and its functionality was demonstrated in several sexually reproducing monocots but not in any dicots. Methods: We introduced the PsASGR-BBML gene regulated by egg cell-specific promoters, either AtDD45 or AtRKD2, into tobacco, and analyzed progeny of the transgenic lines resulting from self-pollination and crossing by flow cytometry. Results: We identified haploid progeny at a frequency lower than 1% in the AtDD45pro lines, while at a frequency of 9.3% for an octoploid (2n = 8x) AtRKD2pro line. Haploid production in the T2 generation, derived from the tetraploid T1 offspring of this original octoploid AtRKD2pro line, was also observed. Pollinated by homozygous transgenic tobacco carrying a DsRed marker gene, 4x progeny of the AtRKD2pro line yielded parthenogenetic embryos identified as DsRed negative. We verified that the DsRed negative seedlings recovered were haploid (2x). Conclusion: The PsASGR-BBML gene regulated by egg cell-specific promoters could enable parthenogenesis in tobacco, a dicotyledon species.

The Efficiency and Effectiveness of Open Pollination in Musa Breeding

Aims: This field experiment was conducted to determine if hand and open pollination methods affected performances of Musa progenies from 4x - 2x crosses and to identify promising progenies for recurrent selection.
 Study Design: The experimental design was a randomized complete block design with two replications of 6 plants per genotype.
 Place and Duration of Study: International Institute of Tropical Agriculture (IITA) High Rainfall Station, Onne (4º51’N, 7º03’E, 10 m above sea level), in Rivers State, South-south Nigeria for 24 months.
 Methodology: Two-month old seedlings of hand pollinated (6 diploid, 6 tetraploid) and open pollinated (6 diploid, 6 tetraploid) progenies, along with parental clones (2x) and (4x) of each genotype were planted at 3 m x 2 m spacing. Data on phenology, vegetative growth, yield and yield characters were collected at flowering and harvest over three crop cycles. Genotypes were partitioned into 5 clusters assayed by means of orthogonal contrasts to compare the performance of progenies from both pollination methods.
 Results: Pollination methods produced no significant (P = .05) differences, unfavourable effects or reduction in performance of economically important yield and yield components of 4x and 2x progenies of similar genotype. Some significant (P = .05) linear correlations and relationships between phenological and vegetative traits; and yield and yield components changed with pollination methods and ploidy levels but did not affect final outcomes. Promising open pollinated diploids include the early maturing TMP2x 2829-62OP; and for high yield and yield components measured, TMB2x 8084-2OP and TMP2x 1448-1OP. Promising open pollinated tetraploids include TMP4x 7002-1OP and TMP4x 2796-5OP.
 Conclusion: Open pollination did not result in unfavorable effects or reduction in performance of economically important yield and yield components in progenies of similar genotypes. Therefore, open pollination could be considered for Musa breeding. This will reduce cost, labour, time and stress involved in Musa improvement.

Transcriptomic analysis reveals differential gene expressions for cell growth and functional secondary metabolites in induced autotetraploid of Chinese woad (Isatis indigotica Fort.).

The giant organs and enhanced concentrations of secondary metabolites realized by autopolyploidy are attractive for breeding the respective medicinal and agricultural plants and studying the genetic mechanisms. The traditional medicinal plant Chinese woad (Isatis indigotica Fort., 2n = 2x = 14) is now still largely used for the diseases caused by bacteria and viruses in China. In this study, its autopolyploids (3x, 4x) were produced and characterized together with the 2x donor for their phenotype and transcriptomic alterations by using high-throughput RNA sequencing. With the increase of genome dosage, the giantism in cells and organs was obvious and the photosynthetic rate was higher. The 4x plants showed predominantly the normal meiotic chromosome pairing (bivalents and quadrivalents) and equal segregation and then produced the majority of 4x progeny. The total 70136 All-unigenes were de novo assembled, and 56,482 (80.53%) unigenes were annotated based on BLASTx searches of the public databases. From pair-wise comparisons between transcriptomic data of 2x, 3x, 4x plants, 1856 (2.65%)(2x vs 4x), 693(0.98%)(2x vs 3x), 1045(1.48%)(3x vs 4x) unigenes were detected to differentially expressed genes (DEGs), including both up- and down-regulated ones. These DEGs were mainly involved in cell growth (synthesis of expansin and pectin), cell wall organization, secondary metabolite biosynthesis, response to stress and photosynthetic pathways. The up-regulation of some DEGs for metabolic pathways of functional compounds in the induced autotetraploids substantiates the promising new type of this medicinal plant with the increased biomass and targeted metabolites.

Ploidy variation of Musa hybrids from crosses

Plantain and banana (Musa spp) breeding involves crossing 3x (triploid) landraces to 2x (diploid) accessions as female and male parents, respectively, selecting 4x (tetraploid) and 2x primary hybrids from the 3x - 2x progenies, and crossing 4x - 2x hybrids to produce secondary 3x hybrids. In these crosses, complex ploidy and genome arrays occur frequently making it difficult to predict the products of crosses. The objective of the study was (a) to determine the ploidy of progenies of 2x – 2x, 2x – 4x and 4x – 2x crosses, and (b) to assess the agronomic characteristics of diploid and triploid progenies recovered from the same or similar crosses. The breeding population involved progenies of interploidy crosses field established in a randomized complete block design (RCBD) with three replications. Ploidy was determined using Flow cytometry analysis of nuclear DNA content and chloroplast characteristics. Results indicated that progenies of 2x–2x crosses were predominantly diploid (99.7%), those of 2x–4x crosses were mainly diploid (96.2%), while the 4x–2x crosses produced predominantly triploid progenies (94.1%). Very highly significant differences (P guard cells of progenies. Diploid hybrids recovered from crosses had shorter days to flowering, plant height and bunch weight compared to triploid hybrids

VNTR-based diversity analysis of 2x and 4x full-sib Musa hybrids

Summary of inheritance of maternal (AAB genome) and paternal (AA genome) VNTR amplification fragments in the full -sib 2x and 4x progeny populations. ____________________________________________________________________________ Maternal fragments Paternal fragments (Obino l’Ewai) (Calcutta 4) # % # % ____________________________________________________________________________ Diploid progeny Segregation 10 46 15 65 No segregation 8 36 3 13 Absent 4 18 5 22 Tetraploid progeny Segregation a 18 82 20 87 b No segregation 4 18 1 4 Absent 0 0 2 9 _________________________________________________________________________ ___ a segregation of 7 fragments solely due to absence of that fragment in TMPx 548 -9 b segregation of 1 fragment solely due to absence of that fragment in TMPx 548 -9 TMPx 7002-1 are most closely related to the maternal genotype (Obino l’Ewai). In cont rast, all the diploid hybrids form two groups associated at the 60% level of similarity, which are distinct from both parental genotypes (Figure 1). The major exceptions are TMPx 548 -9, which is separated at the 20% level of similarity from all other genotypes, tested and hybrid 5860-1 (4x) that is grouped with the diploid progeny. Hybrid 5860-1 (4x) was found to haveCalcutta 4 were very similar; 0.42 (±0.046) and 0.39775% similarity to TMP2x 1549 -5, both of which have 65% similarity to hybrid 1549-7 (2x) (Figure 1). This anomaly may be a consequence of hybr id 5860-1 (4x) possessing a high proportion of bi-allelic loci in common with these particular diploid full -sibs, as compared to multi -allelic loci of the other tetraploid hybrids. Clearly, this type of molecular marker analysis can not be used for inferring hybrid ploidy. Finally, TMP2x 1448 -1 and hybrid 1586 -2 (2x) appear particularly distinct from the majority of their 2x and 4x full -sib hybrids (Table 2). Both diploid and tetraploid hybrids had comparable mean levels of similarity to Calcutta 4 (32.2% and 34.3 %, respectively). However, tetraploid hybrids had more fragments in common with Obino l’Ewai than their diploid full-sibs (56.1% and 44.6%, respectively). This relationship was expected because the tetraploid hybrids have 75% of their genes donated by the maternal plantain 2n gametes (and the remaining 25% from the diploid banana), while their diploid full-sibs inherit 50% of their genes from each parent. The average phenotype of the tetraploid hybrids was also more closely related to Obino l’Ewai than Calcutta 4 (Table 3). The phenotypic distance between the tetraploid hybrids and Obino l’Ewai was 0.322 (±0.027), while the phenotypic distance between the tetraploid hybrids and Calcutta 4 was 0.470 (±0.021). Conversely, the phenotypic distance indic es for comparisons of diploid hybrids with Obino l’Ewai and (±0.043), respectively. This was expected based on the gametic contributions to diploid and tetraploid hybrids described above. For the relationship between hybrids and Obino l’Ewai, there was a significant association between the genetic similarity (based on VNTR) and the phenotypic resemblance (r = -0.546, P = 0.042). The linear relationship (R

Comparison of haploid Tuberosum- Solanum chacoense versus Solanum phureja -haploid Tuberosum hybrids as staminate parents of 4x–2x progenies evaluated under distinct crop management systems

The relative value of haploid Tuberosum-Solanum chacoense (TCH) vs. Solanum phureja-haploid Tuberosum (PTH) hybrids as male parents in 4x–2x crosses was estimated under two distinct crop management systems. The first experiment (E#1) was carried out with supplementary irrigation (three times a week) at Hancock-Wisconsin (WI) with 27 families derived from a sub-set of crosses involving 11 tetraploid (4x) cultivars and 10 diploid (2x) clones (3 TCH; 6 PTH; and one TCH-PTH hybrid clone). A second experiment (E#2) was conducted at Rhinelander-WI with the same group of families but it was only rainfed with no supplementary irrigation being provided during the entire course of the assay. For comparison, a complete set of 4x and 2x clones (used as parents) was also planted in each location. Three traits were evaluated in both locations: total tuber yield (TTY), haulm maturity (HM), and plant vigor (PV). Altogether, 18 out of 27 and 10 out of 27 families outyielded the corresponding 4x parents at E#1 and E#2, respectively. Significant differences among all 4x–2x families and among families of the TCH group were observed for TTY at both locations. No difference was found for TTY among families of PTH and [TCH–PTH] groups at each location. A total of 75% and 50% of the TCH families outyielded their 4x parents at E#1 and E#2, respectively. The percentage of families from the PTH group that outyielded their 4x parents at E#1 and E#2 was 83.3% and 25%, respectively. The TCH group had on average lower HM scores than PTH (at both locations) indicating some segregation for earliness. However, the HM values of the 4x–2x families were, in general, higher than those observed for the 4x parents. For PV, the PTH group mean was significantly higher than TCH group only in E#1. The PV values of the 4x–2x families were higher than the 4x-parent group at both locations. Our results indicated that TCH hybrids might be as good parents as PTH to increase the TTY of 4x–2x progenies. In addition, the TCH families displayed a slight higher level of performance under more stressful growing conditions than PTH hybrids as indicated by the assay at Rhinelander. Therefore, selection of genetic materials with potential broad range of adaptation seems to be feasible with both hybrids via the 4x–2x breeding scheme.

Comparison of FDR- and SDR-derived tetraploid progeny from 2x×4x crosses using haploids of Solanum tuberosum L. that produce mixed modes of 2n eggs

The relationship between heterozygosity and heterosis in tetraploid potato (Solanum tuberosum subsp. tuberosum L., 2n"4x"48) was examined in a series of first-division restitution (FDR)- and second- division restitution (SDR)-derived tetraploid sub- populations. The subpopulations were constructed using two 2n egg-producing, mixed-mode haploids (2n"2x"24) crossed to three tetraploid (2x" 4x"48) potato clones. Half-tetrad analysis using a co- dominant electrophoretic marker (Pgm-2), which is closely linked to the centromere, discriminated between FDR- and SDR-derived 4x progeny. The FDR:SDR ratio of the 4x progeny observed was dependent upon the haploid parent used in the 2x)4x cross. Field studies were conducted between 1992 and 1996 to com- pare the yield and specific gravity of the two sub- populations and their parents from three crosses. There was no di⁄erence in the total tuber yield or specific gravity between the FDR- and SDR-subpopulations based upon family means, despite the expectation that FDR-derived progeny would transmit a greater por- tion of the genome's heterozygosity intact than SDR- derived progeny. The 4x parent in each family had a higher yield than either 4x progeny subpopulation. Inbreeding, as a consequence of the haploidization process and a lack of genetic diversity, may have negated any advantage of the FDR-derived progenies over the SDR-derived progenies.

Effect of selection and temperature stress on the production of 2n gametes in Lotus tenuis

AbstractFive Lotus tenuis genotypes showing 2n pollen production greater than 5% and producing 4x progenies in 4x(male sterile Lotus corniculatus) × 2x crosses were found in different natural populations. Meiotic analyses of these plants showed some second metaphases with disoriented spindles, i.e. parallel and tripolar spindles which resulted in diads and triads, respectively. Because useful genes for winter growth and resistance to powdery mildew from L. tenuis can be introgressed into L. corniculatus via 2n gametes, a programme of selection was started to increase the occurrence of 2n gamete production in L. tenuis. Controlled crosses were performed between genotypes producing 2n gametes and the progenies were checked for 2n pollen production and meiotic aberration, following the same methodologies as used for the parental plants. Although in some progeny plants the frequency of 2n gamete production increased remarkably, the overall progeny level of 2n gamete production remained unchanged. This suggested that different genes for meiotic aberration exist in natural populations and/or differential seed‐setting ability exists between genotypes. Diallel crosses between selected genotypes confinned this hypothesis. Evaluation of 2n pollen production of clones grown under different temperature regimes (warm vs. temperate) showed that, in L. tenuis, the effects of genotype, temperature and genotype × temperature interaction are significant. In particular, high temperature increases the level of 2n gamete production in L. tenuis.

Identification, transmission, and cytological behavior ofSolanum lycopersicoidesDun. monosomic alien addition lines in tomato (Lycopersicon esculentumMill.)

The wild nightshade Solanum lycopersicoides possesses a number of disease- and arthropod-resistance and environmental stress tolerance traits of potential value for the cultivated tomato, Lycopersicon esculentum. F1hybrids are readily obtained by sexual crosses, but backcrosses to tomato are hindered by sterility and incompatibility barriers. In the present studies, intergeneric sesquidiploid hybrids (2 genomes of L. esculentum and 1 of S. lycopersicoides) were pollinated with Lycopersicon pennellii derived bridging lines, yielding diploid, aneuploid (up to 2n + 6), and polyploid (3x, 4x) progeny. After diploids (58% of total progeny), the group most frequently obtained was 2n + 1 (31%), which included all 12 possible monosomic alien addition lines (MAALs) containing a single extra chromosome from S. lycopersicoides in a tomato background. MAAL groups were identified with a combination of allozyme, RFLP, and monogenic visual markers. Transmission rates of the extra chromosomes in MAALs varied from 0%, for chromosome 6, to 24%, for chromosome 10. At diakinesis and metaphase I of meiosis, the alien chromosome was usually unpaired (in an average of 90% of PMCs), but association of homoeologous chromosomes was observed in each of the 12 MAAL groups (in an average of 5.7% of PMCs). The MAALs showed a striking morphological resemblance to the corresponding tomato primary trisomic. In addition, each MAAL expressed S. lycopersicoides specific leaf, flower, and (or) fruit characters. This set of MAALs, the first of its kind in Lycopersicon, is expected to be useful for a variety of gene-mapping and introgression studies.

Cytometric and morphometric identification of diploid, tetraploid and pentaploid plants derived from Actinidia arguta (2n = 4x) crossed with A. deliciosa (2n = 6x)

AbstractInterspecific crosses between Actinidia arguta (In = 4x) and A. deliciosa (2n = 6x) were carried out in order to transfer cold resistance from a wild to the cultivated kiwifruit species. Of the 65 progenies obtained and analysed by flow cytometry, 16 were diploid, one was tetraploid and 48 were, as expected, pentaploid. Morphologically, the diploids can be distinguished from the pentaploids by their longer lamina. The existence of 5x progenies confirms the existence of gene flow between A. arguta and A. deliciosa, whereas the existence of 2x and 4x progenies provides evidence for parthenogenesis within the Actinidia genus following an interspecific cross. The study demonstrates the utility of flow cytometry for genomic analysis of progenies derived from interspecific crosses. Possible uses of both sexual and parthenogenetic progenies in the breeding and genetics of kiwifruit are discussed.

Inheritance of black sigatoka disease resistance in plantain-banana (Musa spp.) hybrids

Black sigatoka (Mycosphaerella fijiensis Morelet), an airborne fungal leaf-spot disease, is a major constraint to plantain and banana (Musa spp.) production world-wide. Gaining further knowledge of the genetics of host-plant resistance will enhance the development of resistant cultivars, which is considered to be the most appropriate means to achieve stable production. Genetic analysis was conducted on 101 euploid (2x, 3x and 4x) progenies, obtained from crossing two susceptible triploid plantain cultivars with the resistant wild diploid banana 'Calcutta 4'. Segregating progenies, and a susceptible reference plantain cultivar, were evaluated over 2 consecutive years. Three distinct levels of host response to black sigatoka were defined as follows: susceptible (< 8 leaves without spots), less susceptible (8-10) and partially resistant (> 10). Segregation ratios for resistance at the 2x level fitted a genetic model having one major recessive resistance allele (bs 1) and two independent alleles with additive effects (bsr 2 and bsr 3). A similar model explains the results at the 4x level assuming that the favourable resistance alleles have a dosage effect when four copies of them are present in their respective loci (bs i (4) ). The proposed model was further validated by segregation data of S 1 progenies. Mechanisms of black sigatoka resistance are discussed in relation to the genetic model.

Male sterility and 2n pollen in 4x progenies derived from 4x×2x and 4x×4x crosses in potatoes

The objectives of these experiments were to evaluate male sterility (MS) and 2n pollen frequency in 77 4x×2x families and 26 4x×4x families. The 2x parents were haploid-species hybrids and the 4x parents mainly clones withS. tuberosum ssp.tuberosum (tbr) andS. tuberosum ssp.andigena (adg) in their genetic background. The female parents had different cytoplasms: adg, tbr,S. demissum (dms), andS. stoloniferum (sto). Families from female parents with adg or dms cytoplasm did not have MS plants. However, families derived from crosses of many tbr females with 2x hybrids and a 4x adg clone had a high percentage of MS plants; all the progenies from cv. Serrana (sto cytoplasm) possessed tetrad sterility. These results can be explained as due to an interaction between a dominant male sterility (Ms) gene with tbr and sto cytoplasms. Some MS 4x hybrids (tbr×adg) used as females had the Ms gene, but male fertility was restored in some plants of their progenies when they were crossed with 4x tbr clones. This indicates that some tbr clones have a fertility restorer gene (Rt). The results from both 4x×2x and 4x×4x families fit the expected ratios assuming chromatid segregation for both the Ms and Rt loci. The gene frequency of parallel spindles was estimated in the 4x population as 0.74.

Flowering characteristics and fertility of interploidy progeny from normal and 2n gametes in Dactylis glomerata L.

We performed an extensive experiment to compare male fertility, flowering period, seed set and germination rate between triploid and tetraploid cross-progeny of Dactylis glomerata L. originating from reciprocal diploid (2x) × tetraploid (4x) crosses. Four hundred and forty-five triploid and tetraploid individuals, constituting the cross progeny of 2x-4x reciprocal crosses between diploids from nine distinct subspecies, and tetraploids from five subspecies, were tested for male fertility and phenology under homogeneous conditions. A significant negative correlation was observed between the parental genetic distance and the frequency of sterile panicles (ST) in 3x progeny, suggesting that selection may occur against fertility in triploid progeny from interploid crosses involving closely related subspecies. Genetic distance between the parental subspecies was not significantly correlated with the frequency of fertile panicles (FT), non-spreading panicles (NT, non-dehiscent anthers with pollen inside), and ST panicles in the 4x progeny, nor with the frequency of FT and NT panicles in the 3x progeny. Among the 4x progeny individuals that produced panicles, more than 60% were fully fertile, but 24% possessed only sterile panicles. In the triploids, 63% of the reproductive plants were male-sterile, but 31% had variable proportions of fertile panicles and 6% possessed only fertile panicles. Male fertility in the progeny was significantly affected by individual ploidy level, parental diploid subspecies identity, and by the interaction between these 2 factors in the case of the fertile progeny plants. The frequency of vegetative plants was higher in 4x progeny than in 3x progeny, more particularly when the seed parent was tetraploid. However, when the seed parent was diploid, the frequencies of vegetative plants were similar in 3x and 4x progeny. Pollen viability was significantly higher in anthers from fertile panicles than in anthers from non-spreading panicles; viability of pollen from non-spreading panicles was, in turn, significantly higher than in sterile panicles. Also, seed set was higher in panicles with fertile anthers than in the two other types. However, seeds from plants with fertile, non-spreading and sterile anthers showed no differences in germination rate. Therefore, in Dactylis glomerata, tetraploid cross progeny arising from 2n gametes possess higher fertility, on average, than triploid cross progeny involving normal gametes. In addition to lower male fertility (non-spreading or atrophied anthers), the triploids also showed significantly lower pollen fertility in anthers and lower seed set in open pollinating conditions, probably because disturbance in meiosis, due to odd chromosome associations, also occurs in eggs. Moreover, triploids were observed to reproduce preferentially with individuals possessing the same ploidy levels as their seed parent. These results suggest that the production of 4x individuals, derived from 2n gametes in interploid crosses, is likely to be the main effective means of direct gene transfer in contact areas of populations or subspecies possessing distinct ploidy levels.

Production of 2n Gametes in Diploid Subspecies of Dactylis glomerata L. 1. Occurrence and Frequency of 2n Pollen

The occurrence of plants producing 2n pollen was ascertained in nine diploid (2n=2x=14) subspecies of the polyploid grass complex Dactylis glomerata by determining the ploidy level of the progeny in 100 tetraploid (4X) × diploid (2X) crosses. The possibility of using pollen size to screen for plants which produce large quantities of 2n pollen was tested by comparing the frequency distributions of pollen diameter in plants which did and did not produce 4X progeny in an interploidy cross. The ploidy level of the progeny was determined by chromosome counts or nuclear DNA quantification using flow cytometry

Development and performance of balck sigatoka-resistant tetraploid hybrids of plantain (Musa spp., AAB group)

A strategy to control the black sigatoka disease (Mycosphaerella fijiensis Morelet) of plantain (Musa ssp., AAB grou) in Africa, targeting the incorporation of durable host plant resistance, was initiated at the International Institute of Tropical Agriculture (IITA). The commonly accepted intractability of plantain to genetic improvement has been challenged by the identification of 37 different, seed-fertile plantain cultivars and by the production of 250 hybrids in four years of breeding work. Twenty tetraploid hybrids have been selected for their increased black sigatoka resistance, high yields, large parthenocarpic fruits and improved ratooning. ‘Calcutta 4’ (Musa acuminata spp. burmannicoides) was the diploid male parent of 17 of the selected hybrids, which indicates that the inferior bunch characteristics of this wild banana were generally not transmitted to its tetraploid progenies. Conversely, the 4x progeny of plantain readily expressed black sigatoka resistance when crossed with ‘Calcutta 4’. Progenies of the triploid plantain cvs. ‘Obino l'Ewai’ and ‘Bobby Tannap’ differed in their black sigatoka breeding values, the former producing larger numbers of promising hybrids. Tetraploids obtained from crosses of plantain cultivars with the homozygous ‘Calcutta 4’ displayed variation in black sigatoka reaction, qualitative morphological traits and growth and yield parameters, suggesting the occurrence of segregation and recombination during the modified megasporogenesis leading to the formation of 2n eggs in the triploid female plantain parents. The variation in black sigatoka reaction among the tetraploid progenies of plantain suggests that resistance could be regulated by recessive, additive genes.

Potato haploid performance in 2x × 4x crosses

Nineteen haploids (2n = 2x = 24) extracted from four tetraploids ofSolanum tuberosum (2n = 4x = 48) with high 2n egg frequencies and known mode of 2n egg formation were evaluated via 4x progeny from 2x × 4x crosses (haploid × cultivar) for total tuber yield, tuber appearance and tuber set. Ninety-eight, 4x families (15-20 genotypes each) were planted in the randomized complete block design with two replications at two locations. Analyses of variance were conducted for all haploids over three common testers, and each of the four groups of haploids individually over a variable number of testers. The performance of haploids, based on general combining ability effects, was consistent for all traits, when tested with either three or more testers. Diploid clones can be evaluated in 2x × 4x crosses using three unrelated adapted 4x testers. Nonorthogonal comparisons indicated no significant differences for all traits among four groups of haploids, and between modes of 2n egg formation. Significant variation within each group of haploids indicates that selection at the haploid level should be carried out among haploids from either the same or different cultivars. Superior haploids can be utilized in 2x × 4x and 2x × 2x breeding schemes to generate 4x clones.

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

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.

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

AbstractA 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.

Evaluation of Potato Hybrids Obtained from Tetraploid‐Diploid Crosses II. Progeny Analysis

AbstractA 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. Means of six traits of the 45 progenies obtained from a two‐year experiment were used in progeny analysis. A new mathematical model is developed for analyzing the incomplete two‐way table. It is modified from the conventional model for factorial mating design. The aim of the model is to evaluate the potential of 2x parents in terms of their mean effects (μi) and sensitivity measures (Bi) to the 4x parents. The 4x parents were treated as testers in the model. They were compared by the estimates of the magnitude of tester effects (gi). The mean effects of 2x parents showed a dominant influence on variation of tuber number and mean tuber weight of the hybrid progenies. They also demonstrated a substantial influence on the variation of total and marketable yield, chipping score and specific gravity. The linear response of 2x parents to 4x parents, however, also showed a substantial influence on the variation of the latter traits. The information on μi and Bi are useful in terms of choosing superior 2x parents and, together with gj of 4x parents, 4x—2x progenies for specific traits. The μi and Bi estimates of the six traits showed a complicated interrelationship to each other.

Efficiency of potato breeding using FDR 2n gametes for multitrait selection and progeny testing

The objective of this research was to compare the efficiency of the 4x×2x breeding scheme with the traditional 4x×4x method with respect to potato improvement. The basis for such a comparison was the parental value of four 2x and four 4x male parents from the International Potato Center (CIP) as measured by multitrait selection and progeny testing. The 2x parents produced 2n pollen by parallel spindles at anaphase II, which is genetically equivalent to a first division restitution (FDR) mechanism. Both 2x and 4x parents were crossed with four common 4x female parents. Thus, 32 families were evaluated over 2 years at four Peruvian locations. A selection index which considered tuber yield, tuber number, average tuber weight and specific gravity was used for multitrait selection. Three FDR 2x parents had better selection index scores than the 4x parents over the four locations. Estimates of broad-sense heritability for total yield using different number of replications and locations were calculated by using the variance components. The 4x × 2x breeding scheme was found to be better than the traditional 4x × 4x method since fewer replications and locations are required to evaluate tuber yield in 4x × 2x progenies than in 4x × 4x progenies. The FDR 2x parents were also better material than the 4x parents for testing combining ability for tuber yield of the 4x progenitors. This could be the result of the mode of FDR 2n pollen formation. The pollen of FDR 2x parents is more heterozygous, but more homogenous than n pollen from 4x parents.