The biological characters and polyploidy of progenies in hybridization in 4×–2× crosses in <Emphasis Type="Italic">Dianthus caryophyllus</Emphasis>

Crosses were carried out between tetraploid Dianthus caryophyllus cv ‘Butterfly’ (2n = 4×= 60) and seven diploid cultivars (2n = 2×= 30). Fewer seeds were obtained and the low seed germination was found which suggested the post-fertilization barrier in 4×–2× crosses. 12 progeny were obtained from 5 crossing combinations. Chromosome analysis revealed that they consisted of 5 triploid hybrid plants and 7 tetraploid hybrid plants, suggesting that unreduced male gamete maybe be involved in polyploid formation. Various flower shapes and colours were observed in the polyploid progenies, showing that sexual polyploidization results in greater variability and fitness. The hybrids obtained by 4×–2× crosses showed the flower-size intermediate between the parents or larger than the parents. Some favourable characters of parents such as flower shape, flower colour and resistance to Fusarium oxysporum, were successfully transmitted to the hybrids. Since polyploid hybrids have some of the profitable characters of the parents, they are expected to be used for future breeding in carnation.     

Ploidy manipulation and introgression breeding in Darwin hybrid tulips

Meiotic polyploidisation via crossing with 2n gamete producing genotypes and interploidy crosses are two of the main methods currently used to obtain polyploid tulips. In our study diploid 2n gamete producing F₁ hybrids of Darwin hybrids (Tulipa gesneriana × Tulipa fosteriana) and triploid hybrid resulting from ‘Rhodos’ × ‘Princeps’ cross were used as pollen donor and crossed with cultivars of T. gesneriana in the following combination: 2x × 2x, 3x × 2x, 2x × 3x, and 3x × 3x. The progenies resulting from crosses at diploid level were mostly diploid, whereas a few seedlings were triploid. In 3x × 2x crosses aneuploids with chromosome constitution in between triploid and tetraploid (43–45 chromosomes) were predominant, but also one tetraploid (2n = 4x = 48) and four pentaploids (2n = 5x = 60) were obtained. In 2x × 3x crosses most progenies were triploid with the exception of a few aneuploids (3x + 1 and 3x ? 1), whereas in 3x × 3x cross diploid and aneuploid genotypes were recorded with chromosome number varied from 27 to 34. These results indicate that triploid parents produced aneuploid as well as euploid (x, 2x, 3x) gametes and that success in ploidy manipulation in tulip depends to a large degree on the ploidy level of the parental genotypes used for hybridization. Genome constitution of selected population of F1 and BC1 hybrids was analyzed through genomic in situ hybridization (GISH). GISH analysis of the BC1 showed a considerable amount of intergenomic recombination which is desirable for introgression breeding.     

Study on reproductive barriers in 4x–2x crosses in Dianthus caryophyllus L

Reproductive barriers often exist in 4x–2x crosses in Dianthus caryophyllus L. and seriously lead to low seed set and breeding efficiency. In this study, we inspected pre-fertilization and post-fertilization barriers in crosses between ‘butterfly’ (2n = 4x = 60) and the fertile breeding line ‘NH10’ (2n = 2x = 30) and ‘NH14’ (2n = 2x = 30) using fluorescence microscopy and paraffin section. The growth of the pollen tubes up to the ovary was observed in crosses of ‘butterfly’ × ‘NH10’ and ‘butterfly’ × ‘NH14’. However, abnormalities of pollen tube including coiled and spiky and callose deposition in incompatible tubes has also been detected. In the cross ‘butterfly’ and ‘NH10’, the number of pollen tubes at the base of the style reached the highest value, 67.2 ± 10.8 at 48 h after pollination, but the number of ovules with micropylar penetration was low, 19 ± 4.5. Similarly, in the cross between ‘butterfly’ and ‘NH14’, pollen tubes at the base of the style at 48 h after pollination was 51 ± 6.2 and the ovules with penetrated micropyles per ovary at 48 h after pollination was 11 ± 2.4. It indicated the pollen tubes difficultly penetrated into the ovules in 4x–2x crosses. In addition, it was observed that non-fused of egg and sperm or non-fused polar nucleus resulted in endosperm without embryos, and embryos without endosperm. Embryos aborted at different developmental stages (prembryo to cotyledon embryo, main globular embryo), which might be related to a deficient endosperm development in reciprocal crosses. The results suggested that pre-fertilization and post-fertilization barriers were detected in these combinations. Due to the post-fertilization barriers in 4x–2x crosses, the embryo rescue might be contributed to obtain hybrid F1 plants.     

Compatibility relations between the edible carrot Daucus carota and D. pusillus, a related wild species from the Argentinian Pampas

To establish the feasibility of hybridization between the wild carrot species Daucus pusillus Michx. (2n = 2x = 22; 2n = 2x = 22 and 20), collected in the pampas grasslands of Argentina, and the edible carrot, Daucus carota L. (2n = 2x = 18), controlled pollinations were attempted on the plant. Due to the difficulties encountered, flowers of 12 accesions and three commercial cultivars were excised from individual plants and pollinated in Petri dishes following an incomplete diallel design. After processing, the pollinated pistils (four to six per genotypic combination) were observed under a microscope with UV light. Pollen tubes reaching the ovaries and/or the ovules (compatible relation) were observed in six out of nine D. pusillus × D. pusillus and seven out of 18 D. pusillus × D. carota genotypic combinations. In the eight D. carota × D. pusillus genotypic combinations, only ungerminated pollen, pollen not adhered to the stigmas or pollen tubes overlapping the stylar tissue were observed. Additional flowers were pollinated in a sample of compatible genotypic combinations and the pollinated pistils were in vitro culture to study embryo and endosperm development. Eight out of nine pistils from the intraspecific and nine out of 13 from the interspecific crosses enlarged to form apparently normal schizocarps. Histological analyses revealed normal development of embryo and endosperm. The breeding barriers between the two species are incomplete, making feasible the obtainment of interspecific hybrids by conventional techniques.     

Interspecific hybridization in <Emphasis Type="Italic">Sarcococca</Emphasis> supported by analysis of ploidy level,genome size and genetic relationships

Knowledge of ploidy level differences, genome size and genetic relationships between species facilitates interspecific hybridization in ornamentals. For Sarcococca (Buxaceae) only limited (cyto)genetic information is available. The aim of this study was to determine the genome size and chromosome number and to unravel the genetic relationships of a breeder’s collection using AFLP marker analysis. Based on these results, interspecific crosses were made and the efficiency and hybrid status was verified. Two groups of diploid plants (2n = 2x = 24) were observed, with either a genome size of 4.11–4.20 or 7.25–9.63 pg/2C. All the tetraploid genotypes (2n = 4x = 48) had genome sizes ranging from 7.91 to 8.18 pg/2C. In crosses between parents with equal ploidy level and genome size a higher crossing efficiency (on average 58% of the hybridizations resulting in fruits) and more true hybrids (on average 96% of the offspring) were obtained compared to crosses between plants with different genome size and ploidy level (on average 23% fruits and 24% hybrids, respectively). In none of the cross combinations, the ploidy level or genome size was found to be a complete hybridization barrier, although unilateral incongruity was found in some cross combinations. Distant genetic relationships did not hamper the hybridization within Sarcococca genotypes. Our findings will contribute to a more efficient breeding program and a faster achievement of hybrids with an added value.     

Overcoming obstacles to interspecific hybridization between <Emphasis Type="Italic">Gossypium hirsutum</Emphasis> and <Emphasis Type="Italic">G. turneri</Emphasis>

Gossypium turneri, a wild cotton species (2n = 2X = 26, D₁₀D₁₀) originating from Mexico, possesses invaluable characteristics unavailable in the cultivated tetraploid cotton gene pool, such as caducous involucels at anthesis, resistance to insects and tolerance to abiotic stresses. However, transferring desired characteristics from wild species into cultivated cotton is often fraught with diverse obstacles. Here, Gossypium hirsutum (as the maternal parent) and G. turneri were crossed in the Hainan Province of China, and the obtained hybrid seeds (2n = 3X = 39, ADD₁₀) were treated with 0.075% colchicine solution for 48 h to double the chromosome complement in order to overcome triploid F₁ sterility and to generate a fertile hexaploid. Chromosome doubling was successful in four individuals. However, the new synthetic hexaploids derived from these individuals were still highly sterile, and no seeds were generated by selfing or crossing. Therefore, an embryo rescue technique was employed in an attempt to produce progenies from the new synthetic hexaploids. Consequently, a total of six large embryos were obtained on MSB2K medium supplemented with 0.5 mg l^?1 KIN and 250 mg l^?1 CH using ovules from backcrossing that were 3 days post-anthesis. Four grafted surviving seedlings were confirmed to be the progenies (pentaploids) of the new synthetic hexaploids using cytological observations and molecular markers. Eight putative fertile individuals derived from backcrossing the above pentaploids were confirmed using SSR markers and generated an abundance of normal seeds. This research lays a foundation for transferring desirable characteristics from G. turneri into upland cotton.     

A tetraploid hybrid plant from 4x × 2x crosses in Vitis and its origin

To study the origin of unreduced (2n) gametes in diploid Vitis cultivars, we surveyed the occurrence of tetraploid hybrid seedlings from 40 interploid crosses with five tetraploid and seven diploid cultivars. A total of 250 seedlings from the interploid crosses were established through embryo culture and by seed sowing. In 20 2x × 4x crosses, no tetraploid hybrid seedlings were derived from 8,902pollinations. In 20 4x × 2x crosses, two tetraploid hybrid seedlings were obtained from 8,057 pollinations. Investigation of isozyme genotypes of the two tetraploid seedlings using three variable enzyme systems indicated that one of the two seedlings resulted from the union of a diploid egg with a 2n male gamete and that failure of second meiotic division resulted in the formation of the 2n male gamete. The percentage of giant pollen grains in `Muscat Bailey A', a pollen parent of the tetraploid seedling, was relatively high(about 5.9%) and 10.9% of the giant pollen grains germinated on agar medium. These results suggested that 2n pollen of diploid cultivars are useful for breeding tetraploid grape. This revised version was published online in August 2006 with corrections to the Cover Date.     

Analysis of endosperm development of allotriploid × diploid/tetraploid crosses in <Emphasis Type="Italic">Lilium</Emphasis>

Lilium are Fritillaria-type plants. Triploid lilies, regardless of their male sterility, can be used as female parents to cross with appropriate males, in contrast to other triploid Polygonum-type plants, which are usually seedless. Up to now, however, little attention has been paid to the reason. Here we made allotriploid × diploid/tetraploid crosses in Lilium. Endosperm and progenies of LAA × AA/AAAA crosses were analyzed for ploidy level and genome composition. The endosperm of some LAA × AA/AAAA ovules developed well but there was little or no development of LAA × LL/OO endosperm. The endosperm genome composition of LAA × AA, LAA × AAAA, LAA × LL, LAA × OO are theoretically derived as 5A + 2L, 6A + 2L, 4A + 3L, 4A + 2L + O, respectively. Genomic in situ hybridization showed that the progenies of LAA × AA/AAAA were aneuploid. Based on the experimental results and analyses, our hypothesis is that five same genomes of endosperms are essential for its development in triploid × diploid/tetraploid crosses of Lilium. The hypothesis can explain the success or failure of 3x × 2x/4x crosses in Lilium and is of importance for lily breeders who need to know the likelihood of success when producing new cultivars of this economically important horticultural crop.     

Intergeneric hybrid between Chrysanthemum × morifolium and Artemisia japonica achieved via embryo rescue shows salt tolerance

Using embryo rescue, we generated an intergeneric hybrid between Chrysanthemum × morifolium ‘Maoyan’ and Artemisia japonica Thunb. Cytological tests confirmed that regenerated plantlets were all genuine hybrids possessing 45 chromosomes, with 27 chromosomes inherited from C. × morifolium (2n = 6x = 54) and the other 18 derived from A. japonica (2n = 4x = 36). Hybrid plant flowered normally. The shape and color of the hybrid flowers and leaves resembled those of chrysanthemum, while leaf width, leaf length, plant height, and inflorescence diameter were intermediate between those of the parents. Hybrid plant had higher levels of chlorophyll and free proline, and lower concentrations of malondialdehyde and Na⁺, than the maternal parent (C. × morifolium), and these levels were correlated with the hybrid’s enhanced salt tolerance. These results clearly demonstrate that intergeneric hybridization is an effective method of cultivar improvement in chrysanthemum.     

Improving seed yield in red clover through marker assisted parentage analysis

Red clover (Trifolium pratense L.) is an important perennial forage crop in Europe. Unfortunately, seed production of modern cultivars is often unsatisfactory and breeding progress for higher seed yield is slow. We evaluated whether marker assisted parentage analysis could identify progeny plants with two high seed-yielding parents in a diploid selection trial. Furthermore, we studied if breeding for high seed yield could be speeded up by selecting progeny plants with two high seed-yielding parents, compared to traditional selection on the mother plant only. In this study, 111 genotypes with excellent vegetative traits from a first cycle selection trial were allowed to pollinate each other in isolation. After seed harvest, ten high seed-yielding plants were identified and their seeds were used to grow ten half-sib progeny populations, with 1,121 plants in total. Two multiplex primer sets targeting 18 SSR loci were designed and used to fingerprint parental and progeny plants. Parentage was reliably determined for 1,083 progeny plants: 135 plants were the result of a cross between two high seed-yielding parents. In a 2-year seed yield trial, the seed yield of these 135 progeny plants was compared to that of a control group of 216 progeny plants from the same ten high seed-yielding mother plants but with no selection on the male contribution, as is current practice. Compared to the control group, progeny plants with two high seed-yielding parents had significantly higher seed yields, 23.0 and 75.9 % higher in 2012 and 2013 respectively, and a significantly higher thousand seed weight, 6.6 and 5.7 % higher in 2012 and 2013 respectively. Seed yield was strongly correlated with flower abundance (r = 0.643) and plant volume (r = 0.593 in 2012 and r = 0.509 in 2013) and negatively correlated with susceptibility to mildew (r = ?0.145). We conclude that breeding programs aiming at increasing seed yield in diploid red clover cultivars would strongly benefit from the integration of marker assisted parentage analysis to identify those progeny plants that are the result of a cross between two high seed-yielding plants. This goal cannot be achieved with traditional family selection where only the seed yield of the mother plant is known.     

Morphological and molecular characterization of the second backcross progenies of Ogu-CMS Chinese kale and rapeseed

Ogura cytoplasmic male sterility (Ogu-CMS) is widely used in the production of commercial hybrids of Brassica oleracea. However, the widespread application of the Ogu-CMS system in B. oleracea has hindered the germplasm innovation of Ogu-CMS resources due to the lack of a natural restorer line. Previously, the Ogu-CMS fertility-restored interspecific hybrids between rapeseed 15Y403 (2n = 38, AACC) and Chinese kale JL1 (2n = 18, CC) have been successfully produced. However, these progenies, which still contained a large proportion of rapeseed genomic components, showed poor fertility and a low seed setting rate under natural pollination. To improve fertility and seed setting, a successive backcross with JL1 was performed to produce BC₂ progenies. Screening with the Rfo-specific marker, five individuals harboring the Rfo gene were identified among 98 BC₂ progenies. These five individuals underwent background marker screening and an evaluation of agronomic traits and fertility. One individual (code: 15Q23) was identified with higher pollen viability, better seed setting under natural pollination, and a closer genetic background to the parent Chinese kale JL1. Many morphological traits showed no significant differences (P < 0.05) between 15Q23 and the backcross parent JL1. However, the average seed setting of 15Q23 under natural pollination was 0.72 seeds per pod, which was 50 times higher than that of BC₁ progenies, and the average pollen viability was 87.07%, which was significantly better than that of the F₁ and BC₁ plants (P < 0.01). The genetic background of 15Q23 was more closer to the parent JL1 than that of BC₁ plants and another BC₂ fertility-restored individual, with 82% of the polymorphic alleles being the same as those of the parent Chinese kale JL1. Thus, the individual 15Q23 could be used as a donor plant for further backcrosses. This study lays the foundation for the development of Ogu-CMS restorer material in B. oleracea.     

Incongruity in the interspecific crosses of Vitis L. Reproductive expression in the F1 progeny

Summary F₁ progeny from three classes of crosses were studied for indications of incongruity in the form of reproductive dysfunction. Percent fruit set was found to be approximately equivalent to the additive inverse of percent aborted flowers, as the effect of shot berries was small and not significant. In interspecific crosses, barriers were manifested as reduced fruit set and seed number per berry. These two characters were used to create an index of reproductive efficiency. Nine F₁s from crosses of most divergent class, V. riparia × V. vinifera (R × V), displayed significantly tower reproductive efficiency, percent fruit set, and seed number per berry. Four female F₁s from the least divergent class, V. riparia × V. riparia (R × R), had the highest reproductive efficiency, percent fruit set and seed number per berry, when pollinated by two V. riparia pollen sources. Seventeen F₁s from the highly-intercrossed class, V. riparia × French Hybrid (R × FH), displayed intermediate levels of reproductive efficiency, percent fruit set, and seed number per berry. The R × FH class involved V. vinifera, V. rupestris, V. aestivalis, V. labrusca, V. berlandieri, V. cinerea, and V. riparia. It is proposed that the complex intercrossing in the pedigrees of the R × FH class ameliorated incongruity. In the most divergent class, R × V, progenies of one of the V. riparia parents displayed fewer incongruity effects than progenies of the other V. riparia parents. Progeny testing of many proposed parental combinations may be used to uncover useful congruent combinations.Minnesota Agricultural Experiment Station, Scientific Journal Series No. 20835     

Cross compatibility between diploid and tetraploid perennial ryegrass (Lolium perenne L.)

Summary A number of crosses were made between diploid and tetraploid Lolium perenne plants in order to determine the degree of cross compatibility between the two different ploidy levels. The range of hybridization undertaken involved diploidxdiploid, and tetraploid x tetraploid control crossings, diploid x tetraploid crosses, tetraploid x diploid crosses and mixed pollinations. The seed set, the germination capacity of resultant hybrid seeds, and the chromosome constitution of derived seedlings was determined. In addition attempts were made to culture 12 day-old hybrid embryos from the diploid x tetraploid and reciprocal crosses.The crossing results obtained indicated no barrier to fertilization between diploids and tetraploids, but the irregular and poor development of the seed accompanied by much reduced germination, indicated unsatisfactory endosperm establishment of large numbers of triploids, as confirmed by the result of culturing embryos dissected from 12-day old seeds.In preliminary studies undertaken to determine the extent to which diploid and tetraploid ryegrass varieties intercross when grown in close proximity for seed production, Griffiths and Pegler (1966) observed a very low incidence of triploids amongst the progenies of diploid plants exposed to the pollen of tetraploid plants and also amongst the progenies of tetraploid material exposed to the pollen of diploid plants.In these and subsequent studies it was noted that only a very small proportion of the so-called fully formed seeds derived from diploid x tetraploid crosses did actually germinate. When, in 1964, propagules of clones of S.24 perennial ryegrass, well separated from one another, were placed in drills of the tetraploid perennial ryegrass variety Reveille, the proportion of florets which formed caryopses, and of caryopses which germinated was as follows (Table 1).The data (Table 1) clearly indicate that a considerable proportion of the S.24 florets had been fertilized by 2n pollen and had formed caryopses, but most of these had proved to be defective. As noted in the previous studies, the frequency of triploid seedlings was low, thus suggesting incompatibility between the n and 2n gametes for the production of viable zygotes.Of the total progenies classed as tetraploids in the early seedling stage, 80% proved to be ryegrass x tall fescue F₁ hybrids. These arose as a result of free crossing with     

Creating cold resistant strawberry via interploidy hybridization between octoploid and dodecaploid

Strawberry cultivars showed limited cold resistance in the Northeast of China, while we obtained a synthetic dodecaploid strawberry hybrid ‘YH15-10’ (2n = 12x = 84) which showed sufficient cold resistance in this area. The reciprocal crosses between F. × ananassa cv. ‘Allstar’ (2n = 8x = 56) and ‘YH15-10’ (2n = 12x = 84) were carried out to select cold resistant strawberry in this study. The 134 seedlings were obtained from the cross of Allstar × YH15-10, while failed in its reciprocal cross. The 30 randomly selected seedlings were examined in terms of morphological characters, chromosome numbers and cold resistance. Most morphological characters were widely separated among F1 progeny with a high broad-sense heritability, which showed that these variations mainly resulted from genetic effect. Some hybrids exhibited heterosis, especially in growth vigor and runner production. Among the 30 tested hybrids, 28 decaploids (2n = 10x = 70), one octoploid (2n = 8x = 56) and one enneaploid (2n = 9x = 63) were observed. The 63.3% hybrids demonstrated higher cold resistance than that of ‘Allstar’ at P < 0.05. These high polyploidy strawberries have potential values in commercial production and modern cultivar improvement.     

Creation of novel interspecific-interploid Hylocereus hybrids (Cactaceae) via embryo rescue

In this study we developed a technique for rescuing embryos at a very early developmental stage following interspecific-interploid crosses between Hylocereus sp. Controlled hand pollinations were performed between the tetraploid H. megalanthus (Vaup.) Bauer as the female parent and either diploid H. monacanthus (Lem.) Britton et Rose or H. undatus (Haw.) Britton et Rose as the male parent. The fertilized ovules were excised from ovaries 10 or 30 days after pollination (DAP). Pollinated ovules containing the funiculi and placental tissue and immature embryos were placed on half-strength basal Murashige and Skoog (MS) medium containing 680 μM glutamine, 0.54 μM α-naphthaleneacetic acid, and 0.45 μM thidiazuron and supplemented with 0.00, 0.09, 0.17 or 0.26 M sucrose concentrations. The best ovule response was recorded at 30 DAP, and the most prolific callus formation was observed at 10 DAP. Callus formation was observed in most of the treatments using whole ovules but not in the isolated immature embryos. The calli were mucilaginous or compact, transparent and friable, but they did not form embryogenic structures. Embryo development was significantly affected by the sucrose concentration, and the best results were obtained with 0.17 M sucrose. More than 70 % of the obtained hybrids were successfully hardened off and transplanted in soil where they grew normally. Ploidy level analyses of 77 putative hybrids exposed diploid, triploid, tetraploid, and higher than tetraploid levels. Among those hybrids studied, 22 progenies were randomly chosen for amplified fragment length polymorphism analysis, and all were identified as genuine hybrids. The technology described here is an additional stage in the breeding program for Hylocereus species resulting in novel, interspecific hybrids obtained using the embryo rescue technique.     

Combining ability of seed vigor and seed yield in soybean

Studies have shown no consensus in relationships between seed yield and vigor in soybean [Glycine max (L.) Merrill]. The lack of information regarding the inheritance of seed vigor prompted this study to determine the types of gene action and combining ability estimates for seed vigor and its related traits. Five high and six low seed vigor soybean genotypes were crossed in a diallel, and selfed to produce 55 F₂ progenies, which were examined, along with the parents, for seed vigor, yield, and seed weight. Significant genotype and environment effects were found for seed vigor and yield. General combining ability (GCA) effects for seed vigor and seed yield were significant (p≤ 0.01) and larger than specific combining ability (SCA) effects. Significant GCA and SCA effects were found for seed weight, indicating that both additive and non additive genetic effects were involved in conditioning seed weight. The ratios of mean square, 2GCA / (2GCA+SCA), were 0.96 for seed vigor and 0.93 for seed yield. These ratios indicated that additive gene effects were more important than non additive gene effects for seed vigor and seed yield in these crosses. Mean seed vigor(83.8%), as determined by accelerated aging germination, and mean seed yield (2,155 kg ha^-1)in high vigor × high vigor crosses were higher than the high vigor × low vigor and low vigor × low vigor crosses. Mean percent accelerated aging germination rates in F₂ populations from diallel crosses were significantly related to mid-parent seed vigor(r² = 0.52^**) and midparent seed size (r² = 0.31^**). These results indicated that levels of seed vigor can be improved through breeding, while maintaining high yields because of the predominance of GCA effects in both seed vigor and seed yield. This revised version was published online in July 2006 with corrections to the Cover Date.     

Introgression of Cajanus platycarpus genome into cultivated pigeonpea, C. cajan

Summary Cajanus platycarpus, an incompatible wild species from the tertiary gene pool of pigeonpea (C. cajan (L.) Millspaugh), has many desirable characteristics for the improvement of cultivated varieties. To necessitate such transfers, embryo rescue techniques were used to obtain F₁ hybrids. The F₁ hybrids were treated with colchicine to obtain tetraploid hybrids, that were selfed to obtain F₂, F₃ and F₄ progenies. All of the hybrids and subsequent progenies had an intermediate morphology between the two parents. Backcrossing of the tetraploid hybrids with cultivated pigeonpea was not possible given embryo abortion, with smaller aborted embryos than those obtained in the F₀ parental cross.As a route of introgression, diploid F₁ hybrids were backcrossed with cultivated pigeonpea and BC₁ progeny obtained by in vitro culture of aborting embryos. BC₂ plants were obtained by normal, mature seed germination. Although embryo rescue techniques had to be used to obtain F₁ and BC₁ plants, it was possible to produce BC₂ and subsequent generations through direct mature seed. Every backcross to cultivated pigeonpea increased pollen fertility and the formation of mature seeds.Special project assistant till December, 2003.     

Progenies of allotriploids of Oriental × Asiatic lilies (Lilium) examined by GISH analysis

With the aim of utilizing allotriploid (2n = 3x = 36) lily hybrids (Lilium) in introgression breeding, different types of crosses were made. First, using diploid Asiatic lilies (2n = 2x = 24), reciprocal crosses (3x − 2x and 2x − 3x) were made with allotriploid hybrids (AOA) obtained by backcrosses of F₁ Oriental × Asiatic hybrids (OA) to Asiatic cultivars (A). Secondly, the AOA allotriploids were crossed with allotetraploid (OAOA, 2n = 4x = 48), in 3x − 4x combination. Finally, the AOA allotriploids where crossed to 2n gamete producer F₁ OA hybrids (3x − 2x (2n)). Two types of triploids were used as parents in the different types of crosses, derived from: (a) mitotic polyploidization and (b) sexual polyploidization. Ploidy level of the progeny was determined by estimating the DNA values through flowcytometry as well as chromosome counting. The aneuploid progeny plants from 3x − 2x and reciprocal crosses had approximate diploid levels and in 3x − 4x crosses and 3x − 2x (2n) the progeny had approximate tetraploid levels. Balanced euploid gametes (x, 2x and 3x) were formed in the AOA genotypes. Recombinant chromosomes were found in the progenies of all crosses, except in the case of 2x − 3x crosses through genomic in situ hybridization (GISH) analyses. Recombinant chromosomes occurred in the F₁ OA hybrid when the triploid AOA hybrid was derived through sexual polyploidization, but not through mitotic polyploidization with two exceptions. Those recombinant chromosomes were transmitted to the progenies in variable frequencies.     

Pattern of genetic inheritance of morphological and agronomic traits of sorghum associated with resistance to sorghum shoot fly, <Emphasis Type="Italic">Atherigona soccata</Emphasis>

Sorghum shoot fly, Atherigona soccata is an important pest of sorghum during the seedling stage, which influences both fodder and grain yield. To understand the nature of inheritance of shoot fly resistance in sorghum, we performed generation mean analysis using two crosses IS 18551 × Swarna and M 35-1 × ICSV 700 during the 2013–2014 cropping seasons. The F₁, F₂, BC₁ and BC₂ progenies, along with the parental lines were evaluated for agronomic and morphological traits associated with resistance/susceptibility to sorghum shoot fly, A. soccata. The cross IS 18551 × Swarna exhibited significant differences between the parents for shoot fly deadhearts (%) in the postrainy season. The progenies of this cross exhibited lower shoot fly damage, suggesting that at least one of the parents should have genes for resistance to develop shoot fly-resistant hybrids. Leaf glossiness, leafsheath pigmentation and plant vigor score during the seedling stage exhibited non-allelic gene interactions with dominant gene action, whereas 100 seed weight showed both additive and dominant gene interactions. Presence of awns showed recessive nature of the awned gene. Generation mean analysis suggested that both additive and dominance gene effects were important for most of the traits evaluated in this study, but dominance had a more pronounced effect.     

Effectiveness of early-generation testing applied to upland rice breeding

A plant breeding program is a long-term investment. Therefore, periodic assessment of the effectiveness of a breeding strategy is essential to maximize genetic gains per unit of time and resource invested. In this work, we assessed the effectiveness of the early-generation testing (EGT) approach used in the upland rice (Oryza sativa L.) breeding program at Embrapa (Brazilian Agricultural Research Corporation), Brazil, estimating the genetic progress achieved for three traits in two distinct phases, spanning 15 years. In the first phase (from 2003 to 2010), it was used the bulk method within F₃ progenies with prior testing of F₂ crosses, while in the second phase (from 2010 to 2017), it was used the bulk method within F₂ progenies. The dataset comprised 70 yield trials, involving 1884 F_3:5 progenies (phase I) and 925 F_2:4 progenies (phase II) from an elite population, and 10 check cultivars, evaluated for grain yield (GY), plant height (PH) and days to flowering (DTF). For estimating the genetic gain, we adapted a generalized linear regression method to compute bi-segmented linear regression coefficients. Desirable genetic gains were achieved only for GY in both phases of the breeding program, with 78.75 kg ha^?1 year^?1 (2.68%) in the first phase, and 53.78 kg ha^?1 year^?1 (1.54%) in the second phase. These results show the effectiveness of EGT, especially via bulk method within F₃ progenies with prior testing of F₂ crosses, applied to upland rice breeding. Some refinements are discussed in the method to make it more cost-effective and more efficient in achieving genetic gains.