Production of two mandarin × trifoliate orange hybrid populations via embryo rescue with verification by SSR analysis

Intergeneric sexual hybridizations were conducted between two genera of Rutaceae, with Satsuma mandarin (Citrus unshiu Marc) and Red tangerine (C. reticulata Blanco) as maternal parents, and Poncirus trifoliata (L.) Raf as the paternal parent, in an effort to generate hybrid populations for both molecular mapping and rootstock breeding. Embryo rescue is important for citrus sexual breeding because polyembryony can interfere with hybrid embryo recovery. Immature embryos of 80, 85, 90 days after pollination (DAP) from the Satsuma mandarin (S) × trifoliate orange (P) cross, 80 and 85 DAP from the Red tangerine (R) × trifoliate orange (P) cross, were cultured on MG1.0 medium consisting of MT basal medium supplemented with 1.0 mg l⁻¹ GA₃ and 4% sucrose. The results showed that 80 DAP was the optimal time for embryo rescue of the tested crosses, as evidenced by embryos at this stage exhibiting high germination rates, 37.3% for S × P and 51.3% for R × P. Among the eight tested media, MT medium supplemented with 0.5 mg l⁻¹ GA₃ was the best one for Satsuma mandarin, and MT plus 1.0 mg l⁻¹ GA₃ for Red tangerine. A total of 85 plants were obtained from S × P, and 340 from R × P. Out of them, 44 progenies from S × P and 111 from R × P were sufficiently confirmed to be hybrids by morphological characterization and SSR analysis. In addition, two hybrid callus lines were obtained from S × P and R × P respectively.     

In vitro rescue of zygotic embryos of sour orange, Citrus aurantium L., and their detection based on RFLP analysis

Embryo development in vivo has been studied in four Citrus aurantium L. polyembryonic genotypes. Seeds were collected 65, 85, 105, 125 and 220 days after pollination (DAP). None of the immature seeds harvested 65 and 85 DAP contained visible embryos. A single embryo at a more advanced developmental stage was observed in the central position at the micropylar apex of the embryo sac in about 74% of seeds harvested at 105 DAP, while at 125 and 220 DAP the majority of seeds had two or more embryos at the same developmental stage crowded together. Restriction fragment length polymorphism (RFLP) analysis of lowand high-copy-number nuclear DNA was used to distinguish zygotic from nucellar seedlings. Analysis of plantlets derived from in vitro culture of the bigger embryos, located in the central position at the micropylar apex of the embryo sac of seeds harvested at 105 DAP, established that the frequencies of zygotic embryos ranged from 82 to 88%. Media for immature embryo germination in vitro were based on the nutrients and vitamins of Murashige and Skoog (MS) and Murashige and Tucker (MT) media supplemented with various concentrations of sucrose and growth regulators. A total of 76% of globular stage embryos (<0.3mm) germinated on MT medium containing 150 mM sucrose and 14.4 μM gibberellic acid. Heart stage embryos (0.3-0.8 mm) germinated at 95% on MT medium supplemented with 150 mM sucrose and 2.9 μM gibberellic acid. The addition of 500 mg/l malt extract to MS medium increased the germination of early cotyledon stage (0.8-2.0mm) embryos to 98%. The optimum sucrose concentration for embryo rescue was 150 mM for the three embryo developmental stages. The ability to form plants in vitro strongly increased with increasing embryo developmental stage.     

Embryo rescue and plant regeneration following interspecific crosses in the genus <Emphasis Type="Italic">Hylocereus</Emphasis> (Cactaceae)

The aim of this study was to develop an efficient methodology to rescue embryos following interspecific crosses in the genus Hylocereus. Crosses between the diploids Hylocereus polyrhizus and H. undatus in both directions were performed. Fertilized ovules carrying embryos at very early pro-embryonic stages were excised from ovaries 5 days after pollination (DAP) and placed on half-strength basal MS medium containing 680 μM glutamine, 0.55 μM α-naphthaleneacetic acid (NAA), 0.45 μM thidiazuron (TDZ) and various concentrations of sucrose. After 30 days in culture, ovules were isolated from the surrounding tissue and transferred to the same fresh medium. Significant differences were found between the main effects (cross and sucrose concentration) in ovule response, i.e., increased ovule size and callus formation. The best responses were obtained in the cross: H. polyrhizus × H. undatus; and sucrose concentration of 0.09 M. In terms of embryo conversion, polyembryony and number of regenerated plants, the highest responses were observed on the culture medium supplemented with 0.17 M sucrose in both interspecific crosses. All tested plants were found to be diploid by flow cytometric analyses. Fluorescent amplified—fragment length polymorphism (fAFLP) confirmed the hybrid origin of the regenerated plants. This study reports on the success of a three-step embryo rescue procedure for Hylocereus species. The procedure developed here provides the means for producing plants from very-early embryo stage, thus expanding the prospects for vine-cactus breeding programs.     

Effect of embryo age and culture media on plant growth and vernalization response in winter wheat

Summary Embryo age and composition of nutrient medium affected plant growth and response to vernalization in winter wheat (Triticum aestivum L.). Root and shoot development was more in older than in younger excised embryos, and more in a medium without kinetin than in one with kinetin. Kinetin (2 mg/l) in the medium did not accelerate vernalization, probably because it tended to inhibit seedling and plant growth.Embryo age and media did not completely replace vernalization. Twenty- and 16-day-old embryos responded by flowering after 4 weeks of vernalization. Among plants raised on a standard medium from 20-day-old embryos and vernalized for 4 weeks, 84.2% flowered by or before 50 days after transplanting. Time from embryo culture to heading for 20-day-old embryos with-4-week vernalization averaged 84.6 days. Immature embryos (16–20 days old) needed only 4 weeks of vernalization compared to 6 weeks for mature embryos. Excised embryos could be vernalized as efficiently as seedlings raised by embryo culture. Embryo culture at 16–20 days after anthesis coupled with 4-week cold treatment shortens generation time of winter wheat by about 40 days.Contribution No. 82-131-j, Department of Plant Pathology, Kansas Agricultural Experiment Station, Manhattan, KS 66506, USA.     

In vitro interspecific fertilization,embryo development and formation of hybrid seedlings between Gossypium hirsutum and G. arboreum

Summary In vitro hybridization between Gossypium hirsutum, and G. arboreum was carried out. Hybrid seedlings were obtained after successive use of the following five kinds of media: 1) pollen grain germination medium, 2) double-fertilization medium, 3) embryo development medium, 4) seedlings formation medium, 5) green seedlings growth medium. The factors affecting in vitro interspecific fertilization, embryogenesis and seedling formation were studied. The key factors were temperature and relative humidity (RH). The optimum RH for in vitro interspecific fertilization was 65%, and a suitable temperature range was 26–30°C. Pollen grain germination ratio decreased rapidly at a RH of 80%. When the temperature and the RH were higher than 32°C and 80%, respectively, the fertilization rate decreased to zero.Effects of petals and calyces of the maternal flowers on in vitro interspecific fertilization and ovule growth were identified. Even though the petals or calyces were excised, hybrid plantlets were also obtained after media were improved and a shake culture was used. In addition, the process of double-fertilization and embryo development were studied cytologically. The developmental characteristics of the hybrid embryos derived from in vitro interspecific fertilization were a later occurrence of the double-fertilization process and a much lesser division of endosperm cells. But the embryo development was not affected by these characteristics, and young hybrid embryos can develop to their cotyledon stage finally on artificial media.     

小麦与玉米杂交产生小麦单倍体与双单倍体的稳定性

小麦与玉米杂交是诱导小麦单倍体最有效的途径之一, 但单倍体和双单倍体产生频率不稳定影响了该技术的应用。选用13个小麦杂种F₁代单交组合与玉米杂交, 研究了不同小麦生长环境、生长素处理、培养基和壮苗处理对单倍体及双单倍体产生频率的影响。小麦生长在大田, 去雄后割穗培养与玉米杂交平均得胚率为23.9%, 每个杂交穗平均得胚数6.8个, 均是返青后从大田移回冷温室盆栽的3倍以上;不同小麦杂交组合间胚产生频率存在明显差异。生长素Dicamba蘸穗处理平均得胚率是21.5%, 与2,4-D处理得胚率(21.1%)无显著差异, 但不同杂交组合间差异显著。B5培养基幼胚萌发率为70.9%~88.3%, 平均82.0%;1/2 MS培养基胚萌发率为70.0%~86.0%, 平均76.6%;两种培养基平均胚萌发率无显著差异。试管苗经壮苗培养基壮苗处理与试管苗经移栽壮苗处理后加倍效率分别是67.6%和8.6%。移栽壮苗处理的苗分蘖少, 生长较弱, 加倍处理后存活率低和加倍率低是其单倍体加倍效率低的原因。     

Development and characterization of interspecific hybrids of Trifolium alexandrinum×T. apertum using embryo rescue

This is the first report on the development of interspecific hybrids between Trifolium alexandrinum and T. apertum using embryo rescue and characterization of F₁ plants. T. apertum was used as the male parent and T. alexandrinum as the female parent. Development of interspecific hybrids under natural conditions is not successful and so embryo rescue was attempted. Of the several combinations tried, pollination 2 days after emasculation and embryos rescued 11 days after pollination was found to be the best. For embryo culture, EC3 medium consisting of MS basal supplemented with 2.3 μM kinetin and 3% sucrose was used. Germinated embryos were transferred to LSP3 medium 25 days after inoculation wherein most of the cultures showed multiple shoots that were split and subcultured on RL1 medium for rooting. After hardening, about 75% of hybrids were successfully transferred to the field. The hybrids, in general, showed morphological traits intermediate between the two parents; however, a few hybrids showed better growth than either parent. Some F₁ plants were almost 3 weeks later in flowering than the female parent. Pollen fertility among these plants ranged from 78 to approximately 100%. Chromosomal associations at diakinesis and isozyme banding patterns for acid phosphatase (ACP), superoxide dismutase (SOD) and peroxidase also confirmed the hybrid nature of the plants.     

Field assessment of a second generation backcross (BC1 × Passiflora edulis) of passion fruit for agronomic performance and resistance to CABMV

The objective of this work was to characterize and identify among the passion fruit hybrids of the second generation backcross (BC2: BC1 × P. edulis), resistance to passion fruit woodiness disease (CABMV) with desirable agronomic attributes. Ninety‐two progeny of BC2 hybrids were evaluated for severity of the virus in conditions of field, physical and chemical characterization of the fruits and precocity of the plant in relation to the vegetative and reproductive phases. The data were analysed using multivariate statistics and mixed models (REML/BLUP) to estimate genetic parameters. The was conditioned by and , for number of fruits, fruit and peel weight and soluble solids and the effect of for virus severity to CABMV and effect of on the variables fruit length, fruit diameter, peel thickness and pulp weight. For the titratable acidity it had no . The progeny BC2‐A, BC2‐C, BC2‐E, BC2‐F and BC2‐H families can be selected for use in passion fruit breeding programmes by adding high vegetative vigour, early fruit production, resistance to CABMV and also desirable agronomic attributes.     

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.     

General combining ability and heterosis regarding the phytochemical properties in strawberry (Fragaria × ananassa) hybrids

Strawberries are a common and important fruit in human diet because of their high content of essential nutrients and beneficial phytochemicals, which have relevant biological activity in human health. In this study, six cultivars and 15 selected F₁ hybrids between S₄ inbred lines and tester (cv. ‘Dukat’) were evaluated in a field trial over a period of two consecutive years. The amounts of total anthocyanin, phenolics and vitamin C in fruits and antiradical activity were analysed. General combining ability (GCA) and mid‐parent heterosis were also determined for those characteristics. Among all the genotypes tested, the inbred lines of ‘Senga Sengana’ 17 showed the highest breeding value based on GCA for all studied traits. Estimated heterosis varied among genotypes tested. The highest heterosis in terms of vitamin C occurred in the offspring of clone 1387 18‐15 × ‘Dukat’, but with regard to phenols and antiradical activity in hybrid ‘Teresa’ 18‐15 with cv. ‘Dukat’. This study revealed that the differentiation in chemical composition of strawberry fruits between genotypes is clearly dependent on individual genotype combinations and demonstrated the presence of heterosis in phytochemical contents in some specific genotypic combinations.     

Reproductive compatibility studies between wild and cultivated strawberries (Fragaria × ananassa) to obtain “bridge species” for breeding programmes

The Fragaria genus has become relevant due to worldwide economic importance of the octoploid hybrid F. × ananassa. Interspecific hybridizations are widely employed in breeding programmes to introduce useful characteristics from wild species to cultivated varieties, although many reproductive barriers prevent gene flow between species. Hybrid genotypes that can act as a bridge between related wild species and cultivated strawberry could be useful to overcome those barriers. The aim of this work was to obtain interspecific hybrids of Fragaria and to analyse possible reproductive barriers. In the Banco de Germoplasma de Frutilla‐UNT, crossings were carried out among wild species of Fragaria with different ploidy levels and cultivated strawberry. To confirm hybrid condition of genotypes obtained, histological techniques and microsatellite markers were used. When wild species were crossed with cultivated strawberry in both directions, there was low production of achenes, while in crosses between wild species of Fragaria genus, achene production was very high. The percentage of germination of achenes was high when crossed species had the same ploidy level, and very low or null when they presented different ploidy levels. Although pre‐zygotic incompatibility associated to the degree of domestication and postzygotic barriers related to different ploidy levels of the progenitors were detected, new hybrid genotypes of Fragaria were obtained. These new hybrids could be used as “bridge species” in breeding programmes, since preliminary results showed no incompatibility barriers when they were crossed with Duchesnea indica.     

冬小麦幼胚一步成苗技术优化研究

为实现冬小麦幼胚一步苗移栽于夏季大田加代,本试验以‘济麦20’、‘烟优361’、‘泰农18’、‘中优9507’以及（‘济麦20’ב烟优361’）F1的幼胚为材料,分别选用11~13天、14~16天和17~19天3种胚龄的幼胚接种在MB、A和B三种培养基上,经离体培养一步成苗。研究结果表明,在9种接种试验处理中,11~13天接种于A培养基上出苗率最高,平均出苗率为98.30%,14~16天幼胚接种于MB培养基上,获得较高的出苗率（平均95.80%）,17~19天幼胚接种于B培养基上,出苗率最高,平均出苗率为95.20%;17~19天幼胚接种于MB培养基上,出苗率最低（平均16.20%）。14~16天幼胚接种于MB培养基上,不仅出苗率较高,而且胚苗根较长,侧根较多,表明14~16天幼胚适宜接种于MB培养基。     

Genetic effects of embryo and endosperm for four malting quality traits of barley

Seed of seven cultivars of two-rowed barley (Hordeum Vulgare L.) and F2 seed from a half-diallel set of crosses among the cultivars were malted in two years to obtain data on diastatic power (DP), alpha-amylase activity (αAA), beta- amylase activity (βAA) and malt nitrogen (N) content. Embryo and endosperm genetic effects on the traits were studied by using a genetic model including genotype × environment interaction for malting quality characters. Variation of the four malting quality traits was affected by gentic effects and environmental interaction. Performance of DP and βAA was mainly controlled not only by endosperm dominance effects but also by embryo genotype × environment interaction and endosperm dominance × environment interaction. Variation of αAA and malt N content was controlled by both embryo and endosperm genetic effects, but the embryo dominance and endosperm additive effects contributed a major part to the total genetic effects. Significant interaction variances (embryo additive × environment and dominance × environment and endosperm dominance × environment) were also observed for αAA and malt N content. Diastatic power was related positively to βAA. Malt N content was associated positively with DP, largely because of the relationship between malt N and βAA. No obvious phenotype association between DP and αAA was found. General narrow-sense heritabilities of αAA and malt N content were 26.1% and 27.8%, respectively. This revised version was published online in July 2006 with corrections to the Cover Date.     

Generation of interspecific hybrids between <Emphasis Type="Italic">Trifolium vesiculosum</Emphasis> and <Emphasis Type="Italic">T. alexandrinum</Emphasis> using embryo rescue

Interspecific hybrids were developed between Trifolium alexandrinum cultivar Wardan × Trifolium vesiculosum and T. alexandrinum cultivar BL1 × T. vesiculosum through embryo rescue, as the crosses failed to set seed under natural conditions. Trifolium vesiculosum was used as a donor/male parent in this study as it is reported to possess tolerance to stem rot and high forage yield. Fertilization in crossed florets of the crosses was manifested from the recovery of swollen ovaries (< 7.80%) and confirmed from the presence of one degenerated ovule in most (> 93.00%) of the swollen ovaries. The hybrid embryos at various developmental stages (heart, torpedo and cotyledonary) were rescued at a frequency of 2.56% from Wardan × T. vesiculosum and 6.12% from BL1 × T. vesiculosum. Differentiation occurred only in the cotyledonary stage embryos, resulting in 17 putative interspecific hybrid plantlets. The assessment of plantlet hybridity through SSR markers (for the alleles inherited from the donor parent), micromorphological leaf traits (leaf texture and stomata) and morphological characters (plant height, leaflet length and width) confirmed production of two interspecific hybrids designated as AV1 and BV3 representing both the crosses. AV1 displayed moderate resistance and BV3 was resistant to stem rot.     

Molecular mapping of soybean seed tocopherols in the cross ‘OAC Bayfield’ × ‘OAC Shire’

Soybean (Glycine max [L.] Merr.) is cultivated primarily for its protein and oil in the seed. In addition, soybean seeds contain nutraceutical compounds such as tocopherols (vitamin E), which are powerful antioxidants with health benefits. The objective of this study was to identify molecular markers linked to quantitative trait loci (QTL) that affect accumulation of soybean seed tocopherols. A recombinant inbred line (RIL) population derived from the cross ‘OAC Bayfield’ × ‘OAC Shire’ was grown in three locations over 2 years. A total of 151 SSR markers were polymorphic of which a one‐way analysis of variance identified 42 markers whereas composite interval mapping identified 26 markers linked to tocopherol QTL across 17 chromosomes. Individual QTL explained from 7% to 42% of the total phenotypic variation. Significant two‐locus epistatic interactions were identified for a total of 122 combinations in 2009 and 152 in 2010. The multiple‐locus models explained 18.4–72.2% of the total phenotypic variation. The reported QTL may be used in marker‐assisted selection (MAS) to develop high tocopherol soybean cultivars.     

Hybrid performance and heterosis in strawberry (Fragaria × ananassa Duchesne), regarding acidity,soluble solids and dry matter content in fruits

A top‐cross‐mating design among 29 S₄ inbred lines and tester (cultivar ‘Dukat’) was carried out to study their breeding value in terms of general combining ability (GCA). The objectives of this study were to evaluate the acidity, soluble solids and dry matter contents in fruits of progeny F₁ in comparison with S₄ inbred lines as well as the cultivars (S₀); identify strawberry genotypes with high value of GCA for use in cultivar development; and determine mid‐parent heterosis regarding S₄ inbred lines and cultivated strawberry. The 2‐year observations showed statistically significant differences between tested genotypes in terms of the studied traits. The highest breeding value based on GCA was estimated for Chandler 123‐5 for soluble solids and dry matter content, and Kent 7‐6 for acidity. Estimated mid‐parent heterosis had positive and negative values. The highest heterosis in terms of extract and dry matter content (26.71% and 17.50%, respectively) occurred in the offspring Chandler 123‐5 × ‘Dukat’, but as regards acidity in hybrid Chandler 123‐22 with cv. ‘Dukat’. The study of genetic divergence by dendrograms may help to identify parents suitable for obtaining hybrids with higher heterosis effects.     

Production of interspecific hybrids between Glycine max and G. tomentella through embryo culture

Summary Interspecific hybrids have been obtained in an incompatible cross between Glycine max and G. tomentella through the in vitro culture of hybrid embryos. The percentage of successful pod setting in the crosses averaged 12.8% but there were marked differences depending on the soybean cultivar used as the female parent. Hybrid embryos at globular to heart stages were extracted from the embryo sac 15–25 days after pollination and cultured in vitro. Hybrid plants were successfully obtained by culturing the embryos on B5 medium supplemented with 0.1 mg/l IBA followed by culture on B5 medium supplemented with 0.1 mg/l TBA plus 0.25 mg/l 2-iP. The F1 plants resembled the wild male parent in growth form, but had an intermediate leaf shape between that of the parents.     

Plant Regeneration from in vitro Cultured Anthers of Black Mustard (Brassica nigra Koch)

Anthers of Brassica nigra, excised from fresh as well as cold-pretreated (3 days at 3 ± 2°C) buds cultivated on modified B₅ medium (Gamborg et al. 1968) containing sucrose level varying from 2 % to 10 %, along with 1O^?6M BAP (benzylaminopurine) and 9 × 10^?6M 2,4-D (2,4-dichlorophenoxyacetic acid), developed calli and/or embryos. The latter response was observed only in anthers reared on media containing 6 % or higher levels of sucrose. On media containing two or four per cent sucrose, the anthers produced calli, exclusively. The growth of embryos was inhibited or else they started callusing if left on the media containing higher levels of sucrose. However, on transfer to MS medium (Murashige and Skoog 1962), containing 2 % sucrose, embryos started callusing and subsequently a few secondary embryos differentiated. Such embryos were sub-cultured on MS + 5 × 10^?6M BAP + 2 % sucrose, wherein numerous shoots developed from embryos. The shoots were rooted by transferring to a medium containing 5 × 10^?6M NAA (naphthalene acetic acid). Within two months of culture, some of these plants started flowering in vitro.     

Mapping QTLs for salt tolerance with additive,epistatic and QTL × treatment interaction effects at seedling stage in wheat

Quantitative trait loci (QTLs) for salt tolerance with additive, epistatic and QTL × treatment interaction effects at seedling stage in wheat were identified. A set of 131 recombinant inbred lines derived from cross Chuan 35050 × Shannong 483 were evaluated under salt stress and normal conditions. Wide variation was found for all studied traits. A total of 18 additive and 16 epistatic QTLs were detected, among which five and 11 were with significant QTL × treatment effects. Ten QTL clusters were identified, and each may represent a single gene or closely linked genes. The locus controlling shoot K⁺/Na⁺ concentration ratio and shoot Na⁺ concentration on chromosome 5A may be identical to Nax2. The interval Xgwm6‐Xgwm538 on chromosome 4B for total dry weight was also identified in a previous study, both near the marker Xgwm6. The marker Xgwm6 may be useful for marker‐assisted selection. Six pairs of homoeologous QTLs were detected, showing synteny among the A, B and D genomes. These results facilitate understanding the mechanisms and the genetic basis of salt tolerance in wheat.     

In vitro embryo germination and interploidy hybridization of rose (Rosa sp)

In roses the problems associated with inter-specific breeding include low percent of seed set and lack or low percent of seed germination. Low seed set is usually due to non-amenable parents, which may have different ploidy level or other divergences that result in embryo abortion at early stages of development. Lack of seed germination is mostly attributed to the mechanical restrictions such as thick pericarp or the regulatory mechanisms such as the hormonal control of dormancy. The aims of the present investigation were to optimize in vitro embryo germination technique in rose and study the ploidy of progenies resulted from interploidy hybridizations. To optimize embryo germination, seeds were surface sterilized, whole pericarp and testa were removed and embryos were placed on half strength Murashige and Skoog media supplemented with different concentrations (0, 1, 2.5, 5 mg l^?1) of benzyladenine (BA) in combination with different concentrations (0, 0.5, 1, 1.5 mg l^?1) of gibberellic acid. The maximum percent of in vitro embryo germination (93.40 %) was observed on medium containing 2.5 mg l^?1 BA. In order to select the most fertile seed parents which could be used in interspecific hybridization, 11 commercial rose cultivars (R. × hybrida) were employed in 36 reciprocal crosses. Three rose cultivars including ‘Golden Celebration’, ‘Tess of the d’Urbervilles’ and ‘Molineux’ were selected as the maternal parents. The selected seed parents were employed in crosses with one rose species from Gallicanae section [R. damascena (2n = 4x)] and four rose species form Caninae section [R. orientalis (2n = 5x), R. iberica (2n = 5x) R. canina (2n = 5x) and R. pulverulenta (2n = 6x)]. The highest percent of hip set and in vitro embryo germination were observed in crosses between tetraploid rose cultivars and R. damascena. In all of the crosses with R. canina, the percent of hip and seed set was 0 %. However, in the crosses between tetraploid rose cultivars and other pentaploid or hexaploid rose species from Caninae section both triploid and tetraploid offsprings were attained. Future morphological analysis of the progenies is necessary to show to what extent progenies demonstrate the characteristics of the pollen parents from the Caninae section. Nevertheless, progenies from interploidy hybridizations would be beneficial in future breeding programs in order to expand the relatively small gene pool of roses.