Production of intraspecific hybrids between wild-type and petaloid-sepal cultivars in Habenaria radiata

Orchids are commercially important plants with flowers that are unique and very specialized in shape and color. The flowers consist mostly of sepals, lateral petals, lip (labellum) and column, and are zygomorphic and resupinate. Whereas most orchid species have petaloid tepals in the first and second whorls, Habenaria radiata has a flower with greenish sepals and white lateral petals and lip. ‘Hishou’, one of the cultivars of H. radiata, is a floral homeotic mutant and has a petaloid median sepal and lip-like lateral sepals in the first whorl. Additionally, this cultivar often has non-resupinate flowers whereas wild-type H. radiata flowers are resupinate. In the present study, we investigated the genetic inheritance of these characters in the ‘Hishou’ cultivar by crossing it with wild-type plants. Some intraspecific hybrids, which were confirmed by PCR-RFLP analysis, had flowers with a petaloid median sepal and lip-like lateral sepals in the first whorl, indicating that these were dominant characters. Since the remainder of the intraspecific hybrids had wild-type flowers, these characters must be heterozygous in ‘Hishou’ plants. Although ‘Hishou’ plants had non-resupinate flowers, intraspecific hybrid flowers were resupinate, even though they had the petaloid median sepal and lip-like lateral sepals. This result indicates that non-resupination must be a recessive character. Since sepal-petalization and triple lip characters of ‘Hishou’ inherited dominantly, these characters can be utilized for the breeding of Habenaria species by intra- and interspecific crosses.     

文心兰开花相关OnAP1-like基因的克隆及表达分析

以南茜文心兰（Gower Ramsey）盛花期花葶提取的总RNA为模板,通过RT-PCR与RACE扩增,获得一个958 bp的AP1（APETALA1）-like基因的cDNA全长序列,其基因编码区690 bp,共编码氨基酸229个,命名为OnAP1-like（登录号：KC426946）。蛋白质二级结构分析表明,该蛋白质48.91%为α螺旋,10.92%为β折叠,40.17%为无规则卷曲,为亲水性蛋白质。蛋白序列比对和进化树分析表明,OnAP1-like 蛋白与蕙兰AP1-like蛋白一致性最高,进化距离最近。利用RT-qPCR对从文心兰不同时期不同器官中的OnAP1-like基因表达量进行分析,结果表明,同一器官不同发育时期比较,在叶中,花蕾期的表达量最高;在根中,随发育时间推移,表达量逐渐升高,至花后期达到最高;在花葶中的表达量的趋势与根相同。同一时期不同器官比较,抽葶前,根中表达量高于叶片;花蕾期,花瓣中的表达量最高;盛花期和花后期,花葶中的表达量最高。推测该基因在花的发育及形成中发挥作用。     

Deferring flowering of nobile dendrobium hybrids by holding plants under low temperature after vernalization

Orchids are currently the most valuable potted crop in the United States. To date, no studies focused on making possible the year-round greenhouse production of flowering nobile dendrobium orchids. This experiment was aimed at developing a strategy to defer flowering of nobile dendrobium orchids by holding them under low temperature. Mature Den. Red Emperor ‘Prince’ and Den. Sea Mary ‘Snow King’ were held at 10 °C for various durations (0, 4, 8, 12 or 16 weeks) after vernalization (4 weeks at 10 °C). Plants were forced in a greenhouse after holding. Time to flower, flower differentiation (flowering node percentage, number of aerial shoot and aborted bud) and flower quality (total flower number, flower diameter, flower number per flowering node and flower longevity) were determined. Increase of low temperature holding duration from 0 to 16 weeks extended time to flower up to 3 months and did not affect parameters of flower except producing larger flowers and reducing flower number per flowering node for Den. Red Emperor ‘Prince’. Notably, the flower longevity was not adversely affected. Defoliation was aggravated in Den. Red Emperor ‘Prince’ by longer duration of cooling and was considered a detrimental effect of low temperature holding.     

茶香月季新品种‘醉红颜’

 ‘醉红颜’是以‘巨型美地兰’ב第一玫瑰红’为亲本育成的茶香月季新品种,花亮粉色, 重瓣,平均花径为13.9 cm,香气浓郁,对黑斑病有一定抗性,适宜以北京气候特点为主的华北地区庭院 栽培。     

Anthocyanins from flowers of Hippeastrum cultivars

The anthocyanins, cyanidin 3-O-(6″-O-α-rhamnopyranosyl-β-glucopyranoside) (1) and pelargonidin 3-O-(6″-O-α-rhamnopyranosyl-β-glucopyranoside) (2), were isolated from the ornamental flowers of a Ugandan Hippeastrum cultivar by a combination of chromatographic techniques, and their structures were elucidated mainly by the use of homo- and heteronuclear nuclear magnetic resonance spectroscopy and electrospray mass spectrometry. The same anthocyanins were found in six different Hippeastrum cultivars purchased in Norway. However, the absolute amount of the anthocyanins (0.08–1.79 mg/g, fresh weight) and the relative proportions of the individual anthocyanins varied from cultivar to cultivar (13.2–96.5% of 1). The colours of the fresh petals of the three cultivars ‘Red Lion’, ‘Royal Velvet’ and ‘Liberty’ were described by the CIELab coordinates L^* (lightness), C^* (chroma) and h_ab (hue angles). All the cultivars showed hue angles corresponding to scarlet nuances (h_ab = 22–35°), with the highest value in ‘Red Lion’. The most reddish petals (in ‘Royal Velvet’) contained the highest relative proportion of 1. Thus, the in vivo colours of these cultivars seem to be correlated with the relative proportions of individual anthocyanin in the petals.     

Chemically induced floral morphological mutations in two cultivars of Ipomoea purpurea (L.) Roth

Two cultivars of Ipomoea purpurea (Morning Glory) differing in flower colour and corolla morphology were investigated for artificial mutagenesis. The violet blue (VB) cultivar bears flowers with a typical gamopetalous corolla. The red purple (RP) cultivar has flowers with an incised corolla at one or more places at the petal union, i.e. incompletely gamopetalous and has petaloid appendages borne on the outer side. It has been described as petalomaniatic. The petaloid appendages were part and parcel of the petal itself. The mutagens used were ethyl methanesulfonate (EMS), N-methyl-N′-nitro-N-nitrosoguanidine (NMG) and sodium azide (SA). Mutants affecting the petal union and petalomaniatic tendency were discussed. It was possible to induce an incompletely gamopetalous petalomaniatic mutant in cultivar VB as well as the typical gamopetalous corolla morphotype in cultivar RP. Preliminary hybridization studies were made using cv. RP and its white colour mutant as one of the parents and crossed with cv. VB. Hybridization studies revealed that independent loci exist for the incomplete gamopetalous nature of the corolla and its petalomaniatic tendency. The mutants obtained were corolla whorl specific and were described as meristic mutations in I. purpurea.     

Identification of S-alleles in pear (Pyrus communis L.) cv. ‘Rocha’ and other European cultivars

In this work we report the cloning and identification of S-RNase alleles responsible for gametophytic self-incompatibility (GSI) of ‘Rocha’ pear and of 13 other European pear cultivars that might be used as its pollinators. Partial sequences of S-RNase alleles were amplified by PCR with specific primers hybridising in conserved regions of previously identified S-RNase alleles of Pyrus communis, cloned and sequenced and the S-genotype of eight pear cultivars was fully determined. Three cultivars (‘General Léclerc’ (SqSl), ‘Tosca’ (SbSl) and ‘Alexandrine Douillard’ (SbSk)) shared no S-alleles with ‘Rocha’ (SaSe) and shall be totally compatible with this cultivar. None of the cultivars analysed showed an identical amplification pattern to the one observed in ‘Rocha’, so the other cultivars shall be at least semi-compatible. One new allele was identified in P. communis cv. ‘Beurré d’Avril’ (designated as St). The determination of both S-RNase alleles of cvs ‘Rocha’, ‘Beurré Precoce Morettini’ (SeSk) and ‘Tosca’ and the identification of one S-RNase allele in cvs ‘Carapinheira’ (Sb), ‘Amêndoa’ (Se), ‘Pérola’ (Sk) and ‘Beurré d’Avril’ (St) are important contributions for the effort recently developed worldwide to establish groups of sexual compatibility among European pears.     

Fruit set and embryo rescue in crosses using parthenocarpic ‘Mopanshi’ persimmon

The ‘Mopanshi’ persimmon is a seedless, astringent parthenocarpic cultivar that does not produce male flowers. After pollination using four non-astringent cultivars (‘Zenjimaru’, ‘Nishimurawase’, ‘Okugosho’ and ‘Hanagosho’), seeds were produced to different degrees. ‘Mopanshi’ fruits pollinated with ‘Zenjimaru’ produced far more seeds than those pollinated with the other three cultivars. The ratio of abnormal seeds obtained from the fruits pollinated with ‘Hanagosho’ was higher than that obtained from the fruits pollinated with the other three cultivars. Most embryos degenerated in the early to late stages of seed development. Immature embryos were cultured in a modified MS medium (half of NO₃ in MS medium + 0.4 μM BA + 0.1 μM IBA) with the greatest success (52–80%) from embryos taken from fruits 60–80 days after pollination. Seedlings failed to initiate radicles so they were transferred to dark culture conditions for 8 days or to a rooting media that contained 3% sucrose and 1% Chinese ink. The seedlings on the medium darkened with ink rooted at greater than 90.83% compared to 75.83% for dark cultured seedlings. This study demonstrated that ‘Mopanshi’ persimmon could be used as a female parent in crosses, but embryos needed to be moved to tissue culture conditions to continue to develop and form plantlets.     

Effect of distance from early flowering pollinizers ‘Maypole’ and ‘Dolgo’ on ‘Fiji’ fruit set

An apple orchard consisting of a single cultivar under the condition of natural pollination must have suitable pollinizers such as Crab apples to ensure stable fruit production. We selected ‘Maypole’ and ‘Dolgo’ as pollinizers for the cultivar ‘Fuji’, and investigated the rate of fruit and seeds in ‘Fuji’ fruits produced by pollen of the pollinizers. We developed a method for tracing pollen flow based on the leaf color of progeny and S-RNase allele of ‘Maypole’, and on Simple Sequence Repeat (SSR) analyses of ‘Maypole’ and ‘Dolgo’. These were powerful tools for determining the distance insects (mainly Osmia cornifrons) carry pollen from the pollinizers to ‘Fuji’. Although the fruit set of ‘Fuji’ apples was not reduced with increasing distance between ‘Fuji’ apple trees and pollinizers (probably due to pollen flow from other commercial cultivars planted outside the area), the rate of ‘Fuji’ apple fruit produced by the pollen of the pollinizers decreased with increasing distance. The rate of fruit produced by the pollinizers was 84% and 77% when ‘Fuji’ was 2.5 m from ‘Maypole’ and ‘Dolgo’, respectively, and 71% and 64% when ‘Fuji’ was 5 m from ‘Maypole’ and ‘Dolgo’, respectively, but was reduced to 47% and 39% when ‘Fuji’ was 10 m from ‘Maypole’ and ‘Dolgo’, respectively. However, the spacing could cause reduced fruit size, and require extra fruit thinning for producing large fruits. It could also cause reduction of the yield for increasing the planting area of pollinizers. We recommend that pollinizers should be planted not more than 10 m from ‘Fuji’.     

Comparison of floral ontogeny in wild-type and double-flowered phenotypes of Syringa vulgaris L. (Oleaceae)

A comparative study of floral ontogeny in normal and double-flowered phenotype of Syringa vulgaris was conducted using the epi-illumination light microscopy. In the wild phenotype, floral differentiation starts with calyx inception and the formation of four sepals in orthogonal positions (two median and two lateral). The corolla emerges as a continuous ring-like structure leading to the appearance of four petal lobes alternating with the sepals. Androecium was formed by initiation of two stamen primordia in transverse plane and finally the bicarpellate gynoecium emerges in median position. In the case of the double-flowered lilac, there are supernumerary petals in an additional whorl. In double-flowered phenotype, a ring meristem is formed acropetally after the first petal whorl initiates. Stamens and carpels initiated similarly in double-flowered as well as in wild-type plants. However, position of stamens deviates from the typical transverse situation. It seems that the appearing of an extra petal whorl did not exhibit any adverse effect on the initiation of other whorls, in terms of organ identity. Therefore, it would be suggested that the double-flowered phenotype of Syringa represents a case of neoheterotopy, with formation of an extra petal whorl, rather than a case of homoheterotopy with transformation of an ancestral androecium whorl into petals.     

‘Swingle’ citrumelo propagation by cuttings for citrus nursery tree production or inarching

‘Swingle’ citrumelo [Citrus paradisi MacFaden × Poncirus trifoliata (L.) Raf.] has been extensively used as a rootstock in several citrus growing regions of the World, including Southern Brazil where ‘Rangpur’ lime (Citrus limonia Osbeck) is still the predominant variety despite being affected by several important pathogens. In this case, ‘Swingle’ citrumelo is used to produce nursery trees to establish new orchards or to be inarched in adult and healthy groves in order to change the rootstock. We report herein a system to produce trees on ‘Swingle’ citrumelo more rapidly by budding onto non-rooted cuttings, as well as assessing potential to rapidly multiply ‘Swingle’ through rooting of non-budded cuttings. Therefore, two potential products are described: budded trees and rooted rootstock cuttings. ‘Valencia’ sweet orange [Citrus sinensis (L.) Osbeck] was budded at different heights on cuttings derived from eight-month old rootstocks. Grafted and additional non-budded cuttings were then treated with indole-3-butyric acid (500 mg L⁻¹) or left untreated before rooting. Three types of cuttings were evaluated: softwood, semi-hardwood and hardwood. The use of nursery trees derived from pre-budded hardwood cuttings of ‘Swingle’ citrumelo is an alternative grafting method on this cultivar. Softwood cuttings with one leaf pair were considered the most adequate material for rapid multiplication of ‘Swingle’ citrumelo by cutting. This could be particularly useful for inarching production or conventional budding after transplant of cutting-derived rootstocks.     

Improvement of flower color by means of leaf treatments in lily

Flower color, an important feature biologically and commercially, is based on four natural pigments – flavonoids, carotenoids, betalains and chlorophylls. Temperature, light, nutrition – as well as additions of sugar, salt, or metals to the conservation water – have an effect on pigmentation. We investigated the effects of K-sulphate and/or sucrose on flower color in leaf treatments applied 30–10 days before harvest to four Asiatic lily (Lilium × elegans Thunb.) cultivars during the winter and summer. Colors of tepals were evaluated by a portable spectrocolorimeter that calculates the standard CIE L*a*b* coordinates and the color differences (E). After leaf treatments during both seasons, cultivars with flowers with high red components (e.g. the purple ‘Fangio’ and the pink ‘Brindisi’) showed significant improvements in color quality. The orange-flowered ‘Tresor’ showed improvement only if K-sulphate and Mix (K-sulphate and sucrose) solution treatments were applied during the summer. The yellow-flowered ‘Menorca’ was not affected by treatments during either summer or winter forcing season. Especially in the winter, a significant reduction in flower abortion was observed for cut flowers of all cultivars. In summer only ‘Fangio’ and ‘Tresor’ showed a reduction in flower abortion. Also, flower size and longevity were improved by the leaf treatment. The results, obtained from a commercial nursery operation, demonstrate that lily growers can adopt a very simple and inexpensive treatment to improve important qualitative traits of their product.     

一个矮牵牛花器官发育突变体aps的表型鉴定及遗传分析

在利用矮牵牛W138株系构建突变体库的过程中,获得了一种花器官发育突变体,该突变体主要表现为花萼和花瓣发育异常,命名为aps(abnormal petals and sepals)。与正常W138植株相比,aps突变体最明显的表型是花冠开裂,通常为3裂,同时5个花萼中有2个出现部分瓣化。扫描电镜观察结果显示,突变体瓣化花萼的表皮细胞表现为正常花瓣的细胞形态。通过突变体自交、与正常姊妹株和矮牵牛W115株系杂交以及F1自交和回交等遗传分析发现,aps突变性状为单基因控制的隐性性状。q PCR分析结果表明,在突变体花萼中,3个B类基因(FBP1、PMADS1和PMADS2)和部分E类基因(FBP2、FBP4和FBP5)出现异位表达或表达量显著升高,而所有A类基因(FBP26、FBP29、PFG、AP2A、AP2B和AP2C)的表达量均明显下降,E类基因FBP9和FBP23的表达量也下调;突变体花瓣中,A类基因除FBP29不表达外,其他所有基因的表达量均显著增加,B类基因FBP1和PMADS2,E类基因FBP2、FBP5、FBP23和PMADS12的表达量上调;C类和D类基因的表达在突变体花萼和花瓣中没有明显变化,但在雄蕊和花柱中C类基因表达量明显下降,D类基因FBP7在子房中显著下调。aps突变体为研究矮牵牛花器官发育的调控机制及相关基因之间的调控关系提供了良好的材料。     

Horticultural performance of ‘Folha Murcha’ sweet orange onto twelve rootstocks

Despite its outstanding position, the Brazilian citriculture is established on a very limited pool of varieties that limits its expansion and restricts the fruit availability throughout the year. This situation determines the urgent necessity of developing alternative scion and rootstock cultivars, with good performance under local conditions. ‘Folha Murcha’ sweet orange (Citrus sinensis (L.) Osbeck) is a late-harvest cultivar, suitable both for the juice processing industry and the fresh fruit market, being described as tolerant to citrus canker (Xanthomonas citri subsp. citri Schaad et al.), and less affected by citrus variegated chlorosis (Xylella fastidiosa Wells et al.). A study was conducted in Bebedouro, São Paulo State, Brazil, to evaluate the horticultural performance of ‘Folha Murcha’ sweet orange budded onto 12 rootstocks: the citrandarin ‘Changsha’ mandarin (Citrus reticulata Blanco) × Poncirus trifoliata ‘English Small’; the hybrid ‘Rangpur’ lime (Citrus limonia Osbeck) × ‘Swingle’ citrumelo (P. trifoliata (L.) Raf. × Citrus paradisi Macfad.); the trifoliates (P. trifoliata (L.) Raf.) ‘Rubidoux’, ‘FCAV’, and ‘Flying Dragon’ (P. trifoliata var. monstrosa); the ‘Sun Chu Sha Kat’ mandarin (C. reticulata Blanco); the ‘Sunki’ mandarin (Citrus sunki (Hayata) Hort. ex. Tanaka); the ‘Rangpur’ limes (C. limonia Osbeck) ‘Cravo Limeira’ and ‘Cravo FCAV’; ‘Carrizo’ citrange (C. sinensis × P. trifoliata), ‘Swingle’ citrumelo (P. trifoliata × C. paradisi), and ‘Orlando’ tangelo (C. paradisi × Citrus tangerina cv. ‘Dancy’). The experimental grove was planted in 2001, using a 7 m × 4 m spacing, in a randomized block design, with five replications and two plants per plot. No supplementary irrigation was applied. Fruit yield, canopy volume, tree tolerance to drought and to citrus variegated chlorosis, and fruit quality were assessed for each rootstock. Trees grafted onto the ‘Flying Dragon’ trifoliate were smaller in size, but had largest yield efficiency when compared to those grafted onto other rootstocks. Lower alternate bearing index was observed on trees budded onto ‘Cravo FCAV’ ‘Rangpur’ lime. Both ‘Rangpur’ lime rootstocks and the ‘Sunki’ mandarin induced higher tree tolerance to drought. The ‘Flying Dragon’ trifoliate induced better fruit quality and higher tolerance to citrus variegated chlorosis (CVC) to ‘Folha Murcha’ trees. A cluster multivariate analysis identified three groups of rootstocks with similar effects on ‘Folha Murcha’ tree performance. Among the 12 evaluated rootstocks, the ‘Flying Dragon’ trifoliate has a unique effect on plant growth, tolerance to drought and CVC, fruit yield and fruit quality of ‘Folha Murcha’ trees, and may be better suited for high-density plantings.     

Identification of organic acid-related genes and their expression profiles in two pear (Pyrus pyrifolia) cultivars with difference in predominant acid type at fruit ripening stage

‘Yandangxueli’ is a pear cultivar with predominant citric acid in the ripe fruit, different from most of pear cultivars such as ‘Gengtouqing’ in which malic acid is the predominant acid type. It was found that ‘Yandangxueli’ accumulated citric acid for three times against that in ‘Gengtouqing’ at fruit ripening stage. To investigate the mechanism of citric acid accumulation in ‘Yandangxueli’, organic acids content, gene expression and enzyme activity were studied in both cultivars. Five genes, Pp:mtCs, Pp:cyAco, Pp:cyIdh, Pp:mtMdh and Pp:cyMe which encoded citric synthase (CS), cytosolic aconitase (cyACO), NADP-dependent isocitrate dehydrogenase (NADP-IDH), NAD-dependent malate dehydrogenase (NAD-MDH) and NADP-dependent malic enzyme (NADP-ME) respectively, were identified from pear fruit. Their expression profiles and the corresponding enzyme activities were determined throughout fruit development in both cultivars. Results from these enzymes indicated that there were no strict relationship between gene expression, enzyme activity and citric acid accumulation. Expression analysis for two Py:vVAtp genes encoding vacuolar H⁺-ATPase A subunit and one Py:vVpp gene encoding Vacuolar H⁺-pyrophosphatase showed that they were all with up-regulated expression at the later development stage of ‘Yandangxueli’ but with down-regulated expression in ‘Gengtouqing’. Therefore, it is concluded that the different ability in citric acid transportation and storage might be involved in the high citric acid content in ‘Yandangxueli’.     

Comparative forcing of Hydrangea macrophylla ‘Bailer’ as a florist's hydrangea

‘Bailer’ (Endless Summer™) is a new, pink- or blue-flowered hydrangea that flowers on new wood continuously throughout the growing season. It is also winter-hardy in northern, temperate climates (USDA Z4). Use of Endless Summer as a florist's potted hydrangea would provide consumers with a dual-use product (flowering potted plant, landscape shrub). The objectives were to determine if Endless Summer could be forced as a florist's hydrangea using two forcing regimes (immediate, standard), two soil pH regimes, and two pinching treatments. ‘Merritt Supreme Pink’ (pink), ‘Blue Danube’ (blue), and Endless Summer (blue, pink) were forced under immediate (no cold treatment, short- and long-day photoperiods) and standard (6 weeks cold, 4 °C to overcome dormancy) conditions. Since commercial liners were not yet available, cuttings of Endless Summer were used in the immediate forcing experiment. Days to visible flower bud, first color, full flower were recorded, as well as height, no. of branches, no. of flowers, and flower size. In the immediate forcing experiment, only Endless Summer produced flowers. Cultivars differed significantly (P < 0.001) for days to visible flower bud, days to first color, days to full flower, and height. The number of flowers and flower size were not significantly different among pink cultivars. In the standard forcing experiment, Endless Summer reached days to visible flower bud, days to first color, and days to full flower significantly earlier than either comparison, although it exhibited weaker stems and less intense flower pigmentation. Pinching (blue pH) had a significant effect on all traits except the days to full flower and flower size, although no pinching treatment was significant for pink pH. Photoperiod was significant only for the days to visible flower bud (pink pH), days to full flower (blue pH), and final height (blue pH). Further research is needed before Endless Summer can be grown as a dual-use florist's potted hydrangea with acceptable quality.     

Syrian pear (Pyrus syriaca) as a pollinator for European pear (Pyrus communis) cultivars

In Israel four European pear cultivars are grown: ‘Spadona’ is the main cultivar and ‘Coscia’, ‘Gentile’ and ‘Spadochina’ are its pollinators. However, molecular S-genotyping revealed that ‘Spadona’ is semi-compatible with its three pollinators. This explains, at least in part, the relatively low pear yield in Israel. The Syrian pear (Pyrus syriaca) grows wild in Israel and blooms intensively, overlapping the blooming of the cultivated European pears. Cross-fertilization between Syrian pear and ‘Spadona’ was shown to be efficient suggesting that Syrian pear might be a potent pollinator for ‘Spadona’. Twenty-six Syrian pear seedlings, from different sites in north-east Israel were S-genotyped identifying 11 that are fully compatible with the four European pear varieties cultivated in Israel. By this screening, 24 different S-RNases were cloned; ten of them are new, whereas the other fourteen had been identified previously. In addition, seedlings of two wild pear species were also S-genotyped. Two seedlings from Pyrus betulifolia and one from Pyrus korshinskii were found to be genetically compatible with the four European pear cultivars. From these seedlings four S-RNases were cloned, two are new, one had been cloned previously and one was identical to an S-RNase allele cloned from Syrian pear in this work.     

Cybrid/hybrid plants regenerated from somatic fusions between male sterile Satsuma mandarin and seedy tangelos

Somatic hybridization provides an alternative for transferring mitochondria-encoded cytoplasmic male sterility (CMS). Herein, symmetric protoplast electrofusion was conducted between embryogenic callus protoplasts of Citrus unshiu Marc. cv. Guoqing No. 1 (G1), a CMS cultivar, and mesophyll-derived protoplasts of seedy ‘Page’ tangelo [C. reticulata Blanco × (C. reticulata Blanco × C. paradisi Macf.)] or ‘Nova’ tangelo [C. reticulata Blanco × (C. reticulata Blanco × C. paradisi Macf.)], to transfer CMS trait. Flow cytometry analysis showed that 14 plants recovered from G1 + ‘Page’ tangelo that displayed typical morphological character of ‘Page’ were diploid, and 6 plants regenerated from G1 + ‘Nova’ tangelo were tetraploid. Genetic compositions of regenerated plants from the two fusions were determined by SSR, CAPS and chloroplast-SSR analysis. Cybrid nature of diploids from G1 + ‘Page’ tangelo with nuclear DNA from ‘Page’, mitochondrial DNA (mtDNA) from the G1 and chloroplast DNA (cpDNA) derived from either parent was confirmed. Tetraploid plants from G1 + ‘Nova’ tangelo were identified as somatic hybrids with random cpDNA inheritance. The regenerated cybrid and hybrid plants hold great potential for Citrus seedless breeding at diploid or triploid levels.