Conversion of a male-specific RAPD marker into an STS marker in Asparagus officinalis L.

Garden asparagus (Asparagus officinalis L.) is an economically important plant. This species is dioecious, and male plants are considered to be more desirable than females due to their higher yields. To reduce the time required for asparagus breeding, molecular marker techniques have been employed to identify sex-linked DNA markers. In the present study, we converted the male-specific random amplified polymorphic DNA marker T35R54-1600 into a sequence tagged site marker. We cloned a male-specific DNA fragment amplified with the T35R54 primer and determined the sequence of the fragment. The size of T35R54-1600 is 1,586 bp, and this fragment is not homologous to known sex-linked BAC sequences, indicating that this fragment is a new sex-linked region. Within this fragment, we designed the primer pair ‘MSSTS710’ to amplify a 710 bp region. This marker could be used to identify the sex of eight cultivars of A. officinalis: ‘Mary Washington 500W’, ‘UC157’, ‘Harumachi Green’, ‘Super Welcome’, ‘F4’, ‘Pacific 2000’, ‘F2’ and ‘Backlim’. We also analyzed the applicability of this marker to two dioecious Asparagus species, A. schoberioides and A. kiusianus, which are cross-compatible with A. officinalis. Although male-specific DNA fragments of two dioecious Asparagus species, A. schoberioides and A. kiusianus, were generated using the existing male-specific marker Asp1T7sp, no amplicon was obtained using the MSSTS710 marker. Since MSSTS710 can be employed for sex identification only in A. officinalis and not in closely related Asparagus species, the DNA region around the MSSTS710 marker must be variable among Asparagus species.     

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.     

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.     

Meiotic chromosome pairing in intergeneric hybrids of colchicaceous ornamentals revealed by genomic in situ hybridization (GISH)

Diploid and triploid intergeneric hybrids obtained by crosses among Gloriosa superba ‘Lutea’ (2n = 2x = 22), G. ‘Marron Gold’ (2n = 4x = 44), Littonia modesta (2n = 2x = 22), and Sandersonia aurantiaca (2n = 2x = 24) were analyzed for their meiotic chromosome pairing in pollen mother cells by genomic in situ hybridization (GISH) with digoxigenin-labeled total DNA of one parent as probe. Chromosomes from each parent could be clearly distinguished in pollen mother cells of all the five intergeneric hybrids by GISH. For three diploid hybrids, L. modesta × G. superba ‘Lutea’ (2n = 2x = 22), L. modesta × S. aurantiaca (2n = 2x = 23) and S. aurantiaca × G. superba ‘Lutea’ (2n = 2x = 23), 0.04?0.27 autosyndetic bivalents (intragenomic pairing of non-homologous chromosomes) and 0.13?0.36 allosyndetic bivalents (intergenomic chromosome pairing) were observed per pollen mother cell, indicating that there are some homologous chromosomal regions within each genome and among the genomes of Gloriosa, Littonia and Sandersonia. Differences in the average number of allosyndetic bivalents per pollen mother cell among different genome combinations may reflect the evolutionary distances among the three genera, and Gloriosa and Littonia may be closely related to each other, while Sandersonia may have relatively distant relationships with Gloriosa and Littonia. For two triploid hybrids, L. modesta × G. ‘Marron Gold’ (2n = 3x = 33) and S. aurantiaca × G. ‘Marron Gold’ (2n = 3x = 34), no allosyndetic bivalents were observed. Based on the results obtained in the present study, possible utilization of the diploid and triploid intergeneric hybrids for further breeding of colchicaceous ornamentals is discussed.     

Genomic in situ hybridization (GISH) analysis of intergeneric hybrids in Colchicaceae

Some intergeneric hybrids produced by the crosses among several colchicaceous ornamentals, Gloriosa superba ??Lutea?? (2n = 2x = 22), G. ??Marron Gold?? (2n = 4x = 44), G. ??Verschild?? (2n = 7x = 77), Littonia modesta Hook. (2n = 2x = 22), and Sandersonia aurantiaca Hook. (2n = 2x = 24), were subjected to genomic in situ hybridization (GISH) analysis in order to clarify their genome constitutions. Chromosome preparation was made from root tip cells of L. modesta × G. superba ??Lutea??, L. modesta × S. aurantiaca, S. aurantiaca × G. superba ??Lutea??, L. modesta × G. ??Marron Gold??, S. aurantiaca × G. ??Marron Gold?? and G. ??Verschild?? × S. aurantiaca. Total DNA of one parent was labeled with digoxigenin or biotin and used as probe, and chromosomes were counterstained with 4??-6-diamidono-2-phenylindole (DAPI). For all the nine intergeneric hybrids, chromosomes from each parent could be clearly distinguished by GISH analysis. Thus GISH analysis is a powerful tool for identifying the genome constitution of intergeneric hybrids in colchicaceous ornamentals. The results obtained by GISH analysis study may be important for further progress in breeding of colchicaceous ornamentals.     

A new DNA marker for sex identification in purple asparagus

Garden asparagus (Asparagus officinalis L.) is an economically-important perennial crop. This plant is dioecious, as there are both male and female individuals; male individuals are preferred over females for agricultural production. To reduce the time required for garden asparagus breeding, various male-specific DNA markers are utilized. Male-specific DNA markers, such as Asp1-T7sp and MSSTS710, are currently available for sex identification in many asparagus cultivars. In the current study, we found that these markers are not suitable for sex identification in the purple asparagus cultivar ‘Pacific Purple’, as male-specific amplification of this marker was detected in some male individuals of this cultivar but not in other males. The Asp1-T7sp marker is suitable for use in sex identification in various Asparagus species related to A. officinalis, indicating that the region around this marker is conserved among these species. Thus, we isolated a DNA fragment around this marker by inverse PCR and produced a new DNA marker, MspHd, based on this sequence. However, like Asp1-T7sp and MSSTS710, MspHd was not suitable for sex identification in the cultivar ‘Pacific Purple’. Since all ‘Pacific Purple’ males have morphologically similar male flowers with functional stamens, we produced a new male-specific marker based on the sex determination gene, MSE1/AspMYB35/AspTDF1, which is responsible for stamen development. This marker, named AspMSD, is suitable for sex identification in ‘Pacific Purple’. In addition, this marker can be utilized for sex identification in various asparagus cultivars and some related Asparagus species.     

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.     

Breakdown of a hybridization barrier between Solanum pinnatisectum Dunal and potato using the S locus inhibitor gene (Sli)

Several attempts have been undertaken to exploit the resistance to late blight [Phytophthora infestans (Mont.) de Bary] of the Mexican diploid Solanum pinnatisectum Dunal (2n = 2x = 24). However, sexual hybridization between S. pinnatisectum and cultivated potatoes is almost impossible because of strict reproductive barriers. In this study, we first induced chromosome doubling of S. pinnatisectum for matching of the species-specific endosperm balance number (EBN), which is a genetic phenomenon responsible for normal endosperm development. We pollinated 4x (2EBN) S. pinnatisectum flowers with pollen from 2x (2EBN) clones carrying the S locus inhibitor gene (Sli), which originally was identified as an inhibitor of S-locus-controlled self-incompatibility in pollen of diploid potatoes. Nine putative hybrid plants were obtained from 613 pollinations, of which two plants were characterized. Both individuals were triploid hybrids with extreme resistance to P. infestans derived from S. pinnatisectum and showed an acceptable level of female fertility for backcrossing. Thus, utilization of the Sli gene, combined with chromosome-doubling for matching of the EBN, is proposed as a technique to overcome barriers to interspecific hybridization between Mexican 2x (1EBN) Solanum species and cultivated diploid potatoes.     

Novel interspecific hybridization between sweetpotato (<Emphasis Type="Italic">Ipomoea batatas</Emphasis> (L.) Lam.) and its two diploid wild relatives

Interspecific hybridization is an important approach to broaden the genetic base and create novel plant forms in breeding programs. However, interspecific hybridization in Ipomoea is very difficult due to the cross incompatibility. Here we report two novel interspecific F₁ hybrids between I. batatas (L.) Lam. (2n = 6x = 90) and two wild species, I. grandifolia (2n = 2x = 30) and I. purpurea (2n = 2x = 30). Hybridization was stimulated by applying plant growth hormones. Morphological, molecular and cytological tests were conducted to confirm their hybridity. We found that the two hybrids were quite distinctive in leaf color and morphology, and yielded intermediate sizes of storage roots compared to their respective parents. Inter-simple sequence repeat analysis showed that the unique DNA bands from the wild parents could be detected in these two hybrids. The cluster analysis also showed that the two F₁ hybrids were closer to I. batatas in phylogeny relationship. The number of chromosomes of the two hybrids was both 60, indicating that the hybrids were tetraploid. The meiotic configuration analysis of the H₁ of I. batatas × I. grandifolia revealed the occurrence of 17.58I + 14.28II + 1.36III + 2.48IV at metaphase I in average chromosome association per pollen mother cells (PMCs), 4.26I + 18.32II + 2.56III + 3.12IV was average meiotic configuration in the H₂ of I. batatas × I. purpurea. Both hybrids appeared to be polyads and multi-microcytes at tetrad phase and differed in their pollen fertility.     

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.     

Creation of long-day adapted tetraploid potatoes from crosses of <Emphasis Type="Italic">Solanum tarijense</Emphasis> females with Andigena males

T-type is a common chloroplast DNA type among modern potato varieties (Solanum tuberosum), their progenitor Chilean tuberosum, and a diploid wild species S. tarijense. To recreate Chilean tuberosum, we made 983 pollinations between 10 accessions of T-type chloroplast DNA-holding S. tarijense used as females and 32 Andean tetraploid landraces (Andigena) used as males, from which 14 tetraploid hybrids were obtained. These interspecific hybrids grew vigorously with long stolons, flowered well with high male and female fertilities, and matured 1 month later than modern varieties. Seedlings of selfed and sib-crossed interspecific hybrids were artificially selected for tuber yield under long days. The selected clones were grown in the field, of which two clones produced over 1 kg of edible tubers per plant. These results lend experimental support to the hypothesis that the Chilean tuberosum originated by selection for long-day adaptability from tetraploid hybrids that occurred by fertilization of a 2n egg of S. tarijense and n pollen of Andigena.     

Interspecific hybridization in sexual diploid Allium senescens var. minor× apomictic tetraploid A. nutans and sexual diploid A. senescens var. minor× apomictic hexaploid A. senescens

Fourteen interspecific hybrids in sexual diploid Allium senescens var. minor× apomictic tetraploid Allium nutans L. crosses, and eight interspecific hybrids in sexual diploid A. senescens var. minor× apomictic hexaploid A. senescens L. crosses were produced. The number of chromosomes was 2n= 24 in interspecific hybrids of diploid × tetraploid, and 2n= 32 in diploid × hexaploid crosses. Triploid and tetraploid interspecific hybrids showed intermediate parental morphological characteristics. Tetraploid interspecific hybrids of A. senescens var. minor×A. senescens crosses formed two groups based on leaf colour and leaf width. Seeds were formed in 11 out of 14 triploid interspecific hybrids under natural conditions. In cytological observations of parthenogenesis, three out of 12 triploid interspecific hybrids and five out of eight interspecific tetraploid hybrids were observed. Parthenogenesis ranged from 26.0% to 86.0% in five tetraploid interspecific hybrids. Non-parthenogenesis to parthenogenesis segregated in a 3:5 ratio in A. senescens var. minor×A. senescens crosses.     

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.     

Potential for analytic breeding in allopolyploids: an illustration from Longiflorum × Asiatic hybrid lilies (<Emphasis Type="Italic">Lilium</Emphasis>)

Ploidy level and intergenomic recombination was studied in interspecific hybrids between Longiflorum × Asiatic lilies (LA hybrid) backcross to Asiatic parents in order to assess the possibility for analytic breeding in lily. By backcrossing the diploid (2n = 2x = 24) F1 interspecific hybrid between Longiflorum × Asiatic lilies to Asiatic parents, 104 BC1 progeny plants were produced. Among these, there were 27 diploids, 73 triploids (2n = 2x = 36) and 4 aneuploids (2x − 1, 2x + 2 or 2x + 3). In addition, by backcrossing triploid BC1 (LAA) plants to diploid Asiatic parents in 2x − 3x and reciprocal combinations, 14 diploid BC2 progenies were produced. Genomic in situ hybridization (GISH) was performed to study the intergenomic recombination and karyotype composition. GISH indicated extensive intergenomic recombination among the chromosomes in LA hybrids. A large number of Longiflorum chromosomes were transmitted to the BC1 progenies from LA hybrids. However, very few Longiflorum chromosomes were transmitted from the BC1 triploid (LAA) plants to the BC2 progenies. The occurrence of diploid plants in the BC progenies of LA hybrids has opened the prospects of analytic breeding in lilies. In this approach, the selection of superior genotypes can be carried out at the diploid level and polyploid forms are synthesized from superior diploid parents. The advantages of analytic breeding are evident: (a) a maximum level of heterozygosity can be attained in the synthetic polyploids and (b) introgression can be achieved with a minimum of linkage drag. Based on GISH results the potential application of analytic breeding in lily allopolyploids has been discussed.     

Meiotic studies of spontaneous hybrids of Amaranthus: genome analysis

The meiotic behaviour of 13 spontaneous interspecific F₁ hybrids of Amaranthus was studied. The hybrids between species with n= 16 chromosomes had 16 bivalents but varied considerably in pollen stainability (0–55%). These results suggest the existence of cryptic structural hybridity. The hybrids involving A. cruentus (n = 17) and species with n = 16 (A. caudatus and A. quitensis) always formed 15II+1 III with very low pollen stainability (5–7%). Further observations indicated that Amaranthus species are allotetraploids with basic numbers of x= 8 and x= 9 but exhibit x= 16 and x= 17 as secondary basic numbers, as demonstrated by (a) the frequent presence of 811 + 171 in the meiosis of the hybrid A. spinosus (n = 17) × A. hybridus (n= 16); and the occurrence of secondary associations between bivalents in MI. Genomic formulae are proposed for each species, on the basis of the meiotic behaviour of the hybrids studied.     

The generation of novel species hybrids between garden dahlias and Dahlia macdougallii to increase the gene pool for variety breeding

To transfer new traits into the gene pool of garden dahlias (Dahlia variabilis), crosses between garden dahlias (2n = 64) and the epiphytic species Dahlia macdougallii (2n = 32) from the section Epiphytum were conducted. Six hybrid plants were obtained. The hybrid status was verified using three SSR markers. In addition, flow cytometry was performed to determine the genome size of the hybrids and to show that the hybrids were hexaploid as expected from crosses between tetraploids and octoploids. The open pollinated progeny of the hybrids produced four progeny with octoploid genomes. The hybrids exhibited indeterminate vegetative growth and the formation of flowers from axillary buds, similar to the father D. macdougallii. This result is of interest for breeding new varieties of dahlia with traits that are not present in the current gene pool.     

Development of a scoring scale for powdery mildew (Golovinomyces cichoracearum (DC.) V.P. Heluta) disease and identification of resistance sources in cultivated and wild sunflowers

Powdery mildew incited by Golovinomyces cichoracearum has become a serious problem on sunflower in India during the past 2–3 years. Genetic resistance in the released cultivars and the parental lines of hybrids is rather limited. Hence, screening of about 420 accessions comprising of wild Helianthus species, interspecific derivatives, core germplasm, inbred lines and few exotic accessions was done under natural field conditions for 2 years and further confirmed by screening under artificial inoculation conditions. PCR analysis using primers specific to powdery mildew causing genera gave a 391 bp band which confirmed the pathogen as G. cichoracearum. Seven different screening methods were tested which induced infection, but dusting of spores on to the healthy leaves proved to be convenient and more effective method of infection. Based on the differential response of the accessions derived from diverse genetic backgrounds, a scale for obtaining reliable estimates of the disease has been devised. Among different cultivar germplasm accessions, the disease severity index (DSI) ranged from 15 to 100 and area under disease progression curve (AUDPC) ranged from 95 to 648. Among the four groups of cultivated sunflower accessions tested, DSI and AUDPC was in the order of exotic lines < interspecific derivatives < inbred lines < core germplasm. Reliable sources of resistance to the pathogen were identified in four annual wild species (H. argophyllus, H. agrestis, H. debilis, H. praecox), six perennials (H. angustifolius, H. atrorubens, H. rigidus, H. salicifolius, H. pauciflorus and H. resinosus), two interspecific derivatives (HIR-1734-2, RES-834-3) and two exotic lines (PI 642072, EC-537925).     

Morphological,cytological and reproductive characterization of tri-species hybrids (GOS) between <Emphasis Type="Italic">Pennisetum glaucum</Emphasis>, <Emphasis Type="Italic">P. orientale</Emphasis> and <Emphasis Type="Italic">P. squamulatum</Emphasis>

New tri-species hybrids (GOS) in the genus Pennisetum involving the cultivated species pearl millet (P. glaucum L.) and two wild species, viz. P. squamulatum Fresen and P. orientale L. C. Rich, are reported. Six hybrid plants were recovered after crossing a backcross hybrid (2n = 3x = 23, GGO) between P. glaucum (2n = 2x = 14, GG) and P. orientale (2n = 2x = 18, OO) with F₁s (2n = 6x = 42, GGSSSS) between P. glaucum (2n = 4x = 28, GGGG) and P. squamulatum (2n = 8x = 56, SSSSSSSS). The hybrids were perennial, morphologically intermediate to their parents, and represented characters from the three contributing species. The hybrids contained 2n = 44 chromosomes (GGGSSO) representing 21, 14 and nine chromosomes from P. glaucum, P. squamulatum and P. orientale, respectively. Meiotic and flow-cytometric analysis suggested origin of these hybrids from unreduced female and reduced male gametes. Average chromosome configuration (8.42_I + 14.32_II + 1.62_III + 0.52_IV) at Meiosis showed limited inter-genomic pairing indicating absence of significant homology between the three genomes. The hybrids were male sterile (except one) and highly aposporous. P. orientale was identified to induce apospory in hybrid background with P. glaucum at diploid and above levels, though it was quantitatively affected by genomic doses from sexual parent. A case of inducible and recurrent apospory is presented whereby a transition from Polygonum-type sexual embryo-sacs to Panicum-type aposporous embryo-sacs was observed in diploid interspecific hybrids. Results supported independent origin and partitioning of the three apomixis-components (apomeiosis, parthenogenesis, and functional endosperm development), reported for the first time in Pennisetum. Potential utilization of GOS hybrids in understanding genome interactions involved in complex traits, such as perenniality and apomixis, is discussed.     

A method for sex identification in asparagus using DNA from seeds

Asparagus (Asparagus officinalis L.) is a dioecious species, with both male and female individuals. Male plants are more desirable to cultivate than female plants because they have higher yields, and, unlike female plants, they do not have a weed problem resulting from fallen seeds. A male-specific DNA marker is currently available to identify the sex of asparagus individuals using total DNA extracted from cladodes and roots. However, no published method is currently available for DNA extraction and PCR amplification from asparagus seeds. In this study, we tested several heat-resistant DNA polymerases for PCR and several methods for extracting DNA from asparagus seeds and successfully established a method for identifying the sex of asparagus seeds using this male-specific DNA marker. We found that PCR amplification of DNA extracted from asparagus seeds using simple methods such as single-step DNA extraction requires the use of high efficiency DNA polymerase. By contrast, many types of heat-resistant DNA polymerases can be used for PCR amplification of high-quality DNA extracted from asparagus seeds using a commercially available DNA extraction kit. Our method for sex identification of asparagus seeds could facilitate quality checking of all-male asparagus seeds and accelerate the screening of super-male asparagus.     

Hybrids of <Emphasis Type="Italic">Passiflora</Emphasis>: <Emphasis Type="Italic">P. gardneri</Emphasis> versus <Emphasis Type="Italic">P. gibertii,</Emphasis> confirmation of paternity,morphological and cytogenetic characterization

Two new varieties of interspecific hybrids of Passiflora have been developed from the cross between P. gardneri versus P. gibertii, both registered under the Passiflora Society International. Twelve putative hybrids were analyzed. Hybridization was confirmed using RAPD and SSR markers. Primer UBC11 (5′-CCGGCCTTAC-3′) generated informative bands. Primer SSR Pe75 has amplified species-specific fragments and a heterozygote status was observed with two parent bands 300 and 350 bp. The molecular markers generated have been analyzed for the presence or absence of specific informative bands. Based on the morphological characterization, we have identified two hybrid varieties: P. ‘Gabriela’ and P. ‘Bella’. P. ‘Gabriela’ produced flowers in bluish tones, bluish petals on the adaxial and abaxial faces, light blue sepals on the adaxial and light green on the abaxial faces, corona with the base of filaments in intense lilac color and white apex. P. ‘Bella’ produced flowers in lilac tones, intense lilac petals on the adaxial and abaxial faces, dark lilac sepals with whitish edges on the adaxial and light green on the abaxial faces, corona with the base of filaments in intense lilac color and white apex. The cytogenetic analysis verified that the hybrids have the same chromosomal number as the parents (2n = 18); the formation of bivalents between the homeologous chromosomes (n = 9) was observad, leading to regular meiosis, which allows the sexual reproduction and use of these hybrids in breeding programs.