Allium sandrasicum is a diploid (2n = 2x = 16) endemic perennial Allium species, which grows naturally on the dry and rocky Mediterranean coast of southwestern Türkiye and the eastern Aegean islands. The bulbs, leaves, and pseudo-stems of A. sandrasicum are collected and consumed fresh or cooked during the spring season. It has potential for use as a food crop and an ornamental plant for landscaping. Experiments were conducted to develop gynogenic and somatic plant regeneration protocols for this endemic species by culturing approximately 16,000 unopened flower buds from six accessions on two BDS (Dunstan and Short 1977) and two MS (Murashige and Skoog 1962)-based induction medium containing 100.0 g L−1 sucrose with or without plant growth regulators (PGRs). Gynogenic plantlet regeneration was induced in the medium with or without PGRs, whereas somatic plantlet regeneration was possible only in the presence of PGRs (2.0 mg L−1 2,4-dichlorophenoxyacetic acid and 2.0 mg L−1 6-benzylaminopurine). Culturing flower buds of accession AS5 in B2 (a PGR-free MS-based medium) resulted in 16.7

Allium cinsi, 750’den fazla türü içeren geniş bir bitki grubudur ve gıda, tıbbi ve süs amaçlı kullanımda önemli bir yere sahiptir. Türkiye, birçok Allium türünün gen merkezi olup, ülkenin farklı bölgelerinde doğal olarak yetişen ya da kültüre alınarak üretilen çok sayıda tür bulunmaktadır. Başlıca ürünler arasında soğan (A.cepa L.) ve pırasa (A.ampeloprasum var. porrum L.) yer almaktadır. Fil sarımsağı (A.ampeloprasum var. holmense), yaygın olarak yetiştirilmese de yenilebilir başları ve süs bitkisi olarak kullanılabilen çiçekleriyle dikkat çekmektedir. Kaya sarımsağı (A.macrochaetum Boiss. & Hausskn.) ve Tunceli sarımsağı (A.tuncelianum Kollman, Özhatay, Mathew, Şiraneci), genellikle doğal ortamlarından toplanmakta veya küçük ev bahçelerinde yetiştirilmektedir. Sirmo (A.vineale L.) ise özellikle otlu peynir üretimi için doğadan toplanmaktadır. Bu çalışmada, incelenen türlerin çiçek sap uzunluğu, umbel çapı, umbel yüksekliği, umbeldeki çiçek tomurcuğu sayısı ve tohum kalite özellikleri değerlendirilmiştir. Türlerin çiçek sap uzunluklarının 79.5-146.15 cm, umbeldeki çiçek tomurcuğu sayısının 29.90-2992.20, umbel genişliklerinin 4.52-15.74 cm ve umbel uzunluklarının ise 4.65-12.83 cm arasında değiştiği tespit edilmiştir. Tohum ile çoğaltılan türlerde 1000 tohum ağırlığı 2.39-3.87 g, 1 gramdaki tohum sayısı 235-458 adet ve çimlenme oranı %72-100 arasında değişim göstermektedir. Bu türlerin, gıda olarak kullanılmalarının yanı sıra çekici çiçek renkleriyle (beyaz, pembe, lila ve mor) süs bitkisi olarak da değerlendirilme potansiyeline sahip olduğu gözlemlenmiştir. Özellikle kentsel tarım kavramının giderek önem kazandığı Türkiye ve dünyada, bu türler hem estetik hem de işlevsel açıdan değerli bir alternatif sunmaktadır.

Drought is widely regarded as the most significant abiotic stressor affecting modern agricultural production. Sweet watermelon (Citrullus lanatus var. lanatus), one of the world’s most cultivated vegetable species, lacks drought resistance and must be grown in dry and semi-arid regions by grafting on tolerant rootstocks. Citron watermelon (Citrullus lanatus var. citroides) is well known for its high drought tolerance and has excellent potential as a watermelon rootstock. There is a high demand for novel citron watermelon cultivars with rootstock potential. Doubled haploid (DH) technology enables the breeding of genetically uniform and fully homozygous plants in a single generation. However, no research has been conducted on the haploidization of citron watermelon using the irradiated pollen technique. The current study aimed to determine the efficiency of gamma-ray sources (Cobalt: Co60 and Cesium: Cs137), irradiation doses (200, 250, and 300 Gy of Co60; 100, 150, 200, and 250 Gy of Cs137), and genotypes (11 citron watermelon lines) on the production of pure citron lines via the irradiated pollen technique (parthenogenesis). The embryos were grown on MS medium supplemented with 0.4 mg/L indole-3-butyric acid. Stomatal observations and flow cytometry were used to conduct ploidy studies on 3–4 weeks old parthenogenic plantlets. The Co60 treatments yielded eight haploid and two mixoploid plants, while the Cs137 treatments yielded 14 haploid and one mixoploid. These findings demonstrated that gamma rays from both radiation sources were effective in parthenogenic embryo induction, and Cs137 was discovered to be a reliable alternative to conventional Co60. The efficiency of gamma-ray sources, irradiation doses, and genotypes were investigated. The parthenogenic embryo induction, and Cs137 was discovered to be a reliable alternative to conventional Co60.

Modern plant breeding studies have focused on the production of F1 hybrid varieties which are tolerant to abiotic stress arising from global climate change. Sweet watermelon (Citrullus lanatus var. lanatus) is susceptible to drought stress, and citron watermelon has unique characteristics, which makes it a potential rootstock for watermelon grafting in water-scarce areas. The present study was conducted to assess whether the production of pure citron lines via anther and ovary culture techniques can improve new F1 hybrid rootstock citron varieties in a short time. However, to date, no successful protocol for anther and ovary culture in citron watermelon has been reported. Therefore, the current study was carried out to shed light on the efficiency of growing media composition in callogenesis, embryogenesis, and plantlet initiation in eleven citron watermelon lines. The combination of plant growth regulator (PGR) and genotype induced varying androgenic and gynogenic responses in terms of the percentages of the induction of callogenesis and embryogenesis. The greatest androgenic and gynogenic potential was observed with the combination of MS + 2 mg/L 2.4-D + 2 mg/L 6-Benzylaminopurine (BAP) + 1 mg/L 1-Naphthaleneacetic acid (NAA) + 30 g/L sucrose (IM1 media) in CW21 and CW4 line, respectively. In total, 11 plants via anther culture and 19 plants via ovary culture were recovered and rooted on the solid MS medium supplemented with 0.4 mg/L indole-3-butyric acid (IBA). The ploidy analysis revealed that all plants were diploid (2n = 22).

Allium tuncelianum (Kollman) Özhatay, Matthew and Şiraneci forms a single-cloved edible white bulb with a mild garlic (A. sativum) odor and taste. Its ability to form seeds makes it suitable for genetic improvement via classical and modern approaches. A detailed study was carried out to determine the gynogenic and somatic plant regeneration potential of two A. tuncelianum accessions (AT1 and AT2). Unopened flower buds of A. tuncelianum accessions were cultured in various BDS (Dunstan and Short, Physiol Plant 41:70-72, 1977) and MS (Murashige and Skoog, Plant Physiol 15:473-497, 1962)-based induction media. Accessions showed slight differences in their responses to gynogenesis and somatic shoot induction cultures. Gynogenic plantlets were obtained in six induction media (T2, T3, T6, T9, T12, and T15). Three of these (T2, T3, and T9) did not contain plant growth regulators (PGRs). Seventeen (0.09

Broccoli (Brassica olearecea var. italica) is a cole crop grown for its floral heads and stalks. It is rich in bioactive chemicals good for human health. Broccoli has been consumed as a vegetable since Roman times, but its production and consumption have increased significantly over the past few decades. Breeders try to develop new broccoli varieties with high yield, improved quality, and resistance to biotic and abiotic stresses. Almost all new broccoli varieties are F1 hybrids. Development of inbred broccoli lines that can be used as parents in hybrid production is a time-consuming and difficult process. Haploidization techniques can be utilized as a valuable support in broccoli breeding programs to speed up the production of genetically pure genotypes. Haploid plants of broccoli can be produced from immature male gametophytes via anther and microspore cultures with similar success rates. The most important parameters affecting the success of haploidization in broccoli are the genetic background (genotype) and the developmental stage of the microspores. Broccoli genotypes differ in their responses to androgenesis induction. The highest androgenesis response could be induced from microspores in late uninucleate and early binucleate stages. Recovery of diploid broccoli plants from haploids is possible via spontaneous and induced doubling. Doubled haploid (DH) broccoli lines are considered to be fully homozygous. Therefore, the production of DH lines is an alternative way to obtain pure inbred lines that can be utilized as parents in the development of new F1 hybrid varieties showing high levels of heterosis, high-quality heads, and uniform harvestable crop. We are using an anther culture-based haploid plant production system to develop DH broccoli lines in our broccoli breeding program. DH broccoli lines are produced from different genetic backgrounds within a year and handed to broccoli breeders.

Allium tuncelianum (Kollman) Özhatay, Matthew & Şiraneci forms a single-cloved edible white bulb with mild garlic (A. sativum) odour and taste. Its ability to form seeds make it suitable for genetic improvement via classical and modern approaches. A detailed study was carried out to determine the gynogenesis potential of two A. tuncelianum (AT1 and AT2) accessions. Unopened flower buds of A. tuncelianum accessions were cultured in various BDS- and MS-based induction media. A total of 17 (0.09%) gynogenic plantlets were obtained from ~20000 flower buds used in gynogenesis induction experiment. Accessions showed slight differences in their responses to gynogenesis induction cultures. The highest gynogenic plantlet production frequency (0.34 %) in AT1 was achieved flower buds cultured in T12 medium (MS with 100 g/L sucrose, 1 mg/L a-naphthalene acetic acid (NAA) and 8 mg/L isopentenyl adenine (2IP). Flower buds of AT2 showed the highest gynogenic plantlet production response (0.44 %) in T2 medium (BDS with 50 g/L sucrose). Eight of 17 gynogenic plantlets continued to grow and became healthy plants with green leaves and well established roots. Flow cytometric (FCM) analysis of well-developed gynogenic plants showed that two were haploid (25 %), four were diploid (50 %), and two were mixoploid (25 %) for haploid and diploid cells. Nine gynogenic plantlets showing abnormal development were diploid. Therefore, formation of gynogenic plantlets with abnormal phenotypes was likely due to genetic factors. Results obtained from this study suggest use of DH technology in the production of homozygous A. tuncelianum inbreeds in variety improvement programs.

Leek (A. ampeloprasum L.) is an economically important vegetable crop from Alliaceae family. It is a non-bulb forming biennial species grown for its pseudostem and leaves. Leek is a tetraploid with one of the largest genomes known among cultivated plant species. It has enormous economic importance all around the world for many purposes such as vegetable, medicinal herb, and food seasoning. Production and consumption of leek is in rise all around the world and breeders are trying to develop new F1 hybrid varieties with desired agronomical traits. Although self-compatible, leek shows high tendency toward outcrossing and display severe inbreeding depression when selfed with its own pollen. Therefore, inbred development through classical breeding techniques is very difficult in this crop. Traditional leek genotypes are highly heterozygous, open pollinated varieties. There is a high demand for F1 hybrid varieties with resistance to biotic and abiotic stresses and high-quality plants. Our group is trying to incorporate gynogenesis-based doubled haploid technology to leek improvement programs. Over the years, many experiments were carried out to determine the gynogenic potential of donor leek genotypes of different genetic backgrounds in different induction media. Here, we report a protocol allowing production of green gynogenic leek plants via single step culture of unopened flower buds. Ploidy levels of gynogenic regenerants are determined by flow cytometry analysis. A majority of the gynogenic leek regenerants produced survived well in vivo.

Aronia (Aronia melanocarpa Elliot, black chokeberry) is an important fruit plant for medical and culinary uses. Its berries are highly nutritive and rich in vitamins and antioxidants. It is a clonally propagated fruit species but there are very few reports of in vitro-based clonal propagation protocols that can be utilized in multiplication of different Aronia cultivars. We developed a highly efficient micropropagation protocol for three Aronia cultivars ('Eastland', 'Viking', and 'Nero'). In vitro cultures were initiated using actively growing shoots where they were collected from the mature field-grown plants. The shoots were washed thoroughly under tap water, cut into similar to 10 cm pieces before sterilization procedure. The shoots were dipped in 70% ethanol for 30 s and sterilized in a sterilization solution for 30 min. Then, the shoots were rinsed three times with sterile double distilled water in a laminar flow hood. In order to initiate in vitro cultures, nodal explants that were obtained from surface sterilized shoots were placed in Magenta boxes containing M1 (MSO) and M2 (MS supplemented with 0.7 mg L-1 6-benzylaminopurine (BAP)) media. After about 10 weeks, shoots developed from these initiation cultures were cut into nodal explants and subcultured in MS2 for further multiplication. For rooting, nodal explants were prepared from in vitro-grown shoots and cultured in R1 (WPM supplemented with 1 mg L-1 indole-3-butyric acid (IBA)) and R2 (1/2 WPM supplemented with 3 mg L-1 IBA) media. Majority of the explants produced shoots and roots in rooting media within three months. All three cultivars were successfully propagated with slight differences. Rooted plants were successfully acclimated and transferred to a greenhouse for further growth. One year-old plants grew up to 55 cm in height and developed multiple shoots. The protocol developed in this study can be used for in vitro clonal propagation of commercially important Aronia cultivars.

A survey for the presence of Grapevine virus E (GVE, genus Vitivirus, family Betaflexiviridae) in vineyards in New York and California was conducted using macroarray hybridization or reverse-transcription polymerase chain reaction (RT-PCR) assays. In New York, GVE was detected in 10 of 46 vines of Vitis labrusca, one V. riparia, and one Vitis hybrid. All GVE-infected New York vines were coinfected with Grapevine leafroll-associated virus-3. In California, GVE was detected in 8 of 417 vines of V. vinifera. All GVE-infected California vines were also coinfected by one of the leafroll-associated viruses and other vitiviruses. In order to assess the genetic diversity among GVE isolates, a viral cDNA was amplified by RT-PCR, and a 675-nucleotide region that included the 3' terminus of the coat protein gene, a short intergenic region, and the 5' terminus of the putative nucleic acid binding protein gene was sequenced. All 20 GVE isolates sequenced in this study were very closely related, with >98% nucleotide identity to the SA94 isolate from South Africa. These findings confirm the presence of GVE in major grape-growing regions of the United States and indicate a very low level of genetic diversity.

This paper presents a new flux-based method for out-of-step protection of synchronous generator. The available measured angular velocity and acceleration data from magnetic flux of the generator, at the location of the relay, are used to detect out-of-step conditions. An out-of-step condition can be distinguished in the point in which the polarity of the angular acceleration changes from a negative to positive value and the angular velocity is greater than the base angular velocity. The basic idea of the approach stems from the fact that the resultant magnetic flux rotates at synchronous speed and cannot change rapidly. In other words, this constant magnetic flux will not be affected by switching transients due to highly inductive characteristics of the machine. Finally, the simulation results verify the straightforward application of the proposed technique even for a multi-machine power system. Therefore, the proposed approach can be directly applied to an interconnected power system without any need to cumbersome network reduction methods. Furthermore, the proposed technique does not require any offline studies and overcomes some of the problems associated with the previous solutions.

A detailed study was conducted to assess the effects of 12 different induction media on the gynogenesis response of four open-pollinated (OP) leek (Allium ampeloprasum L.) genotypes. Gynogenic plant production was induced in 11 of 12 induction media. However, leek genotypes showed substantial differences in their responses to the induction media. Two MS-, two B5-, and one BDS-based media supplemented with auxin- and cytokinin-type plant growth regulators and high concentrations of sucrose (75 or 100 g l−1) provided gynogenic regenerants from all four leek genotypes included in the study. The highest gynogenic plantlet production (4.87 %) was obtained from the buds of an OP selection line (TU) cultured on B5 medium containing 2 mg l−1 dichlorophenoxyacetic acid (2,4-D), 2 mg l−1 6-benzylaminopurine, and 75 g l−1 sucrose. Culture of ~23,000 flower buds in 12 media provided 133 (0.58 %) green gynogenic plants. Flow cytometric analysis of the gynogenic plants showed that the majority (~62 %) of the recovered plants were diploid (1n = 2x) and about one-third was tetraploid (2n = 4x). Acclimated gynogenic plants obtained from the four donor genotypes were grown throughout spring and summer seasons in the greenhouse for morphological evaluations. Diploid gynogenic leek plants were substantially smaller in size (thinner and shorter pseudo-stems) when compared with tetraploid gynogenic and somatic plants. The results of this study show that sufficient numbers of diploid and tetraploid gynogenic leek plants can be produced for use in genetics studies and breeding programs for the improvement of new leek varieties with enhanced agronomical traits.

Polyploid breeding is an effective way to improve ornamental traits of horticultural plants. Colchicine, an anti-mitotic agent, is widely used to induce 2n gametes in horticultural plants, allowing for the production of polyploid plants through sexual hybridization. Based on an understanding of flower bud development and meiosis in pollen mother cells (PMCs) of Paeonia lactiflora cv. 'Fen Yu Nu', 2n pollen were obtained by injecting different concentrations of colchicine (0.1, 0.2, 0.3, 0.4 or 0.5%) into flower buds multiple times (once, twice or three times with a 24-h interval in between). External morphology, viability and possible hybridization of colchicine-induced 2n pollen were observed. Finally, P. lactiflora cv. 'Lian Tai' was pollinated with colchicine-induced pollen from 'Fen Yu Nu'. Polyploids were identified among offspring by flow cytometry, chromosome counting, and molecular assessment using simple sequence repeats (SSR). Flower bud diameter of herbaceous peony was related with meiosis in PMCs. The most suitable period to induce 2n pollen was when most PMCs were in prophase I of meiosis, and when flower buds were about 11-12 mm in diameter. The highest induced incidence (%) of 2n pollen was 47.39% following treatment with 0.4% colchicine solution injected twice. The average diameter of 2n pollen was 46.01 μm, which was 1.53-fold wider than haploid pollen. SEM results indicated that 2n pollen were considerably different from haploid pollen in terms of external shape and surface decoration, and 2n pollen were mostly spherical or with an irregular shape. Induced 2n pollen could germinate in vitro and on the stigma of the female parent, but abnormalities such as pollen tube bifurcation or thickening were observed in both germinated states. The percentage of germinated 2n pollen in vitro was 12.78%. In addition, two tetraploids (2n = 4x = 20) were obtained by 2n pollen hybridization. SSR analysis indicated that tetraploids were produced by the spontaneous fusion of 2n female gametes with colchicine-induced 2n male gametes. This indicates that colchicine did not entirely inactivate 2n pollen. The production of few polyploids may be due to the abnormal germination of 2n pollen and obstacles in ploidy hybridization. Our findings fortify the foundation for Paeonia polyploid breeding.

Crop-specific diagnostics to simultaneously detect a large number of pathogens provides an invaluable platform for the screening of vegetative material prior to its propagation. Here we report the use of what is to-date the largest published example of a crop-specific macroarray for the detection of 38 of the most prevalent or emergent viruses to infect grapevine. The reusable array consists of 1,578 virus-specific 60 to 70mer oligonucleotide probes and 19 plant and internal control probes spotted onto an 18 × 7 cm nylon membrane. In a survey of 99 grapevines from the United States and Europe, virus infections were detected in 46 selections of Vitis vinifera, V. labrusca, and interspecific hybrids. The majority of infected vines (30) was singly infected, while 16 were mixed-infected with viruses from two or more families. Representatives of the four main virus families Betaflexiviridae, Closteroviridae, Secoviridae, and Tymoviridae present in grapevines were found alone and in combination, with a notable bias in representation by members of the family Tymoviridae. This work demonstrates the utility of the macroarray platform for the multiplex detection of viruses in a single crop, its potential for characterizing grapevine virus associations, and usefulness for rapid diagnostics of introduced material in quarantine centers or in certification programs.

Leek (Allium ampeloprasum L.) is one of the economically important members of the genus Allium (family:Alliaceae). It is a self-compatible, outbreeding, tetraploid (2n=4X= 32) crop species that is very difficult to improve through traditional breeding strategies. Entegration of doubled haploid (DH) techniques such as gynogenesis into the leek breeding programs may speed up the variety development. In order to optimize a gynogenesis induction protocol for leek, we carried out experiments by culturing immature flower buds on various tissue culture media. Among the media tested, BDS and MS-based media with various combinations of plant growth regulators provided gynogenic embryos. Frequencies of gynogenic plantlet development were generally low and plantlets were obtained from almost all media included in the experiments. A total of 48 gynogenic plants were produced from approximately 30 thousand immature flower buds from six donor materials included in the experiments. In the DNA amount measurements performed with flow cytometry, it was found that gynogenic A. ampeloprasum plants were mostly diploid (55.56 %) and the others were tetraploid (44.44 %). Diploid and tetraploid gynogenic plants were transferred to in vivo for further evaluation and grown in a greenhouse to produce selfed seed. Our studies with leek and several other Alliums indicate that gynogenesis induction frequencies are generally low due to genotypic effect. In order to obtain sufficient numbers of gynogenic lines from leek breeding populations, high numbers of immature flowers should be cultured on gynogenesis induction media. We suggest use of large size flower buds collected several days prior to anthesis while establishing gynogenesis induction cultures in leek. BDS medium that is commonly used in onion gynogenesis studies can also be used for the production of gynogenic leek lines.

Pandanaceae (screwpines) is a monocot family composed of c. 750 species widely distributed in the Paleotropics. It has been proposed that the family may have a Gondwanan origin with an extant Paleotropical distribution resulting from the breakup of that supercontinent. However, fossils supporting that hypothesis have been recently reassigned to other families while new fossil discoveries suggest an alternate hypothesis. In the present study, nuclear and chloroplast sequences were used to resolve relationships among Pandanaceae genera. Two well-supported fossils were used to produce a chronogram to infer whether the age of major intra-familial lineages corresponds with the breakup of Gondwana. The Pandanaceae has a Late Cretaceous origin, and genera on former Gondwanan landmasses began to diverge in the Late Eocene, well after many of the southern hemisphere continents became isolated. The results suggest an extant distribution influenced by long-distance-dispersal. The most widespread group within the family, the Pandanus tectorius species complex, originated in Eastern Queensland within the past six million years and has spread to encompass nearly the entire geographic extent of the family from Africa through Polynesia. The spread of that group is likely due to dispersal via hydrochory as well as a combination of traits such as agamospermy, anemophily, and multi-seeded propagules which can facilitate the establishment of new populations in remote locations.