Fingerprinting of three typical macrosperma Italian lentil (Lens culinaris Medik.) landraces using fluorescence-based AFLP markers

Italian lentil landraces are principally cultivated for self or local consumption. Most of them are disappearing, particularly macrosperma types by being less required by the market. A pre-requisite for the conservation and the efficient use of genetic resources is the better understanding of the extent and the distribution of the existing genetic variation, useful for future breeding programmes. Our study was undertaken to analyse and quantify the genetic diversity within and among three macrosperma Italian lentil landraces (Onano, Altamura and Villalba), using fluorescent AFLP markers. AFLP markers generated information to differentiate among closely related genotypes and group within the same cluster individuals belonging to the same landrace. The total genetic diversity (H_T), the genetic diversity within population (H_S) and the extent of differentiation between populations (D_ST) were 0.198, 0.155 and 0.043, respectively. The fixation index (G_ST = 0.219) showed that about 78% of the observed total genetic variation can be attributed to within population differences and around 22% is due to differences among populations. The gene flow estimate (N_m = 1.774) and the mean genetic distance value (0.077) suggested narrow genetic base among the analysed populations, confirming the tendency of Italian lentil landraces to group together. The present study showed that fluorescence-based AFLP technique is a biotechnological tool that can provide significant insights for research in genetic diversity of lentil landraces and their subsequent conservation and utilization in breeding programs.     

Genetic characterization of asparagus doubled haploids collection and wild relatives

The genus Asparagus is very large consisting of around 150 species found as herbaceous perennials, tender woody shrubs and vines. The cultivated species (Asparagus officinalis L., diploid) is a highly prized vegetable, grown in different environments ranging from cool temperate zones to deserts, Mediterranean climates and tropical areas. As a consequence, Asparagus breeders have developed different cultivars that differ for their morpho-agronomic traits, habit and ploidic status (few triploid and tetraploid cultivars are used). Several breeding methods are used for developing cultivars, among which a well developed in vitro anther culture technique produces homozygous clones useful for F₁ hybrids constitution. A fluorescent based AFLP (amplified fragment length polymorphism) technique were applied with the aim to assess genetic diversity among a collection of 173 doubled haploid (DH) androgenetic clones, five Asparagus wild species and interspecific hybrids obtained among the cultivated species and two wild relatives. The average number of AFLP fragments generated per primer set was 105, varying in size from 50 to 550 bp. A total of 1054 AFLP fragments were detected, 20% of which were polymorphic. Genetic similarity based on DNA polymorphisms, showed that a few number of AFLP primer combinations are able to distinguish the cultivated DH clones from the wild species. Indeed, from one DH clone for each anther donors and the wild species were used to construct a dendrogram using Dice's coefficient and the unweighted pair group method with the arithmetic mean (UPGMA). Genetic distances among all DH clones were calculated using the C.S. Chord distance; and a neighbour-joining (NJ) consensus tree was constructed in order to support the breeder for parental genotype choice for asparagus hybrid constitution.     

Genetic diversity and population genetic structure of pomegranate (Punica granatum L.) in Iran using AFLP markers

Pomegranate (Punica granatum L.) is one of the oldest known edible fruits. It is native to Iran and spread from Iran to other areas. In this study amplified fragment length polymorphism (AFLP) was used to detect intra- and inter-population genetic diversity of pomegranate. A group of 67 accessions belonged to 4 populations from Iran was studied using eight primer combinations. A total of 221 scorable bands were amplified, of which, 118 (54.13%) were polymorphic. Resolving power (Rp) ranged from 5.70 to 9.21, and the average of polymorphism information content (PIC) per primer pair was 0.40. According to Nei's gene diversity and allelic statistics, Isfahan population had a highest genetic diversity (H = 0.3646, I = 0.5327, N_e = 1.6467). Coefficient of gene differentiation between populations (GST) was 0.124, indicated that mainly proportion of genetic variation (87.6%), was within populations and the remaining (12.4%) of the variation was among populations that, also supported by analysis of molecular variance (AMOVA). The gene flow (Nm) varied from 0.969 to 10.404 between pair-wise populations and was 3.504 among all of the populations. The Jaccard similarity coefficient between individuals ranged from 0.26 to 0.88. The UPGMA dendrogram clustered all 67 accessions into 6 groups. In some cases accessions from same region were grouped together but in most cases, there was gene exchange. To study the genetic relationships among populations, a principal coordinate analysis (PCoA) based on Nei's genetic distances was performed. Results of this study showed that AFLP marker can be a useful tool for investigating the genetic diversity of pomegranate genotypes.     

Identification of parental species of the Alstroemeria cv. ‘Jubilee’ using AFLP marker technique

Twelve Alstroemeria species, two hybrids, one cv. ‘Jubilee’, an anther-cultured plant from cultivar ‘Jubilee,’ and Bomarea salsilla and Leontochir ovallei (the latter two were chosen as outgroup) were evaluated using the AFLP marker technique in order to identify putative parental genotypes of the Alstroemeria cv. ‘Jubilee’ and of known interspecific hybrids, and to estimate their genetic relationships within the genus Alstroemeria. A total of 297 AFLP markers were scored by using the primer combination (E + ACCA/M + CTAG). In order to discriminate all Alstroemeria genotypes, cluster analysis (UPGMA) and principal coordinates analysis were performed. The Alstroemeria cv. ‘Jubilee’, of which the parents are unknown, had genetic distance (GD) 0.54 from Alstroemeria exserens, GD 0.57 from Alstroemeria garaventae, GD 0.62 from Alstroemeria gayana, and GD 0.66 from Alstroemeria hookeri cumminghiana. Thus, these four species are considered as putative parental genotypes. An interspecific hybrid (Alstroemeria aurea × Alstroemeria inodora), showed the smallest genetic distance from A. aurea (GD 0.56) and A. inodora (GD 0.45). The Alstroemeria ligtu group was distantly allocated from other Chilean species. We conclude that the AFLP marker technique appears to be a satisfactory tool for identifying the parental genotypes of interspecific hybrids in Alstroemeria.     

The origin and dissemination of the cultivated almond as determined by nuclear and chloroplast SSR marker analysis

Sixteen nuclear and 10 chloroplast SSR markers were evaluated for 40 almond genotypes including cultivated almond, 18 related species and 5 interspecific-hybrid populations. Results establish the value of SSR (nuclear and chloroplast) markers for distinguishing different genetic lineages and characterize an extensive gene pool available to almond genetic improvement. Hierarchical analysis using integrated nuclear and chloroplast DNA markers support Prunus fenzliana, a species native to the northeast Iran, as a probable ancestor of the cultivated almond. Results also established the importance of interspecific hybridization and subsequent genetic introgression in the development of cultivated almond and demonstrate continuing value of an interspecific gene pool for modern cultivar improvement. Molecular results implicate a dissemination of the cultivated almond from Asia to the Eastern Mediterranean and subsequently the Western Mediterranean and the New World is supported by the molecular analysis of regional germplasm.     

Physical dormancy in myrtle seed

The nature of dormancy in seeds of myrtle (Myrtus communis L.) was investigated. Scarification with cold acid or sand paper and soaking in water increased seed germination, whilst stratification, inserting the seeds in boiling water or hot acid treatment decreased the number of germinating seeds. The highest germination was obtained by treating the seeds with 100 % cold acid for 60 min or 80 % cold acid for 120 min. Hard seed coat was found to be the principal cause of poor seed germination.Scarified and non-scarified seeds were germinated for 2 weeks at 6 temperatures between 5 and 30° C, and 20° C was optimum.     

A SCAR marker linked to the N gene for resistance to root knot nematodes (Meloidogyne spp.) in pepper (Capsicum annuum L.)

The root-knot nematodes (Meloidogyne spp.) are important nematode pests and cause serious diseases in pepper in the world. No molecular markers linked to the nematodes resistance N gene have been reported. In this paper, ‘Carolina Wonder’ (Capsicum annuum L.), a sweet pepper line resistant to root-knot nematode with N gene, ‘20080-5-29’ (C. annuum L.), an inbred line susceptible to root-knot nematode with good horticultural characteristics, and their F₂ progeny with 320 individuals were used as materials. Evaluation of resistance and susceptibility of parental lines, F₁ and F₂ progeny inoculated with root-knot nematodes (Meloidogyne incognita) were carried out. ‘Bulked segregant analysis’ method was used to search for polymorphic markers from 512 pairs of AFLP primers. Based on the assessment of resistance and susceptibility and polymorphism of the AFLP marker in F₂ population, the genetic linkage distance between the AFLP marker and the N gene was estimated. One AFLP marker E39/M41-339 was obtained and transferred to a SCAR marker amplifying a 315 bp DNA fragment linked to the N resistant allele and a 331 bp fragment linked to the N⁺ susceptible allele. The distance between the molecular marker and the nematodes resistance N gene is 6.3 cM. This research delivered a valuable tool for the marker assisted selection of nematodes resistance in pepper.     

Comparative analysis of genetic diversity in Tunisian apricot germplasm using AFLP and SSR markers

Eighty-one accessions representing apricot germplasm in Tunisia were collected from different areas of cultivation and fingerprinted using amplified fragment length polymorphism (AFLP) and microsatellites (SSR) markers. A total of 339 polymorphic markers were revealed using 5 AFLP primers combinations and 24 SSR loci. AFLP and SSR markers expressed a high level of polymorphism allowing the distinction of the accessions with an efficiency coefficient of discrimination of 100% for AFLP and 97% for SSR markers. Genetic diversity structure was assessed with AFLPs and SSRs markers separately then with combined matrix data by the help of hierarchical clustering elaborated using Wards method based on Nei and Li (1979) distances. Comparison of the obtained dendrograms revealed a phylogeographic structure into two major groups with significant conservation between the observed subgroups in relation with the geographic origin of the accessions. The relative efficiency of the markers in determining the genetic relationships among apricot accessions has been assessed and a combination of AFLPs and SSRs markers was the most effective. In addition, Mantel test based on genetic distances indicated highly significant correlation between AFLP-SSR data and each of the AFLP and SSR ones, with Pearson correlation values of r = 0.873 and r = 0.692, respectively, revealing the higher efficiency of the combination of both molecular techniques (AFLP and SSR) to estimate the levels of genetic variability among apricot germplasm.     

Genetic diversity and population's structure in Tunisian strawberry tree (Arbutus unedo L.)

A growing interest in the use of the Strawberry tree (Arbutus unedo L., Ericaceae) has been recently reported for industrial, pharmaceutical and chemical fields. However, the bulk material comes from natural populations because of the lack of selection of interesting cultivars. In Tunisia, A. unedo populations are severely destroyed due to deforestation and over-collecting. The species occurs in small scattered populations decreasing progressively in size. Yet, no conservation or improvement programs are attempted to preserve and promote the potential value of this resource. In this work, we assessed the genetic diversity of nine Tunisian populations of A. unedo L. from different bioclimates, using 65 polymorphic RAPD loci. The analysis of the genetic variation within and among populations is primordial to elaborate conservation and improvement programs. A low genetic diversity within a population estimated by both Nei's (He) and Shannon's diversity (H′) indices (0.155 < He < 0.248; 0.229 < H′ < 0.364) was observed due to genetic drift and selfing. At the species level, the amount of the within population variation estimated by Shannon's index (H_POP/H_SP = 0.686) and the molecular variance (80.67%) was higher than that among populations. A moderate genetic differentiation among populations (Φ_ST = 0.193 and G_ST = 0.314) which could be attributed to the long seed distance dispersal was detected. The UPGMA dendrogram based on Φ_ST values showed three clusters each including populations without relationship to bioclimatic or geographical origin indicating that differentiation occurs at a local space scale. The in situ protection measures should be made appropriately according to a population within bioclimates. The ex situ conservation and the selection of genotypes should involve extensive collection of seeds or cuttings from the within populations rather than among them.     

Molecular analysis of introgression lines from Cucumis hystrix Chakr. to C. sativus L

Two stable introgression lines (ILs), IL56 and IL73 with fourteen chromosomes, derived from a cross between the wild relative Cucumis hystrix Chakr. (2n = 24) and the cultivated cucumber “beijingjietou” (Cucumis sativus L., 2n = 14) were characterized using SSR and AFLP markers. Twenty-three SSR primers were used to detect the introgression from C. hystrix to C. sativus, and one locus at 210 bp was revealed and assigned to introgressive fragment of C. hystrix genome in IL56. Substantial genomic changes in these two lines were detected by AFLP analysis with thirty different primer combinations. C. hystrix-specific bands, novel bands for the two parents and deleted bands in C. sativus were observed. One hundred and forty-seven polymorphic bands were observed, but only four and five introgressed bands were found in IL56 and IL73, respectively. Although representing only a small proportion of the C. hystrix genome, these two introgression lines with downy mildew and fusarium wilt resistance will be valuable to resistant genetics and breeding.     

Analysis of the taxonomic subdivision within the genus Helleborus by nuclear DNA content and genome-wide DNA markers

Helleborus is a genus of herbaceous perennials belonging to the family Ranunculaceae. Within this genus six sections with a total of 22 species are found. The largest section Helleborastrum contains 16 species for which genetic relationships are still unclear. This study represents the first genetic analysis in the genus Helleborus, including the two newly described species H. liguricus and H. abruzzicus based on multilocus amplified fragment length polymorphism (AFLP) markers with a genome-wide distribution in combination with nuclear DNA content data. Chromosome analyses of roots tips revealed a number of 2n = 32 for the selected species, which was congruent with previous observations. The nuclear DNA content of Helleborus was estimated by flow cytometry applying propidium iodide staining and varied between 18 and 33 pg/2C, depending on the species. For AFLP analyses, 19 out of the 22 Helleborus species were studied using 10 AFLP primer combinations, resulting in a total of 1109 polymorphic bands among all species including the outgroup. The genetic distances between species varied between 0.034 and 0.330. Based on genetic distances a phenogram using the Neighbor-joining cluster method with bootstrap analysis was calculated. The results support the previously suggested division of the genus into six sections and thereby approve AFLP data to be applicable for phenetic analyses. Moreover, this genetic information is significant for the development of future Helleborus breeding strategies.     

Genetic relationships among loquat cultivars and some wild species of the genus Eriobotrya based on the internal transcribed spacer (ITS) sequences

The phylogenetic relationships among seven wild species and 25 loquat cultivars of genus Eriobotrya were studied by comparing sequences of the internal transcribed spacer (ITS) region. The length of ITS1 region of Eriobotrya was 223 bp and the lengths of ITS2 were ranged from 201 bp to 203 bp. The G + C contents varied from 64.1% to 65.5% in ITS1 and from 68.5% to 72.1% in ITS2. The aligned sequences were analyzed by neighbor-joining (NJ) and maximum parsimony (MP) methods. The ITS1 spacer region comprised 223 characters, of which 44 were variable sites and 34 were parsimony informative sites. The NJ tree had very similar topology to the MP tree. E. malipoensis and E. seguinii had only one different character in ITS2 region and both of them could be the most primitive species of Eriobotrya genus. ITS2 evolved much faster and had more parsimony informative sites than ITS1. ITS2 presented much more precise information about the phylogenetic relationships. Based on ITS2 regions, the evolution order of the studied taxa in this paper could be concluded, which was E. bengalensis f. angustifolia, E. prinoides var. dadunensis, E. prinoides (E. bengalensis), E. dayaoshanensis and E. japonica Lindl.     

Using bulked AFLP analysis to assess genetic diversity in Echinacea species

Echinacea is an allogamous genus, thus its cultivars or populations are genetically heterogeneous. Using amplified fragment length polymorphism (AFLP) to estimate the genetic diversity of Echinacea is generally limited by the large number of individual plants and the higher cost that need to be processed. In the present study, effectiveness of several sizes of DNA bulking (10, 15, 20, 25 and 30 individuals) with 20, 36 and 55 primer pairs was compared using AFLP in determining the genetic diversity of Echinacea species. The results indicated that the use of bulked DNA-based AFLP analysis by using the selected eight primer pairs was capable of detecting genetic diversity between the tested Echinacea species, provided that the potential presence of low frequency variants was ignored and a possible bias in the estimates of genetic similarity was accepted. The assessments showed that a bulk of 15 individuals could detect sufficient AFLP variations at most genomic sites. Additionally, 20 primer pairs could generate sufficient polymorphic fragments to achieve high resolving power of AFLP for the tested Echinacea species.     

Induction of drought tolerance by paclobutrazol and irrigation deficit in Phillyrea angustifolia during the nursery period

Phillyrea angustifolia is a native Mediterranean species, which has recently been considered suitable for landscaping purposes. We hypothesize that hardening plants in the nursery could increase their tolerance of drought after transplanting. The effects of paclobutrazol (PBZ) and different irrigation regimes applied to seedlings planted in 4.5-L plastic pots were investigated. PBZ was applied as a substrate drench at 0 mL L⁻¹ (untreated control), 30 mL L⁻¹ and 40 mL L⁻¹ per plant and three drip irrigation treatments were used: I100, plants watered at water-holding capacity, I60, plants watered to 60% of I100, and I40, plants watered to 40% of I100. Plants were pot-grown in an unheated greenhouse near the Mediterranean coast of SE Spain. A reduction in plant height and stem diameter was observed one month after being drenched by PBZ. The irrigation regime significantly affected plant height after three months of cultivation and did not affect stem diameter during the nursery period. Significant interaction between the irrigation regime and PBZ dose was evident for plant height during the nursery period. I100 and untreated PBZ plants had the lowest stomata density. PBZ doses significantly reduced canopy weight and leaf area compared with the control. I60 plants showed the greatest leaf area and canopy dry weight, and the highest root length, dry weight, volume and number of forks. Both I60 and I40 treatments showed an equally high water use efficiency (WUE) (calculated as the total plant dry matter divided by the total amount of water supplied by the irrigation treatments). In general, PBZ induced a suite of morphological adaptations (increased root-to-shoot ratio and stomata density, decreased leaf area reduction, fine roots, etc.) that might allow the plants to tolerate drought after transplanting.     

Impacts of changing climate and land use on vegetation dynamics in a Mediterranean ecosystem: insights from paleoecology and dynamic modeling

Forests near the Mediterranean coast have been shaped by millennia of human disturbance. Consequently, ecological studies relying on modern observations or historical records may have difficulty assessing natural vegetation dynamics under current and future climate. We combined a sedimentary pollen record from Lago di Massacciucoli, Tuscany, Italy with simulations from the LandClim dynamic vegetation model to determine what vegetation preceded intense human disturbance, how past changes in vegetation relate to fire and browsing, and the potential of an extinct vegetation type under present climate. We simulated vegetation dynamics near Lago di Massaciucoli for the last 7,000 years using a local chironomid-inferred temperature reconstruction with combinations of three fire regimes (small infrequent, large infrequent, small frequent) and three browsing intensities (no browsing, light browsing, and moderate browsing), and compared model output to pollen data. Simulations with low disturbance support pollen-inferred evidence for a mixed forest dominated by Quercus ilex (a Mediterranean species) and Abies alba (a montane species). Whereas pollen data record the collapse of A. alba after 6000 cal yr bp, simulated populations expanded with declining summer temperatures during the late Holocene. Simulations with increased fire and browsing are consistent with evidence for expansion by deciduous species after A. alba collapsed. According to our combined paleo-environmental and modeling evidence, mixed Q. ilex and A. alba forests remain possible with current climate and limited disturbance, and provide a viable management objective for ecosystems near the Mediterranean coast and in regions that are expected to experience a mediterranean-type climate in the future.     

Genetic diversity in Tunisian olive accessions and their relatedness with other Mediterranean olive genotypes

Eighty-four olive accessions obtained from the National Conservatory of Boughrara-Sfax (Tunisia), previously evaluated for morphological traits, were analysed with 47 random amplified polymorphic DNA (RAPD) markers. They were compared with other olive genotypes originated from Eastern or Western Mediterranean. The highest and lowest similarities between genotypes, estimated by simple matching algorithm, were 0.98 and 0.40, respectively. A dendrogram based on Ward's method and a factorial correspondence analysis (FCA) showed that most of Tunisian accessions are closely related to olive genotypes originating from the Eastern Mediterranean and some are clustering with genotypes originated from the Western Mediterranean. These findings suggested multiple and complex origin of Tunisian olive. A comparative study between a previous morphological analysis and current RAPD assay was carried out and discussed.     

Performance of ‘Coscia’ pear (Pyrus communis) on nine rootstocks in the north of Israel

The vegetative and reproductive performances of ‘Coscia’ pear (Pyrus communis L.) grown on nine rootstocks [OHF 69, OHF 97, OHF 217, OHF 333, OHF 513 and BP 1 (P. communis), clonal seedling (Davis AxB) of Pyrus betulifolia and quince BA 29 and EMA (Cydonia oblonga)] were compared during a 9-year period. The trial was conducted at the Experimental Station ‘Avnei Eitan’ in northern Israel (elevation 400 m above sea level, a.s.l.), on a well-drained soil with pH 7.1. Trees were planted in December 1998, spaced at 4.0 m × 2.0 m and trained with a central axis. The most vigorous trees were on P. betulifolia seedlings, followed by the four OHF rootstock (69, 97, 333, 513) and BP 1 (with no significant difference between them). All the above rootstocks demonstrated greater vigor than OHF 217 and quince BA 29 or EMA. The highest cumulative yields per tree were harvested from trees on the four OHF rootstocks (69, 97, 333, 513) and P. betulifolia followed by the BP 1. The two quince rootstocks, as well as OHF 217, had the lowest cumulative yield and the lowest yield of large fruit. A positive correlation was found between the vigor of the tree, as affected by the rootstock, and both the total yield and the fruit size. We conclude that in a warm climate, yield efficiency is not the only parameter that should be taken into account, and building a strong tree for a weak cultivar is the first requirement for establishing an orchard.     

Molecular phylogeny in Indian Citrus L. (Rutaceae) inferred through PCR-RFLP and trnL-trnF sequence data of chloroplast DNA

The taxonomy and phylogeny of Indian Citrus is revisited using PCR-RFLP of the trnD-trnT and rbcL-ORF 106 regions as well as sequence data analysis of the trnL-trnF intergenic spacer region of cpDNA. The study was based on 50 accessions of Citrus genotypes, collected from wild, semi-wild and domesticated stocks. Of the 13 restriction enzymes (RE) used for restriction digestion of the polymerase chain reaction (PCR) amplicons, four (Hinf I, Msp I, Alu I, Hae III) generated 47 restriction fragments, of which 24 (51%) were polymorphic. PCR-RFLP data showed a genetic distance ranging from 0 to 0.79 among 50 accessions of Citrus, and a cluster analysis, based on Neighbor-Joining (NJ) method, placed all the accessions in eight major clusters. Analysis of trnL-trnF sequences from 23 representative accessions of Citrus showed a pair-wise sequence divergence rate in the range of 0–0.064. NJ, minimum evolution (ME) and maximum parsimony (MP) analyses of trnL-trnF sequences produced phylogenetic trees, which placed all the 23 accessions in five clusters. PCR-RFLP analysis resulted in a well resolved phylogenetic tree with branches supported by moderate to high bootstrap values, while the trnL-trnF sequence-based trees showed only moderate to low bootstrap support for the internal tree branches, indicating uncertain origin of some Citrus genotypes. This study shows that the trnL-trnF spacer sequence data can detect genetic variation in Indian Citrus genotypes, but the utility of the data in inferring phylogeny at intra and inter-specific levels is limited probably by factors such as hybridization, bud mutations, apomixis and polyploidy. However, PCR-RFLP and trnL-trnF data supported the recognition of C. maxima, C. medica, and C. reticulata as the basal species of edible Citrus.     

Growth,yield and essential oil content of three cultivars of basil grown under different levels of nitrogen in the field

A field experiment was conducted in Southern Italy to investigate the effect of different nitrogen (N) rates (0, 100 or 300 kg ha⁻¹) on the number of branchings and leaves per plant, plant height, yield of above-ground fresh biomass, total leaf area, dry weight, leaf-to-stem ratio, leaf essential oil content and the essential oil quality at commercial harvest (full bloom) of the three cultivars of basil (Ocimum basilicum L.): (i) “Mostruoso mammouth” (MM); (ii) “Genovese profumatissimo” (GP); (iii) “Napoletano a foglia di lattuga” (NFL). Nitrogen fertilization up to 300 kg ha⁻¹ increased yield of above-ground and leaf fresh biomass, leaf essential oil yield, but it did not affect leaf-to-stem ratio, plant height and the number of branchings per plant. The increase in essential oil yield induced by N fertilization depended on an increase in both leaf essential oil concentration and leaf biomass. The increase in LAI with increasing N fertilization was due to an increase in leaf number per plant rather than in individual leaf expansion. The cultivar GP was different in morphology from MM and NFL, since it had greater leaf number and plant height, but lower leaf-to-stem ratio and LAI. The physiological efficiency of N use (PE) in producing above-ground biomass was not related to N application whereas PE in producing essential oil (PE-oil) tended to increase with increasing levels of N applied. With regard to the effect of the cultivar, GP was less efficient in N use for oil production than MM and NFL since PE-oil of GP was lower while leaf N and PE-biomass were higher than the other cultivars. NFL was the richest in methyl chavicol but the lowest in linalool, whereas MM and GP contained linalool and eugenol.     

Efficient in vitro multiplication in Ornithogalum ulophyllum Hand.-Mazz. from twin scale explants

Ornithogalum ulophyllum Hand.-Mazz. with beautiful white flowers is an important medicinal and ornamental plant of the Middle Eastern countries and need exploitation for commercial propagation. The study reports in vitro mass proliferation of bulblets achieved from twin scales and “in vitro regenerated bulblet” explants on MS medium supplemented with various concentrations of BAP–NAA. The best regeneration on twin scales and “in vitro regenerated bulblets” was obtained on MS medium containing 2 mg l⁻¹ BAP–0.5 mg l⁻¹ NAA and 2 mg l⁻¹BAP–1 mg l⁻¹ NAA, respectively. However, bulb scales seemed to be more potent for bulblet regeneration. A large number of the developing bulblets rooted on the regeneration medium. Remaining non-rooting bulblets were rooted on MS medium containing 1 mg l⁻¹ NAA. All plants were acclimatized in the environmental chamber for 4 weeks and were transferred to the greenhouse for flowering. Regenerated bulblets developed into morphologically normal plants.