Phenotypic variability and genetic structure in plum (Prunus domestica L.), cherry plum (P. cerasifera Ehrh.) and sloe (P. spinosa L.)

In the present study, phenotypic variability of 80 plum (Prunus domestica L.) varieties maintained in the French National Plum Collection was evaluated with 19 quantitative traits. In addition, genetic diversity and genetic structure was studied in three plum species (P. domestica L., Prunus cerasifera Ehrh. and Prunus spinosa L.) using chloroplast DNA (cpDNA) markers and five single sequence repeat (SSR) loci. Based on phenotypic traits, some varieties, such as mirabelle plums, grouped together. Bayesian structure analysis was used to identify different genetic groups, whereby damson plums were clearly distinguished from greengage plums. When examining the three species together, a higher level of cpDNA allelic richness was found in P. cerasifera and in P. spinosa than in P. domestica where only five cpDNA haplotypes were detected in the national plum collection, with one main haplotype that accounted for 80% of the varieties studied. P. domestica cpDNA haplotypes tended to group together with P. cerasifera haplotypes whereas most of P. spinosa haplotypes formed a separate cluster. SSR markers were somewhat able to distinguish the three species. These results provide some clues as to the origin of plum and the various plum varieties. Our results also provide useful information for the management of plum genetic resources.     

Rose (Rosa hybrida L.) EST-derived microsatellite markers and their transferability to strawberry (Fragaria spp.)

The ‘Genome database for Rosaceae (GDR)’ provides a large collection of expressed sequence tags (ESTs) harboring simple sequence repeats (SSRs) from several Rosaceae genera, including Rosa (rose). Primer pairs flanking SSR were designed for 312 unique Rosa ESTs based on GDR database. Eight rose (Rosa hybrida L.) genotypes were tested for PCR amplification, and 287 (92%) of the primer pairs generated allele-specific PCR bands that were readily scored. From 183 (63.7%) primer pairs that evidenced polymorphic alleles among the eight rose cultivars, 20 pairs evidencing EST sequence homology to known gene functions and high levels of polymorphism were selected and utilized for DNA fingerprinting and genetic diversity assessments of 47 rose hybrids. A total of 202 polymorphic bands were scored and generated unique fingerprints for each rose hybrid. The Nei–Li genetic similarity coefficients among 1081 pair-wise comparisons of 47 cultivars exhibited a broad range of genetic variations from 0.30 (‘Grand King’ and ‘Carnival’) to 0.99 (‘First Red’ and ‘Red Champ’). UPGMA cluster analysis divided 47 hybrids into five major groups and two sub-groups. The cross-species transferability of 273 EST-SSR primer pairs was evaluated using four genotypes of the strawberry, a genus member of the Rosaceae family. PCRs on the DNA samples of strawberry were successful for 165 primer pairs; among these, 123 pairs amplified 243 polymorphic bands. As surrogates of the marker transfer, the phenetic relationship among the four strawberry genotypes was evaluated. Genetic similarity coefficients varied from 0.78 (‘Maehyang’ and ‘Janghyee’) to 0.64 (‘Janghyee’ and ‘Pragana’). The results of cluster analysis showed that the three octaploid strawberry cultivars were quite similar, whereas the diploid ‘Pragana’ was related distantly at the genomic DNA level. The EST-SSR markers developed in the present study can be efficiently utilized for genetic diversity studies in Rosaceae.     

African spider flower (Cleome gynandra L./Gynandropsis gynandra (L.) Briq.) as a red spider mite (Tetranychus urticae Koch) repellent in cut-flower rose (Rosa hybrida L.) cultivation

Companion planting of Cleome gynandra, of Kenyan origin, in beds of cut-flower roses reduces significantly red spider mite (Tetranychus urticae Koch) infestation without any detrimental effect on productivity or flower quality. The level of reduction is dependent upon the density of the C. gynandra plants with 15 plants in a 1.8 m² bed (8.3 plants m²) being the most effective, planted either around the bed perimeter or within the rows of roses. The relatively high density of C. gynandra plants required may limit the direct application of this technology in export-focused, greenhouse rose production yet may be of significant value as a supplement to other mite-control strategies. The potential benefits of such companion planting for growers of field roses and those involved in some domestic markets are also evident. Research into the nature and extraction of the active, volatile mite-repellant components of C. gynandra is indicated.     

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.     

Early detection of graft incompatibility in apricot (Prunus armeniaca L.) using phenol analyses

The study investigated the role of phenols in apricot graft incompatibility. Assays of phloem with cambium from 1-year-old apricot trees of cultivars Marlen, Leskora and Betinka which were grafted on the rootstocks of different genetic origin: M-LE-1, Lesiberian, MY-KL-A, Tetra, Penta, Green Gage, Julior, MRS 2/5 and Isthara were analysed with HPLC (together 23 scion/stock combinations). The phloroglucinol, catechin, p-coumaric acid and further non-identified phenols with the retention time 23–25 and 30 min were determined. The content of individual phenol compounds was related to specific cultivar/rootstock combination. The minimum number of statistical significant differences in the phenol content between tissues above and below graft union was established in homospecific combinations (P. armeniaca/P. armeniaca). Cultivars Marlen, Leskora and Betinka differ in the degree of compatibility or incompatibility with rootstocks. The pattern of non-identified phenol 23 in different graft combinations is similar to catechin and p-coumaric acid.     

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.     

The inheritance of two novel subgynoecious genes in cucumber (Cucumis sativus L.)

Cucumber (Cucumis sativus L.), which is a vegetable crop, has served as the model system for sex expression in flowering plants, and the inheritance of sex expression in cucumber is well documented. However, the genetics of subgynoecism expression in cucumber had rarely been described. In this study, we investigated the inheritance of subgynoecious traits in cucumber plants with the inbred cucumber lines of subgynoecious (C. sativus L. var sativus cv 97-17 and S-2-98) as the materials. Genetic analysis had showed the two subgynoecious inbred lines were controlled by one pair of recessive gene and one pair of incompletely dominant gene, which were designated presently as mod-F2 and Mod-F1, respectively. Furthermore, the mod-F2 and Mod-F1 loci, which enhance the intensity of femaleness, also inherited independently with F and M genes.     

ISSR variations in eggplant (Solanum melongena L.) and related Solanum species

Inter-simple sequence repeat (ISSR) analysis was performed in eight cultivars of eggplant (Solanum melongena) and 12 accessions in eight related Solanum species to evaluate the applicability of this analysis for assessing the phylogenetic relationships and identifying cultivars. A total of 552 polymorphic amplified bands were obtained from 34 of the 100 primers tested, and the percentage of polymorphisms was 99.1%. Cluster analysis based on the ISSR markers classified the Solanum species into seven groups: (i) S. melongena; (ii) S. aethiopicum and S. anguivi; (iii) S. incanum; (iv) S. violaceum and S. kurzii; (v) S. macrocarpon; (vi) S. virginianum and (vii) S. torvum. Combining the ISSR markers obtained by a few of the 34 primers was enough for distinguishing of the eight cultivars of eggplant. This ISSR analysis was demonstrated to be available for the phylogenetic study and the cultivar identification.     

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.     

Mycorrhizal colonization improves onion (Allium cepa L.) yield and water use efficiency under water deficit condition

Most plants benefit from mycorrhizal symbiosis through improvement of water status and nutrient uptake. A factorial experiment with complete randomized blocks design was carried out in greenhouse at Tabriz University, Iran in 2005–2006. Experimental treatments were (a) irrigation interval (7, 9 and 11 days), (b) soil condition (sterile and non-sterile) and (c) arbuscular mycorrhizal fungi (AMF) species (Glomus versiforme, Glomus intraradices, Glomus etunicatum) and non-mycorrhizal (NM) plants as control. Onion (Allium cepa L. cv. Azar-shahr) seeds were sown in sterile nursery and inoculated with fungi species. One nursery left uninoculated as control. Nine weeks old seedlings then were transplanted to the pots. Average pre-irrigation soil water contents reached to about 67, 61.6 and 57.5% of FC corresponding to 7, 9 and 11 days irrigation intervals, respectively. At onion bulb maturity stage (192 days after transplanting), yield, water use efficiency (WUE) and yield response factor (K_y) were determined. The results indicated that AMF colonization increased soil water depletion significantly. G. versiforme under both soil conditions (sterile and non-sterile) and G. etunicatum in sterile soil depleted soil water effectively (P < 0.05). Mycorrhizal fungi improved WUE significantly (P < 0.0001) in both soil conditions. It raised by G. versiforme about 2.4-fold (0.289 g mm⁻¹) in comparison with the control (0.117 g mm⁻¹). G. intraradices and G. etunicatum also had significantly higher WUE than control. Apparently water deficit in 11-day irrigation interval led to lower yield and WUE compared to 9-day interval; the later resulted highest WUE (0.254 g mm⁻¹). Mycorrhizal plants increased seasonal ET significantly due to enhancing in plant growth; G. versiforme in both sterile and non-sterile soil and G. etunicatum in sterile soil had the highest ET. Bulb yield was influenced by irrigation period and fungi species. G. versiforme produced higher yield than other treatments (135.27 g/pot). Mycorrhizal plants in 11-day irrigation interval in spite of suffering from water stress had more bulb yield than non-mycorrhizal plants in all irrigation intervals. Yield in general was higher in 9-day treatments than other irrigating internals significantly (P < 0.05). Onion yield response factor (K_y) was decreased by AMF colonization; implying that symbiosed plants become less responsive to water deficit (longer irrigation interval) compared to the control ones.     

S-RNase based S-genotyping of Japanese plum (Prunus salicina Lindl.) and its implication on the assortment of cultivar-couples in the orchard

Determination of the genetic compatibility between self-incompatible cultivars is crucial in agriculture. The Rosaceae family carries the S-RNase-mediated gametophytic self-incompatibility (GSI) system. Each haplotype is conferred by an S-locus. The S-locus contains two highly polymorphic genes, S-RNase and SFB, which are characteristic of each haplotype and therefore these genes are ideal markers for molecular S-genotyping. In this study 43 Japanese plum cultivars grown in Israel were S-genotyped based on their S-RNase gene sequences. Four alleles, S^b, S^c, S^e and S^h are widespread and together are responsible for 87% of the S-haplotypes therefore many of the cultivar combinations are semi-compatible. In Israel semi-compatibility was shown to correlate with low yield. However, two cultivars, ‘Wickson’ S^fS^k and ‘Shiro’ S^fS^g carry rare S-haplotypes and, therefore, are fully compatible with most of the analyzed cultivars.     

Somatic embryogenesis and plant regeneration in Opuntia ficus-indica (L.) Mill. (Cactaceae)

We present here a method for the regeneration of the prickly-pear (Opuntia ficus-indica (L.) Mill.) through somatic embryogenesis (SE). Direct SE was induced by cultivating shoot apices devoid of leaf primordia on semisolid Murashige and Skoog (MS) basal medium supplemented with 4-amino 3,5,6-trichloropicolinic acid (picloram) at 4 mg l⁻¹. Somatic embryogenesis was influenced by the type, age, physiological and developmental stage of the explants, and also by light conditions, wounding, and the sucrose concentration in the induction medium. A histological analysis confirmed SE by revealing the presence of a closed vascular system in the developing embryos and the absence of a vascular connection between the somatic embryos and the explant.     

Biological activities of essential oil,aqueous and organic extracts of Pituranthos tortuosus (Coss.) Maire

Aqueous and organic extracts, essential oil as well as the powder of Pituranthos tortuosus (Coss.) Maire were tested on the growth of flax (Linum usitatissimum) and turnip (Brassica rapa) to assess its allelopathic potentialities. Besides, biological potentialities of its essential oil were evaluated by estimating its insecticidal (on Cryptolestes ferrugineus and Culex pipiens) and antifungal (on Fusaruim graminearum and Alternaria sp.) activities. Significant growth inhibition of target species was recorded in the presence of the aqueous and acetone extracts, more than 50% in certain cases. P. tortuosus essential oil also induced growth reduction which was almost total in the presence of a concentration of 4000 ppm. In addition, essential oil caused, for C. ferrugineus, a respective mortality of 50% and of 90% at concentrations of 205.68 and 300.37 μl/l air. Otherwise, 50 ppm was sufficient to induce 100% of mortality of C. pipiens larvae, whereas LC₅₀ and LC₉₀ were 15.46 ppm and 23.62 ppm, respectively. A concentration of 4 μl/ml of P. tortuosus essential oil caused a complete inhibition of the mycelial growth of F. graminearum and Alternaria sp. Results show that essential oil as well as organic extracts could be used for the production of natural bio-pesticides which could reduce our dependence on chemical pesticides.     

Cold storage of oil rose (Rosa damascena Mill.) flowers

Oil rose flowers were stored at 0 °C in four different packaging materials [plastic box + stretch film (PB + SF), Xtend^®, Smartbag^® and polyethylene (PE)] for 60 days. During storage, weight loss, O₂ and CO₂ concentrations in the packages, petal color and sensorial attributes were investigated besides essential oil content and composition. Storage duration and packages had significant (p < 0.01) effects on weight loss. At the end of storage, the lowest weight loss was in PE package (1.696%) whereas the highest weight loss was in Xtend^® (10.081%). The essential oil content was significantly (p < 0.01) affected by storage duration and packages. In addition, the essential oil contents obtained from all packages for a storage period of 10 days and the essential oil contents obtained from unstored (control) petals were included in the same group. At the end of storage, the essential oil contents decreased by 91.3, 57.7, 80.0 and 64.3% in PB + SF, Xtend^®, Smartbag^® and PE packages, respectively as compared to control. In addition, storage duration and package types significantly (p < 0.01) affected petal color, O₂ and CO₂ concentrations in the packages and sensorial scores. The concentration of citronellol, a main component of rose oil, increased in all packages during storage of 10 days in comparison to the control group while it varied in other storage durations and package types. However, nerol and geraniol were lower than the control group during storage while concentrations of nonadecane, heneicosane and eicosane were higher. In conclusion, loss of oil yield and quality, due to various reasons and particularly due to fermentation in oil rose from the harvest of petals to their distillation, can be minimized with storage of petals in all package types for up to 10 days.     

Calibration and validation of regression model for non-destructive leaf area estimation of saffron (Crocus sativus L.)

Leaf area measurements are required in several agronomical and physiological studies. Usually, there is an interest for measurement methods that are simple, quick and that will not destroy the leaf. The objectives of this study were to establish equations to estimate leaf area (LA) by using length (L), width (W) and dry weight (DW) of saffron leaves. Leaves of different sizes were collected from the experimental area at different time intervals. Leaf area was measured with an automatic measuring device and leaf dimensions were determined with a ruler. An equation for estimating the leaf area from L and W was developed and validated with the area of leaves collected during different periods. Regression analyses of LA versus L, W, LW and DW led several models that could be used for estimating the area of individual saffron leaves. A exponential model having L as the independent variables [LA = 191.33e^(L)0.0037] provided the most accurate estimate (R² = 0.9373, RMSE = 27.68) of saffron leaf area. Validation of the regression model showed that the correlation between measured and simulated values by the use of this equation was very high.     

The strawberry tree (Arbutus unedo L.) selection in Turkey

These days, the strawberry tree (Arbutus unedo L.), which is a comparatively under-utilized fruit tree species, has attracted attention because of many different possible commercial usages such as for processed and fresh fruit production and ornamental, the pharmaceutical and chemical industrial applications. They are disappearing from the natural flora for many reasons such as their being cut down for fire-wood and structuring on natural vegetation. A selection program was carried out with the aim of selecting strawberry tree genotypes of high quality from the native strawberry tree population grown in the Central Black Sea Region of Turkey. A total of 51 genotypes were evaluated in the program, in 2003 and 2004. The aims of the selection program were focused on fruit pomological characteristics such as fruit weight, total soluble solid, acidity, fruit taste, stoniness, juiciness, roughness and appearance. The data regarding pomological characteristics was evaluated by the Weighted Rankit method. The highest overall score for the characteristics tested was from genotype 57A01 followed by 57A15, 57GÇ08, 57A22 and 57A07, respectively. Their fruit weight was 11.08, 6.17, 6.30, 6.95 and 8.06 g, respectively. TSS contents and acidity of them varied from 21.4 to 30.0% and 0.80 to 1.59%, respectively. Phenological characteristics such as first bloom, fruit set and harvest date and some other fruit, flower and leaf characteristics of selected A. unedo genotypes were also determined. Vitamin C content of the samples varied from 97.83 to 280.00 mg/100 g. Finally, five superior strawberry tree genotypes were selected from the samples we studied for extensive cultivation and thus they were also saved from extinction.     

Root restriction-induced limitation to photosynthesis in tomato (Lycopersicon esculentum Mill.) leaves

Root restriction often depresses photosynthetic capacity and the mechanism for this reduction, however, remains unclear. To identify the mechanism by which root restriction affects the photosynthetic characteristics, tomato (Lycopersicon esculentum Mill.) seedlings were subjected to root restriction stress with or without supplemental aeration to the nutrient solution. With the development of the root restriction stress, CO₂ assimilation rate was decreased only in confined plants without supplemental aeration. There were also significant decreases in leaf water potential, stomatal conductance (g_s), intercellular CO₂ concentration (C_i), and increases in the stomatal limitation (l) and the xylem sap ABA concentration. Meanwhile, the maximum carboxylation rate of Rubisco (V_cmax) and the capacity for ribulose-1,5-bisphosphate regeneration (J_max) also decreased, followed by substantial reductions in the quantum yield of PSII electron transport (Φ_PSII). Additionally, root restriction resulted in accumulation of carbohydrates in various plant tissues irrespective of aeration conditions. It is likely that root restriction-induced depression of photosynthesis was mimicked by water stress.     

Towards crop improvement in bell pepper (Capsicum annuum L.): Transgenics (uid A::hpt II) by a tissue-culture-independent Agrobacterium-mediated in planta approach

Agrobacterium tumefaciens-mediated transformation has been one of the methods used to generate transgenic plants in bell pepper. An alternate transformation method that avoids/minimizes tissue culture would be beneficial for the improvement of bell pepper due to its recalcitrant nature. In this report, transgenic bell pepper plants have been developed by a tissue-culture-independent A. tumefaciens-mediated in planta transformation procedure. In the present study, two open pollinated varieties viz., Arka Gaurav and Arka Mohini were used for transformation. Agrobacterium strain EHA105 harboring the binary vector pCAMBIA1301 that carries the genes for β-glucuronidase (uid A) and hygromycin phosphotransferase II (hpt II) was used for transformation. GUS histochemical analysis of T₀ and T₁ plants at various stages of growth followed by molecular analysis using PCR, Southern analysis and RT-PCR allowed selection of transgenics. The method resulted in 17.8% and 11.4% of the T₀ plants in Arka Gaurav and Arka Mohini being selected as chimeric and 35.0% and 29.7%, respectively, were identified as stable transformants in the T₁ generation based on PCR analysis.