Grafting and calcium cyanamide as alternatives to methyl bromide for greenhouse eggplant production

Eggplant (Solanum melongena L.) seedlings (cv. Tsakoniki) were cultivated in soil artificially infested with V. dahliae Kleb. and then sterilized by either methyl bromide (MB) or calcium cyanamide. Grafted seedlings on the wild species Solanum torvum Sw. and the control seedlings (auto-rooted) were cultivated in soil sterilized by MB and then artificially infested with V. dahliae. The plants grafted on S. torvum and the ones grown in soil treated with calcium cyanamide (2001) exhibited significantly lower leaf symptom index (average value LSI = 1.55 and LSI = 1.00) and disease index (average value DI = 2.05 and DI = 1.20), respectively, as compared to the controls (average value LSI = 3.80 and DI = 5.50). Grafted plants on S. torvum and plants grown in soil treated with calcium cyanamide (2001) were more vigorous, as measured by plant height, stem diameter and root biomass than the controls. This resulted in an increased (over years) early (487.8% and 416.2%, 2001) and late production (277.0% and 241.3%, 2001) and mean fruit weight (over years) in early (93.7% and 49.6%, 2001) and late production (38.3% and 22.8%, 2001) as compared to the controls. In conclusion, grafting of eggplant and soil sterilized by calcium cyanamide had positive effects on growth, production and Verticillium wilt control.     

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.     

Evaluating variability of root size system and its constitutive traits in hot pepper (Capsicum annum L.) under water stress

The importance of root size system has long been recognized as crucial to cope with drought conditions. This investigation was conducted to: (i) evaluate the variability in root size system of hot pepper at maturity; (ii) estimate the effect of root size system on yield under drought conditions; and (iii) effect of water stress on xylem vessel development and total xylem cross-sectional area in roots of hot pepper cultivars. Twelve diverse hot pepper cultivars were grown in wooden boxes with two different water treatments, normal and in 50% water application as water deficit condition. Mean primary root length (PRL) showed a significant positive correlation with final fruit yield at normal as well as stressed condition. Total dry mass of fruit was reduced by 34.7% in drought treatments (DI) compared to full watered treatment (FI). At harvest, water-stressed plants had 21% lower root dry weight mass but higher root:shoot ratio other than FI. PRL, lateral root density, total xylem area per root cross-section showed a significant positive relationship with fruit yield. Also, lateral root density was higher in cultivars with higher xylem density, particularly in tolerant cultivars. Lateral root density (r = 0.847, P < 0.001) and total xylem cross-sectional area in root (r = 0.926, P < 0.001) were tightly related with total biomass production. The importance of root traits contributing to withstand drought in hot pepper is discussed.     

The genotypic effects on the chemical composition and antioxidant activity of sea buckthorn (Hippophae rhamnoides L.) berries grown in Turkey

In this study, chemical composition of berries of 10 sea buckthorn (Hippophae rhamnoides L.) genotypes in Turkey was investigated. The total phenolic content of the berries ranged from 21.31 mg gallic acid equivalents (GAE) per g dry weight basis to 55.38 mg GAE per g. The highest antioxidant activity was 93.54% (similar to the standard BHT at 200 mg/L) and the lowest was 80.38%. There was no correlation (R = 0.688) between the total phenolic content and the antioxidant activity. The major fatty acids in berries were palmitoleic acid (35.48%), followed by palmitic acid (28.13%), oleic acid (22.89%) and linoleic acid (3.96%). Total soluble solid content of sea buckthorn genotypes varied from 10.15 to 14.80%, titratable acidity varied from 2.64 to 4.54%, the pH varied from 2.63 to 2.98 and Vitamin C varied from 19 to 121 mg/100 mL. The average content of minerals in the sea buckthorn berries of different genotypes was 20,800 ppm N, 7100 ppm P, 7260 ppm K, 1960 ppm Ca, 1465 ppm Mg, 32 ppm Zn, 24 ppm Cu, 22 ppm Mn and 7 ppm Fe.     

Comparison of three pea cultivars (Pisum sativum) regarding their responses to direct and bicarbonate-induced iron deficiency

The aim of this work was to explore leaf characteristics underlining the difference in the sensitivity of pea cultivars (cv. Kelvedon, Douce and Lincoln) to Fe deficiency. Plants were grown in a greenhouse under controlled conditions in continuously aerated solution. Three treatments were used: 30 μM Fe (+Fe), 0 μM Fe (−Fe); direct deficiency and 30 μM Fe + 10 mM NaHCO₃ (+Fe+Bic); indirect deficiency for 12 days. Growth parameters, iron status, potassium content, chlorophyll fluorescence and photosynthetic capacity were studied. Our results showed that Fe deficiency led to a significant decrease of chlorophyll index (SPAD readings) and bivalent iron content in all Pisum sativum cultivars. The lower reduction was observed in Fe-deficient plants of Kelvedon and Douce. In addition, shoot length and whole plant dry weight were not affected by Fe deficiency in the latter cultivars. Both tolerant cultivars showed higher accumulation of potassium content in their leaves compared with the sensitive one. Moreover, both chlorophyll fluorescence ratios (F_v/F_m and F_v/F₀) were significantly decreased in all cultivars under both Fe deficiency treatments. The photosynthetic electron transport activity was reduced in the sensitive cultivar, especially in the absence of iron. The adverse effect of bicarbonate-induced Fe deficiency on the above mentioned parameters were more pronounced than that of the direct one. The capacity of both tolerant cultivars to preserve adequate chlorophyll synthesis, photosynthetic capacity and plant growth under iron-limiting conditions is related to the suitable nutrition of their leaves in ferrous iron, due to (at least partially) their higher potassium content.     

Plant spacing and cultivar affect melon growth and yield components

The objective of this study was to evaluate the effect of cultivars and in-row spacing on vegetative growth and yield components in melon (Cucumis melo L.). The cultivars ‘Early Dawn’ (ED), ‘Superstar’ (SS) and ‘Medoro’ (ME) were grown in rows 1.5 m apart at plant spacing 0.6, 0.9, 1.2 or 1.5 m. Cultivars ED and SS were grown in 1998 and 1999, while ED and ME were tested in 2000. The main vine length, diameter, number of leaves and lateral branches were highest at 1.5 m in-row spacing in 1999 and 2000, while there was no influence of plant spacing on observed growth parameters in 1998. ED performed better compared with other two varieties, mostly due to more fruit per ha and per plant. Yield was significantly decreased with increased plant spacing in all 3 years of research. At in-row spacing of 1.5 m yield was in average 25% lower than at 0.6 m spacing. Average fruit weight slightly increased with plant spacing up to 1.2 m, even though the differences were significant only in 1999. Small differences among cultivars were also found. Number of fruit per ha decreased linearly while number of fruit per plant increased linearly as plant spacing increased in all cultivars except SS in 1999.     

Susceptibility of grapevine rootstocks to Cylindrocarpon liriodendri and C. macrodidymum

The susceptibility of the grapevine rootstocks most commonly used in Spain to Cylindrocarpon liriodendri and C. macrodidymum was evaluated. Rooted cuttings of rootstocks 110-R, 1103-P, 140-R, 161-49C, 196-17C, Fercal and SO4 were inoculated by dipping their roots in conidial suspensions (5 × 10⁵ conidia mL⁻¹) of both pathogens and placed in a greenhouse. One month later, each plant was drench inoculated with 20 mL of the corresponding spore solution to guarantee root infection. After four months of incubation, root disease severity index (RSDI) and dry weights of shoot and root were recorded for each plant. RSDI was the best variable to show the effect of the inoculation with C. liriodendi and C. macrodidymum. According to this, all rootstocks inoculated were affected by the disease in some degree, being the rootstock 110-R the most susceptible to both pathogens.     

The influence of flesh colour and growing locality on polyphenolic content and antioxidant activity in potatoes

Compounds contained in potato tubers that may act as antioxidants have been attracting increased attention. The main antioxidants in potatoes are polyphenolic compounds. The total polyphenolic (TP) content and antioxidant activity (AA) in potato tubers of four yellow-fleshed cultivars and two purple-fleshed cultivars from the 2004 harvest were tested. The total polyphenolic content was determined using the Folin-Ciocalteau reagent (FC) and the antioxidant activity, with the DPPH method using stable free radical 1,1-diphenyl-2-picrylhydrazyl (DPPH·). Significant differences were found in total polyphenolic content between the yellow-fleshed cultivars Karin, Impala, Ditta, and Saturna. Significant differences between yellow and purple-fleshed potatoes were found: the yellow-fleshed cultivars had an average of 2.96 g gallic acid equivalents (GAE) per kg DM, the purple-fleshed cultivars 4.68 g GAE per kg DM. Significant differences in AA between the yellow and purple-fleshed cultivars were determined (yellow-fleshed in an average of 139.3 mg equivalents of ascorbic acid [EAA] per kg DM, purple-fleshed 332.3 mg EAA per kg DM) as well as between the purple-fleshed cultivars Valfi (298 mg EAA per kg DM) and Violette (366 mg EAA per kg DM). The major antioxidants in purple-fleshed cultivars were anthocyanidins petunidin (Valfi 92%_rel) and malvidin (Violette 85%_rel). Cv. Violette contained three times higher content of anthocyanidins than cv. Valfi. Among four experimental localities, the higher situated potato-growing region of Stachy (locality with the lowest average annual and daily minimum temperatures and the highest sum of precipitation in the vegetative period) differed significantly from other localities by a higher TP content, AA and yield in the analyzed cultivars. A high positive correlation between antioxidant activity and total polyphenolic content with R = 0.8648, R² = 0.7478 (P = 0.0001) was found.     

Relationship between inbreeding coefficients and plant height of 1-year-old seedlings in crosses among Japanese pear (Pyrus pyrifolia Nakai) cultivars/selections

Plant height, a vigor trait, in 1-year-old seedlings made from Japanese pear (Pyrus pyrifolia) cultivars/selections was measured using 994 individuals from 29 families. The family mean of plant height was negatively correlated (r = −0.72**) to the inbreeding coefficients (F). The regression of the family mean (Fm) on the F value (Fm = 130 − 104F) showed that inbreeding depressions were 8%, 20%, and 40% for F = 0.1, 0.25, and 0.5, respectively. According to the regression, the family mean at F = 0 was estimated at 130 cm. These results showed that the vigor was greatly influenced by inbreeding in Japanese pear. Within-family variances, the genetic segregation of offspring in a family, differed according to family. The proportions of offspring with plant height above 130 cm (estimated Fm for F = 0) were extremely low, i.e., 0–17% for 0.5 ≤ F < 0.60 and 0–8% for F = 0.75.     

胡萝卜镰刀菌根腐病病原菌的鉴定

为明确内蒙古西部地区一种新病害——胡萝卜镰刀菌根腐病病原菌种类,采用组织分离法对病原菌进行分离,用胡萝卜片接种、有伤口和无伤口胡萝卜肉质根接种及盆栽接种进行致病性测定,并通过形态学和分子生物学方法对病原菌进行鉴定。结果表明,分离到的3个菌株F1、F2和F3均表现致病性。经形态学鉴定,F1菌株为尖孢镰刀菌(Fusarium oxysporum),F2和F3为茄病镰刀菌(Fusarium solani)。进一步采用真菌通用引物ITS1/ITS4对病原菌r DNA-ITS进行PCR扩增,并将测序结果在Gen Bank中进行同源性比对分析,鉴定结果和形态学鉴定的结果一致。明确了内蒙古西部地区胡萝卜镰刀菌根腐病的病原菌种类为尖孢镰刀菌(F.oxysporum)和茄病镰刀菌(F.solani)。     

Models for predicting the mass of apricot fruits by geometrical attributes (cv. Shams,Nakhjavan, and Jahangiri)

Fruits with the similar weight and uniform shape are in high demand in terms of marketing value. Therefore, an awareness of grading fruit based on weight is crucial. A part of this research was aimed to present some physical properties of three Iranian apricot cultivars (Shams, Nakhjavan, and Jahangiri). In addition, apricot mass was predicted by different physical characteristics with linear and nonlinear models as three different classifications: (1), single or multiple variable regressions of apricot dimensional characteristics. (2), single or multiple variable regressions of apricot projected areas and (3), estimating apricot mass based on its volume. All properties considered in the current study were found to be statistically significant at the 1% probability level. The highest and the lowest dimensional characteristics were found for Jahangiri and Nakhjavan cultivars, respectively. Sphericity values had significant difference among the tested cultivars. The latter property values were 0.971, 0.917, and 0.973 for Shams, Nakhjavan, and Jahangiri cultivars, respectively. Based on the results, the surface area of the Jahangiri cultivar was found to be 6458.35 mm², followed by the Shams and Nakhjavan cultivars, which had a mean of 6115.36, and 4395.25 mm², respectively. In this paper, the coefficients of static friction of the cultivars on three different surfaces are also reported. The results showed that mass modeling of apricot based on minor diameter and three projected areas are the most appropriate models in the first and the second classifications, respectively. In third classification, the best model was obtained on the basis of the actual volume as M = 0.997V_m + 0301, R² = 0.98, R.S.E. = 1.711 with R² = 0.98. It was concluded that the suitable grading system of apricot mass is based on minor diameter as nonlinear relation: M = 0.0019c^2.693, R² = 0.96, R.S.E. = 2.2.     

Assessment of tolerance to NaCl salinity of five olive cultivars,based on growth characteristics and Na and Cl exclusion mechanisms

Changes caused by NaCl-induced salinity on several growth parameters and ions accumulation have been measured in five olive (Olea europaea L.) cultivars (‘Chemlali’, ‘Chetoui’, ‘Koroneiki’, ‘Arbequina I18’, and ‘Arbosana I43’) growing in a greenhouse in nutrient solution pot experiment. One-year-old plants were transplanted to sand–perlite (1:1) culture, and were irrigated with half-strength Hoagland nutrient solution containing NaCl at various levels (0.5, 50, 100 and 200 mM). Salinity induced significant decrease in growth parameters, but to a different extent in each cultivar. Leaf growth and total leaf area per plant were significantly affected by all salinity treatments in all studied cultivars, being ‘Arbequina I18’ the most sensitive cultivar. Leaf drop phenomenon was observed from 60 days after salt application at high salinity treatments, mainly in Arbequina I18. Contrary to leaf area, leaf thickness increased progressively during the experiment. ‘Chemlali’ developed thicker leaves at the two highest salinity treatments when compared to the other cultivars. Na⁺ and Cl⁻ concentrations were higher in roots than in shoots and leaves in most of the cultivars investigated. The effectiveness of Na⁺ exclusion mechanism in the roots differed significantly among studied cultivars, working effectively in ‘Chemlali’ (by inhibiting translocation of Na⁺ to the aerial part) and being much less efficient in ‘Arbequina I18’. Furthermore, leaf abscission can be considered as an additional tolerance mechanism of olive cultivars allowing the elimination of leaves that had accumulated Na⁺ and Cl⁻ ions. Tolerance to salinity stress was as follows: ‘Chemlali’ > ‘Chetoui’ > ‘Arbosana I43’ > ‘Koroneiki’ > ‘Arbequina I18’. This order of salt tolerance was indicated by lower reduction in plant growth parameters (shoot elongation, trunk diameter, total plant dry weight, internodes length, and total leaf area), the increase of leaf thickness, and by the effectiveness of the exclusion mechanism of Na⁺ and Cl⁻ in the root system.     

Correlation of saponin content and Fusarium resistance in hybrids from different ploidy levels of Lilium Oriental

Lilium Oriental is of great commercial value, but large losses in production can result from its susceptibility to diseases caused by Fusarium ssp. Here we report a mutant of Lilium Oriental resistant to Fusarium, Cai-74, which was generated from crossing tetraploid (from ‘Star fighter’) and diploid (‘Cor. Amore’ × ‘Acapulco’) plants. Hybrid progeny were screened via inoculating Fusarium oxysporum into tissue cultured plantlets in a greenhouse. Although Cai-74 had a saponin content of 3.81 mg g⁻¹, which is much higher than its parents, the highly Fusarium-resistant wild species Lilium dauricum had the highest content of 4.11 mg g⁻¹ among the tested genotypes. Cai-74 had a 3A karyotype rather than 3B found in normal plants of Lilium Oriental. Taken together, our results suggest that Cai-74 bears a chromosomal variation in its structure, and that the saponin content in the lily scales correlate with the Fusarium resistance of Lilium Oriental. Cai-74 could be used as a gene resource for breeding Fusarium-resistant cultivars of Lilium Oriental.     

Gibberellin producing Neosartorya sp. CC8 reprograms Chinese cabbage to higher growth

Gibberellins production by soil fungi received little attention, although substantial work has been carried out on other growth promoting aspects of soil borne fungi. We investigated gibberellins production and growth promoting capacity of a novel fungal strain of Neosartorya, which was isolated from the roots of Chinese cabbage (Bassica rapa L. ssp. pekinensis). Fungal culture filtrates (CF) obtained from pure cultures of 16 endophytic fungi were bioassayed on Waito-C, in order to investigate plant growth promoting capacity of these fungi. The fungal isolate CC-8 induced maximum shoot length of Waito-C (13.0 cm) as compared to control treatments. In a separate experiment, the CF of fungus CC-8 significantly promoted plant length and biomass of Chinese cabbage. The fungal CF also increased endogenous bioactive GA₁ and GA₄ contents of Chinese cabbage. Gibberellin analysis of CF of CC-8 showed presence of both physiologically active and non active gibberellins in higher concentrations (GA₁, 1.42 ng/ml; GA₃, 5.93 ng/ml; GA₄, 11.36 ng/ml; GA₇, 3.25 ng/ml; GA₉, 0.79 ng/ml; GA₁₅, 1.18 ng/ml). The culture filtrate of CC-8 produced higher amounts of GA₃, GA₄, GA₇ and GA₉ than wild type Fusarium fujikuroi, a well known gibberellins producing fungus. The fungal isolate CC-8 was later identified as a new strain of Neosartorya species on the basis of traditional and advance molecular techniques.     

Cultivar and anatomical analysis of corolla enlargement of petunia (Petunia hybrida Vilm.) by cytokinin application

We found that the corolla of petunia (Petunia hybrida Vilm.) could be conspicuously enlarged by the separate application of three cytokinins: forchlorfenuron (CPPU), N⁶-benzylaminopurine (BA), and zeatin. To obtain the same enlargement as that achieved by CPPU, approximately 30 and 900 times the concentration of BA and zeatin, respectively, were required. CPPU at 3.2 μmol/L increased the limb area of the corollas of 15 cultivars to between 1.3 and 2.4 times (1.8 times on average) the size of the control area. The increase was negatively correlated (R = 0.58) with the “genetic” limb area (i.e., that of the untreated plant). The enlargement of the corolla caused by cytokinin application was mainly attributed to an increase in cell number in most cultivars. This increase resulted from a high rate of cell proliferation and from prolongation of the cell proliferation phase during corolla development. This anatomical change caused by cytokinin application was similar to the anatomical difference among cultivars because genetic differences in limb area resulted mainly from differences in cell number.     

Modeling plant morphology and development of petunia in response to temperature and photosynthetic daily light integral

The effects of mean daily temperature (MDT) and mean photosynthetic daily light integral (MDLI) on flowering during the finish stage of two petunia (Petunia × hybrida) cultivars were quantified. Petunia ‘Easy Wave Coral Reef’ and ‘Wave Purple’ were grown in glass-glazed greenhouses at 14–23 °C or 14–26 °C and under 4–19 mol m⁻² d⁻¹ with a 16-h photoperiod. The flower developmental rate was predicted using a model that included a linear MDT function with a base temperature multiplied by an exponential MDLI saturation function. The flower developmental rate increased and time to flower decreased as MDT increased within the temperature range studied. For example, under a MDLI of 12 mol m⁻² d⁻¹, as MDT increased from 14 to 23 °C, time to flower of ‘Easy Wave Coral Reef’ and ‘Wave Purple’ decreased from 51 to 22 d and 62 to 30 d, respectively. Flower developmental rate increased as MDLI increased until saturation at 14.1–14.4 mol m⁻² d⁻¹. Nonlinear models were generated for effects of MDT and MDLI on flower bud number and plant height at flowering. The number of flower buds at flowering increased as MDT decreased and MDLI increased. For example, at an MDT of 14 °C with 18 mol m⁻² d⁻¹, plants had 2.5–2.9 times more flower buds than those grown at 23 °C and 4 mol m⁻² d⁻¹. Models were validated with an independent data set, and the predicted time to flower, flower bud number, and plant height were within ±7 d, ±20 flowers, and ±4 cm, respectively, for 96–100%, 62–87%, and 93–100% of the observations, respectively. The models could be used during greenhouse crop production to improve scheduling and predict plant quality of these petunia cultivars.     

Effect of nitrogen and sulfur interaction on growth and pungency of different pseudostem types of Chinese spring onion (Allium fistulosum L.)

Effects of N and S supply on the growth and pungency (estimated as pyruvic acid levels) of Chinese spring onion (Allium fistulosum L. var. giganteum Makino) were investigated in two pot experiments using soilless growing media. In the first experiment the effects of S supply (0.01 and 4.00 mmol L⁻¹ SO₄²⁻) on the growth and pungency of Chinese spring onion were investigated among four cultivars with fleshy root type or long pseudostem type. In the second experiment the effects of different S (0.01 and 4.00 mmol L⁻¹ SO₄²⁻) and N (1.5, 3.0, 6.0, 12.0 and 24.0 mmol L⁻¹ N) supply levels on the growth and pungency of Chinese spring onion were studied. Fleshy root spring onion had stronger pungency and larger pseudostem diameter than long pseudostem spring onion, and the pungency of fleshy root spring onion was regulated to a greater extent by N and S supply compared with long pseudostem spring onion. Increasing S supply level significantly increased the biomass, N and S uptake and pungency of all cultivars tested. The biomass of Chinese spring onion of fleshy root type (cv Longyao) and long root type (cv Zhangqiu) was more influenced by N supply than it was by cultivar or S supply. Low S supply decreased the pungency of the two cultivars with increasing N supply. No significant differences in N or S uptake or pungency were observed in the two cultivars with different S supply at the N supply level of 1.5 mmol L⁻¹ N, however, cultivar differences in N and S uptake and pungency were investigated at high N supply (12.0 mmol L⁻¹ N) and S supply (4.0 mmol L⁻¹ SO₄²⁻). Excessive N supply (24.0 mmol L⁻¹) significantly inhibited plant growth, retarded S assimilation, and decreased pungency. It is therefore essential to apply the optimum recommended rate of N fertilizer in Chinese spring onion production.     

Genetic assessment of apple germplasm in Bosnia and Herzegovina using microsatellite and morphologic markers

Traditional apple cultivars from Bosnia and Herzegovina (B&H), potentially diverse due to specific geographic location and history of the country, represent a possible source of valuable traits for future breeding efforts and sustainable fruit growing. A total of 39 accessions, 24 traditional B&H cultivars and 15 modern international cultivars, maintained at the ex situ apple collection “Srebrenik” in Northeast Bosnia were, investigated using 10 SSR (simple sequence repeats) markers and 23 morphologic characteristics. All the used primer pairs manage to amplify clearly distinguishable and highly polymorphic SSR alleles, in average 10.4 alleles per locus. More than two different alleles per locus were detected for seven accessions (five traditional B&H cultivars and two international cultivars). Forty one unique alleles were exclusively present within the B&H cultivars, while seven unique alleles were only detected within international cultivars. The differentiation between traditional B&H and international cultivars (Fst = 0.060; P < 0.0001) was significant, also confirmed by analyses of molecular variance (AMOVA) (f_CT = 0.092; P < 0.001). Cluster analyses of 39 apple accessions, based on 10 SSR loci, revealed that only two traditional B&H cultivars grouped tightly with international cultivars (Ljepocvjetka and Bobovec Jon), while the rest formed separate clusters. Multivariate analyses of variance (MANOVA), nonparametric multivariate analyses of variance (NPMANOVA) and analyses of similarity (ANOSIM) showed statistically significant difference in morphologic characteristics between traditional B&H cultivars and the international cultivars. Cluster analyses of 39 apple accessions, based on the morphologic data, displayed less differentiation between traditional and international accessions, in comparison to the cluster analyses based on molecular data. No correlation between the molecular and morphologic data set was detected using the Mantel test. Many of the morphologic characteristics which have been analyzed in this study have significant commercial importance, we can assume that unlike the microsatellites these traits have been under agronomic selection pressure.     

Somatic embryogenesis and Agrobacterium-mediated transformation of Japanese apricot (Prunus mume) using immature cotyledons

This study describes a successful method of somatic embryogenesis and genetic transformation using immature cotyledons of Prunus mume. Immature cotyledons from four different developmental stages of eight different P. mume cultivars were used for the experiments to optimize somatic embryogenesis and genetic transformation protocols. Somatic embryogenesis was induced when the explants were cultured on somatic embryo inducing medium consisting of MS basic medium supplemented with 1 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 1 μM 6-benzyladenine (BA). They were cultured for 30 days and then transferred to somatic embryo propagation medium containing 0.1 μM α-naphthaleneacetic acid (NAA) and 5 μM BA. It appeared that the developmental stage of the immature cotyledons used as explants was the most important factor for somatic embryogenesis; higher frequencies of somatic embryogenesis were observed when the immature cotyledons were less than 5 mm in length regardless of cultivars. For genetic transformation, the immature cotyledons were inoculated with Agrobacterium tumefaciens EHA101 harbouring a binary plasmid vector with neomycin phosphotransferase II and an intron-interrupted β-glucuronidase gene under the control of cauliflower mosaic virus 35S promoter, and three transgenic plant lines were obtained from inoculated “Sirakaga” immature cotyledons. Transgenic somatic embryos and shoots were selected using 25 mg l⁻¹ kanamycin. Integration of transgenes in the genome of GUS-positive putative transgenic shoots was confirmed by PCR and Southern blot analyses.     

High frequency plant regeneration from microspore-derived embryos of ornamental kale (Brassica oleracea L. var. acephala)

The aim of this work was to study the effect of solid medium, developmental stage, embryonic age, cold treatment and additives to the medium on plant regeneration from microspore-derived embryos in four F₁ hybrids of ornamental kale (Brassica oleracea L. var. acephala). The results showed that all of the cultivars responded best when the embryos were cultured in solidified B5 medium with 1% agar. Optimal regeneration was gained when cotyledonary embryos were cultured for 25 days. Cold treatment significantly improved plant regeneration with a frequency of up to 79.0% under 4 °C for 2 d or 5 d. The addition of 3.0 or 5.0 mg/L silver nitrate (AgNO₃) increased the frequency of plant regeneration. In the Zhouyehongxin cultivar, the frequency of plantlet development reached 84.4%. The addition of activated charcoal reduced embryo hyperhydricity.