Identification of Petunia hybrida cultivars that diurnally emit floral fragrances

In order to identify genetic resources for breeding fragrant petunias for use as bedding plants, volatile compounds released by day from the flowers of 40 commercial Petunia hybrida cultivars were analyzed using a solid-phase micro-extraction technique coupled with GC–MS. The three cultivars with solid deep-blue flowers that accumulate malvidin in corollas with high tissue pH were found to emit abundant iso-eugenol as the principal floral fragrance. Several other cultivars that emitted considerable amounts of methylbenzoate and/or benzylbenzoate from the flower were also identified. Association between the floral fragrance and the other floral traits such as floral anthocyanin composition and corolla-tissue pH was discussed.     

Change in flower morphology of Torenia fournieri Lind. induced by forchlorfenuron application

We induced various flower morphologies in torenia (Torenia fournieri Lind.) by the application of forchlorfenuron (CPPU). Those morphologies were the combination of four basic morphological changes, the development of serrate petals, incised petals, a paracorolla, and an increased number of floral organs. These morphological changes occurred systematically depending on the floral stage at the time of CPPU application. Serrate petals were induced when CPPU was applied during the stages of corolla development, whereas application at younger stages induced petal incision. The serrate petal margin resulted from preferential proliferation of cells around the vascular bundles, whereas petal incision likely resulted from the lateral outgrowths of petal. A paracorolla was induced at the adaxial petal face when CPPU was applied between the sepal development stage and early corolla development. The paracorolla appears to have arisen from the lateral outgrowths of the stamen. The numbers of stamens, petals, and sepals increased when CPPU was applied at and before the differentiation of sex organs and the corolla. Enlargement of the floral meristem probably caused this increase. Application of N⁶-benzylaminopurine and zeatin did not induce these morphological changes.     

Application of treated wastewater for cultivation of roses (Rosa hybrida) in soil-less culture

No information is available today concerning the effect of irrigation with secondary treated sewage water on growth, production or quality of roses or other cut flowers. In the present study we investigated the effect of irrigation with treated sewage water on roses cultivated in two soil-less medium, perlite, an inert mineral medium and Choir (coconut fibers), an organic medium of high ion absorption capacity. During 12 months of exposure to the treated water, the visible appearance of the plants, their growth, the quantity and size of the flowering stems and their postharvest performance were not affected by the irrigation treatments. Contents of macroelements in the leaf tissues were unaffected by the irrigation with the secondary treated sewage water. At the same time, Cl contents increased 47% in perlite and 73% in Choir grown plants reaching levels characteristic of exposure to moderate salinity. Mn, Cu and B contents increased as well under cultivation in both perlite and Choir under irrigation with treated sewage water. On the other hand, contents of Fe, Zn, Mo and Al, were similar in all treatments. In all treatments contents of all the examined micro and macroelements were within the range accepted for proper plant function.     

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.     

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.     

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.     

In vitro germination of walnut (Juglans regia L.) embryos

The present studies were undertaken with a view to standardize the medium and culture conditions for embryo culture of five cultivars of walnut viz., ACO 38853, Netar Akhrot, Gobind, Solding Selection and Blackmore. Embryos from mature fruits were aseptically excised and cultured on MS medium supplemented with different combinations of BAP, kinetin and GA₃. Best performing medium was MS with 0.5 mg l⁻¹ kinetin, 0.5 mg l⁻¹ BAP and 2 mg l⁻¹ GA₃ yielding 66.6% germination in Netar Akhrot after 12 days of culturing. Percent germination of excised embryos was higher when GA₃ and cold treatments were simultaneously applied as compared to those when applied separately. Netar Akhrot was found to be the best responding cultivar, which had a range of 25–66.6% embryo germination under different culture conditions. Plantlets with shoots and roots have been obtained in Netar Akhrot and ACO38853 and are transferred to soil after hardening.     

Higher than optimum temperature under CO2 enrichment influences stomata anatomical characters in rose (Rosa hybrida)

There is little available information on the effects of temperature and CO₂ enrichment on stomata anatomical characteristics of plants. Effect of these two microclimates was studied on five rose (Rosa spp.) cultivars, viz. ‘First Red’ (used as check), ‘Arjun’, ‘Raktima’, ‘Raktagandha’ and ‘Pusa Pitamber’. Budded, single-stemmed rose cultivars having five lateral buds were grown in controlled environment growth cabinets under enriched CO₂ (1000 μmol mol⁻¹) and optimum (28/18 °C, T₀) or high (35/25 °C, T₁) temperature for 50 days. All observations were made on the abaxial leaf surface. Significant increases in stomatal density (68.7%), index (29.6%) and epidermal cell density (37.3%) were recorded in plants grown at high temperature over control with CO₂ enrichment. The cultivars responded differently in terms of length and width of guard cell and stoma (pore) under high temperature, however, the values averaged over treatments showed a significant reduction in these parameters. Further, number of stomata per leaf was higher (28.3%) in plants grown at high temperature, except First Red. A reduction in mean leaf area (26.7%) and dry mass (32.0%) was recorded at high rather than optimum temperature. The specific leaf area was maximum in Arjun (87%) while in First Red, a 14% reduction was noted at high temperature.     

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.     

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.     

Leaf anatomical characteristics and physiological responses to short-term drought in Ziziphusmauritiana (Lamk.)

The aim of this work was to study the association between leaf anatomical characteristics and response to short-term drought stress in Ziziphusmauritiana Lamk. Six Z. mauritiana cultivars (Seb, Gola, Umran, Keitly, Q-29 and B-5/4) under field conditions in Israel's Negev desert were studied. Width of palisade mesophyll, spongy mesophyll and epidermis, xylem number and diameter in mid-vein were investigated with light microscopy. Short-term (3 weeks) drought stress tolerance was evaluated by monitoring plant response (leaf transpiration, diffusive resistance, chlorophyll fluorescence, leaf water potential and leaf relative water content). Greater epidermis and mesophyll widths and xylem diameters and densities were associated with increased tolerance to short-term water deficit expressed by preliminary wilting symptoms and proportional differences between initial and final physiological parameters. Significantly larger differences were found in Keitly, Umran and B-5/4 than in Seb, Gola and Q-29, indicating that the former cultivars are more sensitive to drought stress. Our results indicate that tolerance in descending order was Seb, Q-29, Gola, B-5/4, Keitly and finally Umran. The existence of an association between anatomical characteristics and short-term drought stress tolerance based on physiological responses is suggested.     

Development of a plant regeneration system from seed-derived calluses of centipedegrass [Eremochloa ophiuroides (Munro.) Hack]

The objective of this study is to establish plant regeneration system with the seed of the new Chinese selection “E-126”of centipedegrass [Eremochloa ophiuroides (Munro.) Hack] as explant. In present study, the following results were obtained: (1) The medium formulation most suitable for calluses induction was identified to be MS with 1.0 mg l⁻¹ 2,4-D + 30 g l⁻¹ mannitol + 50 ml l⁻¹ coconut milk and the ratio of calluses induction was 96.0%, including 5.2% of yellow granule calluses induction. The above medium formulation was adopted for subculture. (2) The rate of shoot regeneration from yellow granular calluses was 98.0% by MS optimum medium formulation with 2.0 mg l⁻¹ KT + 50 ml l⁻¹ coconut milk. The differentiated rate retained as high as 88.0% even after 5 times of subculture and 18.6% after 15 times of subculture. The optimum medium formulation for shoot growth was identified to be MS medium plus 2.0 mg l⁻¹ BAP, 0.8 mg l⁻¹ NAA and 50 ml l⁻¹ coconut milk. (3) The optimum medium for shoot rooting was identified to be MS medium with 0.6 mg l⁻¹ NAA + 50 ml l⁻¹ coconut milk, and the rooting rate to be 98.0%. The survival rate of transplanted plantlets from tubes to basin with soil was 92.0%. In conclusion, the plant regeneration system was successfully developed in this study, which may provide basic reference for screening of somaclonal variants and genetic transformation of centipedegrass.     

Isoenzyme and microsatellite analysis of Vitis vinifera L. varieties from the Hungarian grape germplasm

The aim of our work was to investigate the genetic diversity of grapevine with biochemical and molecular markers (isoenzyme and SSR). The isoenzyme patterns of 4 enzyme systems (catechol-oxidase, glutamate-oxalacetate-transaminase, acid phosphatase and peroxidase) and the microsatellite profile in 6 loci (VVS2, VVS16, VVMD7, VMC4A1, VMC4G6, VrZag79) of 48 grapevine varieties were analysed. The results with CO, GOT, AcP and PER enzymes were reproducible and the zymograms obtained from the woody stems were independent from the time of sampling during the dormant period of the grape. Based on the isoenzyme patterns of these 4 enzymes most of the investigated varieties (40/48) were identified. A correlation was found between the isoenzyme patterns and the classification to convarietas of the varieties. It was established, that while the varieties of the convarietas pontica differed from those of the convarietas orientalis and occidentalis, the two latter groups could have not been differentiated from each other. Based on the SSR (simple sequence repeat) analyses 46 of the 48 investigated varieties were identified. Even ‘Pinot blanc’ and ‘Pinot gris’ cultivars belonging to the same conculta (Pinot) could be differentiated in their VMC4A1 locus.     

Flower morphologic anatomy and embryological characteristics in Chrysanthemum multicaule (Asteraceae)

Chrysanthemum multicaule is an annual herbaceous ornamental species. The inflorescence is gynomonoecious and consists of bisexual tubular florets and female ray florets. The pistils consist of two stigmas which are of the open type with a hollow stylar canal. At the base of the tubular floret style, the pistil is surrounded by oil gland cells. The anthers are tetrasporangiate and the young anther wall is composed of epidermis, endothecium, middle layer and tapetum. The mature anther wall comprises only thickened endothecium after the release of the pollens. In the tubular florets, simultaneous microsporocyte meiotic cytokinesis results in mostly tetrahedral with a small proportion of decussate tetrads. The mature pollen grain is tricellular. The ovules are unitegmic and tenuinucellate, and the nucellus degenerates during the development of the megasporocyte. The development of the embryo sac follows the Polygonum type. At 4–6 days after blooming, the embryos reached the globular stage, thereafter passing through the heart- and torpedo-shape stages before maturing into the cotyledon embryos. From blooming to seed maturity, it takes about 3–4 weeks under our conditions.     

Chlorocholine chloride application effects on photosynthetic capacity and photoassimilates partitioning in potato (Solanum tuberosum L.)

Treatment of potato (Solanum tuberosum L.) with chlorocholine chloride (CCC) applied twice as a foliar spray 25 and 30 days after planting has shown to decrease shoot and stolon growth but increase tuber yield. However, the regulatory role of CCC on translocation of recently fixed photoassimilates into different parts of potato plants has not been fully illustrated. In this study, ¹⁴C-isotope labelling technique was used to estimate the photosynthetic capacity and photoassimilate partitioning among leaves, stems, roots + stolons, and tubers of potted potatoes treated with 1.5 g l⁻¹ CCC. CCC treatment significantly increased tuber dry mass but reduced leaf dry mass. CCC-treated leaves had significantly higher chlorophyll and carotenoid contents and assimilated 22.0% more ¹⁴CO₂ per leaf dry mass than the controls. Compared with the control, CCC treatment reduced the translocation of ¹⁴C-photoassimilates into leaves, stems and roots + stolons but increased that into tubers. CCC-treated leaves exported 14.6% more ¹⁴C-photoassimilates into other parts of the plants. In addition, CCC treatment reduced ¹⁴C-soluble sugar and ¹⁴C-starch accumulation in leaves and stems but enhanced them in tubers and roots + stolons. Collectively, the results indicate that CCC treatment significantly improves the photosynthetic capacity of potato leaves and promotes photoassimilates partitioning into tubers thereby enhancing tuber growth.     

Chemical compositions and insecticidal effects of essential oils isolated from Achillea gypsicola, Satureja hortensis, Origanum acutidens and Hypericum scabrum against broadbean weevil (Bruchus dentipes)

The essential oils of aerial parts of Achillea gypsicola Hub.-Mor., Hypericum scabrum L., Satureja hortensis L., and Origanum acutidens (Hand.-Mazz.) Letswaart were analyzed in this study by GC and GC–MS and their oils were tested for toxicity against broadbean weevil (Bruchus dentipes). A. gypsicola oil contained camphor (40.17%), 1,8-cineole (22.01%), piperitone (11.29%), borneol (9.50%) and α-terpineol (1.56%) as major components. A total of 74 components were identified by GC–MS in H. scabrum oil, including α-pinene (9.26%), terpinen-4-ol (5.12%), camphor (5.94%), δ-cadinene (4.52%), pulegone (4.45%), γ-muurolene (4.12%), pinocarvone (3.97%) and β-caryophyllene (3.42%) as predominant components. The essential oils of O. acutidens and S. hortensis were characterized by high contents of carvacrol (86.99% and 55.74%), γ-terpinene (0.71% and 20.94%), p-cymene (1.95% and 12.30%), α-terpinene (0.13% and 2.04%) and β-caryophyllene (1.30% and 1.08%). All of the essential oils were toxic to adults of B. dentipes and insect mortality increased with increasing concentration of each oil. The oils (20 μl dose) brought about 100% mortality in 36 h. Although desirable insecticidal activities against the pest were achieved with the oils from all four plant species, S. hortensis and O. acutidens oils were more effective, particularly after 6 h of treatment. The current results concluded that the essential oils, in particular O. acutidens and S. hortensis oils, may be used as potential botanical insecticides against B. dentipes.