Breaking the intergeneric hybridization barrier in Carica papaya and Vasconcellea cauliflora

The present investigation was undertaken to develop PRSV (Papaya ringspot virus) resistant hybrids through intergeneric hybridization. Intergeneric hybridization was done involving nine Carica papaya cultivars as female and Vasconcellea cauliflora as male. To break the intergeneric hybridization barrier, various nutrient combinations were used. Among the combinations used, sucrose 5%, sucrose 5% + boron 0.5% and sucrose 5% + CaCl₂ 0.5% improved the fruit set and seed set percentage. A total number of 1197 flowers were pollinated and 308 fruits were obtained. On extraction, 721 seeds were obtained from CO 7, Pusa Nanha and CP 50. Out of 721 F₀ seeds (crossed seeds) sown, 419 seeds germinated and artificial screening for PRSV was carried out 27 days after sap inoculation. Out of 29 F₁ hybrid plants from CO 7 x V. cauliflora cross, only six plants namely CO 7V1 to CO 7V6 were found free from PRSV symptoms. Similarly, out of 55 F₁ hybrids from cross involving Pusa Nanha x V. cauliflora only 23 plants namely PNV1 to PNV23 were found free from the symptoms and 70 plants namely CPV1 to CPV70 out of 335 plants of CP50 x V. cauliflora cross were found free from PRSV symptoms. Among the crosses, Pusa Nanha x V. cauliflora had higher yield under PRSV infected conditions, however, total soluble solids and total sugars were found lesser than the CO 7 x V. cauliflora cross. The hybridity of the progenies were confirmed by using ISSR (Inter Simple Sequence Repeats) primers by the amplification of DNA from progenies and their parents. ISSR primers UBC 856, UBC807 and ISSR primer combinations UBC 856-817, UBC 810-817, UBC 861-817, UBC 856-810, UBC 861-810 and UBC 856-817 clearly amplified specific bands of the male parent, which were present in F₁ progenies, but it was absent in female parents.     

Quality and volatile attributes of attached and detached ‘Pluk Mai Lie’ papaya during fruit ripening

The ‘Pluk Mai Lie’ papaya (Carica papaya L.) is a promising cultivated fruit for use in fresh and processed products due to its firm flesh, but the aroma it releases is flat. Changes in quality and volatile profiles were analyzed during on- and off-tree fruit ripening. Detached fruit ripened faster than attached fruit, accumulating high internal ethylene levels. Aside from peel color, which was redder in on-tree ripened fruit, most quality attributes changed similarly in the two ripening situations. A slight increase in total soluble solids (TSS) was measured from the onset of the preclimacteric stage, whereas titratable acidity (TA) remained stable throughout the development. Whereas 2-ethyl-1-hexanol was found specifically in green fruit, ethyl octanoate emerged only in fully-ripe fruit. Furthermore, benzyl isothiocyanate was the most abundant volatile and was present in fruit at every stage except full ripening. The production of total esters, highly correlated with a loss of firmness and an increase in cavity ethylene accumulation, was about 10-fold higher in off-tree ripened fruit. The levels of methanol and ethanol sources in fruit increased steadily throughout ripening, with esters formed from ethyl alcohol predominating from the half-ripe through the senescence phases. The alcohol dehydrogenase activity in the mesocarp increased dramatically during the early ripening stages, whereas alcohol acetyltransferase was active throughout ripening. No difference in volatile profiles was found in the papaya fruit during on-tree and postharvest ripening.     

Effect of pollination intensity,frequency and pollen source on fig (Ficus carica L.) productivity and fruit quality

Yield and fruit quality in fig (Ficus carica L.) are highly dependent on cultural practices especially caprification (pollination). However, this technique remains not well controlled in Tunisia. This study was conducted during two successive years, 2009 and 2010, to investigate different pollination conditions, i.e. number of caprifigs and repetitions of caprification on fruiting of two cultivars Bouhouli (San Pedro type) and Zidi (Smyrna type). In addition, the efficiency of four pollen sources was evaluated to identify the most effective pollinator for female cultivar Zidi. The following parameters were recorded: fruit number per shoot, fruit set, productivity, fruit characteristics and vegetative growth.     

Biofertilizers improve plant growth,fruit yield,nutrition, metabolism and rhizosphere enzyme activities of Pomegranate (Punica granatum L.) in Indian Thar Desert

Production of horticultural crops has undergone enormous change in recent years due to development of innovative technologies including integrated nutrient management practices using biofertilizers. The present study represents the positive response of biofertilizers in nursery seedlings followed by their transplantation in harsh field conditions of Indian Thar Desert. Nursery and field experiments were carried out to assess the effectiveness of selected N₂-fixing bacteria and AM fungi alone or in combination, on the growth and biomass production of Punica granatum. In both experiments, the combined treatment of Azotobacter chroococcum and Glomus mosseae was found to be the most effective. Besides enhancing the rhizosphere microbial activity and concentration of various metabolites and nutrients, these bioinoculants helped in better establishment of pomegranate plants under field conditions. A significant improvement in the plant height, plant canopy, pruned material and fruit yield was evident in 5-year-old pomegranate plants in field conditions. In view of the above results, use of biofertilizer technology may be adopted for the establishment and development of other horticultural plant species in arid regions.     

Effect of synthetic auxins on fruit development of ‘Bing’ cherry (Prunus avium L.)

The main cherry cultivar grown in the warm climate of Israel, ‘Bing’, produces relatively small fruit. Over three consecutive years (2003–2005), application of 50 mg l⁻¹ 2,4-dichlorophenoxypropionic acid [2,4-DP; as its butoxyethyl ester (Power™)], 10 mg l⁻¹ 3,5,6-trichloro-2-pyridyloxyacetic acid [3,5,6-TPA; as the free acid (Maxim^®)], or 25 mg l⁻¹ 2,4-dichlorophenoxyacetic acid (2,4-D) plus 30 mg l⁻¹ naphthaleneacetic acid (NAA; 0.3% Amigo™), at the beginning of pit-hardening when fruitlet diameter was ca. 13 mm caused appreciable and significant increases in fruit size and total yield, except when the crop load was heavy. Anatomical studies revealed that the main effect of these synthetic auxins was via direct stimulation of fruit cell enlargement. The above auxins had no negative effect on fruit quality, either at harvest or after 1 month of storage at 0 °C, or on return yield in the following year.     

Selection and testing of epiphytic yeasts to control anthacnose in post-harvest of papaya fruit

Anthracnose, caused by Colletotrichum gloeosporioides, is a major post-harvest disease in papaya fruit. The major objectives of the present work were to isolate, select and test the in vitro and in vivo ability of epiphytic microorganisms, isolated from papaya fruit and leaf surfaces, in controlling anthracnose onset after harvest. A total of 75 bacteria, 67 yeasts and 22 mycelial fungi were isolated. Thirty yeast isolates were able to inhibit the mycelial growth of C. gloeosporioide in vitro and seven of those were used in in vivo assays, resulting in the identification of two very effective isolates. Isolate CEN63, identified molecularly as Cryptococcus magnus, was the most effective in controlling the disease and therefore was studied in more detail. The results of the assays with C. magnus provided evidence that when fruit were treated with the antagonists at concentrations of 10⁷ to 10⁸ cells/ml, as early as 24 h, preferentially 48 h, before inoculation with the pathogen, the development of disease was significantly reduced. C. magnus is a potential antagonist for the development of a commercial product. Additional studies on the modes of action of this yeast isolate, as on its ability to interact with fungicides are being conducted to generate solid basis for the development of an environmentally friendly control agent.     

Agrobacterium × plant factors influencing transformation of ‘Joseph's coat’ (Amaranthus tricolor L.)

A protocol is developed for Agrobacterium-mediated genetic transformation of Amaranthus tricolor via explant co-cultivation with Agrobacterium rhizogenes. Bacteria-plant specific factors which influenced transformation were optimized. Of the two Agrobacterium strains employed, LBA9402 was more infectious compared to A4. Bacterial suspensions grown overnight with 100 μM acetosyringone and experiencing O.D.₆₆₀ = 0.6 followed by dilution to a density of 10⁹ cells ml⁻¹ were the most effective. Explants from garden-grown plants were more responsive than those from in vitro cultures; stem internodes being better than leaves. Immersion of the pre-pricked explants in bacterial suspension resulted in a markedly higher transformation frequency compared to the direct injection method. The infection of internode explants with the LBA9402 strain followed by co-cultivation on growth regulator-free MS medium (MS0) for 5 days resulted in emergence of hairy roots up to a maximum frequency of 97.22%. Roots were individually cultured in MS0, but fortified with bactericidal antibiotic (500 μg ml⁻¹ cefotaxime). Rhizoclones showing prolific growth were renewed through successive subcultures in MS0. Opine gene expression was revealed by positive agropine and mannopine synthesis in all selected transformed rhizoclones. Shoot regeneration from root clones, capable of auxin-independent growth and opine proficiency, was stimulated in MS augmented with 2.0 mg l⁻¹ zeatin. pRi TL–DNA rolB and pRi TR–DNA man2 ORF were detected in leaf tissues of regenerated plants from selected hairy root clones through PCR amplification. The implication of such findings is discussed on the possibility of conferring protection to crop amaranths against biotic stress challenges, particularly due to insects, viruses or fungal pathogens.     

Improving ‘Bing’ sweet cherry fruit quality with plant growth regulators

Final fruit diameter is the prime determinant of sweet cherry fruit value. Previous research has shown that mesocarp cell size accounts predominantly for variability in final fruit size, within a genotype. Our research program evaluated the potential to improve sweet cherry fruit size/weight with growth regulators to affect cell division and/or cell expansion stages. In the current study we screened 8 plant growth regulators (PGRs), including cytokinins, gibberellins, and auxins, and their combinations for their ability to increase ‘Bing’ fruit weight. Each PGR was mixed in lanolin paste and applied to fruit pedicels at 9 or 30 days after full bloom (DAFB), to coincide with estimated peak in cell division and cell expansion activity, respectively. Several cytokinins applied 30 DAFB improved fruit weight significantly (ca. +15%) with N-(2-Chloro-4-pyridyl)-N′-phenylurea (CPPU) and 6-(3-hydroxybenzylamino) purine (mt-Topolin) at 100 mg l⁻¹ being the most effective. Gibberellins, applied alone, improved fruit size and delayed fruit maturation and exocarp coloration. GA₃ at 200 mg l⁻¹ applied at 9 DAFB was the most effective and improved final fruit weight by 15%. Fifty-six percent of the fruit from this treatment were ≥9 g compared to 15% of similar weight fruit from untreated limbs. Both GA₃ and GA_4/7 treatments applied 9 DAFB increased fruit radial expansion. 4-Chlorophenoxyacetic acid, a synthetic auxin, also stimulated higher fruit growth rates at stage I and stage II, and fruit color development, but did not improve final fruit size.     

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.     

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.     

Analysis of highbush blueberry (Vaccinium corymbosum L.) fruit growth with exponential mixed models

Blueberry fruit growth commonly exhibits a double-sigmoid pattern. The goal of this work was to characterize and compare fruit growth of cultivars differing in ripening time using exponential non-linear mixed models. Mixed-effects five-parameter exponential (Gompertz I and II; logistic; monomolecular) models were fitted to fruit diameter data from 2 years and three cultivars grown in the field in a cool temperate environment. Gompertz II mixed model provided the best fit to fruit growth data and was used for further analysis. In later ripening cultivars ‘Cape Fear’ and ‘Herbert’, clear-cut double-sigmoid patterns were observed, absolute growth rate models exhibited two marked peaks, and relative growth rate showed an initial decreasing trend, and a subsequent peak. The earlier cv. ‘O’Neal’ did not exhibit a defined double-sigmoid pattern. The time between relative extremes of absolute growth acceleration is proposed as an objective criterion for fixing stage boundaries within growth curves. Exponential mixed models accurately fitted blueberry growth patterns. These equations highlighted differences in fruit growth patterns between early and late ripening highbush blueberry cultivars.     

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.     

Effect of plant growth temperature on membrane lipids in strawberry (Fragaria × ananassa Duch.)

Changes in membrane lipid composition are important in the acclimation of plants. The influence of four day/night growing temperature combinations (18/12, 25/12, 25/22, and 30/22 °C) on membrane lipids of ‘Earliglow’ and ‘Kent’ strawberry (Fragaria × ananassa Duch.) were studied. The monogalactosyl diglyceride (MGDG) and digalactosyl diglyceride (DGDG) were the major galactolipids in the strawberry leaves. ‘Earliglow’ contained a higher amount of galactolipids in the leaves than ‘Kent’. The major phospholipids in the strawberry leaves, roots, and fruit were phosphatidylcholine (PC), phosphatidylinositol (PI), phosphatidylglycerol (PG), and phosphatidylethanolamine (PE). PC and PE were the two predominant phospholipids in the strawberry. The leaves and fruit of ‘Earliglow’ contained higher amounts of phospholipids compared to those of ‘Kent’, whereas ‘Kent’ strawberry roots had higher phospholipids. Palmitic (C16:0), stearic (C18:0), oleic (C18:1), linoleic (C18:2), and α-linolenic (C18:3) acids were major fatty acids in galacto- and phospholipids of the ‘Earliglow’ and ‘Kent’ strawberry. PC is very rich in linolenic acid in leaves compared to the fruit and root tissues. PC had the highest ratio of unsaturated to saturated fatty acids among all phospholipids. There was a significant increase in the content of galactolipids (MGDG, DGDG) and phospholipids (PC, PI, PG and PE) and unsaturation of their fatty acids in the cool day/night growth temperature. Increasing day/night growth temperatures decreased MGDG/DGDG ratios. The shifts in saturation and composition of fatty acids observed with strawberry may be an adaptation response of plants to the temperature changes.     

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.     

Evaluation of ‘Tonda di Giffoni’ hazelnut (Corylus avellana L.) clones

‘Tonda di Giffoni’ is among the most highly appreciated Italian hazelnut (Corylus avellana L.) cultivars. Due to its round kernels and excellent processing quality, it was awarded a Protected Geographical Indication (PGI) from the European Union. To identify clones expressing improved nut and production qualities, a ‘Tonda di Giffoni’ clonal selection programme was conducted across hazelnut orchards in the Irno valley of Italy from 1995 to 2006. One hundred different clones were selected and propagated in a replicated trial under similar climate, soil, and cultural conditions. From this work, the 29 best clones were identified and from 2006 to 2008 their agronomic and pomological characteristics were observed. Microsatellite or simple sequence repeat (SSR) markers were used to successfully confirm true-to-type identity of the clones. Traits evaluated included flowering time (anthesis), bud break, suckering, trunk diameter, nut and kernel characteristics and productivity (yield). Best linear unbiased predictions for clone means and estimates of intraclass correlation coefficient were obtained using R environment, lme4 and ggplot2 packages. Five clones superior to that of the standard of ‘Tonda di Giffoni’ were identified in this study. Furthermore, yield and number of suckers produced showed sufficient variability to likely be exploited for breeding. The selected clones express features useful for both growers and the processing industry and will be propagated and planted in hazelnut orchards for further study and commercial production.     

A strain of Bacillus subtilis stimulates sunflower growth (Helianthus annuus L.) temporarily

Preliminary studies showed that a Bacillus subtilis strain stimulates plant growth. We investigated how inoculating seeds of a sunflower cultivar (Helianthus annuus L.) with this strain stimulated plant growth, soil properties and emissions of greenhouse gasses, i.e. carbon dioxide (CO₂) and nitrous oxide (N₂O), when cultivated in a greenhouse. Unfertilized sunflowers or fertilized with urea served as controls. After one month, root length and fresh and dry root weight of the sunflower was significantly higher in the bacteria amended plant than in the urea and unfertilized plants. However, at harvest, no positive effect was observed. The number of seeds per plant and seed weight was not significantly different between the treatments, but total plant N was significantly higher in urea-amended plants than in unfertilized plants. The CO₂ production rate was not affected by treatment, but the N₂O emission rate was significantly higher in soil amended with urea plus bacteria soil compared to the unfertilized treatments. It was found that the B. subtilis strain used in this study had a positive, but only temporarily effect on growth of the sunflower cultivar used.     

Impact of iron stress on biomass,yield, metabolism and quality of potato (Solanum tuberosum L.)

In order to study the influence of variable iron on biomass, economic yield and role of iron in potato (Solanum tuberosum) cv. Chandramukhi metabolism, plants were grown in refined sand at variable iron ranging from 0.001 to 2.0 mM. Exposure of potato plants to Fe stress (i.e. a Fe concentration different from 0.1 mM) shows retarded growth, decreased chlorophyll concentration and Hill reaction activity, induced changes in enzyme activities and concentration of Fe and Mn. The visible symptoms of iron deficiency appeared on day 15 at 0.001 mM Fe as chlorosis of young leaves. The excess of iron (at >0.1 mM Fe) appeared later, after 25 days and the chlorosis was observed on older leaves. Both deficiency (0.001 mM) and excess (>0.1 mM) of iron reduced the tuber yield, deteriorating its quality by lowering the concentration of sugars, starch and protein nitrogen and increasing the accumulation of non-protein nitrogen and phenols in tubers.     

H2O2 metabolism during sweet orange (Citrus sinensis L. Osb.) ‘Hamlin’ Xanthomonas axonopodis pv. citri interaction

Sweet orange (Citrus sinensis L. Osb.) ‘Hamlin’ is a canker (Xanthomonas axonopodis pv. citri: Xac) susceptible citrus genotype grown commercially worldwide. Canker causes severe economic losses and restricts the marketability of crop for export. Little is known about the role of oxidative stress in canker development. In the present investigation, sweet orange ‘Hamlin’ leaves were artificially inoculated with Xac to determine the impact of Xac infection on hydrogen peroxide (H₂O₂) metabolism. Characteristic symptoms following artificial inoculation were water soaking of the infiltrated zone between 2 and 8 days after inoculation (dai); raised epidermis accompanying tiny yellow colored bacterial colonies at 8 dai; and yellowing and necrosis of the infected zone by 12–16 dai. In planta Xac population increased 1000 fold by 14 dai from an initial population of 7.3 × 10⁶ cfu cm⁻² (0 dai). Peak concentrations of H₂O₂ were observed at 24 h and between 8 and 10 dai and coincided with higher activity of total superoxide dismutase (SOD). Lower levels of H₂O₂ in infected leaves were maintained by Xac induced higher activities of catalase (CAT), ascorbate peroxidase (APOD), and guaiacol peroxidase (POD). It appears Xac altered H₂O₂ metabolism in C. sinensis L. Osb. ‘Hamlin’ to enhance survival and growth.     

A pilose fruit mutant in banana (Musa spp. ‘Williams’)

A mutant of Musa spp. ‘Williams’ was isolated by screening the plants from tissue culture and rapid propagation. We studied the morphological characteristics of the fruit epidermis by scanning electron microscopy (SEM). The results showed that the pericarp of mutant was covered with trichomes. These trichomes were pointless and the length of the trichomes ranges from 0.02 to 0.37 mm. However, this kind of trichome was not observed on the pericarp of the wild type.