6-Benzylamino purine stimulates in vitro shoot organogenesis in Eucalyptus erythronema, E. stricklandii and their interspecific hybrids

In vitro bud and shoot organogenesis was investigated for the ornamental plants Eucalyptus erythronema var. erythronema, E. stricklandii and their interspecific hybrids cv. ‘Urrbrae Gem’ and ‘Hybrid 2.5’ by using 0.0, 0.1, 0.25, 0.5 or 1.0 μM BAP on apex and leaf explants. Callus developed on all explants and increased with all concentrations of BAP without significant differences between BAP concentrations. Buds formed on apex and leaf explants of E. erythronema and E. cv. ‘Urrbrae Gem’ especially with 1.0 μM BAP, but these buds rarely developed into shoots. Bud clusters formed on E. erythronema and E. cv. ‘Urrbrae Gem’ apex and leaf explants whereas E. stricklandii and ‘Hybrid 2.5’ produced fewer, individual buds on the explant. Shoots regenerated from apex explants of all genotypes with all levels of BAP, whereas few shoots of any genotype regenerated from leaf explants regardless of the number of buds formed. Shoots from apex explants could be multiplied successfully. Light microscopy showed meristems developed within the callus, and at the callus and bud surfaces. However, few shoots developed considering the level of bud and meristem formation. This report is the first for successful shoot organogenesis and multiplication in an ornamental eucalypt.     

Cybrid/hybrid plants regenerated from somatic fusions between male sterile Satsuma mandarin and seedy tangelos

Somatic hybridization provides an alternative for transferring mitochondria-encoded cytoplasmic male sterility (CMS). Herein, symmetric protoplast electrofusion was conducted between embryogenic callus protoplasts of Citrus unshiu Marc. cv. Guoqing No. 1 (G1), a CMS cultivar, and mesophyll-derived protoplasts of seedy ‘Page’ tangelo [C. reticulata Blanco × (C. reticulata Blanco × C. paradisi Macf.)] or ‘Nova’ tangelo [C. reticulata Blanco × (C. reticulata Blanco × C. paradisi Macf.)], to transfer CMS trait. Flow cytometry analysis showed that 14 plants recovered from G1 + ‘Page’ tangelo that displayed typical morphological character of ‘Page’ were diploid, and 6 plants regenerated from G1 + ‘Nova’ tangelo were tetraploid. Genetic compositions of regenerated plants from the two fusions were determined by SSR, CAPS and chloroplast-SSR analysis. Cybrid nature of diploids from G1 + ‘Page’ tangelo with nuclear DNA from ‘Page’, mitochondrial DNA (mtDNA) from the G1 and chloroplast DNA (cpDNA) derived from either parent was confirmed. Tetraploid plants from G1 + ‘Nova’ tangelo were identified as somatic hybrids with random cpDNA inheritance. The regenerated cybrid and hybrid plants hold great potential for Citrus seedless breeding at diploid or triploid levels.     

Effect of NaCl salinity in the genotypic variation of cowpea (Vigna unguiculata) during early vegetative growth

Twenty-five genotypes of cowpea (Vigna unguiculata L. Walp.) were tested for salt-tolerance at the vegetative growth stage in pot in the greenhouse experiments at salinity levels of 0, 85, and 170 mM NaCl. Plant survival was the main criterion for classifying genotypes. Other criteria included the ion concentration (Na⁺ and Cl⁻) in root and shoot and biomass accumulation. Four local accessions (‘Paceño’, ‘Tardón’, ‘Sonorense’, and ‘Cuarenteño’), three accessions from California (‘CB46’, ‘CB27’, and ‘CB3’), and one accession from the International Institute of Tropical Agriculture (IITA) (‘IT82D-889’) survived at concentrations of both 85 and 170 mM NaCl and were classified as salt-tolerant, while ‘IT96D-666’, ‘IT89KD-288’, and ‘IT93K-734’ from IITA were classified as salt-sensitive. One local accession (‘Sesenteño’), three accessions from IITA (‘PEPH-V Wes-85’, ‘IT86D-719’, and ‘IT95K-1090-12’), and one accession from California (‘CB5’) were classified as moderately salt-tolerant. Eight accessions from IITA (‘IT96D-733’, ‘IT90K-277-2’, ‘IT91K-93-10’, ‘IT91K-118-20’, ‘IT90K-284-2’, ‘IT95K-1088-4’, ‘IT89KD-391’, and ‘IT94K-437-1’) and one from California (‘CB88’) were classified as moderately salt-sensitive. Biomass was affected by both 85 and 170 mM NaCl in all groups of genotypes, however, salt-tolerant and moderately salt-tolerant genotypes showed higher biomass than genotypes classified as moderately salt-sensitive and salt-sensitive. In all genotypes Cl⁻ concentration was higher in shoots than roots and increased as salinity increased. Similarly Na⁺ concentration increased with increasing salinity. However, in salt-tolerant and moderately salt-tolerant genotypes, Na⁺ concentration was more in roots than shoots, while in moderately salt-sensitive and salt-sensitive genotypes, Na⁺ was higher in shoots than roots.     

Horticultural performance of ‘Folha Murcha’ sweet orange onto twelve rootstocks

Despite its outstanding position, the Brazilian citriculture is established on a very limited pool of varieties that limits its expansion and restricts the fruit availability throughout the year. This situation determines the urgent necessity of developing alternative scion and rootstock cultivars, with good performance under local conditions. ‘Folha Murcha’ sweet orange (Citrus sinensis (L.) Osbeck) is a late-harvest cultivar, suitable both for the juice processing industry and the fresh fruit market, being described as tolerant to citrus canker (Xanthomonas citri subsp. citri Schaad et al.), and less affected by citrus variegated chlorosis (Xylella fastidiosa Wells et al.). A study was conducted in Bebedouro, São Paulo State, Brazil, to evaluate the horticultural performance of ‘Folha Murcha’ sweet orange budded onto 12 rootstocks: the citrandarin ‘Changsha’ mandarin (Citrus reticulata Blanco) × Poncirus trifoliata ‘English Small’; the hybrid ‘Rangpur’ lime (Citrus limonia Osbeck) × ‘Swingle’ citrumelo (P. trifoliata (L.) Raf. × Citrus paradisi Macfad.); the trifoliates (P. trifoliata (L.) Raf.) ‘Rubidoux’, ‘FCAV’, and ‘Flying Dragon’ (P. trifoliata var. monstrosa); the ‘Sun Chu Sha Kat’ mandarin (C. reticulata Blanco); the ‘Sunki’ mandarin (Citrus sunki (Hayata) Hort. ex. Tanaka); the ‘Rangpur’ limes (C. limonia Osbeck) ‘Cravo Limeira’ and ‘Cravo FCAV’; ‘Carrizo’ citrange (C. sinensis × P. trifoliata), ‘Swingle’ citrumelo (P. trifoliata × C. paradisi), and ‘Orlando’ tangelo (C. paradisi × Citrus tangerina cv. ‘Dancy’). The experimental grove was planted in 2001, using a 7 m × 4 m spacing, in a randomized block design, with five replications and two plants per plot. No supplementary irrigation was applied. Fruit yield, canopy volume, tree tolerance to drought and to citrus variegated chlorosis, and fruit quality were assessed for each rootstock. Trees grafted onto the ‘Flying Dragon’ trifoliate were smaller in size, but had largest yield efficiency when compared to those grafted onto other rootstocks. Lower alternate bearing index was observed on trees budded onto ‘Cravo FCAV’ ‘Rangpur’ lime. Both ‘Rangpur’ lime rootstocks and the ‘Sunki’ mandarin induced higher tree tolerance to drought. The ‘Flying Dragon’ trifoliate induced better fruit quality and higher tolerance to citrus variegated chlorosis (CVC) to ‘Folha Murcha’ trees. A cluster multivariate analysis identified three groups of rootstocks with similar effects on ‘Folha Murcha’ tree performance. Among the 12 evaluated rootstocks, the ‘Flying Dragon’ trifoliate has a unique effect on plant growth, tolerance to drought and CVC, fruit yield and fruit quality of ‘Folha Murcha’ trees, and may be better suited for high-density plantings.     

Eggplant relatives as sources of variation for developing new rootstocks: Effects of grafting on eggplant yield and fruit apparent quality and composition

We propose the utilization of eggplant (Solanum melongena L.) interspecific hybrids derived from crosses with closely related species as an approach for developing new improved rootstocks for eggplant. Here we investigate rootstock effects on fruit yield, apparent quality and proximate and mineral composition of S. melongena ‘Black Beauty’ (BB) scions grafted on interspecific hybrid rootstocks developed from crosses of S. melongena with Solanum incanum L. (SI × SM) and Solanum aethiopicum L. (SM × SA). The results are compared with non-grafted (BB control) and self-grafted (BB/BB) controls and with S. melongena ‘Black Beauty’ scions grafted onto Solanum torvum Sw. (STO) and Solanum macrocarpon L. (SMA) rootstocks. All treatments were grown in a soil naturally infested with root-knot nematodes (mostly Meloidogyne incognita (Kofoid and White) Chitwood). SI × SM and SM × SA interspecific hybrids had high germination (≥90%) and total graft success (100%). Contrary to what occurred with all other treatments, no plants from scions grafted onto these hybrid rootstocks died during the experiment. In particular, the SI × SM hybrid rootstock conferred the highest vigour to the scion, which resulted in the highest values for fruit earliness and early and total yield. Little difference was observed among treatments for apparent fruit quality traits, except for a greater fruit calyx length and prickliness of fruit grafted onto SMA rootstocks. A similar result was obtained for fruit composition where phenolics content was higher in fruit from plants grafted onto SMA rootstocks. Grafting eggplant onto interspecific eggplant hybrids, especially on the SI × SM hybrid, has proved advantageous for eggplant production, as the high vigour and good compatibility of the rootstock with scion results in improved early and total yield without negative effects on apparent fruit quality or composition. Interspecific hybrids represent an alternative to the commonly used STO rootstock, which is a wild species with irregular germination.     

Differential structure and physiological response to canker challenge between ‘Meiwa’ kumquat and ‘Newhall’ navel orange with contrasting resistance

Asiatic citrus canker caused by Xanthomonas citri subsp. citri (Xcc) is one of the most detrimental diseases affecting citrus production. ‘Meiwa’ kumquat (Fortunella crassifolia) and ‘Newhall’ navel orange (Citrus sinensis) are two genotypes with contrasting Xcc resistance. In the current study, efforts were made to perform pairwise comparison between them in terms of structural difference and biochemical response to Xcc attack. Pinprick inoculation demonstrated that ‘Meiwa’ had lower disease incidence rate and smaller lesion size, concurrent with significantly less bacterial number at the inoculation sites relative to ‘Newhall’. Microscopic observation demonstrated that stomatal density and size of ‘Meiwa’ were significantly smaller than ‘Newhall’. Stomatal apertures of ‘Meiwa’ were smaller than ‘Newhall’ measured at 2 h after their leaves were immersed in Xcc suspension, consistent with the bacteria number in the internal tissues counted at 4 d. Epicuticular wax content of ‘Meiwa’ was significantly higher than that of ‘Newhall’, while no difference in palisade and spongy tissue was observed between them. Enzymatic activity assay showed that after Xcc inoculation, ‘Meiwa’ exhibited significantly higher activities of catalase (CAT), peroxidase (POD) and phenylalanine ammonia-lyase (PAL) in comparison with ‘Newhall’ at most of the sampling time points. Our data showed that the two genotypes with contrasting Xcc resistance exhibited clear difference in the structure and physiological response, which may provide information to facilitate our understanding of Xcc resistance.     

Tree performance and fruit yield and quality of ‘Okitsu’ Satsuma mandarin grafted on 12 rootstocks

The citriculture in Brazil, as well as in other important regions in the world, is based on very few mandarin cultivars. This fact leads to a short harvest period and higher prices for off-season fruit. The ‘Okitsu’ Satsuma (Citrus unshiu Marc.) is among the earliest ripening mandarin cultivars, and it is considered to be tolerant to citrus canker (Xanthomonas citri subsp. citri Schaad et al.) and to citrus variegated chlorosis (Xylella fastidiosa Wells et al.). Despite having regular fruit quality under hot climate conditions, the early fruit maturation and absence of seeds of ‘Okitsu’ fruits are well suited for the local market in the summer (December through March), when the availability of citrus fruits for fresh consumption is limited. Yet, only a few studies have been conducted in Brazil on rootstocks for ‘Okitsu’. Consequently, a field trial was carried out in Bebedouro, São Paulo State, to evaluate the horticultural performance of ‘Okitsu’ Satsuma mandarin budded onto 12 rootstocks: the citrandarin ‘Changsha’ mandarin (Citrus reticulata Blanco) × Poncirus trifoliata ‘English Small’; the hybrid Rangpur lime (Citrus limonia Osbeck) × ‘Swingle’ citrumelo (P. trifoliata (L.) Raf. × Citrus paradisi Macfad.); the trifoliates (P. trifoliata (L.) Raf.) ‘Rubidoux’, ‘FCAV’ and ‘Flying Dragon’ (P. trifoliata var. monstrosa); the mandarins ‘Sun Chu Sha Kat’ (C. reticulata Blanco) and ‘Sunki’ (Citrus sunki (Hayata) Hort. ex. Tanaka); the Rangpur limes (C. limonia Osbeck) ‘Cravo Limeira’ and ‘Cravo FCAV’; ‘Carrizo’ citrange (Citrus sinensis × P. trifoliata), ‘Swingle’ citrumelo (P. trifoliata × C. paradisi), and ‘Orlando’ tangelo (C. paradisi × Citrus tangerina cv. ‘Dancy’). The experimental grove was planted in 2001, using a 6 m × 3 m spacing, in a randomized block design. No supplementary irrigation was applied. Fruit yield, canopy volume, and fruit quality were assessed for each rootstock. A cluster multivariate analysis identified three different rootstock pairs with similar effects on plant growth, yield and fruit quality of ‘Okitsu’ mandarin. The ‘Flying Dragon’ trifoliate had a unique effect over the ‘Okitsu’ trees performance, inducing lower canopy volume and higher yield efficiency and fruit quality, and might be suitable for high-density plantings. The ‘Cravo Limeira’ and ‘Cravo FCAV’ Rangpur limes induced early-ripening of fruits, with low fruit quality. ‘Sun Chu Sha Kat’ and ‘Sunki’ mandarins and the ‘Orlando’ tangelo conferred lower yield efficiency and less content of soluble solids for the latter rootstock.     

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.     

Hybrids sugary × sugary enhancer of sweet corn: A valuable option for cool environments

Sugary (Se1Se1) sweet corn hybrids do not have a high quality for human consumption according to the preferences of today's consumers, although they have a good stand establishment in cool environments. On the contrary, sugary enhancer (se1se1) hybrids have a high table quality, but a poor stand establishment in areas with a cool growing season. Sugary × sugary enhancer (Se1se1) sweet corn genotypes could be a valuable alternative for cool environments, but no information is available about the performance of sugary × sugary enhancer hybrids under such conditions. The objective of this paper is to evaluate the potential of sugary × sugary enhancer sweet corn genotypes in cool environments in comparison with sugary and sugary enhancer genotypes. Eight sugary lines and eight sugary enhancer lines were used as the parental lines of 15 sugary, 12 sugary enhancer, and 57 sugary × sugary enhancer hybrids that were evaluated in three different cool environments. The results of this work show that sweet corn cultivars with the Se1se1 genotype are an appropriate alternative for cool areas because they have better plant and ear types and a better eating quality than the Se1Se1 genotypes, and better emergence, early vigor, and shorter flowering time than se1se1 genotypes.     

Yield performance,quality characteristics and fruit storability of winter squash cultivars in sub-humid areas

This research investigated the quality traits of eight winter squash cultivars (Cucurbita maxima, C. moschata, C. pepo and interspecific hybrids of C. moschata × C. pepo) during three years, but only ‘Tetsukabuto’ and ‘Violina’ were tested each year. In 2005 these two varieties were compared to ‘Butternut’; in 2006 to ‘Red Kury’, ‘Tan Cheese’ and ‘Kabosha’; in 2007 to ‘Red Kury’, ‘Mooregold’, and ‘Winter Luxury’. The characteristics recorded were: fruit yield, storage ability, chemical composition and sensory quality. Following harvest and sorting of marketable winter squash, 50 fruits for each cultivar were stored at 12 °C for 12 weeks the 1st year, and for over 20 weeks in 2nd and 3rd years, during which rotten fruits were counted. Compositional analyses regarding sugar concentration, starch, carotenoids and dry matter content at harvest and after storage were carried out; a panel test was organised to assess sensory traits. The most interesting cultivars were ‘Tetsukabuto’ for yield, ‘Tetsukabuto’ and ‘Mooregold’ for storability, ‘Red Kury’ and the same ‘Tetsukabuto’ particularly for soluble sugars and carotenoids and sensory appreciation.     

Evaluation of citrus somatic hybrids for tolerance to Phytophthora nicotianae and citrus tristeza virus

Somatic hybridization is a biotechnology tool that can be used in citrus breeding programs to produce somatic hybrids with the complete genetic combination of both parents. The goal of this work was to test the reaction of citrus somatic hybrids that may be useful as rootstocks to trunk and root infections caused by Phytophthora nicotianae van Breda de Haan (P. parasitica Dastur) and to citrus tristeza virus (CTV). The somatic hybrids evaluated were ‘Caipira’ sweet orange (Citrus sinensis L. Osbeck) + ‘Rangpur’ lime (C. limonia Osbeck), ‘Caipira’ sweet orange + ‘Cleopatra’ mandarin (C. reshni hort. ex Tanaka), ‘Caipira’ sweet orange + ‘Volkamer’ lemon (C. volkameriana V. Ten. & Pasq.), ‘Caipira’ sweet orange + rough lemon (C. jambhiri Lush.), ‘Cleopatra’ mandarin + ‘Volkamer’ lemon, ‘Cleopatra’ mandarin + sour orange (C. aurantium L.), ‘Rangpur’ lime + ‘Sunki’ mandarin (C. sunki (Hayata) hort. ex Tanaka), ‘Ruby Blood’ sweet orange (C. sinensis L. Osbeck) + ‘Volkamer’ lemon, ‘Rohde Red’ sweet orange (C. sinensis L. Osbeck) + ‘Volkamer’ lemon, and ‘Valencia’ sweet orange + Fortunella obovata hort. ex Tanaka. For P. nicotianae trunk and root infection assays, plants of the somatic hybrids, obtained from 9-month semi-hardwood cuttings, were evaluated and compared with diploid citrus rootstock cultivars after mycelia inoculation in the trunk or spore infestation in the substrate, respectively. ‘Cleopatra’ mandarin + sour orange, ‘Rangpur’ lime + ‘Sunki’ mandarin, ‘Cleopatra’ mandarin + ‘Volkamer’ lemon, ‘Ruby Blood’ sweet orange + ‘Volkamer’ lemon, ‘Rohde Red’ sweet orange + ‘Volkamer’ lemon, and ‘Caipira’ sweet orange + ‘Volkamer’ lemon had less trunk rot occurrence, whereas the somatic hybrids ‘Cleopatra’ mandarin + ‘Volkamer’ lemon, ‘Cleopatra’ mandarin + sour orange, ‘Caipira’ sweet orange + ‘Volkamer’ lemon, and ‘Caipira’ sweet orange + ‘Rangpur’ lime were tolerant to root rot. For CTV assays, plants of the somatic hybrids along with tolerant and intolerant rootstocks were budded with a mild strain CTV-infected or healthy ‘Valencia’ sweet orange budwood. Differences in average scion shoot length indicated that the hybrids ‘Cleopatra’ mandarin + sour orange and ‘Valencia’ sweet orange + Fortunella obovata were intolerant to CTV.     

Effect of distance from early flowering pollinizers ‘Maypole’ and ‘Dolgo’ on ‘Fiji’ fruit set

An apple orchard consisting of a single cultivar under the condition of natural pollination must have suitable pollinizers such as Crab apples to ensure stable fruit production. We selected ‘Maypole’ and ‘Dolgo’ as pollinizers for the cultivar ‘Fuji’, and investigated the rate of fruit and seeds in ‘Fuji’ fruits produced by pollen of the pollinizers. We developed a method for tracing pollen flow based on the leaf color of progeny and S-RNase allele of ‘Maypole’, and on Simple Sequence Repeat (SSR) analyses of ‘Maypole’ and ‘Dolgo’. These were powerful tools for determining the distance insects (mainly Osmia cornifrons) carry pollen from the pollinizers to ‘Fuji’. Although the fruit set of ‘Fuji’ apples was not reduced with increasing distance between ‘Fuji’ apple trees and pollinizers (probably due to pollen flow from other commercial cultivars planted outside the area), the rate of ‘Fuji’ apple fruit produced by the pollen of the pollinizers decreased with increasing distance. The rate of fruit produced by the pollinizers was 84% and 77% when ‘Fuji’ was 2.5 m from ‘Maypole’ and ‘Dolgo’, respectively, and 71% and 64% when ‘Fuji’ was 5 m from ‘Maypole’ and ‘Dolgo’, respectively, but was reduced to 47% and 39% when ‘Fuji’ was 10 m from ‘Maypole’ and ‘Dolgo’, respectively. However, the spacing could cause reduced fruit size, and require extra fruit thinning for producing large fruits. It could also cause reduction of the yield for increasing the planting area of pollinizers. We recommend that pollinizers should be planted not more than 10 m from ‘Fuji’.     

Responses of eight chile peppers to saline water irrigation

Salt tolerance of five cultivars of Capsicum annuum L. Early Jalapeno, Golden Treasure, NuMex Sweet, NuMex Joe E. Parker, and Santa Fe Grande, two cultivars of C. chinense Jacq. Habanero and Pimienta De Chiera, and one accession of C. annuum, NMCA 10652, were evaluated in a field study. Seedlings were transplanted in late May to field raised beds containing loamy sand soils in a semi-arid environment. Plants were well irrigated throughout the experiment. Three saline solution treatments, prepared by adding NaCl, MgSO₄, and CaCl₂ to tap water at different amounts to create three salinity levels of 0.82 dS m⁻¹ (control, tap water), 2.5 dS m⁻¹, and 4.1 dS m⁻¹ electrical conductivity (EC), were initiated on 15th June and ended in late August. Among the eight varieties, NMCA 10652 had the highest survival percentage at 100% in the 4.1 dS m⁻¹ treatment, followed by ‘Early Jalapeno’, ‘NuMex Sweet’, ‘Pimienta De Chiera’, ‘Santa Fe Grande’, ‘Golden Treasure’, and ‘NuMex Joe E. Parker’. ‘Habanero’ had the lowest survival at 28%. Compared to control, final shoot dry weight of the plants irrigated with saline solution at 4.1 dS m⁻¹ was reduced by 92% in ‘Habanero’, followed by ‘Golden Treasure’ at 80%. For fruit fresh weight in 4.1 dS m⁻¹ vs. control, ‘Habanero’ had the highest reduction at 86%, followed by ‘Golden Treasure’ at 74%, while NMCA 10652 and ‘Santa Fe Grande’ had the least at 26% and 19%, respectively. NMCA 10652, the most tolerant to salinity, had the lowest leaf Na⁺ accumulation, while ‘Habanero’, the most sensitive to salinity, had the highest Na⁺ in the leaves. For leaf Cl⁻, ‘Early Jalapeno’ had the highest, while ‘Habanero’ had the lowest Cl⁻ accumulation in the leaves. Generally, sensitive varieties accumulated more Na⁺ and/or Cl⁻ in leaves, except for ‘Early Jalapeno’, which was relatively tolerant to salinity but had high Na⁺ and Cl⁻ accumulation in leaves.     

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.     

A protocol to assess heat tolerance in a segregating population of raspberry using chlorophyll fluorescence

A detached leaf protocol was developed to assess heat tolerance in raspberry (Rubus L. subgenus Idaeobatus) by measuring chlorophyll fluorescence. In order to establish the heat screen protocol for field measurements in a segregating population, several baseline experiments were performed on genotypes with known heat tolerance based on visual observations in multiple climates. ‘Mandarin’ and ‘Qualicum’ were used as heat tolerant and heat susceptible controls, respectively. We determined that: optimum collection time was in the morning before ambient temperatures reached 28 °C, critical temperature (Tc) was 45 °C, minimum duration of dark adaptation was 15 min, and optimum heat shock duration was 30 min. Parameters established in baseline experiments were subsequently used for screening a large population (196 genotypes) planted in the field, where warm conditions commonly occur (maximum temperatures ranging between 30 and 35 °C). A significant difference in chlorophyll fluorescence was found between the two controls; ‘Qualicum’, the heat sensitive cultivar, had the lowest Fv/Fm (variable fluorescence/maximum fluorescence) value, while 18.8% of the segregating population had a higher Fv/Fm value than ‘Mandarin’, the heat tolerant cultivar. We have identified 106 individuals with an Fv/Fm above 0.600 (representing 52% of the population), and 7 individuals above 0.680 (4% of the population). The distribution of heat tolerance within the population was essentially normal, and thus quantitative trait analysis is a plausible approach for studying heat tolerance. This protocol has potential application to other Rosaceae species, and is not dependent on visual assessment.     

In vitro germination and seedling development of cryopreserved Dendrobium hybrid mature seeds

In vitro germination and seedling development from Dendrobium Swartz. hybrid ‘Sena Red’, ‘Mini WRL’, ‘Jaquelyn Thomas’, and ‘BFC Pink’ seeds cryopreserved through vitrification (PVS2) were evaluated. Germination percentages after cryopreservation (LN) were variable among different controls and treatments, despite the initial high seed viability for all hybrids. Seeds exposed to PVS2 at ice temperature from 1 to 3 h prior to LN exhibited significantly higher germination than seeds exposed to PVS2 at room temperature for the same time periods. No significant differences in germination percentages were observed for time exposure to PVS2 at 1, 2 or 3 h for ‘Sena Red’, ‘Mini WRL’, and ‘BFC Pink’. Seeds of ‘Jaquelyn Thomas’ exposed to 1 h prior to LN showed higher germination percentage than for exposure to 2 or 3 h. The combination of a pre-cooling treatment (ice) with a dehydration treatment (PVS2) for a period of time of 1–3 h was essential to allow proper germination of cryopreserved seeds. Although variability in seed germination among different hybrids and treatments was observed, germination was above 50% of the controls and all germinated seeds developed into normal seedlings with healthy shoot and root formation. No abnormalities, nutritional deficiencies, or diseases were observed in developed seedlings and no significant differences were observed for seedling growth and development from germinated seeds among the different hybrids. Seedlings transplanted to pots acclimatized well and developed into normal plants within 6–8 months in greenhouse. Transplanted seedlings exhibited 100% survival for all hybrids.     

Selection of potential pollinizers for ‘Hass’ avocado based on flowering time and male–female overlapping

Avocado production is dependent on the singular synchronous protogynous dichogamy of the species that promotes outcrossing. With the objective of selecting potential pollinizer avocado genotypes for ‘Hass’, the most important avocado cultivar worldwide, we have monitored during two consecutive years the flowering phenology of 27 avocado genotypes in South-eastern Spain. The average length of the flowering season was 45 days ranging from 18 days for ‘Harvest’ to 50 days for ‘Fuerte’. The earliest genotypes to flower were ‘Fuerte’ and ‘Shepard’ that started blooming during the third week of March. The latest genotypes to flower were ‘Colin V-33’, ‘Adi’, ‘OA184’ and ‘Harvest’, which started blooming in the second week of April. ‘Hass’ blooming lasted 30 days, from the first week of April until the second week of May. Since a good pollinizer must present not only an overlapping in the flowering season but also an overlapping in sexual stages with the pollinated cultivar, a group of 12 genotypes (‘Hass’, ‘Fuerte’ and 10 genotypes producing ‘Hass-like’ fruit with good overlapping in the flowering season with ‘Hass’) was studied with more detail determining daily the stages of male and female overlapping every 2 h. Results herein indicate that ‘Marvel’ and ‘Nobel’ showed a high sexual overlapping with ‘Hass’. Taking into account the flowering phenology, the overlapping in sexual stages and the fruit set obtained with hand-pollinated flowers in the field, those two genotypes could be an interesting alternative to the current use of ‘Fuerte’ as pollinizer for ‘Hass’ in South-eastern Spain.     

Effect of synthetic auxins on fruit size of five cultivars of Japanese plum (Prunus salicina Lindl.)

Most of the Japanese plum (Prunus salicina) cultivars grown in Israel produce relatively small fruit. Application of 2 l solution tree⁻¹ of 25 mg l⁻¹ 2,4-dichlorophenoxypropionic acid (2,4-DP) as butoxyethyl ester (Power™), 15 mg l⁻¹ 3,5,6-trichloro-2-pyridyloxyacetic acid (3,5,6-TPA) as free acid (Maxim^®), or 25 mg l⁻¹ 2,4-dichlorophenoxyacetic acid (2,4-D) + 30 mg l⁻¹ naphthaleneacetic acid (NAA) (0.3% Amigo™) at the beginning of pit-hardening, when fruitlet diameter was ca. 22 mm, caused an appreciable and significant increase in fruit size. The yield of large fruit per cv.: ‘Kesselmen’ (100% increase), ‘Songold’ (100%), ‘Black Diamond’ (800%), ‘Royal Diamond’ (160%) and ‘Royal Zee’ (100%). As a result, the total yield of all five cultivars was also increased dramatically. Anatomical studies with ‘Songold’ revealed that the main effect of these synthetic auxins was via direct stimulation of fruit cell enlargement. The above auxins had no negative effect either on fruit quality at harvest (and after 1 week in shelf-life), or on return yield in the following year.     

Pollination with laser-irradiated pollens breaks cross-incompatibility between zicaitai (Brassica campestris var. purpurea) and ornamental kale (Brassica oleracea var. acephala) to produce hybrids with the aid of ovule culture

The objective of this study was to produce interspecific hybrids between an Ogura-cytoplasmic male sterile (CMS) line of zicaitai (Brassica campestris var. purupurea, 2n = 20) and cultivars of ornamental kale (Brassica oleracea var. acephala, 2n = 18) to develop a CMS system for hybrid seed production. Pollination with pollen grains of ornamental kales irradiated at a power output of 9.0 mW with a He–Ne laser for 3 min could overcome the cross-incompatibility between the species concerned. Intact hybrids could be efficiently produced from ovules cultured on Murashige and Skoog media supplemented by 0.2 mg l⁻¹ 6-benzyladenine. Chromosome number of hybrids was confirmed to be 2n = 19. Hybrids resembled ornamental kales in leaf morphology and in vernalization response. Pollens of hybrids had a sterile appearance. Moreover the hybridity of the putative hybrids was confirmed by RAPD data on a DNA fragment of 820 bp.     

Evaluation of ‘Hamlin’ sweet orange + ‘Montenegrina’ mandarin somatic hybrid for tolerance to Xanthomonas axonopodis pv. citri and Xylella fastidiosa

Asiatic citrus canker (ACC), caused by Xanthomonas axonopodis Starr & Garces pv. citri (Hasse) Vauterin et al., and citrus variegated chlorosis (CVC), caused by Xylella fastidiosa Wells et al., are considered the main diseases affecting sweet orange scion varieties in Brazil. Among commercial varieties, mandarins and tangerines are recognized as tolerant to these pathogens. We report herein the production of ‘Hamlin’ sweet orange (Citrus sinensis L. Osbeck) + ‘Montenegrina’ mandarin (Citrus deliciosa Ten.) allotetraploid somatic hybrid plants by protoplast fusion with improved disease tolerance that could be used as a donor of resistance genes in interploid hybridisation. Somatic hybridisation was confirmed by leaf morphology, flow cytometry and RAPD analyses. The somatic hybrid was propagated by grafting and cultivated in a screenhouse for tolerance assays. For X. axonopodis pv. citri assays, buds were collected from both ‘Hamlin’ sweet orange and the somatic hybrid and grafted onto ‘Cleopatra’ mandarin (Citrus reshni hort. ex Tanaka). As a negative control, buds from ‘Mexerica Tardia’ mandarin (C. deliciosa) were collected and grafted onto ‘Cleopatra’ mandarin. Two-month old plants with at least one young vegetative flush were individually spray-inoculated with a 10⁶ CFU mL⁻¹X. axonopodis pv. citri suspension and incubated in a growth chamber, at 27 °C, under 16-h photoperiod. The somatic hybrid showed a statistically significant reduction in susceptibility to ACC 30 days after inoculation. Compared to ‘Hamlin’ sweet orange, disease severity was reduced by 70%, with similar tolerance to that of the mandarin negative control. For X. fastidiosa assays, buds were collected from the somatic hybrid and its parental plants and grafted onto ‘Rangpur’ lime (Citrus limonia Osbeck). The developed plants were needle-inoculated with a X. fastidiosa suspension (8.7 × 10¹⁰ CFU mL⁻¹) into the new growth flush stem. Bacterial population was quantified both at 4 (at the inoculation point) and 8 months (50 cm above the inoculation point) after inoculation. The first evaluation detected X. fastidiosa in 63% of ‘Hamlin’ sweet orange and ‘Hamlin’ + ‘Montenegrina’ mandarin samples. In the second evaluation, X. fastidiosa was detected in 47.4% of ‘Hamlin’ sweet orange and 10.5% of ‘Hamlin’ + ‘Montenegrina’ somatic hybrid samples, suggesting that bacterial movement was restricted in the somatic hybrid. X. fastidiosa was not detected in both evaluations in samples collected from leaves of ‘Montenegrina’ mandarin. These results indicate that the ‘Hamlin’ sweet orange + ‘Montenegrina’ mandarin somatic hybrid has potential for improved disease tolerance that should enhance its value regarding future use in citrus breeding programs.