Field performance and molecular diversification of lemon selections

Lemon (Citrus limon (L.) Burm. f.) is one of the most important Citrus fruit for Turkey because of its great amount of production and export. It has been cultivated for a long time in Turkey, and therefore variations for agronomical traits are likely among cultivated lemons due to bud mutations and, hybridizations. The objectives of this study were to determine variations for some selected agronomical traits and genetic markers among 12 new lemons derived from selections. Tree growth, yield, fruit quality, and molecular diversification of these clones were determined. After four years of evaluation, ‘Kutdiken’ M-51 indicated the highest canopy volume. For yield per tree, the best clone was ‘Kutdiken’ M-51. After five years of evaluation, ‘Kibris’ M-54 had the highest fruit weight and acidity. ‘Italian Memeli’ M-56 contained the lowest seed number and the highest total soluble solids. Molecular analysis, as assessed with 22 random amplification of polymorphic DNA (RAPD) and 11 inter simple sequence repeats (ISSR) primers, indicated that seven of twelve clones were separated with RAPD markers, whereas four were distinguished with ISSR markers. Combined analysis of RAPD and ISSR data detected that similarity values among the lemons clones were between 0.97 and 1.00. It can be concluded that variations in orchards are abundant and mainly due to mutations.     

Cross-pollination improves ‘Orri’ mandarin fruit yield

‘Orri’, a selection of ‘Orha’ mandarin [Temple (Citrus temple hort. ex Y. Tanaka) × Dancy (Citrus tangerina hort. ex Tanaka)], is a new high-quality Israeli mandarin which, in the last decade, has become one of the leading varieties in Israel. ‘Orri’ has an excellent taste, the rind is deep orange in color and easily removed, and it contains few or no seeds. However, ‘Orri’ grown in Israel suffers from inadequate yield and no published studies have yet addressed this problem. In the present study we determined that ‘Orri’ productivity depended on conditions being favorable to cross-pollination. Under cross-pollination conditions a positive correlation (R² = 0.97) was found between yield per tree and number of fruits per tree, and more than 90% of the fruits exceeded 60 mm: the most profitable size range. These data suggest that the number of fruits per tree, and not fruit size, is the limiting factor for yield improvement in ‘Orri’ orchards. Studying seed set showed that ‘Michal’ mandarin (Citrus reticulata Blanco) is a compatible pollenizer for ‘Orri’ flowers: the number of seeds per ‘Orri’ fruit increased as the distance from ‘Michal’ trees decreased. The present study demonstrated that cross-pollination of ‘Orri’ resulted in yield improvement, yet at the price of increased seed set.     

Uniconazole is an effective blossom thinner for ‘Bing’ cherry (Prunus avium)

Small fruit size is one of the limiting factors in marketing cherries (Prunus avium L.). Several techniques have been used to improve fruit size; among them, blossom and fruit thinning in order to reduce crop load have proved effective. Unfortunately, hand thinning is very costly, and chemical thinning has proved ineffective. In the present study, carried out in two consecutive years (2006–2007), application of uniconazole (E-1-(4-chlorophenyl)-4,4-dimethyl-2-(1,2,4-triazol-1-yl-1-penten-3-ol)) at 100–125 mg l⁻¹, in the form of 0.2–0.25% Magic™, at 30% full bloom, reduced the fruit set of ‘Bing’ cherry fruit. As a result time spent in hand thinning was reduced by about 50% and the crop load by about 40%, and there was a considerable shift to larger fruit size (>26 mm), provided the crop load was not too low. No negative effects were observed on the return bloom and yield in the following year.     

Effect of explants source on shoot proliferation and adventitious regeneration in 10 Buddleia cultivars

Viable shoot cultures and weaned plants were obtained from cultured apical meristems with 10 Buddleia cultivars giving viabilities of 32–72%. The number of shoots produced, the micropropagation rate and the root number produced in vitro was higher in meristem derived shoots compared to those derived from shoot-tips. The subsequent growth rate of meristem derived plants, in the greenhouse, was also greater. The number of roots produced by conventional cuttings collected from meristem derived plants was significantly higher than in cuttings which were collected from plants derived from shoot-tips or from the original stock plants.Endogenous bacteria were not detected in either shoot cultures derived from meristems or in 10-week-old weaned plants derived from meristems whereas those derived from shoot-tips showed the presence of endogenous bacteria when sterilized explants were cultured on nutrient agar or on tryptic soy broth.Factors affecting adventitious bud and shoot production in leaf and internode explants was determined for ‘Lochinch’, ‘Border Beauty’, ‘Ile de France’ and ‘Pink Delight’ using meristem derived shoot cultures. Adventitious shoots appeared after 4 weeks of culture, in both types of explant when cultured on MS supplemented with 0.5–5.0 μM TDZ. The highest percentage regeneration was achieved from bisected internode explants cultured on 0.5 μM TDZ, with 93–100% regeneration among the cultivars whereas BA was less effective. The best response was obtained using 5.0 μM TDZ which gave over 10–11 shoots per explant in all bisected explants for all cultivars.     

Chill unit models for blackcurrant (Ribes nigrum L.) cultivars ‘Ben Gairn’, ‘Ben Hope’ and ‘Ben Tirran’

Temperate-zone crops require a period of winter chilling to terminate dormancy and ensure adequate bud break the following spring. The exact chilling requirement of blackcurrant (Ribes nigrum), a commercially important crop in northern Europe, is relatively unknown. Chill unit models have been successfully utilized to determine the optimum chilling temperature of a range of crops, with one chill unit equating to 1 h exposure to the optimum temperature for chill satisfaction. Two-year-old R. nigrum plants of the cultivars ‘Ben Gairn’, ‘Ben Hope’ and ‘Ben Tirran’ were exposed to temperatures of −10.1 °C, −3.4 °C, 0.1 °C, 1.5 °C, 2.1 °C, 3.4 °C or 8.9 °C (±0.7 °C) for durations of 0, 2, 4, 6, 8 or 10 weeks and multiple regression analyses used to determine the optimum temperature for chill satisfaction.     

Study of long-distance movement of Plum pox virus (Sharka) as an alternative resistance-evaluation method in Prunus

During the breeding programs for Plum pox virus (PPV, Sharka) resistance in Prunus, the evaluation of the new releases through symptoms observation on leaves has been contradictory and represents one of the main handicaps in these programs. In order to increase the accuracy of this traditional evaluation method, we here analyze an alternative method based on the study of the ability of a genotype to allow the long-distance movement of the virus through its vascular vessels. Two different plant models have been assayed: (a) in Model I, the inoculation was performed in the ‘GF305’ rootstock with a later grafting of the genotype under evaluation and a scion of healthy control ‘GF305’, to evaluate the long-distance movement through the studied genotype from the rootstock to the scion (xylem transport), and (b) in Model II, the inoculation with ‘GF305’ diseased scions was performed by grafting these diseased scions onto the studied genotypes, which were grafted onto healthy ‘GF305’ peach seedlings, to evaluate the long-distance movement through the studied genotype from the scion to the rootstock (phloem transport). The results show that, regardless of the presence of symptoms, susceptible genotypes allowed the movement of the virus through their vascular vessels in both directions studied. However, the resistant apricot ‘Stark Early Orange’ did not allow this movement. We propose the study of the ability of a genotype to allow the long-distance movement of the virus as an alternative and more accurate method for the evaluation of PPV resistance. However, this protocol is much more tedious than the traditional one and could be used mainly in the evaluation of a reduced number of more interesting genotypes.     

Morphogenic gradients of adventitious bud and shoot regeneration in epicotyl explants of Citrus

The in vitro responses of epicotyl explants from ‘Cravo’ rangpur lime (Citrus limonia Osb.), ‘Foster’ grapefruit (C. paradisi Macf.), and ‘Pera’ sweet orange (C. sinensis (L.) Osb.) were characterized for the first time. Further analysis was performed in ‘Cravo’ rangpur lime and ‘Foster’ grapefruit aiming to verify the in vitro morphogenesis of five distinct regions of the epicotyl under different treatments. It was observed the same general pattern of morphogenic gradient along the epicotyl axis in both citrus cultivars, with greater organogenic response as the distance of the explants from the cotyledonary node increased. This morphogenic gradient was influenced by factors related to plant material, composition of the culture medium, and conditions of incubation. The regions of the epicotyl farthest from cotyledons could be used as a source of explants in experiments of genetic transformation of the genotypes evaluated aiming to improve the efficiency of production of transgenic Citrus plants.     

Immunolocalization and quantification of IAA after self- and free-pollination in Olea europaea L.

In order to characterise the self-compatibility in ‘Pisa 9’, that is an improved clone of cultivar Leccino (Olea europaea L.), biological, histological and biochemical analyses were carried out. Observations were also focused on ‘Pendolino’, the main pollinator for Italian cultivars considered as being self-incompatible. After self-pollination, in ‘Pisa 9’ the majority of the pollen tubes reached the distal part of the style, entered the ovule leading to a high fruit-set percentage. These same processes were also noted in the Pendolino cultivar, though in a lesser degree. The indole acetic acid (IAA) concentration and its immunocytochemical localization were determined in pollinated flowers after self- and free-pollination and in unpollinated flowers. In both cultivars the highest IAA levels and signal intensity were observed in free-pollinated pistils, whereas after self-pollination a different behaviour emerged. In ‘Pendolino’ the immunostaining was not uniformly distributed and stopped at the style level. This finding sustained the low percentage of fruit-set obtained in the cultivar. In ‘Pisa 9’ the auxin distribution was similar to the free-pollinated pistils where the immunolocalization staining reached the ovary. The correspondence between the IAA signal in the ovary and the high percentage of fruit-set in ‘Pisa 9’ self-pollinated pistils, confirms the role of this hormone in the induction and regulation of the fertilisation process.     

Hardy framework influence (1969–1983) on yield, tree size and yield efficiency of ‘Wellspur’ and ‘Goldspur’ apple

Cumulative yield, trunk cross-sectional area, and yield efficiency were measured between 1969 and 1983 in an experimental planting (established in 1964 and top-worked in 1965) of ‘Wellspur’ and ‘Goldspur’ apple top-worked on frameworks of 19 cold-hardy cultivars (‘Alnarp 2’, ‘Antonovka’, ‘Beacon’, ‘Canada Baldwin’, ‘Delcon’, ‘Dr. Bill’, ‘Duchess’, ‘Haralson’, ‘Hawkeye Greening’, ‘Heyer 12’, ‘Heyer 20’, ‘J. Luke Seedling’, ‘McIntosh’, ‘Minnesota 447’, ‘Ottawa 271’, ‘Ottawa 292’, ‘Red Astrachan’, ‘Robusta 5’, ‘Yellow Transparent’) that were grafted on domestic seedling roots. There were significant differences in yield, trunk diameter and yield efficiency related to the interstock. ‘Duchess’ and ‘Heyer 12’ ranked highest in yield efficiency and lowest in tree size. ‘Red Astrachan’ ranked lowest in efficiency and largest in tree size. ‘McIntosh’ also tended to be low in efficiency. No severe winter temperatures occurred to test differential cold hardiness.     

柑橘品种对不同基因型柑橘衰退病毒的抑制及变异的影响

为研究不同柑橘品种对柑橘衰退病毒（Citrus tristeza virus,CTV）不同基因型的抑制效果,将T36、T30、VT和T3等基因型CTV的毒源分别接种于‘赛蒙斯’甜橙、‘邓肯’葡萄柚、‘墨西哥莱檬’和‘强徳勒柚’,运用RT-qPCR技术检测被接种植株中CTV复制水平的差异。结果表明,‘墨西哥莱檬’最适于CTV各基因型的复制。‘邓肯’葡萄柚中VT、T3和T36基因型的复制水平最低。4种柑橘品种对T36基因型CTV都有明显的抑制作用。通过对CTV的CP基因进行分析发现,VT基因型CTV在‘赛蒙斯’甜橙中发生了较大变异。     

不同类型柑橘中黄脉病毒衣壳蛋白基因的保守性及复制水平

为研究不同柑橘品种对柑橘黄脉病毒(Citrus yellow vein clearing virus,CYVCV)变异的影响,将CYVCV毒株CQ01嫁接接种于4类共35个柑橘品种,对不同植株中CYVCV的衣壳蛋白(CP)基因进行分析,结果发现仅有少数位点发生了变异。运用RT-qPCR技术检测不同柑橘品种中CYVCV的相对含量,发现甜橙类品种中CYVCV的相对含量最高,墨西哥来檬中CYVCV的相对含量最低。植株中CYVCV的含量与其症状不存在明显的关联。     

The effect of hydrogen cyanamide (Dormex) on flowering and cropping of different apple cultivars under tropical conditions of sub-optimal winter chilling

The effects of pre-bud-break application of 1.5% hydrogen cyanamide (3% Dormex) on the flowering and cropping of 12 apple cultivars were determined over the years 1988/89 to 1991/92 in tropical Zimbabwe, at a site with less than 300 h of winter chilling below 7.2 °C per year. Control trees, other than those of ‘Mutsu’ and ‘Goldjon’ which were indistinguishable from the treated trees, showed varying degrees of delayed flowering and low yield in comparison with the cyanamide-treated trees. The latter, irrespective of cultivar, attained full bloom within 4–5 weeks of the common date of treatment, so that blossoming of the different cultivars, as well as that of the different trees of each cultivar, was synchronized, and all cropped heavily in relation to tree size. Very satisfactory yields of ‘Mollies Delicious’, ‘Canvada’, ‘Drakenstein’, ‘Golden Delicious’, ‘NJ46’, ‘Marjorie Pye’, ‘Spartan’, ‘Starking’, ‘Granny Smith’ and ‘Ohinemuri’ were thus obtained by the use of cyanamide in an environment otherwise unsuitable for them, especially for the later cultivars on this list.     

Development of seedless and Mal Secco tolerant mutant lemons through budwood irradiation

Mal secco (caused by Phoma tracheiphila (Petri) Kantsch. and Gik.) is the most destructive fungal disease of lemon plantations worldwide and seedless lemons would be preferred by most consumers. Five dosage levels, 0, 3, 5, 7, and 9 kiloradian (krad), of cobalt (⁶⁰Co) gamma irradiation were applied to budstick of ‘Kutdiken’ lemon (Citrus limon (L.) Burm. f.) clone KT-2A. Mutations were stabilized in three vegetative generations. Three hundred fifty-eight and 478 M1V3 (mutation one and vegetation three) plants were evaluated for seed number and mal secco tolerance in the field and the greenhouse, respectively. LD₅₀ was around 5 krad gamma irradiation for ‘Kutdiken’ lemon. The seed number varied from 0 to 34 per fruit. The level of mal secco tolerance also varied significantly among the plants from 1.0 (no symptom) to 4.3 (high level of disease occurance). The stable seedless and mal secco tolerant plants were obtained from 5 and 7 krad irradiation: the three mutants from 5 krad irradiation gave more lemon-like fruits, while 7 krad irradiation caused altered tree morphology and early maturation of fruits. This study shows considerable potential for lemon cultivar improvement aiming to obtain seedless and mal secco tolerant lemons.     

Flavonol and phenolic acid composition of sweet cherries (cv. Lapins) produced on six different vegetative rootstocks

The effect of rootstock (‘MaxMa 14’, ‘Weiroot 13’, ‘PiKu 1’, ‘Weiroot 158’, ‘Gisela 5’ and ‘F12/1’) on phenolic acid and flavonol content of “Lapins” sweet cherry was investigated. Phenolic acids and flavonols were isolated from sweet cherries and analyzed by using reversed phase high-performance liquid chromatography (RP-HPLC). The major phenolic acids in sweet cherries were neochlorogenic acid (18–50 mg kg⁻¹), chlorogenic acid (19–62 mg kg⁻¹) and p-coumaric acid derivatives (15–125 mg kg⁻¹). The amount of flavonol quercetin-3-rutinoside (8–37 mg kg⁻¹) was significant as well. There are significant variations in the phenolic compound content among sweet cherry fruits grown on trees grafted on different vegetative rootstocks. The significantly higher chlorogenic acid, neochlorogenic acid, p-coumaric derivative and quercetin-3-rutinoside contents were found in sweet cherry fruits grown on trees grafted on ‘Weiroot 13’ and ‘PiKu 1’ rootstocks. Sweet cherries produced on trees grafted on other rootstocks had significantly lower phenolic compound content.     

Identification and investigation of relationships of mandarin types using isozyme analysis

Sixteen enzyme systems with 19 loci were used to discriminate 19 mandarin cultivars and hybrids. Variability was observed at 12 loci, and all but 3 mandarin types could be discriminated, 2 of which were probably identical. Relatedness within the tangelo and tangor groups was high, probably reflecting their recent origin. Relatedness within the common mandarin group was low, reflecting their multiple origins and long cultivation. Exceptions were cultivars ‘Algerian’ and ‘Beauty of Glen Retreat’, which differed at only 1 locus. Relatedness between the groups was generally low, with the least relatedness between the tangelos and the other groups, probably owing to their grapefruit parent. The ‘Ellendale’ cultivars formed a particularly cohesive group, but with 2 isozyme genotypes, differing at 2 loci, both marketed as ‘Ellendale’. Three of the ‘Ellendale’-type cultivars probably arose by self-pollination of ‘Ellendale’, the rest either by self-pollination or by mutation.     

Analysis of genetic diversity in Portuguese Ceratonia siliqua L. cultivars using RAPD and AFLP markers

Although carob (Ceratonia siliqua L.) is of great economic importance little is still known about the pattern of genetic variation within this species. Morphological characteristics based on 31 fruit and seeds of continuous characters determinant for agro-industrial uses, were compared with RAPD and AFLP markers for assessing genetic distances in 68 accessions of carob trees, from different cultivars, varieties and eco-geographic regions of Algarve. Eighteen selected RAPD primers applied to the 68 accessions produced a total of 235 fragments ranging from 200 to 2000 bp, of which 93 (40%) were polymorphic. Four AFLP selective primer combinations generated a total of 346 amplification fragments of which 110 were polymorphic. The average level of polymorphism based on four primer combinations was 31.8%. The phenetic trees based on RAPD and AFLP analyses gave high co-phenetic correlation values, and were found to be consistent in general with the analysis of morphological data, carried out on the same accessions. A number of RAPD and AFLP markers were found to be diagnostic for ‘Canela’ cultivar and 13 wild ungrafted trees.     

柑橘果实糖积累与蔗糖代谢酶活性的研究

研究了脐橙、柠檬果实发育过程中汁胞和果皮内糖含量与糖代谢酶活性的变化。结果表明：不同柑橘品种、不同部位糖积累和相关酶活性变化不同。柑橘汁胞和果皮中还原糖含量均高于蔗糖含量。随着脐橙汁胞中糖分的积累,酸性转化酶(SAd)活性持续下降,至采收时几乎为零,说明SAd 的下降与脐橙汁胞糖分积累有很大关系。柠檬汁胞中的中性转化酶(NI)活性随着果实成熟而下降,在成熟时保持极低水平,而此时SAd 活性则较高,是其它酶活性的6倍以上,表明柠檬汁胞中糖分的利用在果实成熟前与NI活性有极大的关系,在成熟期则与高SAd 活性有关。脐橙和柠檬汁胞间糖组分的明显差异和相关酶的变化动态表明蔗糖合成酶(SS)和转化酶在柑橘汁胞糖代谢中可能起到重要作用。     

The inhibition of flowering by means of gibberellic acid application reduces the cost of hand thinning in Japanese plums (Prunus salicina Lindl.)

The application of gibberellic acid during flower bud induction significantly reduced flowering of ‘Black Diamond’ and ‘Black Gold’ Japanese plums. The response depended on the concentration applied and on the type of shoot. Mixed shoots had a similar response in both varieties, flowering being reduced by 40% for GA₃ 50 mg l⁻¹ and by 75–90% for GA₃ 75 mg l⁻¹ or higher concentration. With regard to spurs, GA₃ 50 mg l⁻¹ reduced flowering intensity by 40% and 25% in ‘Black Gold’ and ‘Black Diamond’, respectively, and GA₃ 75 mg l⁻¹ or higher concentration reduced flowering by 70% and 50%, respectively. This partial inhibition of flowering significantly reduced the cost of manual thinning. The best GA₃ concentration was found to be 50 mg l⁻¹, since it reduced the cost of thinning by 45–47% and increased final fruit weight by 7–33% for ‘Black Diamond’ and ‘Black Gold’, respectively. Not significant differences in yield and in mature fruit characteristics of treated trees were found compared to untreated trees.     

Fruit quality parameters of ‘Pioneer’ Japanese plums produced on eight different rootstocks

The influence of Marianna GF 8-1, GF 677, Julior-Ferdor, Nemared, Monegro (G × N 9), Felinem (G × N 22), Adesoto 101 and Cadaman rootstocks on fruit quality of ‘Pioneer’ Japanese plum was analysed. The study was performed during three consecutive years in the Guadalquivir Valley (Seville, Spain), on a silt loam and calcareous soil. Colour, fruit size, fruit and stone weights, hardness, soluble solid concentration and acidity were measured for fruit quality evaluation. In general, the effect of the different rootstocks on most of the analysed quality parameters was variable because a strong interaction rootstock × year was observed. However, several parameters as fruit shape, soluble solid concentration, acidity and maturity seem to be harder conditioned in some rootstocks, did not show significant differences year-by-year. Fruits produced on scions grafted on Marianna and Nemared showed the highest soluble solid concentration and Cadaman the lowest, the last also originated the hardest fruits and the lowest maturity.