Syrian pear (Pyrus syriaca) as a pollinator for European pear (Pyrus communis) cultivars

In Israel four European pear cultivars are grown: ‘Spadona’ is the main cultivar and ‘Coscia’, ‘Gentile’ and ‘Spadochina’ are its pollinators. However, molecular S-genotyping revealed that ‘Spadona’ is semi-compatible with its three pollinators. This explains, at least in part, the relatively low pear yield in Israel. The Syrian pear (Pyrus syriaca) grows wild in Israel and blooms intensively, overlapping the blooming of the cultivated European pears. Cross-fertilization between Syrian pear and ‘Spadona’ was shown to be efficient suggesting that Syrian pear might be a potent pollinator for ‘Spadona’. Twenty-six Syrian pear seedlings, from different sites in north-east Israel were S-genotyped identifying 11 that are fully compatible with the four European pear varieties cultivated in Israel. By this screening, 24 different S-RNases were cloned; ten of them are new, whereas the other fourteen had been identified previously. In addition, seedlings of two wild pear species were also S-genotyped. Two seedlings from Pyrus betulifolia and one from Pyrus korshinskii were found to be genetically compatible with the four European pear cultivars. From these seedlings four S-RNases were cloned, two are new, one had been cloned previously and one was identical to an S-RNase allele cloned from Syrian pear in this work.     

Identification of RAPD marker linked to fruit skin color in Japanese pear (Pyrus pyrifolia Nakai)

In Japanese pear, fruit skin color is a very important trait for growers because the russet skin protects the fruit against external stress caused by disease, insects, bad weather, and shipping. This report describes the development of a randomly amplified polymorphic DNA (RAPD) marker linked to major genes controlling the fruit skin color in Japanese pear. Two F₁ progenies from the cross of ‘Kousui’ × ‘Kinchaku’ and ‘Niitaka’ × ‘Chikusui’ segregated by fruit skin color were used for bulked segregant analysis. Four kinds of bulked DNA were constructed and used for the polymerase chain reaction in RAPD analysis. After 200 random primers were screened against four bulks, a band named OPH-19₄₂₅ was selected in cooperation with green bulks. The recombination rate between OPH-19₄₂₅ and the green skin phenotype was 7.3%. The RAPD marker (OPH-19₄₂₅) could select green fruit with probability as high as approximately 92%. The marker was apparently useful for the selection of green-skinned individuals in a breeding program. This is the first report on developing a DNA marker closely linked to the fruit phenotype in Japanese pear.     

Identification of seven haplotype-specific SFBs in European pear (Pyrus communis) and their use as molecular markers

The European pear (Pyrus communis) carries the S-RNase-mediated gametophytic self-incompatibility (GSI) system. The S-haplotype is conferred by an S-locus, which contains the style-specific expressed S-RNase, and the pollen-specific expressed F-box genes (SFB). Both the S-RNase and the SFB genes are multi-allelic and each is characteristic of one of the S-haplotypes. Therefore, they are ideal markers for molecular S-genotyping. In this work, for the first time, seven haplotype-specific SFBs were isolated from European pears. Particular primers for each of these SFBs were generated, thus providing an additional tool for S-genotyping of European pear cultivars.     

Spray pollination as a labor-saving pollination system in Japanese pear (Pyrus pyrifolia (Burm.f.) Nakai): Development of the suspension medium

Effects of suspension media used for spray pollination on pollen grain viability were investigated in Japanese pear (Pyrus pyrifolia (Burm.f.) Nakai ‘Kosui’). The suspension media tested in this study consisted of pectin methylesterase (PME) and polygalacturonase (PG) combined with either 0.1% agar or 0.04% xanthangum (XG). We also evaluated the influences of spray pollination on fruit set and fruit quality. Pollen grain viability, as measured by germination and pollen tube growth rates, was maintained in media supplemented with 0.1 mg L⁻¹ PME or 0.1 mg L⁻¹ PG. The level of fruit set after spray pollination using media containing PME or PG in combination with either 0.1% agar or 0.04% XG was almost the same as the level after artificial pollination by hand (hand pollination). The media containing XG combined with either PME or PG seemed to show better results for pollen grain viability and fruit set, although the results were variable from year to year. With regard to fruit size, shape and other parameters for fruit quality, spray pollination and hand pollination gave comparable results, irrespective of the medium composition. The time required for spray pollination was less than half of that required for hand pollination, and the amount of pollen grain required for spray pollination was one-third or less than the amount required for hand pollination. Thus, spray pollination may be a time- and labor-saving pollination system for the cultivation of Japanese pear.     

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.     

Performance of ‘Coscia’ pear (Pyrus communis) on nine rootstocks in the north of Israel

The vegetative and reproductive performances of ‘Coscia’ pear (Pyrus communis L.) grown on nine rootstocks [OHF 69, OHF 97, OHF 217, OHF 333, OHF 513 and BP 1 (P. communis), clonal seedling (Davis AxB) of Pyrus betulifolia and quince BA 29 and EMA (Cydonia oblonga)] were compared during a 9-year period. The trial was conducted at the Experimental Station ‘Avnei Eitan’ in northern Israel (elevation 400 m above sea level, a.s.l.), on a well-drained soil with pH 7.1. Trees were planted in December 1998, spaced at 4.0 m × 2.0 m and trained with a central axis. The most vigorous trees were on P. betulifolia seedlings, followed by the four OHF rootstock (69, 97, 333, 513) and BP 1 (with no significant difference between them). All the above rootstocks demonstrated greater vigor than OHF 217 and quince BA 29 or EMA. The highest cumulative yields per tree were harvested from trees on the four OHF rootstocks (69, 97, 333, 513) and P. betulifolia followed by the BP 1. The two quince rootstocks, as well as OHF 217, had the lowest cumulative yield and the lowest yield of large fruit. A positive correlation was found between the vigor of the tree, as affected by the rootstock, and both the total yield and the fruit size. We conclude that in a warm climate, yield efficiency is not the only parameter that should be taken into account, and building a strong tree for a weak cultivar is the first requirement for establishing an orchard.     

Regeneration and characterization of plants derived from leaf in vitro culture of two sweet orange (Citrus sinensis (L.) Osbeck) cultivars

In the current work attempts were made to investigate culture of leaf explants derived from in vitro seedlings of two sweet orange (Citrus sinensis (L.) Osbeck) cultivars, Bingtangcheng and Valencia. Effects of several factors, including culture medium, lighting condition, explant age and genotype on regeneration response were examined based on three parameters, percentage of explants producing shoots, mean number of shoots per explant and shoot forming capacity. Culture of the explants on shoot-inducing media (SIM) composed of MT salts supplemented with different growth regulators gave rise to disparate shoot regeneration, in which SIM₁ (MT + 0.5 mg L⁻¹ BA + 0.5 mg L⁻¹ Kinetin + 0.1 mg L⁻¹ NAA + 3% sucrose + 0.8% agar, pH 5.8) was shown to be the most effective medium for direct induction of shoots from leaf explants. Highly significant difference in the response of shoot bud regeneration was noted between the two cultivars, with Bingtangcheng being more responsive than Valencia. Culture of explants from fully developed leaves led to better shoot regeneration capacity in comparison to undeveloped ones. However, the two lighting conditions used herein did not cause significant difference in shoot regeneration. Phenotypic observation and randomly amplified polymorphic DNA (RAPD) analysis confirmed that all the regenerated plants from both genotypes were genetically identical to their donor plants, suggesting absence of detectable genetic variation in the regenerated plants. The data presented here demonstrated that direct initiation of plants from leaf explants has been successfully accomplished. To our knowledge, this is the first report on direct regeneration of shoots from leaf explants in Citrus, which will provide an alternative source for citrus genetic manipulation in the future.     

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

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

Effects of post-harvest foliar boron and calcium applications on subsequent season's pollen germination and pollen tube growth of pear (Pyrus pyrifolia)

The primary objective of this study was to evaluate the effects of foliar boron and calcium application after harvest on the quantity and activity of pollen in the ‘Housui’ and ‘Wonwhang’ pears on a subsequent year. Pollen grains of the ‘Housui’ pear were cultured on germination medium, to which had been added boric acid (0, 25, 75, 100, 200, 300, 400, and 500 mg L⁻¹) and calcium nitrate (0, 10, 25, 50, 100, 150, 200, 250, 300, 400, and 500 mg L⁻¹). Boric acid, which was added to the germination media, exerted a significant stimulatory effect on both pollen germination and pollen tube growth, although pollen tube growth was inhibited at higher concentrations than 300 mg L⁻¹. Calcium nitrate addition stimulated pollen germination, except at concentration of 500 mg L⁻¹. However, pollen tube growth was significantly inhibited with increasing concentrations of calcium nitrate. In the orchard experiment, boron and calcium were sprayed at concentrations of 0, 100, 200, 500 or 1000 mg L⁻¹ onto leaves after harvest, respectively. Boron and calcium content in the tissues as well as pollen production and growth were determined after these treatments. The foliar application of boron mainly resulted in an increase of boron concentration in buds. It also induced an increase in the weight of the anther and pollen in the following year. On the other hand, the foliar application of calcium resulted in an increase of calcium concentration mainly in the leaves, but pollen weight was decreased at high concentration treatment in the following year. The germination rate and tube growth of collected pollen were highest in the trees which had received boron treatment at a concentration of 200 mg L⁻¹. In contrast, the germination rate and tube growth of collected pollen were decreased by calcium application at concentrations of 500 and 1000 mg L⁻¹ without significant increase at lower concentrations. Consequently, the accumulation of boron in the developed buds of pear trees subjected to post-harvest foliar boron application generated positive effects on both the quantity and quality of pollen in the following year.     

S-RNase based S-genotyping of Japanese plum (Prunus salicina Lindl.) and its implication on the assortment of cultivar-couples in the orchard

Determination of the genetic compatibility between self-incompatible cultivars is crucial in agriculture. The Rosaceae family carries the S-RNase-mediated gametophytic self-incompatibility (GSI) system. Each haplotype is conferred by an S-locus. The S-locus contains two highly polymorphic genes, S-RNase and SFB, which are characteristic of each haplotype and therefore these genes are ideal markers for molecular S-genotyping. In this study 43 Japanese plum cultivars grown in Israel were S-genotyped based on their S-RNase gene sequences. Four alleles, S^b, S^c, S^e and S^h are widespread and together are responsible for 87% of the S-haplotypes therefore many of the cultivar combinations are semi-compatible. In Israel semi-compatibility was shown to correlate with low yield. However, two cultivars, ‘Wickson’ S^fS^k and ‘Shiro’ S^fS^g carry rare S-haplotypes and, therefore, are fully compatible with most of the analyzed cultivars.     

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.     

Identification of organic acid-related genes and their expression profiles in two pear (Pyrus pyrifolia) cultivars with difference in predominant acid type at fruit ripening stage

‘Yandangxueli’ is a pear cultivar with predominant citric acid in the ripe fruit, different from most of pear cultivars such as ‘Gengtouqing’ in which malic acid is the predominant acid type. It was found that ‘Yandangxueli’ accumulated citric acid for three times against that in ‘Gengtouqing’ at fruit ripening stage. To investigate the mechanism of citric acid accumulation in ‘Yandangxueli’, organic acids content, gene expression and enzyme activity were studied in both cultivars. Five genes, Pp:mtCs, Pp:cyAco, Pp:cyIdh, Pp:mtMdh and Pp:cyMe which encoded citric synthase (CS), cytosolic aconitase (cyACO), NADP-dependent isocitrate dehydrogenase (NADP-IDH), NAD-dependent malate dehydrogenase (NAD-MDH) and NADP-dependent malic enzyme (NADP-ME) respectively, were identified from pear fruit. Their expression profiles and the corresponding enzyme activities were determined throughout fruit development in both cultivars. Results from these enzymes indicated that there were no strict relationship between gene expression, enzyme activity and citric acid accumulation. Expression analysis for two Py:vVAtp genes encoding vacuolar H⁺-ATPase A subunit and one Py:vVpp gene encoding Vacuolar H⁺-pyrophosphatase showed that they were all with up-regulated expression at the later development stage of ‘Yandangxueli’ but with down-regulated expression in ‘Gengtouqing’. Therefore, it is concluded that the different ability in citric acid transportation and storage might be involved in the high citric acid content in ‘Yandangxueli’.     

Relationship between the oil and sugar content in olive oil fruits from Moraiolo and Leccino cultivars during ripening

In our previous work (Cherubini et al., 2009), sugar concentration was proposed as an accurate, reproducible index for technological ripening of olive oil fruits. During 2009 crop season, sugar and oil content of olive oil fruits from Moraiolo and Leccino cultivars were measured during ripening. The sugar content of olive oil fruits was determined both by titration and by using a portable refractometer.     

Identification of S-genotypes and novel S-RNase alleles in Japanese apricot cultivars native to China

Information on S-genotypes is essential for designing orchards of Japanese apricot (Prunus mume Sieb. et Zucc.) and for hybridization. However, this information is lacking for most cultivars grown in China. Thus, in this work, the S-genotypes of 24 Japanese apricot cultivars native to China were identified by sequencing the PCR products obtained from allele-specific polymerase chain reaction (AS-PCR). Seventeen S-RNase alleles were amplified, ten of them for the first time. The new S-RNase alleles were submitted to GenBank and denoted them as S¹⁷, S¹⁸, S¹⁹, S²⁰, S²¹, S²², S²³, S²⁴, S²⁵ and S²⁶. Furthermore, the S-genotypes of four Japanese apricot cultivars were confirmed by field-testing cross-pollination.     

Alteration of tomato fruit quality by root inoculation with plant growth-promoting rhizobacteria (PGPR): Bacillus subtilis BEB-13bs

In this study the effect of inoculation of tomato (Lycopersicon esculentum Mill.) roots with plant growth-promoting rhizobacteria (PGPR) on yield and fruit quality was evaluated. The control treatment was non-inoculated (CTL) and the PGPR treatment was inoculated with Bacillus subtilis BEB-lSbs (BS13). Yield per plant and marketable yield, as well as fruit weight and length were increased by the BS13 treatment when compared to the CTL treatment. Texture of red fruits was also enhanced by the BS13 treatment compared to that in the CTL treatment. These results demonstrated that PGPR have positive effects on tomato fruit quality attributes, particularly on size and texture.     

Amplified fragment length polymorphism (AFLP) markers for fingerprinting of Argyranthemum frutescens cultivars

Argyranthemum frutescens is a commercially important ornamental species with extensive breeding programmes in several countries. Since it is vegetatively propagated there is a great need for tools for identification of cultivars. Vegetatively propagated species require clean-up from diseases, often performed through meristem-tip cultures. Forty-three cultivars of A. frutescens propagated by meristem-tip culture and traditional cuttings were analyzed for genetic relatedness and possible somaclonal variation using AFLP. Five primer combinations resulted in a relatively high degree of polymorphism. Ten molecular markers generated by one primer combination could distinguish between all 43 cultivars. Differences in fingerprints between meristem-tip culture and cuttings from the same cultivars were found, but the proportion of fragments being specific for either tissue culture or cuttings was relatively low. Some cultivars that did not display somaclonal variation as judged by the AFLP-fingerprints may still be genetically unstable since some morphological variation was observed in the true to type test.     

Photosynthesis of field-grown Arracacha (Arracacia xanthorriza Bancroft) cultivars in relation to root-yield

There has been limited research on measuring potential differences in leaf gas exchange of Arracacha (Peruvian parsnip, Arracacia xanthorriza Bancroft) cultivars, as affected by different environments, as well as its relation to storage root-yield. The present paper reports field measurements of leaf CO₂ assimilation rates (A) for five contrasting cultivars grown at two different high-altitude locations. Using a design of plots chosen at random with three repetitions, commercial root production was determined in the two locations at different altitude (1580 and 1930 m). Daily leaf gas exchange was repeatedly monitored with a portable open-mode infrared gas analyzer at different times in both locations during the growth cycle. Root-yield, leaf area and dry weight were measured. Significant differences in leaf photosynthetic rate and in specific leaf area (SLA) were observed among cultivars. Cultivars with high SLA, had high CO₂ assimilation. Mean (A_n) and total (A_tot) of CO₂ assimilation and SLA were significantly correlated with storage root-yield across cultivars and locations. The three cultivars with the greatest commercial root production also had the highest maximum values for A and the highest specific leaf area, indicating that these two parameters can be used to select for highly productive cultivars of A. xanthorriza.     

Production of intraspecific hybrids between wild-type and petaloid-sepal cultivars in Habenaria radiata

Orchids are commercially important plants with flowers that are unique and very specialized in shape and color. The flowers consist mostly of sepals, lateral petals, lip (labellum) and column, and are zygomorphic and resupinate. Whereas most orchid species have petaloid tepals in the first and second whorls, Habenaria radiata has a flower with greenish sepals and white lateral petals and lip. ‘Hishou’, one of the cultivars of H. radiata, is a floral homeotic mutant and has a petaloid median sepal and lip-like lateral sepals in the first whorl. Additionally, this cultivar often has non-resupinate flowers whereas wild-type H. radiata flowers are resupinate. In the present study, we investigated the genetic inheritance of these characters in the ‘Hishou’ cultivar by crossing it with wild-type plants. Some intraspecific hybrids, which were confirmed by PCR-RFLP analysis, had flowers with a petaloid median sepal and lip-like lateral sepals in the first whorl, indicating that these were dominant characters. Since the remainder of the intraspecific hybrids had wild-type flowers, these characters must be heterozygous in ‘Hishou’ plants. Although ‘Hishou’ plants had non-resupinate flowers, intraspecific hybrid flowers were resupinate, even though they had the petaloid median sepal and lip-like lateral sepals. This result indicates that non-resupination must be a recessive character. Since sepal-petalization and triple lip characters of ‘Hishou’ inherited dominantly, these characters can be utilized for the breeding of Habenaria species by intra- and interspecific crosses.     

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.