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Richard P. Buchner, William H Olson, Vito S. Polito, and Katherine Pinney

Walnut Blight caused by the bacteria Xanthomonas campestris pathovar juglandis is a very destructive disease for California walnut production. Streptomycin is an effective disease control material; however, Streptomycin sprays can result in significant nut drop 3 to 5 weeks after spray application. We investigated the basis for walnut drop following applications of Streptomycin (Agrimycin) for walnut blight control. Flowers and developing nuts were collected from four treatments, plus an unsprayed control. 200 ppm Streptomycim was applied at 1) budbreak; 2) pre, full, and post-bloom; 3) postbloom; 4) budbreak and postbloom; 5) untreated control. Samples were collected regularly beginning at the first budbreak spray and extending through the period of nut drop. Samples were fixed and prepared for histological examination. In treatments with a high incidence of nut drop, the embryo failed to develop. Examination of the stigma and style in flowers from these treatments showed inhibited pollen tube growth. Results indicate that Streptomycin inhibits pollen tube growth, which precludes fertilization. This pattern of development and timing of nut drop following Streptomycin application at full bloom is similar in all ways to unpollinated walnut flowers. Nut growth and development appear normal for 3 to 5 weeks; then nuts abort. If Streptomycin became available for walnut blight control, sprays timed to coincide with pistillate bloom and pistillate flower receptivity should be avoided.

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Rebecca J. McGee and James R. Baggett

There was no difference in percentage in vitro germination of pollen from stringless pea (Pisum sativum L.) cv. Sugar Daddy and stringy `Oregon Sugarpod II' (OSP) and `OSU 705' (705). However, pollen tubes of `Sugar Daddy' grew more slowly in vitro than those of OSP or 705. Differences in pollen tube growth rate were demonstrated in vivo following time-course pollinations involving reciprocal crosses of `Sugar Daddy' with OSP and 705, along with the selfed parents. After 8 hours, pollen tubes from stringless peas (“stringless” pollen) had entered 13% of the ovules compared with 51% for those from stringy peas (“stringy” pollen). Stringless pollen tubes entered 29% and stringy pollen tubes 66% of the ovules after 10 hours. The slower growth of stringless compared with stringy pollen tubes is a plausible explanation for previously observed deficiencies of stringless plants in segregating populations.

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Yasumasa Takatsu, Masakazu Kasumi, Toru Manabe, Mikio Hayashi, Eiichi Inoue, Wataru Marubashi, and Masaru Niwa

Interspecific hybridization between a modern cultivar of Gladiolu×grandiflora hort. (2n = 60) and the wild species G. tristis L. (2n = 30) was made to introduce characteristics of the wild species into the cultivated one. Gladiolus ×grandiflora is a summer-flowering species, and G. tristis flowers in winter. The effect of storage temperature on pollen viability was tested, as long-term storage of pollen was necessary to facilitate crossing these two species. Pollen of G. tristis could be stored at -20 °C for ≈1 year, and was more practical than storage at -80 °C. Air temperature affected pollen tube growth, fertility, and fruit set in the cross between G. ×grandiflora and G. tristis, and low temperatures (15 to 20 °C) were best. The morphological data and flow cytometric analysis showed that the F1 plants were hybrids between G. ×grandiflora and G. tristis.

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R. Paul Baker, Karl H. Hasenstein, and Michael S. Zavada

In order to characterize the self-incompatibility system in Theobroma cacao, the levels of ethylene, indole-3-acetic acid (IAA), and abscisic acid (ABA) were determined after pollination with compatible and incompatible pollen and in unpollinated flowers. Pollen tube growth rates after incompatible and compatible pollinations were identical, and the majority of the pollen tubes reached the ovules between 12 and 20 hours after pollination. ABA levels rose in incompatibly pollinated flowers, and fell in compatibly pollinated flowers, prior to pollen tube—ovule contact. Ethylene evolution remained stable in compatibly pollinated flowers and rose in incompatibly pollinated flowers. IAA concentrations increased in compatibly pollinated flowers, and remained stable in incompatibly pollinated flowers after pollination and subsequent to pollen tube—ovule contact.

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Julián Cuevas and Vito S. Polito

We investigated pollination and fruit set parameters in `Manzanillo' olive (Olea europaea L.) following self-pollination and pollination with `Sevillano', `Ascolano', and `Mission' pollen. Results of analyses and experiments conducted over 2 years in central California indicated that `Manzanillo' behaves as a self-incompatible cultivar (index of self-incompatibility = 0.22 to 0.24). Pollination with `Sevillano' resulted in a more than 4-fold increase in fruit set over self-pollination. When `Mission' or `Ascolano' pollen was used, there was no increase over self-pollinated samples. Analyses of pollen tube growth, fertilization, initial fruit set, and final fruit set were consistent with `Manzanillo' being considered as a self-incompatible cultivar cross-incompatible with `Mission' and `Ascolano'. Our results indicate that `Manzanillo' is likely to be more productive when interplanted with `Sevillano' rather than when planted without a pollinizer or with `Mission' or `Ascolano'.

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A.M.S. Nyomora, P.H. Brown, K. Pinney, and V.S. Polito

The effect of boron (B) on in vivo and in vitro development of almond [Prunus dulcis (Mill.) D.A. Webb (syn. P. amygdalus Batsch)] pollen and pollen tubes and the resultant effect on fruit set was studied in mature trees. The cultivars Mono (pistil donor) and Butte (pollinizer) in an orchard with low soil B in Fresno, California were sprayed with B at 0, 0.8, 1.7, or 2.5 kg·ha-1 during Fall 1993. Pollen viability as indicated by the fluorescein diacetate method (FDA) was >85% and was not affected by field-applied B, however, in vivo pollen germination and tube growth were enhanced by foliar-applied B. More effect of applied B on in vivo growth appeared as pollen tubes progressed toward the ovary. For in vitro germination, foliar-applied B reduced bursting of tubes, and addition of B to the culture media significantly increased pollen germination and pollen tube growth.

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Maria Victoria González, Manuel Coque, and Maria Herrero

The effective pollination period was determined in kiwifruit [Actinidia deliciosa (Chev.) Liang and Fergusonl and the factors affecting it were evaluated. The effective pollination period, measured as the capability to set fruit after hand-pollinating flowers of different ages, was 4 days; 5 days after anthesis fruit set decreased and 2 days later it was nil. Pollen tube growth did not appear to he a limiting factor since pollen tubes grew quickly and reached the base of the style 2 days after pollination and reached the ovules 1 day later. Ovules appeared viable for the 7 days following anthesis, and visibly degenerated within the following 3 days. Stigmatic receptivity was determined by the ability to sustain pollen germination after hand pollinating flowers of different ages. The duration of stigmatic receptivity closely fit the effective pollination period determined through fruit set. Thus, it appears that stigma receptivity is the main factor responsible for the short effective pollination period.

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Hector G. Nunez-Palenius, Daniel J. Cantliffe, Harry J. Klee, and Don J. Huber

Pollen germination timing has a paramount role in fertilization of a flower. Rapid germination and outgrowth of a pollen tube that penetrates the stigma is required. Physical and biological factors can affect pollen germination timing. The objective of this study was to determine if ACC oxidase antisense gene expression could influence in vitro pollen germination and in vitro pollen tube length growth. A transgenic (ACC oxidase antisense) `Galia' male parental line had a reduced fruit set compared to its wild type. Likewise, embryo abortion and empty seeds after self-pollination in a `Galia' male parental line were observed. Wild type and transgenic `Galia' male parental line melon plants were grown in a greenhouse according to the practices of Rodriguez (2003). Male flowers were collected from these plants between 10 to 12 am; pollen was obtained by dipping the anther in germination medium (10.25% sucrose, 0.031% calcium nitrate, 0.015% boric acid, 0.0075% KNO3, and 0.016% MgSO4) at 25 °C and analyzed immediately, either for total percentage of germination after 5 minutes of incubation or to measure pollen tube growth rate every 5 minutes during 1 hour. Each flower provided an average of 250 pollen grains. Assays were conducted by using the “Hanging Drop Method” (Okay and Ayfer, 1994). Percentage of pollen germination in WT `Galia' male parental line was greater than the transgenic line. Likewise, in vitro pollen tube growth in wild type `Galia' melon was greater than pollen from the transgenic line. Possibly the ACC oxidase antisense gene expression in `Galia' male parental line may have had an influence on the reduced fruit set observed.

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Javier Sanzol, Pilar Rallo, and María Herrero

Apples and pears are fruit crops particularly susceptible to cropping irregularities. A strong relationship has been observed between the effective pollination period (EPP) and the general cropping of the orchard. The EPP concept has also been proven to be a useful parameter to establish a relationship between the variation in the reproductive process and cropping behaviors. For apples and pears, a slow pollen tube growth has been shown to be the main limiting factor of the EPP in the traditional cooler temperate cultivation regions. However, while higher temperatures speed up the pollen tube growth, the expansion of these crops into warmer areas often results in failures of fruit set. Thus, with the aim to ascertain the main limiting factor responsible for fruit set failures in Mediterranean conditions we have evaluated the EPP for two consecutive years in `Agua de Aranjuez' pear, the main Spanish cultivar, by studying the stigmatic receptivity, pollen tube kinetics, and ovule development. Complete flower fertility was maintained for just 2 days after anthesis in both years. Pollen tube kinetics and ovule degeneration do not appear to limit flower receptivity. However, the stigmatic receptivity expressed as flowers with at least one receptive stigma, closely matches the duration of the EPP evaluated from fruit set experiments. This was consistent over the 2 years of experiments, in spite of the differences recorded in the EPP, suggesting that stigmatic receptivity is clearly the limiting factor of flower receptivity. This is the first report for stigmatic receptivity limiting the EPP in pears and suggests that stigmatic receptivity could be an important factor limiting pear flower receptivity and hence cropping performance under warmer conditions.

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Hisayo Yamane, Ryutaro Tao, Akira Sugiura, Nathanael R. Hauck, and Amy F. Iezzoni

This report demonstrates the presence of S-ribonucleases (S-RNases), which are associated with gametophytic self-incompatibility (SI) in Prunus L., in styles of self-incompatible and self-compatible (SC) selections of tetraploid sour cherry (Prunus cerasus L.). Based on self-pollen tube growth in the styles of 13 sour cherry selections, seven selections were SC, while six selections were SI. In the SI selections, the swelling of pollen tube tips, which is typical of SI pollen tube growth in gametophytic SI, was observed. Stylar extracts of these selections were evaluated by two-dimensional polyacrylamide gel electrophoresis. Glycoproteins which had molecular weights and isoelectric points similar to those of S-RNases in other Prunus sp. were detected in all selections tested. These proteins had immunological characteristics and N-terminal amino acid sequences consistent with the S-RNases in other Prunus sp. Two cDNAs encoding glycoproteins from `Erdi Botermo' were cloned. One of them had the same nucleotide sequence as that of S4-RNase of sweet cherry (Prunus avium L.), while the amino acid sequence from the other cDNA encoded a novel S-RNase (named Sa-RNase in this study). This novel RNase contained two active sites of T2/S type RNases and five regions conserved among other Prunus S-RNases. Genomic DNA blot analysis using cDNAs encoding S-RNases of sweet cherry as probes indicated that three or four S-RNase alleles are present in the genome of each selection regardless of SI. All of the selections tested seemed to have at least one S-allele that is also found in sweet cherry. Genetic control of SI/SC in tetraploid sour cherry is discussed based on the results obtained from restriction fragment length polymorphism analysis.