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Neil O. Anderson and Peter D. Ascher

The snail flower, V. caracalla, was a popular ornamental flowering plant in conservatories at the turn of the century. It's popularity was due, in part, to its showy, orchid-like flowers whose fragrance rivals Stephanotis The indeterminate, vining growth habit can produce plants > 20 feet in height. V. caracalla is of interest for genetic and evolution studies since it is an ancestral species and possesses diagnostic traits of both Phaseolus (coiled style, leaf length/width ratios) and Vigna (> 10 seeds/ovary, long seed pods). However, its reproductive biology and use as an intergeneric hybrid bridge is unknown. Plants were examined for male and female fertility, self compatibility, and cross compatibility. Genotypes were self-incompatible; with one exception, self seed set did not occur following artificial manipulation. Selfed flowers abscised within 1-2 days post-pollination. Accessions were cross-compatible and highly fertile. To date, intergeneric hybridizations performed with P. coccineus --theancestral Phaseolus --have aborted following fertilization.

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John M. Capik and Thomas J. Molnar

are monoecious, wind-pollinated, and self-incompatible. Reproduction is restricted by a sporophytic self-incompatibility system, which is controlled by a single locus with various S-alleles determining compatibility ( Mehlenbacher, 1997 ; Olsen et al

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Cecil Pounders, Sandra Reed, and Margaret Pooler

Crapemyrtle (L. indica and L. indica × L. fauriei hybrids) is one of the most popular flowering landscape plants in the U.S. Although many cultivars have been developed through breeding efforts, little has been published on the reproductive biology of the genus. The objective of this study was to evaluate barriers to successful self-seed production in crapemyrtle. Self-compatibility was assessed by comparing pollen tube growth, fruit and seed production, and seed germination following controlled self- and cross-pollinations. Observations of pollen tube growth at intervals up to 24 hours after self- and cross-pollination indicated no barriers to self-fertilization acting at the stigmatic or stylar level in L. indica, L. fauriei or cultivars derived from inter-specific hybrids of these two species. Self-pollinations of `Catawba', `Whit IV', `Tonto' and `Tuscarora' had lower percent seed pod set and seed germination than did cross-pollinations of these cultivars. The number of seeds per pod was lower when `Catawba', `Whit IV' and `Tuscarora' were self-rather than cross-pollinated, but no difference between `Tonto' self- and cross-pollinations was observed. When decreased pod set is combined with much lower seed germination for self-pollinations, selfing of crapemyrtle is extremely unproductive when compared to cross-pollination. A late-acting self-incompatibility system or inbreeding depression is indicated for L. indica and inter-specific crosses with L. fauriei.

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Aurora Díaz, Antonio Martín, Pilar Rallo, Diego Barranco, and Raúl De la Rosa

We studied the self-incompatibility of two main Spanish olive (Olea europaea L.) cultivars, `Picual' and `Arbequina', by testing the selfing of the seeds with microsatellites. For this purpose, we used a rapid single-seed DNA extraction method and four highly polymorphic microsatellites. We analyzed seeds produced in branches bagged for selfing from mono- and multi-cultivar orchards in 2002 and 2003. We did not find any seed coming from selfing in the bagged branches, for either cultivar, in the two types of orchards. Additionally, we tested seeds coming from free pollination in mono-cultivar orchards from different locations. In the case of `Picual' olive, only three seeds out of the 70 collected were the product of selfing, although they came from mono-cultivar orchards located in areas where the cultivar used as the female parent was predominant. From the 20 seeds of `Arbequina' olive harvested in the middle of two high-density plantations, not one was a product of selfing. According to this, olive would behave as an allogamous species in mono-cultivar growing conditions and the pollen coming from long distances would be able to produce a normal bearing. Therefore, there is strong evidence to support the idea that the cultivars studied could be self-incompatible. Future experiments in self-compatibility should include a paternity check of the possible self seeds obtained.

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Sandra M. Reed

The objectives of this study were to evaluate self-incompatibility in Hydrangea paniculata Sieb. and H. quercifolia Bartr. and to determine optimum time for pollination of these two species. Flowers from three genotypes of each species were collected 1, 2, 4, 8, 24, 48, and 72 hours after cross- and self-pollination, stained with aniline blue and observed using a fluorescence microscope. In both species, pollen germination was observed on stigmas of over half of the flowers collected 4 to 72 hours after cross- or self-pollination. Differences in pollen tube length between cross- and self-pollinated flowers were noted from 8 to 72 hours after pollination in H. paniculata and from 24 to 72 hours after pollination in H. quercifolia. By 72 hours after pollination, most self-pollen tubes had only penetrated the top third of the style but cross-pollen tubes had grown to the base of the style and entered 40% to 60% of the ovules. Stigmas of H. paniculata were receptive to pollen from anthesis to 5 days after anthesis, while stigmas of H. quercifolia were receptive from 1 to 5 days after anthesis. This study provides evidence of a gametophytic self-incompatibility system in H. paniculata and H. quercifolia. Occasional self-seed set previously observed in these species was theorized to have been due to pseudo-self compatibility.

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Shawn A. Mehlenbacher and David C. Smith

The cutleaf hazelnut [Corylus avellana L. f. heterophylla (Loud.) Rehder] is an ornamental form with strongly dissected leaf morphology. Its stigmas express incompatibility allele S20 but none of the other 25 S-alleles was detected with fluorescence microscopy. Three seedlings from a cross of the cutleaf hazelnut and VR6-28 lacked S20 and were investigated further. Each expressed an allele from the parent VR6-28 (S2 S26), S26 in OSU 562.031 and OSU 562.048 and S2 in OSU 562.049. S2 and S26 are low in the dominance hierarchy, so we expected the new allele from the cutleaf hazelnut to be expressed in their pollen. Unexpectedly, fluorescence microscopy showed that pollen of all three selections was compatible on their cutleaf parent and on each other, and furthermore, self-pollinations showed the excellent germination and long parallel tubes in the styles that are typical of a compatible pollination. Controlled self- and cross-pollinations in the field verified the self-compatibility of two selections. Cluster set for self-pollinations was very high (75-90%) and within the range observed for compatible cross-pollinations. Furthermore, the frequency of blank nuts was low (<10%). The second allele in the cutleaf hazelnut is designated S28, and its presence in seedlings of `Cutleaf' is indicated by the absence of S20. Controlled pollinations in the field also showed that selection OSU 562.069 (S2 S28) from the cross `Cutleaf' × `Redleaf #3' was self-compatible. Fluorescence microscopy showed that two additional seedlings were self-incompatible [OSU 367.052 (S1 S28) and OSU 367.076 (S6 S28)] while a third [OSU 706.071 (S9 S28)] was self-compatible. Self-compatibility may be limited to genotypes that combine S28 with a second allele that is low in the dominance hierarchy.

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Attila Hegedüs, Zoltán Szabó, József Nyéki, Júlia Halász, and Andrzej Pedryc

The most commercially grown peach [Prunus persica (L.) Batsch.] cultivars do not require cross-pollination for reasonable fruit set; however, self-incompatibility is a well-known feature within the Prunoideae subfamily. Isoelectric focusing and native polyacrylamide gel electrophoresis of S-ribonucleases; PCR analyses of S-RNase and S-haplotype-specific F-box genes as well as DNA sequencing were carried out to survey the self-(in)compatibility allele pool and to uncover the nature of self-compatibility in peach. From 25 cultivars and hybrids with considerable diversity in phenotype and origin, only two S-haplotypes were detected. Allele identity could be checked by exact length determination of the PCR-amplified fragments and/or partial sequencing of the peach S 1-, S 2-, and Prunus davidiana (Carr.) Franch. S 1-RNases. S-RNases of peach were detected to possess ribonuclease activity, and a single nucleotide polymorphism in the S 1-RNase was shown, which represents a synonymous substitution and does not change the amino acid present at the position in the protein. A 700-bp fragment of the peach SFB gene was PCR-amplified, which is similar to the fragment size of functional Prunus L. SFBs. All data obtained in this study may support the contribution of genes outside the S-locus to the self-compatible phenotype of peaches.

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Sandra M. Reed

Breeding efforts in Clethra alnifolia L., an ornamental shrub native to the Eastern U.S., are hindered by a lack of information on the reproductive behavior of this species. The objective of this study was to evaluate self-compatibility, time of stigma receptivity, and the relationship between time of pollen shed and stigma receptivity in C. alnifolia. Stigma receptivity and changes in floral morphology were monitored over a 7-day period beginning at flower opening. Pollen germination and pollen tube growth in styles were examined following self- and cross-pollinations using fluorescence microscopy. Seed set and germination were compared following self- and cross-pollinations. Anthers began to dehisce in `Hummingbird' and `Ruby Spice' the day after flowers opened, but stigmas did not become fully receptive to pollen until 2 days later. An increase in the length of pistils was observed following flower opening. Maximum elongation of pistils occurred at approximately the same time stigmas became receptive and could be utilized as an indicator of receptivity. While self-pollen tubes appeared to grow slightly slower than cross-pollen tubes, there was no indication of a self-incompatibility system acting at the stigmatic or stylar level in C. alnifolia. Self-pollinations of `Hummingbird' and `Ruby Spice' produced fewer seeds than did cross-pollinations of these cultivars. Germination of all seed obtained from this study was too poor to allow a comparison of germination rates of the self- and cross-pollinated seed. However, because a few self-progeny were obtained, emasculation is recommended when making controlled pollinations. The presence of a late acting self-incompatibility system or early-acting inbreeding depression was proposed as being responsible for the lower seed set following self-pollination.

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H. Yamane, R. Tao, A. Sugiura, N. Hauck, and A. Iezzoni

Most fruit tree species of Prunus exhibit gametophytic self-incompatibility, which is controlled by a single locus with multiple alleles (S-alleles). One interesting aspect of gametophytic self-incompatibility is that it commonly “breaks down” as a result of polyploidy, resulting in self-compatible individuals. This phenomenon is exhibited in the diploid sweet cherry (P. avium) and the tetraploid sour cherry (P. cerasus), in which most cultivars are self-compatible. Recently, S-gene products in pistil of Prunus species were shown to be S-RNases. As sour cherry is one Prunus species, it is likely to possess S-alleles encoding pistil S-RNases. To confirm this, we surveyed stylar extracts of 11 sour cherry cultivars, including six self-compatible and five self-incompatible cultivars, by 2D-PAGE. As expected, all 11 cultivars tested yielded glycoprotein spots similar to S-RNases of other Prunus species in terms of Mr, immunological characteristics, and N-terminal sequences. A cDNA clone encoding one of these glycoproteins was cloned from the cDNA library constructed from styles with stigmas of a self-compatible cultivar, `Erdi Botermo'. Deduced amino acid sequence from the cDNA clone contained two active sites of T2/S type RNases and five conserved regions of rosaceous S-RNases. In order to determine the inheritance of self-incompatibility and S-allele diversity in sour cherry, we conducted genomic DNA blot analysis for sour cherry germplasm collections and mapping populations in MSU using the cDNA as a probe. To date, it appears as if self-compatibility in sour cherry is not simply controlled by a self-fertile allele as demonstrated in other Prunus species.

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R.J. Knight Jr.

When Passiflora incarnata L. was crossed with P. edulis f. flavicarpa Degener, all plants of the diploid hybrid were pollen-sterile and nonfruitful. Doubling the chromosome number of emergent FI seedlings with colchicine restored fertility in some individuals, but all plants were strongly self-incompatible and many showed low pollen viability. Crossing colchicine-treated plants that had been converted to amphiploids produced a tetraploid hybrid group of four seedling progenies that had some degrees of cross-compatibility. Juice of the amphiploid hybrid is lighter in color than that of P. edulis, but is sweet, strongly aromatic, and may have use alone or, typically, as a blend with other juices.