Fruit set variation associated with apozygotic reproduction in sugar beet <Emphasis Type="Italic">Beta vulgaris</Emphasis> L.

Fruit set upon apozygotic reproduction was studied for four years in the msSOAN-5 pollen-sterile inbred sugar beet line. The progenies obtained from pollen-sterile plants by apozygotic reproduction had both fruits with normal seeds and parthenocarpic fruits without seeds, which was not an occasional event. Growth conditions proved to strongly affect the fruit set and seed quality. For instance, water deficiency during early plant development increased the proportion of parthenocarpic seeds. Water deficiency combined with a lower temperature during flowering additionally caused a substantial decrease in the total number of fruits. Under the same growth conditions, related accessions did not differ in seed productivity, but varied in the proportion of normal (with seeds) and parthenocarpic (without seeds) fruits.     

Parthenocarpy and seed predation by insects in Bursera morelensis

Background and Aims

While parthenocarpy (meaning the production of fruits without seeds) may limit fecundity in many plants, its function is not clear; it has been proposed, however, that it might be associated with a strategy to avoid seed predation. Bursera morelensis is a dioecious endemic plant that produces fruits with and without seeds, and its fruits are parasitized by insects. Its reproductive system is not well described and no published evidence of parthenocarpy exists for the species. The purpose of this work was to describe the breeding system of B. morelensis and its relationship to seed predation by insects.

Methods

The breeding system was described using pollination experiments, verifying the presence of parthenocarpic fruits and apomictic seeds. Reproductive structures from flower buds to mature fruits were quantified. For fruits, an anatomical and histological characterization was made. The number of fruits in which seeds had been predated by insects was correlated with parthenocarpic fruit production.

Key Results

The major abortion of reproductive structures occurred during fruit set. The results discard the formation of apomictic seeds. Flowers that were not pollinated formed parthenocarpic fruits and these could be distinguished during early developmental stages. In parthenocarpic fruits in the first stages of development, an unusual spread of internal walls of the ovary occurred invading the locule and preventing ovule development. Unlike fruits with seeds, parthenocarpic fruits do not have calcium oxalate crystals in the ovary wall. Both fruit types can be separated in the field at fruit maturity by the presence of dehiscence, complete in seeded and partial in parthenocarpic fruits. Trees with more parthenocarpic fruits had more parasitized fruits.

Conclusions

This is the first time the anatomy of parthenocarpic fruits in Burseraceae has been described. Parthenocarpic fruits in B. morelensis might function as a deceit strategy for insect seed predators as they are unprotected both chemically and mechanically by the absence of calcium oxalate crystals.Key words: Parthenocarpy, Bursera morelensis, predation, seeds, insects, breeding system, calcium oxalate crystals     

Expression of Mdh1 locus alleles in apozygotic progenies of sugar beet (Beta vulgaris L.)

Expression of Mdh1 alleles has been studied in 60 apozygotic (agamospermic) sugar beet progenies. Seed progenies were obtained by uniparental (pollen less) mode of seed reproduction: selfing of pollen-sterile plants isolated with paper bags. The apozygotic seed progenies demonstrate a disomic gamete autosegregation, i.e., the ratio between genotypes in the progenies correspond to the gamete segregation in a duplex heterozygote of an autotetraploid. It was shown that the ratio between the Mdh1 phenotypes in apozygotic progenies is strongly affected by spontaneous inactivation of one of the alleles. In most progenies, the excess of FF phenotypes and the deficit of SS phenotypes were observed. In our opinion, such deviations in genotype and phenotype frequencies result from conversion of the active Mdh1-S into the inactive Mdh1-S0 allele (epigenetic gene inactivation). The spontaneous inactivation of one allele results in extremely variable frequencies of heterozygous Mdh1-F/Mdh1-S genotypes and phenotypes in the apozygotic seed progenies. The empirical distribution of the frequencies of heterozygous genotypes in the apozygotic seed progenies is given by a negative binomial distribution describing the expected time of occurrence of random events.     

Expression of Alleles of the Mdh1Locus in Apozygotic Progenies of Sugar Beet Beta vulgarisL.

Expression of Mdh1alleles has been studied in 60 apozygotic (agamospermic) sugar beet progenies. Seed progenies were obtained by uniparental (pollenless) mode of seed reproduction: selfing of pollen-sterile plants isolated with paper bags. The apozygotic seed progenies demonstrate a disomic gamete autosegregation, i.e., the ratio between genotypes in the progenies correspond to the gamete segregation in a duplex heterozygote of an autotetraploid. It was shown that the ratio between theMdh1phenotypes in apozygotic progenies is strongly affected by spontaneous inactivation of one of the alleles. In most progenies, the excess of FF phenotypes and the deficit of SS phenotypes were observed. In our opinion, such deviations in genotype and phenotype frequencies result from conversion of the active Mdh1-Sinto the inactive Mdh1-S ⁰allele (epigenetic gene inactivation). The spontaneous inactivation of one allele results in extremely variable frequencies of heterozygous Mdh1-F/Mdh1-Sgenotypes and phenotypes in the apozygotic seed progenies. The empirical distribution of the frequencies of heterozygous genotypes in the apozygotic seed progenies is given by a negative binomial distribution describing the expected time of occurrence of random events.     

Analysis of serial sequences of seeds in sugar beet plants by apozygotic reproduction

Dichotomic serial sequences of fruits of two types (with and without seeds) on shoots were analyzed in sugar beet upon apozygotic reproduction. Serial sequences were analyzed using serial statistical tests. The distribution of fruits of the two types was nonrandom in most cases. The variation of serial sequences is considered in terms of the epigenetic variation of discrete characters in plants.     

Analysis of serial sequences of seeds in sugar beet plants upon apozygotic reproduction

Dichotomic serial sequences of fruits of two types (with and without seeds) on shoots were analyzed in sugar beet upon apozygotic reproduction. Serial sequences were analyzed using serial statistical tests. The distribution of fruits of the two types was nonrandom in most cases. The variation of serial sequences is considered in terms of the epigenetic variation of discrete characters in plants.     

The role of endogenous gibberellin in seed and fruit development of tomato: Studies with a gibberellin-deficient mutant

The role of endogenous gibberellin (GA) in seed and fruit development was studied with the use of the GA-deficient ga-1 mutant of tomato ( Lycopersicon esculentum Mill. cv. Moneymaker). Flowers of the ga-1 mutant were abnormal and sterile, but parthenocarpic fruit development was observed occasionally on the dwarf plants. A single application of GA₄₊₇ restored the fertility of the mutant flowers and resulted in seed set. Development of GA-producing and GA-deficient seeds in GA-deficient fruits was compared by pollination of ga-1/ga-1 flowers with wild-type or ga-1 pollen, respectively. In ga-1/ga-1 seed dehydration started about 1 week earlier than in Ga-1/ga-1 seeds. Ultimate fresh and dry weights of mature Ga-1/ga-1 seeds were higher than those of ga-1/ga-1 seeds and showed negative correlations with the total number of seeds per fruit. Total content and composition of seed proteins were not influenced by the GA-deficiency. Germination of the mature seeds depended on embryonal GA synthesis and was not influenced by maternal GA production. Final fresh weight of the ga-1/ga-1 fruits was positively correlated with the number of seeds per fruit. In these fruits, the minimum number of seeds for growth above the parthenocarpic level was about 10 or 35 in the presence of Ga-1/ga-1 or ga-1/ga-1 seeds, respectively. Fruits containing GA-producing seeds reached a higher fresh weight than those containing GA-deficient seeds, and their ripening was delayed by one week. It is concluded that gibberellin is indispensable for the development of fertile flowers and for seed germination, but only promoting in later stages of fruit and seed development.     

Cloning of gibberellin 3 beta-hydroxylase cDNA and analysis of endogenous gibberellins in the developing seeds in watermelon

We have isolated Cv3h, a cDNA clone from the developing seeds of watermelon, and have demonstrated significant amino acid homology with gibberellin (GA) 3 beta-hydroxylases. This cDNA clone was expressed in Escherichia coli as a fusion protein that oxidized GA(9) and GA(12) to GA(4) and GA(14), respectively. The Cv3h protein had the highest similarity with pumpkin GA 2 beta,3 beta-hydroxylase, but did not possess 2 beta-hydroxylation function. RNA blot analysis showed that the gene was expressed primarily in the inner parts of developing seeds, up to 10 d after pollination (DAP). In the parthenocarpic fruits induced by treatment with 1-(2-chloro-4-pyridyl)-3-phenylurea (CPPU), the embryo and endosperm of the seeds were undeveloped, whereas the integumental tissues, of maternal origin, showed nearly normal development. Cv3h mRNA was undetectable in the seeds of CPPU-treated fruits, indicating that the GA 3 beta-hydroxylase gene was expressed in zygotic cells. In our analysis of endogenous GAs from developing seeds, GA(9) and GA(4) were detected at high levels but those of GA(20) and GA(1) were very low. This demonstrates that GA biosynthesis in seeds prefers a non-13-hydroxylation pathway over an early 13-hydroxylation pathway. We also analyzed endogenous GAs from seeds of the parthenocarpic fruits. The level of bioactive GA(4 )was much lower there than in normal seeds, indicating that bioactive GAs, unconnected with Cv3h, exist in integumental tissues during early seed development.     

Fruit characteristics, seed production and pollen tube growth in the wild chilli pepper Capsicum flexuosum

Fruits formed after different pollination regimes (flowers hand pollinated, unpollinated, and open pollinated) and the seeds obtained were characterized in the wild chilli pepper Capsicum flexuosum Sendtn. Pollen tube development in vivo and ovary growth were also analyzed. Seedless fruits and empty seeds were abundant among the fruits from hand pollinated and open pollinated flowers, while no more than one seed with embryo was found in a low percentage of fruits from such pollination treatments. Parthenocarpic fruits were formed from unpollinated flowers. Pollen tube growth was arrested in the upper third of the style for almost all pollen tubes except for a single one that may continue elongating occasionally. The ovary size increased continuously after pollination, even without fertilization. The sum of the evidence registered may help to explain the low number of seeds with embryo harvested, the abundance of seedless fruits formed from pollinated flowers (possibly parthenocarpic), and the high rate of parthenocarpic fruits formed from unpollinated flowers.     

Temporal variation in the effects of habitat fragmentation on reproduction of the Mediterranean shrub <Emphasis Type="Italic">Colutea hispanica</Emphasis>

Habitat fragmentation poses a major threat to the viability of plant populations. However, the intensity of fragmentation effects may vary among years. We studied two possible effects of habitat fragmentation (patch size and isolation) on the reproduction and proportion of damaged fruits in 24 patches of the self-compatible shrub Colutea hispanica for three consecutive years with different climate conditions. We also studied the effect of fragmentation on the incidence of two main pre-dispersal seed predators, the butterflies Iolana iolas and Lampides boeticus. High between-year variability was found in number of viable seeds per fruit, number of fruits per plant, total number of viable seeds per plant and proportion of damaged fruits. In 2003, small, isolated patches had a higher fruit set and number of fruits per plant. The proportion of damaged fruits was significantly lower in isolated populations in 2003, while it was very high in all patches in 2004 and 2005. High between-year variability was also found in the proportion of fruits per plant with I. iolas eggs. In 2003 isolated patches had a lower proportion of fruits with I. iolas eggs, but no significant effect of patch size and isolation was found in 2004 or 2005. The proportion of fruits with L. boeticus eggs was similar in the three years of study, although it was slightly higher in large, non-isolated patches in 2003. Thus, the effects of fragmentation on plant reproduction cannot be generalized from one single-year survey. In contrast to the generally accepted idea that fragmentation reduces plant reproduction, plant fitness may increase in isolated patches in years with high fruit production and low seed predation.     

Empty seeds reduce seed predation by birds in Juniperus osteosperma

Utah juniper (Juniperus osteosperma) is one of many plant species that produce large numbers of fruits containing parthenocarpic or otherwise empty or inviable seeds. We tested the hypothesis that production of empty fruits in this species results in reduced levels of predation on fertile seeds. In a population in west-central Utah, we estimated the proportion of fruits with filled seeds in trees suffering high levels of fruit destruction by the seed-eating bird Parus inornatus and in neighbouring trees similar in crown and fruit-crop size but suffering negligible predation. We found that the heavily attacked trees had higher proportions of filled seeds. Thus, juniper may benefit from producing fruits that contain no offspring. This is the first study to demonstrate that empty seeds may reduce predation by vertebrate seed eaters and the first to demonstrate discrimination based on seed filling at the level of whole plants.     

Induced parthenocarpy—a way of changing the levels of endogenous hormones in tomato fruits (Lycopersicon esculentum Mill.) 1. Extractable hormones

Ethylene, indol-3-acetic acid (IAA), gibberellin-like substances (GAs) and abscisic acid (ABA) were analysed in extracts from normal, seed-containing and parthenocarpic tomato fruits throughout fruit development. Parthenocarpic fruit growth was induced with an auxin (4-CPA), morphactin (CME) or gibberellic acid (GA₃) and compared with that of pollinated control fruits. Fruit growth was only affected by the treatment with GA₃, decreasing size and fresh weight by 60%. The peak sequence of hormones during fruit development was ethylene-GAs-IAA-ABA. Seeded fruits contained the highest levels of IAA and ABA but the lowest levels of GAs. Also, in seeded fruits, a high proportion of IAA and ABA was found in the seeds whereas this was not the case for GAs.Hormone levels of tomato fruits may be successfully, easily and reproducibly altered by inducing parthenocarpic fruit growth and thus eliminating development of seeds which are a major source of hormone synthesis. In spite of markedly changed hormone levels, there was no obvious relationship between fruit growth and extractable hormones per se. However, the results indicate that a high ratio of GAs: auxins is unfavourable for growth of tomato fruits.     

Genetic engineering of parthenocarpic fruit development in tomato

Parthenocarpy was engineered in two genotypes of Lycopersicon esculentum Mill. by using the DefH9-iaaM chimeric gene. The parthenocarpic trait consists of fruit set and growth in the absence of fertilization. Seedless parthenocarpic fruits were obtained from emasculated flowers, and fruits with seeds from pollinated flowers. All parthenocarpic tomato plants analysed expressed the DefH9-iaaM gene during flower development. The fruit set percentage of emasculated transgenic flowers was similar to that of control plants. In 7 out of 8 independent transgenic plants, the fresh weight of fruits derived from pollinated or emasculated flowers did not significantly differ from that of fruits obtained by pollination of the control plants. The pH of the parthenocarpic fruit was generally unaffected and the soluble solid concentration was either unchanged or increased. Thus, the DefH9-iaaM gene is a genetic tool that might be used to improve tomato productivity.     

Parthenocarpic fruits in wild parsnip: Decoy defence against a specialist herbivore

Summary Parthenocarpy, the production of fruits without viable seeds, is a widespread phenomenon in plants. While failure to effect pollination or fertilization is often cited as the cause of parthenocarpy, this explanation alone is inadequate to explain why plants produce, maintain and further develop fruits. Wild parsnips (Pastinaca sativa) frequently produce parthenocarpic fruit. When parsnip webworms (Depressaria pastinacella), specialist feeders on wild parsnip, were given choices between normal fruit and parthenocarpic fruit, they exhibited a strong preference for parthenocarpic fruit. However, on parthenocarpic fruit, insects fed less efficiently and grew more slowly than insects fed normal fruit. Parthenocarpic fruits, then, may act as decoys that divert herbivores away from fruits that contain plant offspring.     

Early anther ablation triggers parthenocarpic fruit development in tomato

Fruit set and fruit development in tomato is largely affected by changes in environmental conditions, therefore autonomous fruit set independent of fertilization is a highly desirable trait in tomato. Here, we report the production and characterization of male‐sterile transgenic plants that produce parthenocarpic fruits in two tomato cultivars (Micro‐Tom and Moneymaker). We generated male‐sterility using the cytotoxic gene barnase targeted to the anthers with the PsEND1 anther‐specific promoter. The ovaries of these plants grew in the absence of fertilization producing seedless, parthenocarpic fruits. Early anther ablation is essential to trigger the developing of the transgenic ovaries into fruits, in the absence of the signals usually generated during pollination and fertilization. Ovaries are fully functional and can be manually pollinated to obtain seeds. The transgenic plants obtained in the commercial cultivar Moneymaker show that the parthenocarpic development of the fruit does not have negative consequences in fruit quality. Throughout metabolomic analyses of the tomato fruits, we have identified two elite lines which showed increased levels of several health promoting metabolites and volatile compounds. Thus, early anther ablation can be considered a useful tool to promote fruit set and to obtain seedless and good quality fruits in tomato plants. These plants are also useful parental lines to be used in hybrid breeding approaches.     

Composition and seasonal dynamics of seed rain in broad-leaved Korean pine (Pinus koraiensis) mixed forest in Changbai Mountain, China

To explore the composition and spatio-temporal dynamics of seed rain in broad-leaved Korean pine (Pinus koraiensis) mixed forest, 150 seed traps were set up in a 25 hm2 plot in Changbai Mountain. Seeds, fruits, anthotaxy and others in seed traps were collected, identified and divided into 4 types. From 2005 to 2006, we collected 47 different types. Total number of seeds and fruits was 121291, including 23147 mature seeds and fruits (19.1% of the total). Tilia amurensis and Fraxinus mandshurica, with the most seeds and fruits, accounted for 90% of the total. The analysis on seasonal dynamics of seed rain showed that there were the largest number of seeds and fruits between July and October, which were composed of immature seeds and fruits. In mid-October, mature seeds and fruits reached their peak, but immature seeds and fruits still accounted for high proportion. There were 91 traps that contained 100–200 mature seeds and fruits, and one trap without any mature seed or fruit. The largest number of species found in a trap was 7, and usually 3 or 4 species were found in most of the traps. There were obvious relationships between spatial patterns of mature seeds and fruits and their parent trees, indicating that their mature seeds and fruits were not dispersed far from their parent trees.     

Effects of self-pollination and maternal resources on reproduction and offspring performance in the wild lupine, Lupinus perennis (Fabaceae)

We examined the effects of self-pollination and resource addition to maternal plants of Wild Lupine on seed production in the field, and on offspring performance in the greenhouse. Although 24% of flowers set fruits when open-pollinated, only 11% of flowers set fruits when self-pollinated. Self-pollination significantly reduced fruit and seed production per inflorescence and increased aborted seeds per fruit. Resource addition to maternal plants significantly increased fruit and seed number in the field. Moreover, selfed plants exhibited greater variability in seed production in the absence of resource addition to the maternal plant. We planted a total of 1,306 of the seeds from this experiment in the greenhouse. While self-pollination did not affect the proportion of seeds emerging, it slowed seedling emergence by 5–10%, and reduced offspring biomass by 25–35%. Interestingly, resource addition to the maternal plants significantly decreased proportion of seedlings surviving after 5 months. Moreover, offspring from maternal plants with resource addition expressed more inbreeding depression in the seedling stage compared to offspring from maternal plants without resource addition, for which more inbreeding depression occurred during seed maturation and emergence. These results indicate that conservation efforts using benign environments to increase number of seeds or offspring may face compensating reductions in survivorship at other life stages.     

A Role for Auxin and Auxin Transport Inhibitors on the Ca Content of Artificially Induced Parthenocarpic Fruits

Artificially induced parthenocarpic fruits of apples, pears and tomatoes, as well as seeded fruits treated with 2,3,5-triiodobenzoic acid, frequently show symptoms of Ca deficiency and a low Ca content. It was concluded that auxins, probably produced by the seeds, play a significant role in Ca translocation into fruits. Exogenous indoleacetic acid but not 4-chlorophenoxyacetic acid applications could replace the effect of seeds in this respect. Auxin transport, rather than auxin accumulation, seems to be necessary for Ca transport, as can be concluded from the effect of auxin transport inhibitors.