The effect of heat stress on tomato pollen characteristics is associated with changes in carbohydrate concentration in the developing anthers

Continuous exposure of tomato 'Trust' to high temperatures (day/night temperatures of 32/26 degrees C) markedly reduced the number of pollen grains per flower and decreased viability. The effect of heat stress on pollen viability was associated with alterations in carbohydrate metabolism in various parts of the anther during its development. Under control, favourable temperature conditions (28/22 degrees C), starch accumulated in the pollen grains, where it reached a maximum value 3 d before anthesis; it then diminished towards anthesis. During anther development, the concentration of total soluble sugars gradually increased in the anther walls and in the pollen grains (but not in the locular fluid), reaching a maximum at anthesis. Continuous exposure of the plants to high temperatures (32/26 degrees C) prevented the transient increase in starch concentration and led to decreases in the concentrations of soluble sugars in the anther walls and the pollen grains. In the locular fluid, however, a higher soluble sugar concentration was detected under the high-temperature regime throughout anther development. These results suggest that a major effect of heat stress on pollen development is a decrease in starch concentration 3 d before anthesis, which results in a decreased sugar concentration in the mature pollen grains. These events possibly contribute to the decreased pollen viability in tomato.     

Comparing heat stress effects on male-fertile and male-sterile tomatoes

To separate the effects of heat stress on male and female reproductive tissues, male-sterile (MSs) and male-fertile tomatoes (MFs) were placed in growth chambers at 12 h day/12 h night temperatures of 28/22, 30/24 or 32/26 °C from flower appearance to seed maturation (daily mean temperatures of 25, 27 or 29 °C). Pollen from MFs was applied individually to MS flowers. As MFs were self-pollinated, heat stress was experienced by both male and female tissues. At growth temperatures of 29 °C fruit number, fruit weight per plant, and seed number per fruit were only 10%, 6·4% and 16·4%, respectively, compared with those at 25 °C. Heat stress also adversely affected fruitset in MSs, especially when experienced by donor pollen. No fruit at all developed on MSs receiving pollen produced at 29 °C, even when ovule development, pollen germination and subsequent embryo development all took place at 25 °C. Effects on fruitset in MSs were reduced if donor pollen had not experienced heat stress. MSs grown at 29 °C but receiving pollen developing at 25 °C produced 73% as much fruit (both on number and weight basis), had 40% as high fruitset and produced 87% of the seed per fruit as MSs grown at 25 °C. This use of male-sterile and male-fertile lines of tomato provides new evidence that impairment of pollen and anther development by elevated temperature will be an important contributing factor to decreased fruit set in tomato, and possibly other crops, with global warming.     

Physiological factors limit fruit set of tomato (Lycopersicon esculentum Mill.) under chronic, mild heat stress

The effects of chronic, mild heat stress on fruit set, fruit production, release of pollen grains, photosynthesis, night respiration and anther dehiscence were examined in tomatoes (Lycopersicon esculentum Mill.) differing in high‐temperature sensitivity. Plants were grown under three temperature regimes: (1) 28/22 or 26/22 °C (optimal temperature); (2) 32/26 °C (high temperature); and (3) 32/26 °C day/night temperatures relieved at 28/22 °C for 10 d before anthesis, then returned to 32/26 °C (relieving treatment). FLA 7156 was the only cultivar with fruit set at 32/26 °C. All five cultivars, however, had fruit set under the relieving treatment (RT). The longer the relief, the higher the percentage of fruit set. Longer periods of relief also increased the number of pollen grains released, and linear regression analysis showed a significant relationship between the number of pollen grains released and the percentage of fruit set. Germination of pollen grains was also lowered in high‐temperature‐grown plants. The number of pollen grains produced, photosynthesis and night respiration did not limit fruit set under chronic, mild heat stress, however. This suggested that cultivar differences in pollen release and germination under heat stress are the most important factors determining their ability to set fruit.     

缙云山四川大头茶花粉游离脯氨酸含量与生殖产量特征研究

四川大头茶（Gordonia acuminata）是缙云山亚热带常绿阔叶林的优势树种之一,在缙云山的很多群落类型中均有分布。对缙云山四川大头茶的花粉游离脯氨酸含量的影响因素以及花粉游离脯氨酸含量与四川大头茶生殖产量之间的关系进行了研究。研究表明,缙云山四川大头茶的花粉游离脯氨酸含量不受植株年龄大小的影响,但与植株所处的群落类型有很大关系。常绿阔叶中四川大头茶的花粉游离脯氨酸含量最高,四川大头茶纯林中次之,针阔混交林中四川大头茶花粉游离脯氨酸含量最低。四川大头茶花蕾产量和果实产量随植株年龄的增大而增大,但座果率与植株年龄大小无关。对花粉游离脯氨酸含量与花蕾产量、果实产量和座果率之间的相关关系分析,花粉游离脯氨酸含量与花蕾产量不具有相关关系,与果实产量具有正相关关系,与座果率具有极显著正相关关系。这证实了与受精作用无关的生殖产量特征不受花粉游离脯氨酸含量影响,而与受精作用有关的生殖产量特征将受花粉游脯氨酸含量影响的研究假设。由于与花粉游离脯氨酸含量呈极显著正相关关系,座果率具有和花粉游离脯氨酸含量一样的群落间变化模式。     

Movement herkogamy in Kosteletzkya virginica: Effect on reproductive success and contribution to pollen receipt and reproductive assurance

Herkogamy may act as a mechanism to reduce interference between the reproductive functions of female and male organs, but too great a distance between stigmas and anthers may also decrease the outcross pollen transfer and male and female fitness. Our goal was to quantify the effect of movement herkogamy in Kosteletzkya virginica on reproductive success and its contribution to pollen receipt and reproductive assurance. Field investigations and hand-pollination experiments were performed to test: (1) the relationship between herkogamy before curvature of un-pollinated lobes and reproductive success under pollinator exclusion; (2) the contribution of movement herkogamy to pollen receipt; and (3) the relationship between pollen load and seed set. We noted that variation in herkogamy before curvature of un-pollinated lobes did not affect the outcome of autogamous self-pollination measured by fruit set and mean number of seeds per fruit. Under natural conditions, 39% of pollen grains deposited per flower were from autogamous self-pollination via movement herkogamy resulting in contact between anthers and stigmas. The relationship between the amount of pollen grains and the number of seeds per capsule was highly significant. Flowers producing four or five seeds received more pollen grains than those producing only one seed. Our data provides experimental evidence that movement herkogamy, which decreases the separation between anthers and stigmas and results in delayed self-pollination, contributes to pollen receipt and further enhances seed set when pollinators are scarce or absent.     

Movement herkogamy in Kosteletzkya virginica: Effect on reproductive success and contribution to pollen receipt and reproductive assurance

Herkogamy may act as a mechanism to reduce interference between the reproductive functions of female and male organs, but too great a distance between stigmas and anthers may also decrease the outcross pollen transfer and male and female fitness. Our goal was to quantify the effect of movement herkogamy in Kosteletzkya virginica on reproductive success and its contribution to pollen receipt and reproductive assurance. Field investigations and hand-pollination experiments were performed to test: (1) the relationship between herkogamy before curvature of un-pollinated lobes and reproductive success under pollinator exclusion; (2) the contribution of movement herkogamy to pollen receipt; and (3) the relationship between pollen load and seed set. We noted that variation in herkogamy before curvature of un-pollinated lobes did not affect the outcome of autogamous self-pollination measured by fruit set and mean number of seeds per fruit. Under natural conditions, 39% of pollen grains deposited per flower were from autogamous self-pollination via movement herkogamy resulting in contact between anthers and stigmas. The relationship between the amount of pollen grains and the number of seeds per capsule was highly significant. Flowers producing four or five seeds received more pollen grains than those producing only one seed. Our data provides experimental evidence that movement herkogamy, which decreases the separation between anthers and stigmas and results in delayed self-pollination, contributes to pollen receipt and further enhances seed set when pollinators are scarce or absent.     

Response of Tomato Sugar and Acid Metabolism and Fruit Quality under Different High Temperature and Relative Humidity Conditions

The combined stress of high temperature and high relative air humidity is one of the most serious agrometeorological disasters that restricts the production capacity of protected agriculture. However, there is little information about the precise interaction between them on tomato fruit quality. The objectives of this study were to explore the effects of the combined stress of high temperature and relative humidity on the sugar and acid metabolism and fruit quality of tomato fruits, and to determine the best relative air humidity for fruit quality under high temperature environments. Four temperature treatments (32°C, 35°C, 38°C, 41°C), three relative air humidity (50%, 70%, 90%) and four duration (3, 6, 9, 12 d) orthogonal experiments were conducted, with 28°C, 50% as control. The results showed that under high temperature and relative air humidity, the activity of sucrose metabolizing enzymes in young tomato fruits changed, which reduced fruits soluble sugar content; in addition, enzyme activities involved phosphopyruvate carboxylase (PEPC), mitochondria aconitase (MDH) and citrate synthetase (CS) increased which increased the content of organic acids (especially malic acid). Eventually, vitamin C, total sugar and sugar-acid ratio decreased significantly, while the titratable acid increased, resulting in a decrease in fruit flavor quality and nutritional quality in ripe fruit. Specifically, a temperature of 32°C and a relative air humidity of 70% were the best cultivation conditions for tomato reproductive growth period under high temperature. Our results indicating that fruit quality reduced under high temperature at the flowering stage, while increasing the relative air humidity to 70% could alleviate this negative effect. Our results are benefit to better understand the interaction between microclimate parameters under specific climatic conditions in the greenhouse environment and their impact on tomato flavor quality.     

Morph-biased male sterility in the tropical distylous shrub Erythroxylum havanense (Erythroxylaceae)

This study explores whether ecological factors, such as pollinators and pollen flow, or variation in pollen and ovule development account for the observed differences (approximately twofold) in the reproductive output of pin and thrum individuals of Erythroxylum havanense. The importance of ecological factors was assessed by means of comparison of the identity of pollinators and the rates of flower visitation, and by performing controlled hand pollinations and measurements of fruit set. In addition, we described the pollen and ovule development of thrum and pin individuals. Our results indicate that pollinators of E. havanense do not distinguish between floral morphs. The differences in fruit set between pin and thrum plants held even after hand pollination and, therefore, the observed differences in reproductive output between floral morphs of E. havanense cannot be explained in terms of asymmetrical pollen flow. There were no differences in the pattern of gynoecium development between the pin and thrum morphs, however androecium development showed marked differences between the morphs, and there was a resemblance between the developmental pathways leading to male sterility of the thrum morph of E. havanense with that of species with cytoplasmic male sterility (CMS).     

Mapping quantitative trait loci for heat tolerance of reproductive traits in tomato (<Emphasis Type="Italic">Solanum lycopersicum</Emphasis>)

Global warming has become a worldwide concern due to its adverse effects on agricultural output. In particular, long-term mildly high temperatures interfere with sexual reproduction and thus fruit and seed set. To uncover the genetic basis of observed variation in tolerance against heat, a bi-parental F₂ mapping population from two contrasting cultivars, i.e. Nagcarlang and NCHS-1, was generated and phenotyped under continuous mild heat conditions for a number of traits underlying reproductive success, i.e. pollen viability, pollen number, style length, anther length, style protrusion, female fertility and flowering characteristics, i.e. inflorescence number and flowers per inflorescence. Quantitative trait loci (QTLs) were identified for most of these traits, including a single, highly significant one for pollen viability, which accounted for 36% of phenotypic variation in the population and modified pollen viability under high temperature with around 20%. QTLs for some traits colocalised, indicating trait dependency or pleiotropic-effect loci. We conclude that a limited set of major genes determines differences in performance of reproductive traits under continuous mild heat in tomato. The results contribute to our fundamental understanding of pollen thermotolerance and may support development of more heat-tolerant tomato varieties.     

Floral biology and reproductive system of enantiostylous Senna corymbosa (Caesalpiniaceae)

The genus Senna (K. Bahuin) Miller (Cassieae) is represented in Argentina by 35 species and several varieties distributed in temperate, tropical and subtropical regions, and presents a high degree of endemism. Some taxa are used for medicine, animal foraging and ornamental purposes. Floral morphology, phases, rewards, attractants, visitors, pollen, reproductive system, P/O ratio. OCI and ISI indexes of enantiostylous Senna corymbosa were analyzed for morphological androecial differentiation and possible related functional differences between stamens groups. The study was carried out over three consecutive flowering seasons in March of 1999, 2000 and 2001, in two populations near the city of Buenos Aires, Argentina. The species has a buzz-pollination syndrome. The pollinators are Bombus atratus that vibrate the stamens, leading to nothotribic or stemotribic deposition of pollen. P/O ratio and pollen production were high thanks to the precise pollinating mechanism that needs a large delivery of pollen to ensure effective pollen deposition. P/O ratio also indicates that the species is xenogamous, although geitonogamy and autogamy (only induced, not spontaneous) were also recorded. The species is self-compatible. There were differences in hand-pollination treatments between long and medium stamens in fruit set, as well as in vitro differential fertility between their pollen grains. Hence, there are morphological and functional androecium differences: the medium stamens play a nutritional role while the long ones play a reproductive one. These differences are reflected in the breeding system. Moreover, differential fertility and enantiostyly may diminish the effects of self-compatibility by partially reducing the contribution of geitonogamy to selfing.     

Flowering and morphological characterization of Dendrocalamus asper androecium and pollen grains

Abstract

Little is known about the reproduction of Dendrocalamus asper because it flowers only every 100 to 120 years. In the present work we describe some reproductive features of this bamboo and characterise flowers and pollen at various developmental stages. Number of pollen grains and ovules per flower, pollen/ovule ratio, in vitro twinning and pollen grain viability in vivo were evaluated and the different stages of floral development identified. Further, we performed a morphological analysis of androecium and pollen development. Seven distinct stages of flower development were identified; four initial stages, a pre-anthetic stage, and two stages of anthetic. Dendrocalamus asper pseudospikelets avoid inbreeding by means of protogyny. The floral and pollen characteristics suggest that the species is anemophilous. The ultrastructural characteristics of free microspores (stage two of floral development), vacuolated microspores (stage five) and mature pollen (anthetic) were analysed. During maturation, pollen grains accumulate larger and more numerous amyloplasts and organelles such as mitochondria. Pollen disperse in the tricellular development stage. Pollen is monoporate with an operculum-like pore, with a rugulate structure and a spinose tectum.     

Pollination intensity and potential seed set in Passiflora vitifolia

Summary Initial seed set and fruit set were pollen-limited in a Costa Rican population of Passiflora vitifolia, a self-incompatible species with 200–350 ovules per flower. Pollination intensity was measured by counting the number of allogamous pollen grains on stigmas of the large one-day flowers. Hand-pollinations demonstrated that 25–50 pollen grains are required for fruit set, and >450 are needed for maximum seed set, with a pollen:seed ratio of about 1.6:1.0. Hummingbirds (Phaethornis superciliosus) delivered sufficient allogamous pollen for maximum seed set to only 28% of the flowers examined. Naturally pollinated flowers yielded fewer fruits and fewer seeds per fruit than those pollinated by hand. Most pollen transferred by humming-birds was self-incompatible; emasculated flowers yielded higher seed set than flowers with intact anthers. Visitation rates did not provide a good index of effective pollination.There were significant differences in ovule number, maximum seed set, and maximum per cent seed set among individual vines. More than half of an individual's flowers failed to set fruit, whether pollinated by birds or by hand. In this population, maximum reproductive potential may be limited by maternal resources for fruit development, but seed set varies with pollination intensity. Pollen-limited seed set may be a disadvantage of self-incompatibility, especially in species with many-seeded fruits.     

An overview of heat stress in tomato (Solanum lycopersicum L.)

Heat stress has been defined as the rise of temperature for a period of time higher than a threshold level, thereby permanently affecting the plant growth and development. Day or night temperature is considered as the major limiting factor for plant growth. Earlier studies reported that night temperature is an important factor in the heat reaction of the plants. Tomato cultivars capable of setting viable fruits under night temperatures above 21 °C are considered as heat-tolerant cultivars. The development of breeding objectives is generally summarized in four points: (a) cultivars with higher yield, (b) disease resistant varieties in the 1970s, (c) long shelf-life in 1980s, and (d) nutritive and taste quality during 1990s. Some unique varieties like the dwarf “Micro-Tom”, and the first transgenic tomato (FlavrSavr) were developed through breeding; they were distributed late in the 1980s.High temperature significantly affects seed, pollen viability and root expansion. Researchers have employed different parameters to evaluate the tolerance to heat stress, including membrane thermo stability, floral characteristics (Stigma exertion and antheridia cone splitting), flower number, and fruit yield per plant. Reports on pollen viability and fruit set/plant under heat stress by comparing the pollen growth and tube development in heat-treated and non-heat-stressed conditions are available in literature. The electrical conductivity (EC) have been used to evaluate the tolerance of some tomato cultivars in vitro under heat stress conditions as an indication of cell damage due to electrolyte leakage; they classified the cultivars into three groups: (a) heat tolerant, (b) moderately heat tolerant, and (c) heat sensitive.It is important to determine the range in genetic diversity for heat tolerance in tomatoes. Heat stress experiments under field conditions offer breeders information to identify the potentially heat tolerant germplasm.     

An Examination of the Function of Male Flowers in an Andromonoecious Shrub Capparis spinosa

The pollen donor and pollinator attractor hypotheses are explanations for the functions of the male flowers of andromonoecious plants. We tested these two hypotheses in the andromonoecious shrub Capparis spinosa L. (Capparaceae) and confirmed that pollen production and cumulative volume and sugar concentration of nectar do not differ between male and perfect flowers. However, male flowers produced larger anthers, larger pollen grains and smaller ovaries than perfect flowers. Observations on pollinators indicated that two major pollinators (Xylocopa valga Gerst and Proxylocopa sinensis Wu) did not discriminate between flower morphs and that they transferred pollen grains a similar distance. However, there were more seeds per fruit following hand pollination with pollen from male flowers than from perfect flowers. Individuals of C. spinosa with a larger floral display (i.e. bearing more flowers) received more pollen grains on the stigma of perfect flowers. Female reproductive success probably is not limited by pollen. These results indicate that male flowers of C. spinosa save resources for female function and that they primarily serve to attract pollinators as pollen donors.     

Abnormal development of floral meristem triggers defective morphogenesis of generative system in transgenic tomatoes

Parthenocarpy and fruit malformations are common among independent transgenic tomato lines, expressing genes encoding different pathogenesis-related (PR) protein and antimicrobal peptides. Abnormal phenotype developed independently of the expression and type of target genes, but distinctive features during flower and fruit development were detected in each transgenic line. We analyzed the morphology, anatomy, and cytoembryology of abnormal flowers and fruits from these transgenic tomato lines and compared them with flowers and fruits of wild tomatoes, line YaLF used for transformation, and transgenic plants with normal phenotype. We confirmed that the main cause of abnormal flower and fruit development was the alterations of determinate growth of generative meristem. These alterations triggered different types of anomalous growth, affecting the number of growing ectopic shoots and formation of new flowers. Investigation of the ovule ontogenesis did not show anomalies in embryo sac development, but fertilization did not occur and embryo sac degenerated. Nevertheless, the ovule continued to differentiate due to proliferation of endothelium cells. The latter substituted embryo sac and formed pseudoembryonic tissue. This process imitated embryogenesis and stimulated ovary growth, leading to the development of parthenocarpic fruit. We demonstrated that failed fertilization occurred due to defective male gametophyte formation, which was manifested in blocked division of the nucleus in the microspore and arrest of vegetative and generative cell formation. Maturing pollen grains were overgrown microspores, not competent for fertilization but capable to induce proliferation of endothelium and development of parthenocarpic ovary. Thus, our study provided new data on the structural transformations of reproductive organs during development of parthenocarpic fruits in transgenic tomato.     

Elucidating the unusual floral features of Swartzia dipetala (Fabaceae)

In this study, we evaluated the floral ontogeny of Swartzia dipetala, which has peculiar floral features compared with other legumes, such as an entire calyx in the floral bud, a corolla with one or two petals, a dimorphic and polyandrous androecium and a bicarpellate gynoecium. We provide new information on the function of pollen in both stamen morphs and whether both carpels of a flower are able to form fruit. Floral buds, flowers and fruits were processed for observation under light, scanning and transmission electron microscopy and for quantitative analyses. The entire calyx results from the initiation, elongation and fusion of three sepal primordia. A unique petal primordium (or rarely two) is produced on the adaxial side of a ring meristem, which is formed after the initiation of the calyx. The polyandrous and dimorphic androecium also originates from the activity of the ring meristem. It produces three larger stamen primordia on the abaxial side and numerous smaller stamen primordia on the adaxial side. These two types of stamens bear morphologically similar ripening pollen grains. However, prior to the dehiscence of thecae and presentation of pollen in the anther, only the pollen grains of the larger stamens contain amyloplasts. Two carpel primordia are initiated as distinct protuberances, alternating with the larger stamens, in a slightly inner position in the floral meristem, constituting the bicarpellate gynoecium. Both carpels are able to form fruit, although only one fruit is generally produced in a flower. The increase in gynoecium merism probably results in an increase in the surface deposition of pollen grains and consequently in the chance of pollination. This is the first study to thoroughly investigate organogenesis and the ability of the carpel to form fruit in a bicarpellate flower from a member of Fabaceae, in addition to the pollen ultrastructure in the heteromorphic stamens associated with the ‘division of labour’ sensu Darwin. © 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 173 , 303–320.     

Pollen load size, reproductive success, and progeny kinship of naturally pollinated flowers of the tropical dry forest tree Pachira quinata (Bombacaceae)

Several studies have demonstrated, using controlled pollinations, that the number and identity of pollen grains deposited onto a flower's stigma affect the reproductive success of plants. However, few studies have shown this relationship under conditions of natural pollination. Using the tropical dry forest tree Pachira quinata, we evaluated the relationship between the number of microgametophytes per pistil and the number of sires with respect to the production of fruits and seeds in a natural population of Pachira quinata. Our study demonstrates that fruit and seed production are directly related to the number of microgametophytes per pistil in natural populations of P. quinata. Only 6% of the marked flowers developed into mature fruits and 10% of the marked flowers initiated fruits but later aborted them. A mean of 23 pollen grains were required to produce a seed. Flowers with >400 pollen grains on the stigma always developed into mature fruits, whereas flowers that received <200 grains never matured fruits. Half of the pollen grains transferred to a flower stigma germinated and developed pollen tubes to the base of the style. The number of pollen grains on a stigma explained 34% of the variation in seed number per fruit, and the number of seeds produced per fruit is positively correlated with the size of the seeds. The population of P. quinata studied is predominantly outcrossing, and seeds within fruits are sired by one or a few donors. The total seed crop within trees was sired by three to five donors. Our study examined the implications of the variation in size of microgametophyte loads per pistil with respect to the breeding system and the paternity of progeny under natural conditions. The competitive ability of pollen and pollen tube attrition are important factors regulating fruit production in P. quinata.     

Multifunctional bracts enhance plant fitness during flowering and seed development in Rheum nobile (Polygonaceae), a giant herb endemic to the high Himalayas

Specialized bracts are thought to be important for the successful reproduction of some plants and are regarded as adaptations to diverse driving forces. However, few empirical studies have quantified the adaptive significance of bracts within a cost–benefit framework. We explored the adaptive significance of large and showy bracts for reproduction in Rheum nobile, a giant herb endemic to the high Himalayas. We examined whether the bracts enhance reproductive success during flowering and seed development. Bracts increased flower and fruit temperature on sunny days, greatly decreased the intensity of ultraviolet-B (UV-B) radiation reaching flowers and fruits, and prevented pollen grains being washed away by rain. Experiments indicated that high temperature could promote pollen germination, while pollen grains exposed to rain and UV-B radiation at ambient levels were seriously damaged. Furthermore, bract removal decreased the number of pollinators visiting flowers. When bracts were removed before or after flowering, fecundity and progeny quality were adversely affected, but seed predation by larvae of pollinators decreased. A cost–benefit analysis demonstrated that the cost of bracts, i.e., increased seed predation, is modest. Our results suggest that the bracts of R. nobile promote pollen germination, protect pollen grains from rain and intense UV-B radiation, enhance pollinator visitation during flowering, and facilitate the development of fertilized ovules during seed development. We conclude that multifunctional bracts of R. nobile are an effective adaptive strategy in alpine environments and might have been selected for because of abiotic environmental conditions as well as for enhancing pollination success.