Effect of source–sink ratio on seed set and filling in sunflower (Helianthus annuus L.)

Poor seed development in sunflower may result from insufficient assimilate supply (source limitation). To test this hypothesis, the effects of changed source–sink ratio on seed set (measured as percentage of empty achenes) and seed filling (measured as dry mass per filled achene) in individual plants were investigated. Source–sink ratio, defined as leaf area per floret (LAF), was experimentally altered using invasive (floret removal, defoliation) and non‐invasive (pulse of chilling, short days or shading during leaf or floret initiation) treatments. Shading at floret initiation proved the most effective non‐invasive method. Generally, an increase, or decrease, in LAF improved, or impaired, both seed set and filling. Increasing LAF by 2.0 cm²[95% confidence interval (1.5, 2.5)] decreased the percentage of empty achenes by 36.9%‐points (?41.9, ?30.9) and increased dry mass per filled achene by 20.1 mg (13.6, 26.7) in the capitulum centre. The effect of source–sink ratio on seed set was always strongest in the centre, whereas peripheral whorls were not affected. Achene mass was affected in all parts of the capitulum. It is concluded that source limitation is a major cause for empty achenes in sunflower plants grown under non‐stress conditions.     

Polyamines in normal and auxin-induced strawberry fruit development

The possible involvement of polyamines during strawberry ( Fragaria x ananassa Duch.) fruit development was investigated. Putrescine, spermidine, and spermine were identified in strawberry receptacles and achenes at all stages of development. Total (free) polyamine levels decreased from a maximum of 485 nmol g⁻¹ fresh weight at pollination to a minimum of 55 nmol g⁻¹ fresh weight in ripe receptacles. Total polyamine concentrations during corresponding stages of development were consistently higher in achenes than in receptacles, and ranged from 891 to 203 nmol g⁻¹ fresh weight. Removal of achenes from the surface of developing receptacles 10 days after pollination reduced receptacle growth, and re-initiation of growth by application of 1 m M α-naphtaleneacetic acid (α-NAA) was accompanied by a rapid increase in polyamine concentrations 24 h after treatment. Polyamine content per receptacle increased >3-fold in normally developing receptacles and in de-achened, auxin-treated receptacles 10 days after removal of achenes, but did not increase during this period in de-achened receptacles not treated with exogenous auxin. α-NAA increased growth and polyamine levels to a greater extent than the structurally related, but less effective auxin, β-NAA. Polyamine concentrations in receptacles with intact achenes remained similar to those of auxin depleted (de-achened) receptacles, implying that the concentration of these compounds may not be limiting following achene removal.     

Polyamines in normal and auxin-induced strawberry fruit development

The possible involvement of polyamines during strawberry ( Fragaria × ananassa Duch.) fruit development was investigated. Putrescine, spermidine, and spermine were identified in strawberry receptacles and achenes at all stages of development. Total (free) polyamine levels decreased from a maximum of 485 nmol g⁻¹ fresh weight at pollination to a minimum of 55 nmol g⁻¹ fresh weight in ripe receptacles. Total polyamine concentrations during corresponding stages of development were consistently higher in achenes than in receptacles, and ranged from 891 to 203 nmol g⁻¹ fresh weight. Removal of achenes from the surface of developing receptacles 10 days after pollination reduced receptacle growth, and re-initiation of growth by application of 1 m M α-naphtaleneacetic acid (α-NAA) was accompanied by a rapid increase in polyamine concentrations 24 h after treatment. Polyamine content per receptacle increased >3-fold in normally developing receptacles and in de-achened, auxin-treated receptacles 10 days after removal of achenes, but did not increase during this period in de-achened receptacles not treated with exogenous auxin. α-NAA increased growth and polyamine levels to a greater extent than the structurally related, but less effective auxin, β-NAA. Polyamine concentrations in receptacles with intact achenes remained similar to those of auxin depleted (de-achened) receptacles, implying that the concentration of these compounds may not be limiting following achene removal.     

Morphological variability of Tragopogon pterodes Pančić ex Petrović achenes: taxonomic evaluation of heterocarpy

This study presents the carpological heterogeneity of Tragopogon pterodes, an endemic plant species from Balkan Peninsula. The goal of this paper was to determine the variability of morphological characters of achenes and assess the degree of their morphological differentiation relative to the position they occupy in the receptacle. Analysis included four different populations of T. pterodes and examination of 18 (8 qualitative and 10 quantitative) characters of achenes in total. Data analysis employed basic statistic, univariate (ANOVA, Tukey HSD) and multivariate (CDA, CA) analyses to check the level of heterocarpy in this species. The results indicate possible morphological differentiation of three groups of achenes (outer, median and inner achenes), depending of their position on the receptacle. The members of all three types were described, illustrated and compared. The variability of investigated carpological characters in this species is providing important information prior to their further taxonomic treatment.     

Morphometric Analysis of Sunflower (<Emphasis Type="Italic">Helianthus annuus</Emphasis> L.) Achenes from Mexico and Eastern North America<Superscript>1</Superscript>

Morphometric Analysis of Sunflower ( Helianthus annuus L.) Achenes from Mexico and Eastern North America. Sunflower (Helianthus annuus L.) has played a major role in the evolution of agricultural systems in the Americas. The discovery of ancient domesticated remains from archaeological deposits in pre-Columbian Mexico offers new dimensions to widely accepted viewpoints on the domestication pattern of H. annuus. Although American sunflower populations north of Mexico have been examined extensively, Mexican indigenous domesticated landraces have not been studied in any detail. In this study, we morphologically assessed wild and domesticated sunflower achenes from Mexico and compared them to similar datasets from eastern North America. Additionally, we evaluated the utility of four computer-assisted shape measurements in discriminating between wild and domesticated sunflower achenes (fruits) and compared variation in achene size among modern wild and cultivated populations from both Mexico and the U.S. We found that, of the shape parameters tested, none were informative in distinguishing wild achenes from domesticated varieties. Subsequent size analysis, using conventional parameters of length, width, and thickness, showed that modern wild populations from Mexico had smaller achenes compared to modern populations from eastern North America. Domesticated achenes unearthed from Mexican archaeological sites, however, were significantly larger than the early domesticated specimens recovered from eastern North America. Our methodological results indicate that variation in archaeological sunflower achenes is better described by conventional size parameters rather than computerized shape analysis.     

Accumulation of a glycine rich protein in auxin-deprived strawberry fruits

Growth of strawberry (Fragaria ananassa Duch. cv. Ozark Beauty) receptacles is regulated by auxin supplied from the achenes. The receptacle growth can be stopped at any stage by deachening the fruits, and can be resumed by exogenous application of auxin. In our earlier studies we demonstrated auxin regulated polypeptide changes at different stages of strawberry fruit development. Removal of achenes from fruits and growing the receptacles without auxin resulted in the time-dependent accumulation of 52,000 Mr polypeptide. Amino acid analysis revealed that the protein is rich in glycine. Our studies, with normal and variant strawberry receptacles, indicate that the synthesis and accumulation of this glycine-rich protein correlates with cessation of receptacle growth. These results suggest a possible role for the glycine-rich protein in cessation of growth.     

Multi-substrate flavonol O-glucosyltransferases from strawberry (Fragaria x ananassa) achene and receptacle

In an effort to characterize fruit ripening-related genes functionally, two glucosyltransferases, FaGT6 and FaGT7, were cloned from a strawberry (Fragaria x ananassa) cDNA library and the full-length open reading frames were amplified by rapid amplification of cDNA ends. FaGT6 and FaGT7 were expressed heterologously as fusion proteins in Escherichia coli and target protein was purified using affinity chromatography. Both recombinant enzymes exhibited a broad substrate tolerance in vitro, accepting numerous flavonoids, hydroxycoumarins, and naphthols. FaGT6 formed 3-O-glucosides and minor amounts of 7-O-, 4'-O-, and 3'-O-monoglucosides and one diglucoside from flavonols such as quercetin. FaGT7 converted quercetin to the 3-O-glucoside and 4'-O-glucoside and minor levels of the 7- and 3'-isomers but formed no diglucoside. Gene expression studies showed that both genes are strongly expressed in achenes of small-sized green fruits, while the expression levels were generally lower in the receptacle. Significant levels of quercetin 3-O-, 7-O-, and 4'-O-glucosides, kaempferol 3-O- and 7-O-glucosides, as well as isorhamnetin 7-O-glucoside, were identified in achenes and the receptacle. In the receptacle, the expression of both genes is negatively controlled by auxin which correlates with the ripening-related gene expression in this tissue. Salicylic acid, a known signal molecule in plant defence, induces the expression of both genes. Thus, it appears that FaGT6 and FaGT7 are involved in the glucosylation of flavonols and may also participate in xenobiotic metabolism. The latter function is supported by the proven ability of strawberries to glucosylate selected unnatural substrates injected in ripe fruits. This report presents the first biochemical characterization of enzymes mainly expressed in strawberry achenes and provides the foundation of flavonoid metabolism in the seeds.     

Effects of pollen donors on seed formation inEspeletia schultzii (Compositae) populations at different altitudes

The influence of different pollen donors on seed formation was investigated in three populations ofEspeletia schultzii that differ in environmental conditions and life history characteristics. Self pollen and pollen from different donors (< 15m apart) within each population was used in a diallel design in order to test the genetic base of seed set variation. Three measures of seed formation were used: (1) achene number; (2) proportion of filled achenes (fruits) that distinguishes between achenes with seeds and empty achenes; (3) proportion of aborted seeds that distinguishes between viable and aborted seeds. Self-pollinations resulted in empty achenes. Achene number did not vary between the different pollen donors. A bimodal pattern of filled achenes was found in two populations in two consecutive years. On the other hand, a unimodal pattern was found in crosses between more distant donors (> 30m). These patterns seems to be the results of a sporophytic incompatibility system. Seed abortion was highest at the higher elevations and seems to be correlated with elevation rather than with any genetic effect.     

Pericarp strength of sunflower and its value for plant defense against the sunflower moth,Homoeosoma electellum

Sunflower pericarps provide a barrier against seed feeding by larvae of the sunflower moth, Homoeosoma electellum. Pericarp hardening is thought to be accelerated by a phytomelanin layer beneath the hypodermis, but among germplasm with phytomelanin, broad variation in sunflower pericarp strength exists. To facilitate the use of pericarp strength in sunflower breeding, resistance to mechanical puncture was assessed for diverse sunflower germplasm, and feeding tests were used to evaluate whether differences in pericarp strength prevent H. electellum larvae from penetrating achenes. Test on field-grown sunflowers indicates that public restorer lines have lower pericarp strengths compared with maintainer lines and commercial hybrids at 14 days after the start of anthesis. Interspecific crosses or plant introduction (PI) accessions believed to be resistant to H. electellum, including PI 170415, comprised a group with exceptionally high pericarp strength relative to other germplasm. In subsequent tests on greenhouse-grown sunflowers, overall results were similar, but using field-grown plants provided greater statistical power. In choice tests with achenes that differed in pericarp strength, 7- to 9-day-old sunflower moth larvae fed more often on seed protected by a weaker pericarp, at a more than 5-to-1 ratio, while 10-day-old larvae fed indiscriminately. Pericarp strength data contradict previous published results for individual entries and heterotic groups, but support the generalization that improved physical resistance to the sunflower moth is possible. To use pericarp strength in PI 170415 or similar sources, the inheritance of high pericarp strength and potential trade-offs between pericarp strength and other agronomic traits need to be understood.     

Effects of foliar applied gibberellic acid and benzyladenine upon yield components in sunflower (Helianthus annuus L.)

Sunflower yield is determined by seed number/m^–2 and by achene weight. Frequently, a high percentage of empty achenes in the inner portion of the capitulum, probably due to a reduced vascularization of that section of the flower head, decreases final yield. The objective of the present research is to determine if foliarly applied gibberellic acid (GA) and benzyladenine (BA) can enhance the vascularization in the inner portion of the capitulum, improving photoassimilate translocation. Field experiments were conducted during 1989/90 with hybrid SPS 894 and during 1990/91 with hybrid ACA 882. GA (150 mg/l^–1), BA (150 and 250 mg/l^–1) and GA 150+BA 150 mg/l^–1 each were foliarly applied 20, 40, or 60 days after emergence. For both seasons and hybrids plant growth regulator (PGRs) applications significantly reduced the percentage of empty achenes, increased achene weight, achene weight (× 1000) and achene number in the inner portion of the capitulum and in the middle and outer portion during 1990/91. A 25% increase in seed yield was achieved due to PGR application and the capitulum partition index (achene weight/receptacle weight^–1, CPI) was significantly increased due to an improvement in photoassimilate distribution. A distribution model was derived showing that preferential allocation of photoassimilates in the outer portion of the capitulum can be modified by PGR application, demonstrating that photoassimilate distribution is under hormonal control.Consejo Nacional de Investigaciones Científicas y Tecnológicas (CONICET)     

Cloning and molecular characterization of a strawberry fruit ripening-related cDNA corresponding a mRNA for a low-molecular-weight heat-shock protein

We have isolated and characterized a cDNA from a strawberry fruit subtractive library that shows homology to class-I low-molecular-weight (LMW) heat-shock protein genes from other higher plants. The strawberry cDNA (clone njjs4) was a 779 bp full-length cDNA with a single open reading frame of 468 bp that is expected to encode a protein of ca. 17.4 kDa with a pI of 6.57. Southern analysis with genomic DNA showed several high-molecular-weight hybridization bands, indicating that the corresponding njjs4 gene is not present as a single copy in the genome. This strawberry gene was not expressed in roots, leaves, flowers and stolons but in fruits at specific stages of elongation and ripening. However, a differential pattern of mRNA expression was detected in the fruit tissues achenes and receptacle. The njjs4 gene expression increased in achenes accompanying the process of seed maturation whereas in the receptacle, a high mRNA expression was detected in the W2 stage, during which most of the metabolic changes leading to the fruit ripening are occurring. Our results clearly show a specific relationship of this njjs4 strawberry gene with the processes of seed maturation and fruit ripening, and strongly support that at least some of the class-I LMW heat-shock protein-like genes have a heat-stress-independent role in plant development, including fruit ripening.     

The domestication of Helianthus annuus L. (sunflower)

All modern domesticated sunflowers can be traced to a single center of domestication in the interior mid-latitudes of eastern North America. The sunflower achenes and kernels recovered from six eastern North American sites predating 3000 b.p. that document the early history of this important crop plant are reanalyzed, and two major difficulties in the interpretation of archaeological sunflower specimens are addressed. First, achenes and kernels obtained from a modern wild sunflower population included in a prior genetic study because of its minimal likelihood for crop-wild gene flow, and its close genetic relationship to domesticated sunflowers, provide a new and more tightly drawn basis of comparison for distinguishing between wild and domesticated achene and kernel specimens recovered from archaeological contexts. Second, achenes and kernels from this modern wild baseline population were carbonized, allowing a direct comparison between carbonized archaeological specimens and a carbonized modern wild reference class, thereby avoiding the need for the various problematic shrinkage correction conversion formulas that have been employed over the past half century. The need for further research on museum collections is underscored, and new research directions are identified.     

REGENERATION OF THE SUNFLOWER CAPITULUM AFTER CYLINDRICAL WOUNDING OF THE RECEPTACLE

Using the young capitulum of Helianthus annuus L., a cylindrical plug of undifferentiated receptacle tissue, 1 mm in diameter, was isolated from lateral communication with the rest of the receptacle surface by a vertical circular wound cut, while retaining continuity with the subapical meristem. Within 24 hr, active cell division was induced at the inner and outer surfaces of the wound and in the receptacle epidermis bordering the wound edges, creating a rounded rim at the top of the wound. Within 3–6 days, floral initials, spaced 133–166 μm apart appeared on the flanks of both rims and later on the top of the plug and surrounding receptacle surface. The first formed initials developed into involucral bracts or ray florets and the later ones into disc florets which were organized into contact parastichies, the number of which did not conform with the Fibonacci series. The base of the plug developed into a stem-like structure completing the regeneration of a fully formed functional capitulum. This operation was demonstrated for two sunflower cultivars and occurred in both long and short daylengths.     

Effects of nitrogen supply on growth, yield and yield components of safflower and sunflower

Safflower (Carthamus tinctorius L.) represents an important oil crop internationally and may have a certain production potential under low input conditions, particularly in organic farming systems, where the putatively low nutrient requirement would be an advantage. However, little is known about the nutrient requirements of safflower. This study was undertaken to determine the growth and yield response of safflower as compared to sunflower (Helianthus annuus L.) under different N supplies. Safflower and sunflower plants were grown in Mitscherlich pots containing equal volumes of sand, nutrient-poor limed soil, and perlite. Nitrogen supply was the same for both species (0.25, 0.5, 1.0, 1.5, and 2.0 g per pot) in the first year, but was raised for sunflower (0.5, 1.0, 2.0, 3.0, and 4.0 g per pot) in the second. Increased N supply enhanced plant growth and yield for both species. Growth and yield of safflower increased up to 1.0 g pot⁻¹, while the optimum for sunflower was 2.0 g pot⁻¹. Safflower out-yielded sunflower at low N supply, while at high N level the opposite occurred. Functional analysis according to Michaelis–Menten revealed that – in terms of yield formation – safflower is superior to sunflower under N-limited conditions. Safflower is generally more efficient than sunflower in concentrating N in their shoots. N concentration in the photosynthetically active youngest mature blade of both species exhibited the same functional relationship with the N supply, but safflower required a much lower leaf N concentration to produce optimal yield as compared to sunflower, indicating the higher efficiency of the former in terms of NUE. Yield components analysis revealed that in safflower, yield is tightly correlated with the number of capitula per plant and the mass per achene, both being strongly correlated, too. On the contrary, sunflower yield was merely determined by the number of achenes per capitulum, followed by the mass per achene. Path coefficient analysis showed that in safflower, the direct effects of the achene and leaf N content as well as the leaf dry matter on oil yield are small, and mediated principally via indirect effects on the number of achenes per capitulum, while for sunflower the number of achenes per capitulum exerts a strong direct effect. Dedicated to the late Prof. Dr. Burkhard Sattelmacher, who passed away on November 21, 2005.     

Escape of Lygus hesperus (Heteroptera: Miridae) Eggs from Parasitism by Anaphes iole (Hymenoptera: Mymaridae) in Strawberries: Plant Structure Effects

Herbivorous insects in natural and agricultural systems experience variation in parasitoid attack on different plant species due to direct and indirect plant influences on parasitoids. Lygus hesperus is a native polyphagous mirid that suffers up to 100% parasitism by the native egg parasitoid Anaphes iole in certain weed hosts, but with inundative releases in commercial strawberries, we achieve <65% L. hesperus suppression. We examined L. hesperus egg distribution in individual strawberry plants and parasitism by A. iole of eggs in different strawberry plant structures to determine whether plant-related factors affected parasitoid performance in strawberries. L. hesperus laid more eggs (46.5% of all eggs laid) in the fruit (between the achenes [seeds] in the fleshy receptacle) than in the petiole (23.3%), leaflet (20.3%), peduncle (6.2%), or calyx (3.7%). In a no-choice test, parasitism by A. iole was higher in the petiole (96.7%), calyx (91.9%), and leaflet (85.2%) than in the fruit (51.8%), in which the achenes appeared to hinder parasitoid access to host eggs. In addition, in young fruits in which the interachene distance was minimum, parasitism was considerably lower (25.4%) than in fruits in which receptacle swelling had resulted in interachene distances that were medium (65.7% parasitism) or large (77.1% parasitism). Our results suggest that strawberry fruits can provide refugia from parasitism by A. iole and that maximum protection occurs when the achenes are contiguous. The presence of refugia in strawberries limits the impact of augmentative biological control with A. iole, highlighting the need for its integration with other strategies to effectively suppress L. hesperus in strawberries.     

Reconfiguration of the achene and receptacle metabolic networks during strawberry fruit development

The anatomy of strawberry (Fragaria x ananassa) fruit, in which the achene is found on the outer part of the fruit, makes it an excellent species for studying the regulation of fruit development. It can provide a model for the cross talk between primary and secondary metabolism, whose role is of pivotal importance in the process. By combining gas chromatography-mass spectrometry and liquid chromatography-mass spectrometry with the aim of addressing the metabolic regulation underlying fruit seed development, we simultaneously analyzed the composition of primary and secondary metabolites, separately, in achene and receptacle during fruit ripening of strawberry cultivar Herut. The results from these analyses suggest that changes in primary and secondary metabolism reflect organ and developmental specificities. For instance, the receptacle was characterized by increases in sugars and their direct derivatives, while the achene was characterized by a major decrease in the levels of carbon- and nitrogen-rich compounds, with the exception of storage-related metabolites (e.g. raffinose). Furthermore, the receptacle, and to a lesser extent the achene, exhibited dynamic fluctuations in the levels and nature of secondary metabolites across the ripening process. In the receptacle, proanthocyanidins and flavonol derivatives characterized mainly early developmental stages, while anthocyanins were abundant in the mature red stage; in the achene, ellagitannin and flavonoids were abundant during early and late development, respectively. Correlation-based network analysis suggested that metabolism is substantially coordinated during early development in either organ. Nonetheless, a higher degree of connectivity within and between metabolic pathways was measured in the achenes. The data are discussed within the context of current models both of the interaction of primary and secondary metabolism and of the metabolic interaction between the different plant organs.     

A growth-controlled model of the shape of a sunflower head

A mathematical model is presented which predicts the shape of a sunflower receptacle (or the compact receptacle of various other taxa) and the pattern of its floral parts (florets) from the time of their initiation to maturity. The model assumes that the expansion and curving of the receptacle surface is just sufficient to accommodate the development of the florets, thus minimizing the quantity of plant tissue involved. The model assumes a fixed angular separation (divergence) between successive florets, an S-shaped (sigmoidal) growth function followed by each florets, and a fixed time delay (period) between the initiation of successive florets. It is further assumes that the shape and relative position occupied by the florets on the receptacle surface are invariant in time. By this theory, the shape of the receptacle surface is fully determined once the mathematical form of the growth function is specified. Using the logistic growth function, the theory is tested against the measured shapes of plant receptacles from different taxa at various points in their development. The least-squares adjusted fits to the theory are, in most cases, very good indeed.     

Wound-Induced Initiation of Involucral Bracts and Florets in the Developing Sunflower Inflorescence

The production of additional floral organs by the inflorescenceof Helianthus annuus as a boron deficiency symptom was examinedand found to be related to the occurrence of minute splits inthe receptacle of the young capitulum. Wounding the receptacleby puncturing or cutting at an early stage when the receptacledome was forming (floral stage 3) or later when the receptaclehad either become saucer-shaped (floral stage 4) or the firstrows of disc floret primordia were appearing on the rim of thereceptacle (floral stage 5) resulted in the initiation of involucralbracts, ray and disc florets in the wound area, reproducingthe symptoms of boron deficiency. When the receptacle was woundedat later floral stages, when the receptacle was partially orwholly covered by disc floret primordia, involucral bracts andray florets were not formed in the proximity of the wound, leadingto the conclusion that the commitment of floral organ primordiabegins at the time of their initiation. The inductive effectsof wounding could not be reproduced by indoleacetic acid, naphthaleneaceticacid or benzyladenine applied to the receptacle surface in lanolinepaste. The results are related to the normal development ofthe sunflower inflorescence and it is concluded that the firstfloral organ primordia that appear in floral stage 3 and developinto involucral bracts during floral stage 4 may determine thepositions of subsequent primordia and establish the radial symmetryof the inflorescence. ¹ Supported by a grant from the Australian Research Grants Scheme. (Received August 10, 1982; Accepted October 12, 1982)     

COMPARATIVE MORPHOLOGY OF THE CAPITULUM OF ENCELIOPSIS (ASTERACEAE: HELIANTHEAE)

Features of the capitula of all four species of Enceliopsis were examined with light and scanning electron microscopy, measured, and subjected to univariate analysis. Differences were found in the complement, distribution, and density of trichomes, the size of the receptacle, ray ligule, and disk corolla tube, and the size and shape of the phyllaries, paleae, disk corolla throats, and achenes. Morphology of trichomes, phyllaries, and achenes, as well as other vegetative characteristics, show that E. nudicaulis, E. covillei and E. argophylla form a group separate from E. nutans. Features of the achene suggest affinity of the former group with Geraea, whereas features of the trichomes and achene of E. nutans support affinity with Encelia. Characters of the receptacle, paleae, ray and disk corollas, and achene clearly show that E. nudicaulis, E. covillei, and E. argophylla are distinct.