Seed yield and plant biomass increases in rice are conferred by deregulation of endosperm ADP-glucose pyrophosphorylase

In this work we test the hypothesis that yield of rice ( Oryza sativa L.) can be enhanced by increasing endosperm activity of ADP-glucose pyrophosphorylase (AGP), a key enzyme in starch biosynthesis. The potential for increases in yield exist because rice initiates more seeds than are taken to maturity and possesses excess photosynthetic capacity that could be utilized if there were more demand for assimilate. Following an approach already shown to be successful in wheat, experiments were designed to increase demand for assimilate by increasing the capacity for starch synthesis in endosperm. This was accomplished by transforming rice with a modified maize AGP large subunit sequence ( Sh2r6hs) under control of an endosperm-specific promoter. This altered subunit confers upon AGP decreased sensitivity to allosteric inhibition by inorganic phosphate (Pi) and enhanced heat stability, potentially leading to higher AGP activity in vivo. The Sh2r6hs transgene increased AGP activity in developing endosperm by 2.7-fold in the presence of Pi. Increases in AGP activity in transgenic seeds compared with controls were maximal between 10-15 days after anthesis. Starch content of individual seeds at harvest was not increased, but seed weight per plant and total plant biomass were each increased by more than 20%. Increased endosperm AGP activity thus stimulates setting of additional seeds and overall plant growth rather than increasing yield of seeds already set. Results demonstrate that deregulation of endosperm AGP increases overall plant sink strength, leading to larger, more productive plants in a manner similar to that in wheat having similar genetic modification.     

Over-expression of AGPase genes enhances seed weight and starch content in transgenic maize

Cereal crops accumulate starch in the seed endosperm as an energy reserve. ADP-glucose pyrophosphorylase (AGPase) plays a key role in regulating starch biosynthesis in cereal seeds. The AGPase in the maize endosperm is a heterotetramer of two small subunits, encoded by Brittle2 (Bt2) gene, and two large subunits, encoded by the Shrunken2 (Sh2) gene. The two genes (Bt2, Sh2) from maize were introduced into two elite maize inbred lines, solely and in tandem, and under the control of endosperm-specific promoters for over-expression. PCR, Southern blotting, and real-time RT-PCR analysis indicated that the transgenes were integrated into the genome of transgenic plants and were over-expressed in their progeny. The over-expression of either gene enhanced AGPase activity, seed weight and starch content compared with the WT, but the amounts were lower than plants with over-expression of both Bt2 and Sh2. Developing seeds from co-expression transgenic maize plants had higher cytoplasmic AGPase activity: the 100-grain weight increased 15% over the wild type (WT), and the starch content increased to over 74% compared with the WT of 65%. These results indicate that over-expression of the genes in transgenic maize plants could improve kernel traits. This report provides a feasible approach for increasing starch content and seed weight in maize.     

Increasing maize seed weight by enhancing the cytoplasmic ADP-glucose pyrophosphorylase activity in transgenic maize plants

ADP-glucose pyrophosphorylase (AGPase) plays a key role in regulating starch biosynthesis in cereal seeds and is likely the most important determinant of seed strength. The Escherichia coli mutant glgC gene (glgC16), which encodes a highly active and allosterically insensitive AGPase, was introduced into maize (Zea mays L.) under the control of an endosperm-specific promoter. Developing seeds from transgenic maize plants showed up to 2–4-fold higher levels of AGPase activity in the presence of 5 mM inorganic phosphate (Pi). Transgenic plants with higher cytoplasmic AGPase activity under Pi-inhibitory conditions showed increases (13–25%) in seed weight over the untransformed control. In addition, in all transgenic maize plants, the seeds were fully filled, and the seed number of transgenic plants had no significant difference compared with that of untransformed control. These results indicate that increasing cytoplasmic AGPase activity has a marked effect on sink activity and, in turn, seed weight in transgenic maize plants.     

A shrunken-2 transgene increases maize yield by acting in maternal tissues to increase the frequency of seed development

The maize (Zea mays) shrunken-2 (Sh2) gene encodes the large subunit of the rate-limiting starch biosynthetic enzyme, ADP-glucose pyrophosphorylase. Expression of a transgenic form of the enzyme with enhanced heat stability and reduced phosphate inhibition increased maize yield up to 64%. The extent of the yield increase is dependent on temperatures during the first 4 d post pollination, and yield is increased if average daily high temperatures exceed 33 °C. As found in wheat (Triticum aestivum) and rice (Oryza sativa), this transgene increases maize yield by increasing seed number. This result was surprising, since an entire series of historic observations at the whole-plant, enzyme, gene, and physiological levels pointed to Sh2 playing an important role only in the endosperm. Here, we present several lines of evidence that lead to the conclusion that the Sh2 transgene functions in maternal tissue to increase seed number and, in turn, yield. Furthermore, the transgene does not increase ovary number; rather, it increases the probability that a seed will develop. Surprisingly, the number of fully developed seeds is only ～50% of the number of ovaries in wild-type maize. This suggests that increasing the frequency of seed development is a feasible agricultural target, especially under conditions of elevated temperatures.     

Arabidopsis sucrose synthase 2 and 3 modulate metabolic homeostasis and direct carbon towards starch synthesis in developing seeds

Two genes encoding sucrose synthase (SUS), namely SUS2 (At5g49190) and SUS3 (At4g02280), are strongly and differentially expressed in Arabidopsis seed. Detailed biochemical analysis was carried out in developing seeds 9–21 days after flowering (DAF) of wild type and two knockouts. SUS2 and SUS3 are not redundant genes since single knockouts show a phenotype in developing seeds. The mutants had 30–50% less SUS activity and therefore accumulated 40% more sucrose and 50% less fructose at 15 DAF. This did not affect the hexose-P pool, but led to 30–70% less starch in embryo and seed coat. Lipids were 55% higher in both mutants at 9–15 DAF. It seems that sucrolysis via SUS is not required for oil or protein synthesis but rather for channeling carbon toward ADP-glucose and starch in seeds. Metabolite profiling with GC–TOF revealed specific downstream changes in primary metabolism as a consequence of signaling or regulatory fine-tuning. While sucrose increased, hexoses and specific amino acids decreased reciprocally. There was a developmental shift regarding an earlier timing of dry weight accumulation, germinative maturity, oil deposition, sugar levels, transient starch buildup, and protein storage. Nevertheless, final seed size and composition were unaltered due to an earlier cessation of growth, thus giving rise to an apparent silent phenotype of mature mutant seeds. We conclude that SUS is important for metabolite homeostasis and timing of seed development, and propose that an altered sucrose/hexose ratio can modify carbon partitioning and the pattern of storage compounds in Arabidopsis.     

Can the <Emphasis Type="Italic">Giberella zeae</Emphasis> toxin zearalenone affect the photosynthetic productivity and increase yield formation in spring wheat and soybean plants?

The seeds of soybean cv. Aldana and spring wheat cv. Torka were soaked for 24 h in solution of zearalenone [ZEN, 2,4-dihydroxy-6-(10-hydroxy-6-oxo-trans-1-undecenyl)-benzonic acid lactone, 4 mg dm⁻³] and then they were sown in the pot experiment in an open vegetation hall. The after-effects of ZEN on growth of plants, net photosynthetic (P _N) and transpiration (E) rates, stomatal conductance (g _s), photochemical efficiency of photosystem II (PSII) and on final seeds yield, were determined. A significant increase of seeds yield was revealed in plants of both cultivars i.e. by 22% and 19% of seed (grain) number and by 28 and 24% of seed (grain) mass, in soybean and in wheat, respectively. The photosynthetic rate (P _N) was stimulated during the juvenile and final phase by about 13.6% (average) in soybean plants. During other developmental stages, assimilation of CO2 was retarded. The response of CO₂ assimilation in wheat plants was less pronounced as compared to that in soybean, but an increase of P _N by over 24% near the final stage of development was observed. The quantum yield of PSII electron transport (Φ_PSII) in soybean plants was changed after the treatment of seeds by ZEN similarly as for the rate of CO₂, whereas in wheat it continued to gradually increase i.e. during the whole growth period. Changes of Φ_PSII both in soybean and in wheat plants, as the response to ZEN treatment, were accompanied with an increase in the efficiency of changes occurring within the antenna (F_v′/F_m′) as well as within centres of photochemical reactions (q_p). The conclusion is that ZEN can affect plant growth and development in many ways, as well as in the status and functioning of the photosynthetical apparatus. Some of the effects can be very longlasting, as e.g. stimulation of production of seed yield in response to treatment of seeds with this substance.     

COMPARISON OF INTRASPECIFIC VARIATIONS IN THE REPRODUCTION AND PHOTOSYNTHESIS OF AN UNDERSTORY HERB,ARNICA CORDIFOLIA

Intraspecific variations in the reproduction of individual ramets and monospecific patches of the understory herb Arnica cordifolia Hook. were compared with variations in photosynthesis and understory light conditions. Ramets and patches were compared from three microhabitats (open, intermediate, and shaded) that differed in daily integrated irradiance. Individual ramets from open microhabitats (> 12 MJ m⁻² d⁻¹) had 23% more total dry wt and produced twice as many seeds, when compared to ramets from shaded locations (< 5 MJ m⁻² d⁻¹). In addition, monospecific patches from open locations were 63% more dense, and estimates of seed and vegetative patch reproductive effort were 4 and 2 times greater, respectively, when compared to shaded plant patches. For all measurements, ramets and patches from intermediate understory locations (6–10 MJ m⁻² d⁻¹) were intermediate in reproductive capacity between those of open and shaded locations. In addition, A. cordifolia seeds from open microhabitats germinated significantly better (45%) than either intermediate or shaded location seeds under high light and only seeds from shaded microhabitats germinated (14%) in the dark. Compared to shaded location plants, the greater total dry weight and seed production of individual ramets and the greater estimated reproductive effort of patches from open locations corresponded to a greater maximum photosynthetic rate (16.9 μmoles m⁻² s⁻¹) and daily carbon gain (12.2 g m⁻² d⁻¹). Possibly, a greater photosynthetic capacity may make more photosynthetic resources available for reproduction by A. cordifolia plants in open locations. Thus, intraspecific variation in physiology may contribute to intraspecific variation in reproduction.     

Overwintering and growth regulator treatment effects on celery (Apium graveolens L.) flowering and seed production

Celery (Apium graveolens L.) plants cv. Jason overwintered in a polythene tunnel flowered earlier and grew taller than similar plants given a 10-week cold-treatment at 5°C prior to transplanting in the same tunnel in mid-February. However, there was no significant difference in the yield of seeds obtained from both treatments, plants grown at a density of 4m^-2 yielded less seeds than those at 2m^-2, though the yield per unit area was slightly higher from the high density treatment. Treatment with 100 mgl^-1 GA₃ applied twice just prior to flowering and during anthesis increased flower stalk, flower pedicel and stamen length but delayed flower opening and seed ripening and decreased seed set and seed yield. Treatment with a mixture of 1000 mgl^-1 GA₄ and GA₇ plus 1000 mgl^-1 ethephon on three occasions during seed ripening decreased seed yield and reduced seed germination though those seeds capable of germinating were less dormant than seeds from untreated plants. The size distribution of seeds was unaffected by any treatment other than the preseeding spray with GA₃ which reduced the percentage of medium-size seeds.     

Arabidopsis thaliana mutants lacking ADP-glucose pyrophosphorylase accumulate starch and wild-type ADP-glucose content: further evidence for the occurrence of important sources, other than ADP-glucose pyrophosphorylase, of ADP-glucose linked to leaf starch biosynthesis

It is widely considered that ADP-glucose pyrophosphorylase (AGP) is the sole source of ADP-glucose linked to bacterial glycogen and plant starch biosynthesis. Genetic evidence that bacterial glycogen biosynthesis occurs solely by the AGP pathway has been obtained with glgC? AGP mutants. However, recent studies have shown that (i) these mutants can accumulate high levels of ADP-glucose and glycogen, and (ii) there are sources other than GlgC, of ADP-glucose linked to glycogen biosynthesis. In Arabidopsis, evidence showing that starch biosynthesis occurs solely by the AGP pathway has been obtained with the starchless adg1-1 and aps1 AGP mutants. However, mounting evidence has been compiled previewing the occurrence of more than one important ADP-glucose source in plants. In attempting to solve this 20-year-old controversy, in this work we carried out a judicious characterization of both adg1-1 and aps1. Both mutants accumulated wild-type (WT) ADP-glucose and approximately 2% of WT starch, as further confirmed by confocal fluorescence microscopic observation of iodine-stained leaves and of leaves expressing granule-bound starch synthase fused with GFP. Introduction of the sex1 mutation affecting starch breakdown into adg1-1 and aps1 increased the starch content to 8-10% of the WT starch. Furthermore, aps1 leaves exposed to microbial volatiles for 10 h accumulated approximately 60% of the WT starch. aps1 plants expressing the bacterial ADP-glucose hydrolase EcASPP in the plastid accumulated normal ADP-glucose and reduced starch when compared with aps1 plants, whereas aps1 plants expressing EcASPP in the cytosol showed reduced ADP-glucose and starch. Moreover, aps1 plants expressing bacterial AGP in the plastid accumulated WT starch and ADP-glucose. The overall data show that (i) there occur important source(s), other than AGP, of ADP-glucose linked to starch biosynthesis, and (ii) AGP is a major determinant of starch accumulation but not of intracellular ADP-glucose content in Arabidopsis.     

Mapping of the ADP-glucose pyrophosphorylase genes in barley

cDNA probes encoding the barley endosperm ADP-glucose pyrophosphorylase (AGP) small subunit (bepsF2), large subunit (bepl10), and leaf AGP large subunit (blpl) were hybridized with barley genomic DNA blots to determine copy number and polymorphism. Probes showing polymorphism were mapped on a barley RFLP map. Probes that were not polymorphic were assigned to chromosome arms using wheat-barley telosomic addition lines. The data suggested the presence of a single-copy gene corresponding to each of the cDNA probes. In addition to the major bands, several weaker cross-hybridizing bands indicated the presence of other, related sequences. The weaker bands were specific to each probe and were not due to cross-hybridization with the other probes examined here. The endosperm AGP small subunit (bepsF2) majorband locus was associated with chromosome 1P and designated Aga1. The endosperm AGP large subunit (bepl10) major-band locus was mapped to chromosome 5M and designated Aga7. The endosperm AGP large-subunit minor bands were not mapped. The leaf AGP large-subunit major band was associated with chromosome 7M and designated Aga5. One of the leaf AGP large-subunit minor bands was mapped to chromosome 5P and designated Aga6. A clone for the wheat endosperm AGP large-subunit (pAga7) hybridized to the same barley genomic DNA bands as the corresponding barley probe indicating a high degree of identity between the two probes.     

Parsnip (Pastinaca sativa) seed production: effects of seed crop plant density, seed position on the mother plant, harvest date and method, and seed grading on embryo and seed size and seedling performance

Parsnip seeds from four years of seed production (1980–83) were used to examine the effects of position of the seed on the parent plant and several cultural factors on seed and embryo growth and seedling performance. Maximum seed dry weight was reached c. 50 days after flowering (DAF) but embryos continued to grow for a further 7–15 days. The coefficient of variation (C.V.) of seed weight fell from c. 35–40% at 40 DAF, to about 20–25% at maturity. The corresponding figures for C.V. of embryo length were 25–35% at the early harvest and 12–15% at maturity. The effects of seed position, plant density, harvest method, seed grading after harvest on seedling emergence and weight characters were small. Harvesting seeds c. 60 DAF gave, on average, 3.3 days earlier emergence, c. 3.6 days lower ‘spread’ of emergence time and c. 9% higher seedling emergence than harvesting 2 wk earlier. Late harvesting of the seed crop reduced the C.V. of seedling weight from 53 to 46%, on average, but there were variations in response from year to year. Seedling emergence and seedling weight characters were more closely related to embryo length than to seed weight characters.     

Optimum leaf excision increases the biomass accumulation and seed yield of maize plants under different planting patterns

Without developing new agronomic practices, present rates of improvement in seed yields of cereal crops globally are insufficient to fulfil the estimated increasing food demand for 2050 and beyond. Intercropping is one of the agricultural practices that can lead to greater crop yields. However, there exists leaf redundancy for maize in intercropping systems, and the top canopy leaves shade more competent leaves at middle strata of maize plants. Therefore, this work aimed to elucidate the effect of leaf excision treatments in maize to understand the optimum leaf area of maize plants under a maize–soybean relay‐intercropping system (MS_R) and a sole cropping system (SM). The effects of four‐leaf excision treatments (T₁, 0; T₂, 2; T₃, 4; T₄, 6 leaves excised from the top of maize plants until 7 days after silking) on light interception, leaf area index (LAI), photosynthetic characteristics, total biomass accumulation at blistering stage (BS), dough stage (DS) and physiological maturity (PM), and seed yield of maize were investigated through field experiments for 2 years under MS_R and SM. Results showed that, under MS_R and SM, as compared to control (T₁), optimum excision of leaves (T₂) from the top of maize plants significantly improved the light interception (by 25, 18 and 16% at BS, DS and PM, respectively) to lower strata leaves and accelerated the biomass partitioning to maize seeds (by 13 and 12% at DS and PM, respectively). Importantly, plants under T₂ exhibited higher green leaf area than control, that is, excision the top two leaves led to an increase in LAI at PM by 10%, suggesting that leaf senescence under T₂ was delayed which enhanced the photosynthetic rate at PM by 7% in 2017 and 6% in 2018. Relative to T₁, maize under T₂ produced 19 and 13% higher maize yield under MS_R and SM, respectively, and relay‐cropped maize had 90% of SM seed yield. These results suggest that by manipulating the canopy structure of maize plants we can enhance the biomass accumulation and seed yield of maize crops under MS_R and SM.     

Non-lethal freezing effects on seed degreening in Brassica napus

The effects of a non-lethal freezing stress on chlorophyll content, moisture level and distribution, and abscisic acid (ABA) levels were examined in siliques and seeds of Brassica napus (canola). A non-lethal freezing stress resulted in the retention of chlorophyll in seed at harvest that was most pronounced for seeds 28, 32 and 36 days after flowering (DAF). This increase was primarily due to an increased retention of chlorophyll a relative to chlorophyll b. Chlorophyll retention in seeds exposed to a non-lethal freezing stress correlated with an increased ABA catabolism, as measured 1, 3 or 7 days after the stress treatment. Although the non-lethal freezing stress had no significant effect on moisture content in seeds of siliques stressed at 28–44 DAF, moisture distribution, as viewed by magnetic resonance imaging, showed an uneven drying of 32 and 40 DAF siliques after exposure to the non-lethal freezing stress. Moisture was initially lost more rapidly from the silique wall between seeds, than in control non-stressed siliques. Increased moisture loss was not due to structural changes in the vasculature of the silique/seed of stressed tissues. These results are consistent with the hypothesis that a non-lethal freezing stress-induced decrease in ABA level, during seed maturation, effects an inhibition of normal chlorophyll a catabolism resulting in mature but green B. napus seed.     

Fruit Fate, Seed Germination and Growth of an Invasive Vine – an Experimental Test of ‘sit and Wait’ Strategy

Oriental bittersweet (Celastrus orbiculatus Thunb.) is a non-indigenous, invasive woody vine in North America that proliferates in disturbed open sites. Unlike most invasive species, C. orbiculatus exhibits a sit and wait strategy by establishing and persisting indefinitely in undisturbed, closed canopy forest and responding to canopy disturbance with rapid growth, often overtopping trees. We compared fruit fates of C. orbiculatus and native American holly (Ilex opaca). We also explored mechanisms for this sit and wait invasion strategy by testing the effect of C. orbiculatus fruit crop density on removal rates and by examining the influence of seed treatment and light intensity on seed germination and seedling growth. More C. orbiculatus than I. opaca fruits became damaged, and damage occurred earlier. More fruit fell from C. orbiculatus than I. opaca, but removal rates by frugivores did not differ (76.0 ± 4.2% vs 87.5 ± 3.7%, respectively). Density (number of fruits in a patch) of C. orbiculatus did not influence removal rates. Scarification (bird-ingestion) of C. orbiculatus seed delayed germination but seeds germinated in similar proportion to manually defleshed seeds (sown either singly or all seeds from a fruit). Germination of seeds within intact fruits was inhibited and delayed compared to other treatments. Seed treatment did not affect seedling growth. The proportion of seeds germinating and time until germination was similar among five light intensity levels, ranging from full sun to closed-canopy. Seedlings in >70% photosynthetically active radiation (PAR) had more leaves, heavier shoots, and longer, heavier roots than seedlings at lower PAR levels. Results show that most (>75%) C. orbiculatus seeds are dispersed, seedlings can establish in dense shade, and plants grow rapidly when exposed to high light conditions. Control strategies for this highly invasive species should likely focus on minimizing seed dispersal by vertebrates.     

Seed yield,head characteristics and oil content in sunflower varieties as influenced by seeds from single and multiple headed plants under humid tropical conditions

Field trials were conducted during 2004 and 2005 to determine the effect of sowing seeds from plants with multiple heads and seeds from single headed plants of sunflower on seed yield, head characteristics and oil content of three widely grown open pollinated varieties (Funtua, Record and Isaanka) in the humid forest—savanna transition zone which is outside the current growing areas with a view to improving stability and sunflower productivity in this region. Seeds from multiple headed plants produced plants that flowered and matured 2–3 days later than plants from single headed plants. Apart from days to flowering in 2004, number of days to maturity and plant height were affected independently by variety and seed source factors. However, the seeds from single headed plants produced plants that recorded significantly (P < 0.01) higher head weight, head diameter, achene weight and number per head than plants from seeds of plants with multiple heads. Seed source had little effect on sunflower seed yield and oil content. However, Funtua produced significantly (P < 0.05) high seed yield (1956.0 kg ha^?1 ± 76.06) when seeds from plants with multiple heads were sown, while Isaanka recorded comparatively high seed yield from seeds of plants from either multiple (1221.0 kg ha^?1 ± 165.90) or single heads (1388.0 kg ha^?1 ± 135.84) and Record (1201.0 kg ha^?1 ± 96.97) when seeds from single headed plants were sown. Therefore, it is recommended that prospective sunflower growers who wish to cultivate Isaanka, can sow seeds from either the multiple or single head and preferably the multiple head for Funtua, and single head for Record.     

Flavonoid compounds related to seed coat color of wheat

In red wheat, reddish-brown pigments accumulate in testa of mature seeds. Half-cut wheat seeds were immersed in p-dimethylaminocinnamaldehyde (DMACA) reagent that stains flavanol structures blue. Testa of 10–40 days after flowering (DAF) in red wheat (“Norin 61” and “Satonosora”) seeds were stained blue and the reagent color changed to blue with 10–25 DAF seeds. No blue staining was observed in white wheat (“Tamaizumi”) seeds during maturation. “Norin 61” seed coats at 10 DAF contained dihydroquercetin, dihydromyricetin, (+)-catechin, procyanidin B3, and prodelphinidin B3, which were identified by HPLC-diode array detector and LC-MS/MS analyses. These five components began accumulating 7 DAF, reached maxima at 10 or 15 DAF, and then decreased in red wheat seeds, but were not detected in white wheat seeds. These results suggest that flavanol and proanthocyanidins are possible precursors of the reddish-brown pigments of red wheat seeds, and are converted to insoluble compounds as the seeds mature.     

Rice calcium-dependent protein kinase isoforms OsCDPK2 and OsCDPK11 show different responses to light and different expression patterns during seed development

We investigated the spatial and temporal expression patterns of two rice calcium-dependent protein kinases (CDPKs), OsCDPK2 and OSCDPK11, using isoform-specific antisera. Bands of the expected molecular sizes for OsCDPK2 (59 kDa) and OsCDPK11 (61 kDa) were detected on western blots. OsCDPK2 and OsCDPK11 mRNA and protein levels increased in unison during flower development. However, at the onset of seed development, the protein expression profiles diverged significantly. OsCDPK2 protein was expressed at low levels during early seed development, but increased to high levels that were maintained in later stages (20 days after fertilisation, DAF). Conversely, OsCDPK11 protein levels were high at the beginning of seed development, but fell rapidly from 10 DAF onwards. This decrease in the level of OsCDPK11 protein was associated with the abundant synthesis of a truncated mRNA species. OsCDPK2 expression was also closely associated with light perception. OsCDPK2 protein was barely detectable in green leaves exposed to light, but levels increased sharply when plants were shifted to darkness. Initially, this increase reflected a rapid elevation in the levels of OsCDPK2 mRNA, which was normally located in the mesophyll. Conversely, OsCDPK11 mRNA and protein levels were unaffected by light. These data strongly indicate that two rice CDPK isoforms have different functions in seed development and in response to light in leaves.     

Seed set in Guayule (Parthenium argentatum,asteraceae) in relation to insect pollination

Guayule (Tarthenium argentatum, Asteraceae) is one of two major plant species grown for natural rubber. Studies were conducted to determine the effect of honey bee (Apis mellifera) pollination and season on seed set and total seed yield/ha. The experiments involved four pollination treatments: plants caged with bees; plants caged without bees; plants open pollinated (uncovered); and plants individually covered. Seeds were harvested monthly July–September 1984, and May–September 1985. Plots with bees produced at least 150% more seeds than plots without bees, and there were no qualitative differences in the seed weights among treatments. Highest seed yield was in May and September. Results indicate that (1) insect pollination in guayule increases seed yield and (2) fewer seeds are produced in the warmest months.     

Freezing injury to ovules, pollen and seeds in winter rape

Winter rapeseed (Brassica napus, cv. Samouraï) flowersearly in spring and, under field conditions, short freezingperiods can occur. Unacclimatized plants were freeze-stressed(–3°C for 4 h) at different developmental stages ofbuds, open flowers and seeds. The dissection of pistils from stressed plants showed that freezingresults in shrivelled ovules. We assessed freezing injury onthe basis of per cent of shrivelled ovules: ovule sensitivitybegins early (8 d before anthesis) but increases up to anthesis.Crosspollination of stressed pistils with non-stressed pollenshowed that recording of freeze-injured ovules is a good methodfor early estimation of the effect of stress on seed yields. On the other hand, stress does not reduce the viability of pollen,except when it was applied at the binucleate pollen stage. Useof frozen pollen x nonstressed pistils has little effect onseed yields. Freeze injury on seeds was assessed by seed filling:seeds are very susceptible just after fertilization until 20d after fertilization (DAF). Freezing seems to inhibit seedfilling. A germination test of stressed seeds during their developmentindicated that embryo viability is not affected if the stressoccurs after 35 DAF. As the embryos develop, resistance to stressincreases. Key words: Brassica napus, rapeseed, freeze injury, pollen and ovule, seed filling     

Seed production of a subalpine Festuca nigrescens–Agrostis capillaris semi-natural grassland in the eastern Italian Alps

Studying the seed production of semi-natural grasslands helps to understand their regeneration processes and to evaluate the possibility of collecting seed useful for ecological restoration. In a subalpine Festuca nigrescens–Agrostis capillaris meadow, the development of the main grasses' seed production and the standing seed yield at meadow maturity were studied. The inflorescence size was in the lower part of the variation range reported in previous studies. The seed viability was average to high and durable. The biological floret site utilization was mostly above 50%. With 458 fertile stems m^? 2, the seeds produced (all seeds produced up to the time of the meadow maturity) by the main grasses were 21,000 m^? 2, of which 77% were of A. capillaris and 81% were viable. On a 3-year average, the standing seed yield (seeds present on the plants, i.e. produced minus shed seeds) at meadow maturity was 6953 seeds m^? 2. Amount and composition of the produced seeds were affected mainly by the variability among years of the fertile stems density of the perennial grasses and the annual hemiparasite Rhinanthus freynii. The percentage of produced seeds which formed the standing seed yield was affected mainly by the collecting date.