Dynamics of structural traits in two competing C3 grass species: influence of neighbours and nitrogen

This work analysed the regulatory structural mechanisms involved in the competitive interactions between the annual grass Bromus willdenowii Kunth. (BW = prairie grass) and the perennial C₃ grass Dactylis glomerata L. (DG = orchardgrass) during pasture establishment. Four combinations of species (pure BW, pure DG, DG flanked by BW and BW flanked by DG plants), with and without winter nitrogen fertilization, were factorially arranged in a randomized complete block design. Data were recorded at two organization levels: tillers (three tiller age cohorts) and target plants. Annual neighbours caused a decrease in the number of living leaves in tillers of intermediate age of both species. This structural regulatory mechanism led to a decrease in tiller number per plant and, therefore, restricted the development of horizontal space occupation. Annual neighbours did not cause an increase in tiller size, measured as lamina length or pseudostem height, but decreased root biomass. As a consequence, annual neighbours did not lead the hierarchy in light capture, but limited species radical colonization and competitive ability for soil resources. Winter nitrogen fertilization only affected tiller size in older tillers. These findings emphasize the importance of the cultural decisions, as sowing densities and nitrogen fertilization, to optimize pasture floristic composition.     

The effects of clipping height and frequency on net primary production of Andropogon gerardii (C4 grass) and Bromus inermis (C3 grass) in greenhouse experiments

There are potential agronomic and environmental benefits from incorporating warm‐season (C₄) grasses into temperate pasture systems, usually dominated by cool‐season (C₃) grasses, but there is a lack of information on how frequency and height of defoliation affects C₄ grasses. Three greenhouse experiments were conducted under (i) spring, (ii) summer and (iii) spring + summer clipping regimes. In each experiment, the effects of clipping frequency (weekly and monthly) and clipping height (clipped to 5 and 10 cm) were determined on above‐ and below‐ground net primary production (ANPP and BNPP) and total and seasonal dry matter (DM) yield for Andropogon gerardii Vitman (big bluestem, C₄ grass) and Bromus inermis Leyss (smooth brome, C₃ grass). Six replicates per treatment were used. In all experiments, ANPP and BNPP of smooth brome was greater than that of big bluestem although during late summer months big bluestem had higher DM yields of herbage than smooth brome. There were different effects of frequency and height of clipping for both species on two similar measurements: total annual DM yield and ANPP, indicating that the ability to generalize about the effects of defoliation from ecological and agronomic grassland standpoints is questionable. Clipping effects on ANPP and BNPP were different for summer‐clipped pots than for spring, and spring + summer‐clipped pots, indicating that management could be tailored to meet specific agronomic or conservation goals.     

Shade reduces growth and seed production of Echinochloa colona,Echinochloa crus-galli,and Echinochloa glabrescens

Echinochloa species are problematic weed species in direct-seeded rice systems in Asia. Because of concern about the continuous use of single herbicides, cultural weed management strategies need to be developed to maintain the sustainability of direct-seeded rice systems. However, the design of such strategies requires an understanding of the differential responses of weeds to shade caused by crop interference. The effects of shade on growth and seed production of Echinochloa colona, Echinochloa crus-galli, and Echinochloa glabrescens were determined. Weeds of three Echinochloa species were grown continuously in full sunlight or in 50% or 25% of full sunlight, or started in full sunlight and transferred to 50% or 25% of full sunlight at 21 days after sowing. The results suggested that changes in shade regime did not affect the plant height of E. colona and E. glabrescens; however, shade reduced the height of E. crus-galli. Compared with the plants grown in full sunlight, 75% of continuous shade reduced E. crus-galli height by 22%. Shade reduced leaf, total shoot, and root biomass and seed production in all the weed species, if occurred during the early growth of the weeds. The weeds responded with increased leaf biomass ratio when grown in shade. Compared with full sunlight, continuous shade of 75% increased leaf biomass ratio by 90% in E. colona and this value was 25% in the other two species. The results of this study show that shade can reduce weed growth and seed production of Echinochloa species but it should not be considered as a stand-alone strategy to manage these weeds in rice. This highlights the need for the integration of other weed management strategies to achieve complete control of these species.     

Light and plant growth‐promoting rhizobacteria effects on Brachiaria brizantha growth and phenotypic plasticity to shade

This is the first report on the effect of light intensity and plant growth‐promoting rhizobacteria (PGPR) on the growth of a tropical forage grass, being a relevant study to improve pasture management in conventional farming and integrated crop‐livestock‐forestry systems. In this study, our aim was to evaluate the effects of light intensity and Burkholderia pyrrocinia and Pseudomonas fluorescens inoculation on Brachiaria brizantha cv. BRS Piatã growth, and phenotypic plasticity to shade. The experiment was conducted in a semi‐controlled environment. Seedlings of B. brizantha were allocated to full sun and shade. P. fluorescens and B. pyrrocinia were inoculated individually or co‐inoculated by soil drench, 14 days after seedling emergence. We evaluated morphogenesis, structural and growth parameters. Irrespective of the light regime, co‐inoculated plants had greater leaf area and SPAD index (chlorophyll content). Increase in total biomass production in co‐inoculated plants was over 100% and 300%, under full sun and shade respectively. Co‐inoculated P. fluorescens and B. pyrrocinia increased shade tolerance in B. brizantha, improving plant performance. Co‐inoculation promoted growth in B. brizantha under both sun and shade, indicating its potential as a bio‐fertilizer in conventional and integrated systems, especially in silvopastoral systems, where light availability to pasture growth may be limited.     

Ultrastructure of Hybrid Callus between C3 and C4 Species of Amaranthaceae

Summary

Unlike the parent line, the hybrid calli between C₃ and C₄ species of Amaranthaceae have poor division capability. Therefore, in this study the fine structures of hybrid callus derived from protoplasts of Celosia cristata L. cv. Pink Charm (C₃ species) cell suspension and Amaranthus tricolor L. cv. Perfecta (C₄ species) cotyledon callus were investigated by light and electron microscopy, and compared with the parental. All callus lines were composed of parenchymatous cells possessing a voluminous vacuole. Parental callus lines contained organelles with a relatively normal structure. The peculiar feature characterizing the C₃/C₄ hybrid callus was the presence of highly lobed nucleus with multinucleoli and numerous small vacuoles having autophagic activity scattered in the cytoplasm. The autophagic activity is apparently related to the existence of several inclusions such as cytoplasmic organelles in the central vacuole. It might also be related to the pronounced expansion of central vacuole and the reduced cytoplasm. The failure to sustain proliferation related to some atypical features of the organelles is discussed.     

Rubisco small subunits of C4 plants,Napier grass and guinea grass confer C4-like catalytic properties on Rubisco in rice

ABSTRACT

Overexpression of Rubisco small subunit (RbcS) of C₄ plant, sorghum (sorghum bicolor) was shown to enhance the catalytic turnover rate (k _cat) of Rubisco in rice (Oryza sativa). In this study, the effects of other Rubisco small subunits of C₄ plants, Napier grass (Pennisetum purpureum) and guinea grass (Megathyrsus maximus) on kinetic properties of Rubisco in rice were studied. The expression levels of Napier grass RbcS (NgRbcS) and guinea grass RbcS (GgRbcS) proteins accounted for 41% and 45% of total RbcS, respectively in homozygous overexpression lines. The k _cat and K _m for CO₂ (Kc) of Rubisco were increased in all transgenic lines. Interestingly, the k _cat was markedly higher in NgRbcS homozygous line, whereas K _c was notably higher in GgRbcS homozygous line. Although its effects depend on species, these results suggest that the introduction of C₄ RbcS are effective approaches to alter the catalytic properties of Rubisco in rice.     

Effect of drought on biomass allocation in two invasive and two native grass species dominating the mixed‐grass prairie

Control of exotic plant species invading the native prairie relies on our understanding of the eco‐physiological mechanisms responsible for the spread of these species as they compete with native plants for soil resources. We used a greenhouse pot experiment to study vegetative biomass allocation in response to drought stress in two exotic grass species, Kentucky bluegrass (Poa pratensis L.) and smooth brome (Bromus inermis Leyss), and two native species, western wheatgrass (Pascopyrum smithii (Rydb.) A. Löve) and green needlegrass (Stipa viridula Trin.). The experiment was conducted over 3 months in 2010 and again in 2011 in a factorial design of four species and two drought treatments. The proportional data of biomass allocation to shoots, roots, rhizomes and crowns (shoot base) of grass seedlings were analysed by both the nonparametric Mann–Whitney U‐test on the original data and one‐way anova on the arcsine‐transformed data. Our data suggest a clear distinction between the two invasive and two native species in potential competitiveness in soil resource use, with the two exotic species having higher biomass allocation to roots than the two native species and the native species having a higher biomass allocation to crowns than the two exotic species. It is interesting to note that the strongly rhizomatous smooth brome did not produce rhizomes in the first season's growth, regardless of the water stress level. The effect of drought stress on biomass allocation manifested itself more on rhizomes or crowns than on roots or shoots of the four studied grass species, with the effects species‐specific in nature.     

Interactive Effects of CO2 and O3 on the Growth of Trisetum flavescens and Trifolium pratense Grown in Monoculture or a Bi-Species Mixture

SUMMARY

Golden oat grass (Trisetum flavescens L.) and red clover (Trifolium pratense L.) were grown as monocultures or bi-species mixtures under controlled conditions and exposed to ambient (350 ppm) or elevated (580 ppm) CO₂, with or without addition of O₃ (diel profile with 150 ppb maximum). Shoot biomass measurements after the initial growth and two re-growth periods were used to determine the specific responses of both species, and the difference in the specific response between monocultures and mixtures. T. pratense was much more responsive to CO₂, O₃, and their combination, compared to T. flavescens. In the case of O₃ but not of CO₂, the difference in sensitivity between species was larger in mixture than in monoculture. In contrast to elevated CO₂, O₃ significantly reduced the root:shoot ratio in the mixture, which could explain the increasing negative effect of O₃ on clover with progressing harvests.

The relative CO₂ stimulation of T. pratense and of the cumulative mixture shoot biomass was larger in the presence than in the absence of O₃, which was due to an almost complete protection from O₃ stress by elevated CO₂. In the mixture, the fraction of T. flavescens was small and increased during the experiment; this increase was most pronounced with O3, but any change in mixture biomass was dominated by the response of T. repens.

The results confirm that in grass/legume mixtures legumes are most sensitive to elevated CO₂ and O₃, but the magnitude of specific responses depends on canopy structure and of plant development. Elevated CO₂ minimizes the negative impacts of O₃ stress on above- and below-ground plant growth.     

Growth, photosynthesis and root reserpine concentrations of two Rauvolfia species in response to a light gradient

Growth and secondary metabolites responses to light environment can be useful measurements to determine favourable habitat conditions for the cultivation and conservation of medicinal plants. We analyzed the growth, photosynthesis and root reserpine concentrations in seedlings of Rauvolfia vomitoria Afzel and Rauvolfia verticillatae (Lour.) Baill, two important medicinal plants yielding anti-hypertension alkaloids, at four different light levels (20%, 52%, 75% and 100% of full sunlight) in a shade house. Across all light intensities, seedlings of R. vomitoria grew faster with higher relative growth rate (RGR) than R. verticillatae, attributed to its higher photosynthetic capacities (light-saturated photosynthetic rate, A_max) and leaf area ratio (LAR). Typical shade-sun morphological responses to increasing light levels included decreased specific leaf area and LAR, whereas RGR and A_max was highest at median light levels for both species. R. vomitoria allocated higher leaf mass fraction and fine-root mass fraction, similar coarse root mass fraction, but lower stem biomass fraction than R. verticillatae. For fine roots, R. vomitoria had greater specific root length and small diameter than R. verticillatae, indicating its higher resource (water and nutrition) capture abilities. Both species had higher nitrogen concentration and lower reserpine concentration in fine roots than those of coarse roots. Neither reserpine concentration nor nitrogen was affected by light intensities. The reserpine concentration in coarse roots of R. vomitoria increased, whereas that of R. verticillatae decreased with increasing irradiance. Although not significantly, reserpine concentrations in coarse roots were positively correlated with A_max and RGR for both species across all light intensities. Shade or photosynthesis inhibition apparently did not increase alkaloid synthesis, which contrasted with the carbon/nutrient balance theory of plant defense. These results suggested that R. vomitoria and R. verticillatae could attain high biomass and yield of reserpine in high-light habitats (with max. in 75% sunlight), and intermediate-light habitats (with max. in 25–52% sunlight), respectively.     

The effect of Calluna vulgaris cover on the performance and intake of ewes grazing hill pastures in northern Spain

The effect of the proportion of Calluna vulgaris cover on diet composition, intake and performance of sheep grazing hill vegetation communities in northern Spain is examined. A total of 591 non‐lactating Gallega ewes grazed for five consecutive grazing seasons (June to September) on replicated plots of hill pastures (1700 m.a.s.l.) composed principally of Festuca, Agrostis, Nardus and Calluna spp. but with different proportions of Calluna vulgaris cover, either 0·3 (C_0·3) or 0·7 (C_0·7) of the total area. In 1 year, twenty‐eight ewes suckling single lambs also grazed the plots. The mean stocking density over the 5 years was 8·7 ewes ha^–1. On treatment C_0·3, daily liveweight gains (33 g d^–1) of non‐lactating ewes were significantly (P < 0·001) greater than on treatment C_0·7 (12 g d^–1). Likewise in lactating ewes the difference in mean daily liveweight change was 40 g d^–1 (–5 vs. –45 g d^–1 for C_0·3 and C_0·7 treatments respectively; P < 0·001). Liveweight gains of lambs were only 80–100 g d^–1 from June to August and lambs only maintained live weight during August and September. The effect of lactational status on liveweight changes was not significant. Liveweight gains of non‐lactating ewes increased significantly (P < 0·001) from the first to the last year of the experiment on both treatments. The composition of the diet was significantly affected by treatment (P < 0·001), with a higher proportion of grass species on the C_0·3 treatment and a higher digestibility of the diet in the first half of the grazing season (P < 0·001). The proportion of C. vulgaris in the diet was significantly (P < 0·001) higher on the C_0·7 treatment and increased significantly (P < 0·001) from July to September on both treatments. There were no significant differences in the composition of the diet selected by lactating and non‐lactating ewes. The results demonstrate that on hill vegetation communities, in which the grass components (Festuca rubra, Agrostis capillaris, and Nardus stricta) cover at least 0·3 of the area and on which the preferred grass component (Festuca and Agrostis spp.) is maintained at a sward height of at least 2·5 cm, non‐lactating ewes can increase their live weight and body condition, but this increase is influenced by the proportion and quantity of species of grass in the diet, which is affected in turn by the species of grass available and their nutritive quality. However, ewes suckling lambs were not able to maintain their live weight and body condition except when Calluna cover was 0·3 and grass height was more than 3·5 cm. It is concluded that these indigenous vegetation communities can be used in sheep production systems to complement the use of improved pastures at other times of year. In particular, they can be utilized during the non‐lactating period (summer) to increase body condition before the beginning of the mating period in autumn.     

Forage biomass,soil cover,stability and competition in perennial grass–legume pastures with different Paspalum species

The short life span, irregular forage production and susceptibility to weed colonization of cool‐season grass–legume pastures are serious problems in grazing dairy systems in warm‐temperate regions. The inclusion of warm‐season species has the potential to mitigate these problems. In this study, we evaluated the effect of the inclusion of two warm‐season grasses with different growth habits on seasonal forage biomass, soil cover and weed colonization. Three different pasture mixtures were evaluated under grazing: conventional pasture (CP) [tall fescue (Festuca arundinacea), white clover (Trifolium repens) and birdsfoot trefoil (Lotus corniculatus)], CP with Paspalum dilatatum and CP with Paspalum notatum (CP + Pn). Forage biomass and soil cover were sampled thirteen times during a 3‐year trial, and sampling times were grouped by season for the analyses. The mixtures with Paspalum showed higher soil cover in the autumn, while in the winter CP had higher soil cover than CP + Pn. Competition with tall fescue was similar between mixtures with Paspalum, when considering biomass, but it was higher in CP + Pn when considering soil cover. The inclusion of P. notatum increased biomass during the autumn but decreased the mixture performance during winter by reducing tall fescue soil cover. The addition of a warm‐season grass species with a moderate competing ability like P. dilatatum is likely to avoid a negative impact on the cool‐season component of the pasture.     

Changes in soil organic carbon pools in a long‐term trial with perennial fodder crops in acid soils of north‐east India

Efficient estimation of soil organic carbon (SOC) is vital for understanding and monitoring the effect of perennial fodder crops in conserving SOC. In subtropical regions, there is limited information on SOC accumulation and its allocation into different pools under long‐term grasses and legumes. Therefore, we investigated the dynamics of SOC in a 20‐year‐old field trial with seven perennial grass species and a legume in a Typic Paleudalf soil under subtropical climate in north‐east India by analysing oxidizable organic C (C_oc) and its fractions of very labile (C_VL), labile (C_L), less labile (C_LL) and non‐labile (C_NL), microbial biomass C (C_mic) and mineralizable C (C_min). Growing perennial fodder crops increased SOC in the 0–0.60 m soil depth from 19.9%–39.6% compared with the conventional cultivation with maize (Zea mays). The relative efficacy of the fodder species to SOC accumulation was Setaria sphacelata = Brachieria rosenesis > Panicum maximum cv. Makunia = Arachis pintoi > Panicum maximum cv. Hamil > Paspalam conjugalum = Pennisetum purpureum > Thysanolaena maxima. Among the analysed fractions, C_VL, C_L, C_LL and C_mic were influenced most by the fodder crops and the active pools (C_VL+C_L) constituted 71.6% of the SOC. The results indicate that under the tested subtropical climate, soil under perennial grasses and legumes conserves organic C and that most of the SOC is in labile pools of short residence time.     

Calibration and validation of optical chlorophyll‐measuring devices for use in predicting crude protein concentration in tropical grass herbage

The objective of this experiment was to evaluate the Fieldscout CM 1000 NDVI and Yara N‐Tester as easy‐to‐use and cost‐effective tools for predicting foliar chlorophylls (a, b and total) and crude protein (CP) concentrations in herbage from three tropical grass species. Optical chlorophyll measurements were taken at three stages (4, 8 and 12 weeks) of regrowth maturity in Guinea grass (Panicum maximum) and Mulato II (Brachiaria hybrid) and at 6 and 12 weeks maturity in Paspalum spp (Paspalum atratum). Grass samples were harvested subsequent to optical measurements for laboratory analysis to determine CP and solvent‐extractable chlorophylls (a, b and total) concentrations. Optical chlorophyll measurements and CP concentrations were highly correlated (Yara N‐Tester: r² = 0·77–0·89; Fieldscout CM 1000 NDVI: r² = 0·52–0·84). Crude protein prediction models from the Yara N‐Tester and Fieldscout CM 1000 NDVI accounted for 70–89% and 44–73% CP variability, respectively, in Mulato II and Guinea grass. The Yara N‐tester produced more accurate and reliable CP estimates based on very high concordance correlation coefficient [CCC (0·73–0·91)] and low rMSPE, mean and regression bias. It is concluded that the Yara N‐Tester produces more accurate and reliable CP estimates of tropical pastures.     

Effect of thinning and shade removal on green stem disorder in soybean

Green stem disorder (GSD) in soybean (Glycine max (L.) Merrill) retains green stems and leaves as the pods mature, thereby reducing the harvest efficiency and impairing seed quality. In order to elucidate the causes of GSD, the factors that promote GSD need to be identified. In our experiments, we adjusted plant density at the developmental growth stage R1 (the beginning of flowering) or at R5 (the beginning of seed filling), from dense (22.2 plants m^–2) to sparse (5.56 plants m^–2) by thinning. We found that GSD occurrence was increased when plant density was changed, compared to the treatments that were maintained under either dense or sparse conditions. GSD was promoted more strongly when thinning was conducted at R5 than at R1 stage. Shading equipment surrounding plants, except for their upper-most leaves, was implemented to determine the association of shading and GSD. The results of the shade experiment revealed that GSD occurrence generally increased in treatments subjected to shade removal, compared to those that were shaded until R8 stage (full maturity) or never shaded since the time of sowing. GSD was strongly promoted by shade removal at R5 than at R1 stage. The shading results coincide with the results of the plant density experiment, indicating that an increase in light availability enhances source activity relative to sink at R5 stage, thereby promoting GSD occurrence in soybean. Thinning is expected to be used as an easy experimental method to create GSD for research purpose.     

Dual and Triple Intercropping: Potential Benefits for Annual Green Manure Production

Abstract

Greater species diversity in natural ecosystems increases plant biomass production and stability. Intercropping is an agricultural practice that aims to accrue the benefits of species diversity by growing two or more species simultaneously in the same space. Functional group diversity is considered important for enhancing the beneficial effects of species diversity, but most previous intercropping studies used combinations of only two functional groups. Thus, we used three green manure species from different functional groups: sorghum (Sorghum bicolor (L.) Moench.), a C4 grass; crotalaria (Crotalaria juncea L.), a legume; and sunflower (Helianthus annuus L.), a forb. We examined the effects of intercropping on biomass, nutrient uptake, and their stability using a proportional replacement series in a field experiment for three years with four trials. The aboveground biomass was higher with dual and triple-component intercrops compared with sole crops; however, there were no superior effects of triple-component intercropping over dual-component intercropping. There were also no clear advantages of intercropping in terms of the nutrient uptake amount and stability.     

Seasonal patterns of diet composition, herbage intake and digestibility identify limitations to performance of weaner sheep grazing native pasture in the Falkland Islands

The botanical composition, intake and digestibility of the diet consumed by 1‐year‐old weaner sheep grazing a native white grass plant community in the Falkland Islands was measured in five periods between June 1998 and March 1999. Five different sheep were used in each period. Two methods were used for estimating the botanical composition of the diet: plant cuticle patterns in the faeces of the sheep and the patterns of concentrations of n‐alkanes in the faeces of sheep. These methods were used to predict the concentrations of C₃₂‐ and C₃₃‐alkanes in the herbage to allow the estimation of herbage intake and digestibility using the n‐alkane technique. It was concluded that the n‐alkane technique gave more accurate estimates of diet composition based on the comparison with estimates of nutrient intake derived from liveweight gains of sheep. The dominant pasture species, Cortaderia pilosa, was the predominant species consumed during the cooler periods of the year. In the summer, when the highest liveweight gains of sheep occur, the proportion of fine grass species, including Poa spp., Festuca magellanica and Agrostis capillaris, and herbs and sedges in the diet was highest. Herbage intake was the highest during the summer periods when digestibility was also at its peak. Estimated nutritional deficiencies of metabolizable energy, crude protein, phosphorus and vitamin D₃ that limit the growth and development of weaner sheep were evident for up to 9 months of the year. Targeted supplementation regimes that counter specific nutrient deficiencies could be developed on the basis of these results to address critical periods in the growth of young sheep.     

Changes in plant population density, composition and sward structure of a hill pasture during a pastoral fallow

Appropriate pre-sowing methods for the introduction of improved forage legume and grass germplasm are an important issue for hill pasture improvement in New Zealand. A pastoral fallow, which involves not defoliating pasture for a period generally from late spring/early summer to autumn, could create a potentially favourable environment for introducing improved germplasm. A field study was conducted on two aspects (shady and sunny) of moist, low-fertility hill country with or without added fertilizer (phosphorus and sulphur) in the southern North Island of New Zealand, to investigate the changes in plant population density and sward structure during a full or partial pastoral fallow, compared with a rotationally grazed pasture. A 7-month (October to May) pastoral fallow dramatically decreased the densities of grass tillers by 72% (P < 0·01), white clover (Trifolium repens L.) growing points by 87% (P < 0·01) and other species by 87% (P < 0·05). The decline in tiller density by pastoral fallow was enhanced on the shady aspect. Fertilizer application increased white clover growing-point density on the shady aspect (P < 0·05) and grass tiller density on the sunny aspect (P < 0·05). Decreased plant density during pastoral fallowing was attributed to aboveground biomass accumulation, which altered sward structure, leading to interplant competition and mortality by self-thinning and completion of the life cycle of some matured plants. The plant size-density relationship during pastoral fallowing in this mixed-species sward followed the serf-thinning rule, particularly when the calculation was based on all plant species rather than grass alone. There was no significant (P > 0·05) difference in final plant population density between the 7-month pastoral fallow and a shorter term (October to December) pastoral fallow. It is concluded that pastoral fallowing effectively reduced the plant population density and altered sward structure of a hill pasture. Such changes create a more favourable environment for the introduction of improved forage species.     

Ultrastructure of Fusion Product between Protoplasts from C3 and C4 Species of Amaranthaceae

Summary

This paper describes the ultrastructure of the electric field-induced fusion products of C₃ and C₄ species of Amaranthaceae at the early developmental stage. Protoplasts of C₃ species were isolated from a Ceiosia cristata L. cell suspension and, those of C₄ species were isolated from an Amaranthus tricolor L. cotyledon. Incompatibility occurred in the C₃/C₄ hybrid. The incompatibility reactions were detected in the newly formed hybrid cells accompanied with significant changes in the nucleolus (segregation of nucleolar components) and plastids (cup-like shape or amoeboid plastid enclosing cytoplasmic materials) of C₃ species parent. The structural changes in the organelles of the C₄ partner were less marked. After 5 days of culture, most organelles showed high cellular activity, and a normal dedifferentiation process of mesophyll chloroplasts was observed. At this stage nucleolar segregation was not detected and the C₃ species plastids were difficult to distinguish from the proplastids formed from mesophyll chloroplasts. In addition, some mitochondria showed bursting-like structure. However, under the culture condition used these somatic incompatibility did not seem to impair further growth of fusion products since they were still proliferating well resulting in callus formation.     

Forage production and nutritive value of Dactylis glomerata and Trifolium subterraneum mixtures under different shading treatments

Production and nutritive value of forage in silvopastural systems can be improved by introducing shade‐ tolerant grass and legume species in appropriate mixtures. The management of these systems can present a challenge regarding the selection of the proper grass and legume species as well as the maintenance of the optimum balance between the two species in the grass–legume stand. The objectives of the present study were to evaluate the performance of pure stands and mixtures (75:25, 50:50, 25:75) of Dactylis glomerata:Trifolium subterraneum under full sun, 60% shade and 90% shade. Dry matter production of D. glomerata was not affected by shading, while that of T. subterraneum was drastically reduced. Shading increased the crude protein (CP) and acid detergent lignin content, but did not affect the acid detergent fibre and neutral detergent fibre content. The dry matter production of the 75:25 and 25:75 mixtures was higher compared to the one of the 50:50 mixture, while the CP content of the 75:25 mixture was the highest under moderate shading. However, relative yield, aggressivity index and competitive balance index indicated higher competitive ability for D. glomerata compared with T. subterraneum especially under shade. Therefore, the 25:75 mixture is suggested as the most suitable under moderate shade (60%), to perpetuate the stand and avoid the frequent re‐sowing of T. subterraneum.     

Impact of land‐use intensity on the relationships between vegetation indices,photosynthesis and biomass of intensively and extensively managed grassland fens

Vegetation indices are widely used as model inputs and for non‐destructive estimation of biomass and photosynthesis, but there have been few validation studies of the underlying relationships. To test their applicability on temperate fens and the impact of management intensity, we investigated the relationships between normalized difference vegetation index (NDVI), leaf area index (LAI), brown and green above‐ground biomass and photosynthesis potential (PP). Only the linear relationship between NDVI and PP was management independent (R² = 0·53). LAI to PP was described by a site‐specific and negative logarithmic function (R² = 0·07–0·68). The hyperbolic relationship of LAI versus NDVI showed a high residual standard error (s.e.) of 1·71–1·84 and differed between extensive and intensive meadows. Biomass and LAI correlated poorly (R² = 0·30), with high species‐specific variability. Intensive meadows had a higher ratio of LAI to biomass than extensive grasslands. The fraction of green to total biomass versus NDVI showed considerable noise (s.e. = 0·13). These relationships were relatively weak compared with results from other ecosystems. A likely explanation could be the high amount of standing litter, which was unevenly distributed within the vegetation canopy depending on the season and on the timing of cutting events. Our results show there is high uncertainty in the application of the relationships on temperate fen meadows. For reliable estimations, management intensity needs to be taken into account and several direct measurements throughout the year are required for site‐specific correction of the relationships, especially under extensive management. Using NDVI instead of LAI could reduce uncertainty in photosynthesis models.