Surveys of pest incidence on oil-seed rape in south central England

Surveys of pest incidence on oil-seed rape in the south of England in 6 yr have shown that Meligethes aeneus can cause a considerable loss of pods on some fields. There is some evidence that control measures are often applied too late to obtain the best results; preliminary observations on the probable effect on yield are also presented. Ceuthorhynchus assimilis was recorded on the majority of fields with an average of 19% pods infested on spring-sown rape. Very heavy pod infestations were found on farms where rape had been grown regularly for some years. Dasyneura brassicae is at present not an important pest with an average of less than 1% pods infested. However, in each year infestations of up to 11% pod attack were recorded on a few fields.     

Macro-invertebrate populations and production on a salt-marsh in east England dominated by Spartina anglica

Summary The populations and production of the macroinvertebrates of a Spartina anglica salt-marsh in eastern England were studied over two years. A total of fifteen species were recorded in the sediments, of which twelve species were of regular occurrence, and the total population density recorded ranged from 3,481 m^-2 to 11,444m^-2 over the twenty-four sampling occasions.The four most abundant species were Nereis diversicolor, Tubifex costatus, Corophium volutator and Hydrobia ulvae. Thirteen further taxa were associated with the canopy of Spartina, with a total population density ranging from 0 to 1,149 m^-2. Total monthly standing crop ranged from 1.8 to 8.5 g C m^-2 with peaks in July/August in both years. Nereis diversicolor contributed 55% to 86% of total biomass in each month.Production and respiration for each species was determined and annual assimilation calculated. The total annual production was 16 g C m^-2 a^-1 in both 1979 and 1980, with a corresponding assimilation of 60 g C m^-2 a^-1 Nereis diversicolar accounted for >80% of production and assimilation in both years, and the species is clearly of considerable potential importance in the dissipation of Spartina material. The canopy dwelling species accounted for about 1% of the total annual production and assimilation.     

Population dynamics of Tenagomysis tasmaniae Fenton,Anisomysis mixta australis (Zimmer) and Paramesopodopsis rufa Fenton from south-eastern Tasmania (Crustacea:Mysidacea)

The population dynamics of a coastal mysid community was examined at One Tree Point in southern Tasmania over a 12-month period, including three 24-hour sampling sessions. Fourteen mysid species were recorded. Zonation patterns, life-history and diel activity of the three most abundant species, Tenagomysis tasmaniae, Anisomysis mixta australis and Paramesopodopsis rufa were examined. These three species were found in large numbers throughout the year associated with the macroalgal fringe on either side of the bay. Habitat partitioning within this zone of occurrence was evident. T. tasmaniae was the most abundant species (annual mean density 32.4 m^–3), followed by A. mixta australis (annual mean density 20.5 m^–3 ) and P. rufa (annual mean density 11.3 m^–3). The major peaks of abundance of these three species were temporally separate. Breeding was intensive for the three species from spring till late autumn. A cessation of breeding in winter was evident for A. mixta australis and to a lesser extent for P. rufa but T. tasmaniae bred throughout the year although at a reduced rate during winter. Examination of the diel behaviour identified peaks of abundance at sunrise and sunset for T. tasmaniae; P. rufa was caught in greater numbers at night, but no consistent pattern was observed for A. mixta australis.     

2011-2020年长江口大型底栖动物群落结构变化趋势

对2011—2020年夏季长江口48个站位的大型底栖动物定量监测数据进行统计分析,研究长江口海域底栖生物群落时空结构演变特征。结果表明,近10年长江口海域共鉴定大型底栖动物284种,其中多毛类128种,占总种数的45.1%,甲壳类64种占22.2%,软体动物56种占19.7%,棘皮动物16种占5.6%,其他类合计20种。平均生物密度为(79.5±45.9)个/m²(年份变幅14.7—195个/m²)。平均生物量为(5.20±3.25)g/m²(年份变幅1.01—10.11g/m²),多毛类、软体动物、甲壳类是生物密度和生物量组成的主要类群。十年期间种类数、生物密度和生物量均呈现明显上升趋势,口外区贡献最突出。四个监测区域(南支、北支、杭州湾、口外)的优势种差异大。丝异须虫Heteromastus filiformis在各年份的优势种中出现的频度显著最大。总体来看,长江口监测区域大型底栖动物群落自然演变趋势向好。三项多样性指数统计结果表明,口外区大型底栖动物种类组成多样性水平显著高于口内三个区。生物群...     

Phragmites australis makes valuable floating mat biotopes under oligotrophic conditions

To understand the mechanism of how Phragmites australis makes valuable floating mat biotopes under oligotrophic conditions, we investigated the environmental (water chemistry) and vegetational characteristics (growth, plant species richness, and floristic composition) of a floating mat consisting of three main mat-forming species with a zonal distribution (P. australis on the land side of the floating mat, Zizania latifolia on the middle area, and Typha angustifolia on the water side). Although they showed relatively low growth in the floating mat, compared to those in land-based wetlands, P. australis grew better than other mat-forming species in terms of shoot height and biomass production. Specifically, P. australis made more below-ground parts (593?±?38 g/m²) than other mat-forming species (Z. latifolia, 100?±?10 g/m²; T. angustifolia, 167?±?8 g/m²) and more companion species were found in P. australis-dominated plots (8.5?±?1.0 species/m²) than other plots (Z. latifolia-dominated plots, 2.7?±?0.6 species/m²; T. angustifolia-dominated plots, 1.0?±?0.0 species/m²). The larger amount of below-ground P. australis parts could contribute to thicker and denser mat structures, possibly providing more favorable habitats for neighboring plant species, thus facilitating more companion species within the P. australis-dominated area of the mat.

     

The transition from invasive species control to native species promotion and its dependence on seed density thresholds

Question: Does the seed density of invasive species affect establishment by native species in a bare ground context (following invasive species control efforts), and is it possible to promote transition to a native species dominated state by manipulating sowing density of the native community? Location: Experimental wetland basin in Chanhassen, Minnesota, USA. Methods: A mesocosm experiment investigated the influence of Phalaris arundinacea (invasive species) propagule pressure on establishment of native wet meadow species in the context of a newly restored wetland. Mesocosms were sown with P. arundinacea (0, 10, 50, 100, or 500 seeds/m²) and a mix of native species (3000 or 15000 seeds/m²). Results: When planted at densities > 100 seeds/m², P. arundinacea increased suppression of native species. Also, high native seed density suppressed P. arundinacea biomass production. This effect was more pronounced when P. arundinacea seed density was high (> 100 seeds/m²), but high native seed density (15000 seeds/m²) did not suppress recruitment of P. arundinacea from seed. Conclusions: The transition from post‐control bare ground (a common result of efforts to control invasive species) to native species establishment depends on both native species and invader seed density. These results suggest that a threshold of P. arundinacea propagule pressure exists, beyond which transition to a native community is less likely without management intervention. P. arundinacea can establish in the presence of a newly developing native plant community, even at very low densities of P. arundinacea seed. Invader control (following initial site clearing efforts) is essential to native species establishment.     

Population density,microhabitat use,and size characteristics of Pseudothelphusa dugesi,a threatened species of freshwater crab from Mexico (Brachyura: Pseudothelphusidae)

The brachyuran crab Pseudothelphusa dugesi, or cangrejito barranqueño, is an endangered species endemic to Cuernavaca, Morelos, in central Mexico. Individuals of P. dugesi inhabit freshwater springs, which are affected by human actions through wastewater drainage, eutrophication, exploitation, and invasive predators such as rainbow trout (Oncorhynchus mykiss). In this study, we evaluated the population density, microhabitat use, and size characteristics of P. dugesi. Sampling was conducted in May, July, and December of 2018 and March of 2019, during the dry and rainy seasons in a natural protected area, the Barranca de Chapultepec. Population density was 0.082 crabs/m², female density was 0.052 crabs/m², and male density was 0.112 crabs/m²; ovigerous females were not observed. Males were more abundant than females; the female: male ratio was 1:2.14. Males and females had similar carapace widths and lengths, left and right chelae lengths, and mass. Individuals of P. dugesi were found mostly under or between rocks and under leaf litter. Individuals were more active at night. An understanding of the biology and population characteristics of P. dugesi will help conservationists to develop protective measures and a conservation strategy for this endangered Mexican freshwater crab species.     

The effect of the cassava green mite Mononychellus tanajoa on the growth and yield of cassava Manihot esculenta in a seasonally dry area in Kenya

The effect of the cassava green mite Mononychellus tanajoa on the growth and yield of cassava Manihot esculenta was studied over a 10-month period in two field trials near Lake Victoria in Kenya. One plot was maintained free of mites by means of acaricide, while the other was artificially infested.The highest population density of M. tanajoa occurred during the dry season. A maximum leaf area index (LAI) of about 2 was reached at the onset of the dry season. The total leaf area of mite infested plants was reduced compared with uninfested plants during the dry spell. During the following rainy season infested plants recovered and attained the same leaf area as uninfested plants. A multiple regression model predicting the leaf area showed that 58% of the seasonal variation could be explained by plant age, soil water, and leaf injury.The net growth rate of infested plants was lower than that of uninfested plants. Maximum values of 21 (infested plants) and 49 (uninfested plants) g m^-2 week^-1 were attained at the onset of the second rainy season. No difference was found between uninfested and infested plants with respect to net assimilation rates per unit leaf area during the dry season. The net assimilation rates reached a maximum almost at the same time as the growth rates, but the infested plants peaked slightly earlier and at a lower level than the uninfested plants. M. tanajoa did not affect the relative allocation of dry matter into stems and storage roots, but the absolute allocation of dry matter declined with increasing mite injury. Thus, after 10 months the dry matter of infested plants was reduced by 29% and 21% for storage roots and stems, respectively, compared with the uninfested plants.     

Searching and Oviposition Behavior of Anagrus atomus L. (Hymenoptera: Mymaridae) on Four Host Plants of Its Host, the Green Leafhopper Empoasca decipiens Paoli (Homoptera: Cicadellidae)

Anagrus atomus L. is an important egg parasitoid of the green leafhopper Empoasca decipiens Paoli. In this study the ability of the parasitoid to locate and parasitize its host was investigated on four host plants, i.e., broad beans (Vicia faba L.), sweet pepper (Capsicum annuum L.), cucumber (Cucumis sativus L.), and French beans (Phaseolus vulgaris L.). For each plant species, the behavior of the parasitoid was observed on E. decipiens infested and noninfested plants. Searching and oviposition behavior were characterized by drumming, probing, and resting. Parasitoids spent significantly less time on non-infested than infested plants, 274.5 and 875.7 s, respectively, and no probing behavior was observed on non-infested plants. Frequency of resting behavior was significantly greater on non-infested than on infested plants. Total foraging time was significantly longer on infested than on non-infested plants, indicating that A. atomus females can efficiently discriminate between leaves with and without infestation. Parasitism of A. atomus was influenced by parasitoid density, with the highest parasitism rate (64.0%) obtained at a density of 10 A. atomus females/0.1356 m² but the number of parasitized eggs per female and the searching efficiency decreased with increasing parasitoid density.     

Dormant egg bank characteristics and hatching pattern of the Phallocryptus spinosa (Anostraca) population in the Makgadikgadi Pans (Botswana)

The anostracan Phallocryptus spinosa has an almost exclusively palearctic distribution. The Makgadikgadi Pans area in Botswana represents the only distribution record south of the Sahara. In this ephemeral wetland, it is an important food item in the diet of migrating birds. By studying egg bank characteristics (such as depth and density) and hatching requirements, we investigated the persistence of this sub-Saharan population. At localities in the middle and north basin of Sua Pan, sediment cores were taken along a transect, and dormant eggs were isolated. Densities of the active dormant egg bank ranged from 833 to 31449 dormant eggs m⁻², indicating that this species is well established. Most of the dormant eggs were found in the top 4 cm of the sediments, and densities decreased to zero at a depth of 13 cm. Considering the expected low sedimentation rate, the presence of dormant eggs down to 13 cm indicates long-time occurrence of P. spinosa in the Makgadikgadi Pans area. We observed high hatching fractions (up to 85%) at a temperature of 22 °C and a salinity of 5 ppt. A second anostracan species, Branchinella ornata, co-occurred with P. spinosa in our study site. This population also had a large active dormant egg bank (ranging from 6634 to 50557 dormant eggs m⁻²) with dormant eggs present until a depth of about 11 cm. This pattern indicates a long-time co-occurrence of P. spinosa and B. ornata.     

How small is too small for small animals? Four terrestrial arthropod species in different-sized remnant woodlands in agricultural Western Australia

Island biogeography theory, and the 50/500 rule of genetics, have effectively devalued small habitat fragments for species conservation. Metapopulation theory has given new value to small remnants but data on species persistence are scarce. This study examined the capacity of very small and sheep-grazed remnants of eucalypt woodland in agricultural Western Australia to support remnant-dependent terrestrial arthropods. We surveyed 53 sheep-grazed remnants of wheatbelt wandoo Eucalyptus capillosa for the presence of four species of arthropod with different dispersal strategies (terrestrial versus aerial) and diet (predaceous vs. herbivorous): the harvester and mound-building termite Drepanotermes tamminensis, the wood-eating and mound-building termite Amitermes obeuntis, the predaceous and burrowing scorpion Urodacus armatus and the predaceous 'bull' ant Myrmecia nigriceps. All species with the exception of the scorpion disperse aerially, and all construct above-ground structures that are easily recognized. Remnants ranged in size from 50 m² to 21 000 m² (mean 1791 m²), in spatial isolation (distance to the nearest vegetation remnant) from 10 m to 500 m (mean 123 m) and in a length-to-width ratio (shape) from circular (mean ratio 1.0) to linear (mean ratio 4.0). Observations in small and grazed remnants were compared with observations made in six wandoo woodland sites within a large (1040 ha) and ungrazed remnant. The total number of target species was highly correlated with remnant area (r = 0.68). Remnant isolation and remnant shape had no apparent influence on the total number of target species. The minimum area of grazed remnants in which individual species were recorded followed the large predator Urodacus armatus (4515 m²) > smaller predator Myrmecia nigriceps (300 m²) > harvester termites Drepanotermes tamminensis (102 m²) > wood-eating termites Amitermes obeuntis (50 m²). With the exception of U. armatus which occurred only in three of the four largest grazed remnants, the occurrence of all other species increased from small to large grazed remnants, suggesting a remnant-size effect for all species. Remnant isolation or remnant shape had no apparent influence on the occurrence of any one species. The terrestrially dispersing scorpion persisted in remnants despite their isolation from other remnants from 200 m to 500 m. For both termite species, mound heights were significantly greater in large, ungrazed woodlands than in small and grazed woodlands. The incidence of mound abandonment in smaller and grazed remnants was considerably higher for harvester than for wood-eating termite colonies. This suggests differences in spatial requirements and possibly diet-related susceptibilities to fluctuations in food availability. The diameter of Myrmecia nigriceps nests showed no relationship with remnant size or isolation. This study demonstrated that even very small remnant woodlands on farms may play an important role in sustaining small native animals, either as stepping-stones for dispersing individuals (termites, ants) or in providing adequate habitat to sustain populations for longer periods (all four species).     

Using the Mahalanobis distance statistic with unplanned presence-only survey data for biogeographical models of species distribution and abundance: a case study of badger setts

Aim Project‐specific data for biogeographical models are often logistically impractical to collect, forcing the use of existing data from a variety of sources. Use of these data is complicated when neither absence nor an estimate of the area sampled is available, as these are requirements of most analytical techniques. We demonstrate the Mahalanobis distance statistic (D²), which is a presence‐only modelling technique and does not require information on species absence or the sampled area. We use badger (Meles meles) setts as the basis for this investigation, as their landscape associations are well understood, and survey data exist against which to compare estimates of sett distribution and abundance. Location  England and Wales (151,403 km²). Methods We used stratified random samples of sett locations, and landscape variables that are known to be important for choice of badger sett location within a geographic information system at a cell resolution of 100 × 100 m. Landscape conditions at two scales were extracted, at and around sett locations, and the D² was used to classify all cells in England and Wales into a sett suitability model. Comparison of this sett suitability model with known main sett densities allowed estimates of main sett density to be made across England and Wales, with associated uncertainty. Results The sett suitability model was shown through iterative sampling and model evaluation using independent data to be stable and accurate. Main sett density estimates were biologically plausible in comparison with previous field‐derived estimates. We estimate 58,000 main setts within England and Wales, with 95% confidence intervals suggesting a value between 31,000 and 93,000. Main conclusions The D², which could be applied to other species and locations, proved useful in our context, where absence data were not available and the sampled area could not be reliably established. We were able to predict sett suitability across a large area and at a fine resolution, and to generate plausible estimates of main sett density. The final model provides valuable information on probable badger sett distribution and abundance, and may contribute to future research on the spatial ecology of badgers in England and Wales.     

Weed seed banks in arable fields under contrasting pesticide regimes

Viable weed seed banks on a clay soil were estimated for five years from 11 arable fields receiving one of three levels of pesticide application. Low seed densities, averaging 2000 m^-2, were recorded, but dicotyledonous weed seed numbers usually increased following oilseed rape crops, reaching 26 000 m^-2 in one field. Field-to-field variability was large. Samples could be grouped using multivariate techniques into those dominated by grass weed seeds and those containing mainly dicotyledonous weed seeds. Within TWINSPAN classifications, most fields maintained similar seed banks during the study period, indicating relative stability of communities. Differences present at the outset of the experiment, including variation between fields in the same treatment area, were maintained for the five years. Patterns were not associated with straw disposal or spring herbicide application. Oilseed rape crops did not cause qualitative change in seed bank communities, but allowed certain species, notably Stellaria media and Sonchus asper, to increase seed numbers. Smaller and less diverse seed banks in the area receiving most herbicide were not statistically significant. Seed banks gave poor predictions of following weed populations.     

First Quantitative Study of Rove Beetles in Oklahoma Winter Wheat Fields

Adult rove beetles (Staphylinidae) were sampled every 7–14 days from one winter wheat field located in each of the four major wheat growing regions of Oklahoma during the 1999–2000 and 2000–2001 growing seasons. The number of cereal aphids per tiller, wheat plant growth stage, and wheat tiller density also were estimated. A total of 12 genera representing 13 species of beetles were collected from the field. The density of rove beetles was generally low, ranging from 0.003 beetles per m² in fall to 0.106 beetles per m² in spring. Rove beetle communities differed among seasons. After accounting for the effect of season, there was no statistically significant association between rove beetle community structure and field location, aphid density, wheat plant growth stage, or wheat plant density. Most rove beetle species showed no association with a particular season, however, Aleochara notula Erichson, Lathrobium sp., and Oxypoda sp. were present predominantly in fall, while Bisnius inquitus Erichson was associated with winter. Oxypoda sp. was the most abundant rove beetle in winter wheat fields in spring and was relatively abundant in winter, but was not collected from wheat fields in fall. Tachyporus jocosus Say was present in wheat fields during all seasons. T. jocosus was the most abundant rove beetle species in the winter wheat fields in fall and winter and was the second most abundant species during spring.     

Estimating biophysical parameters of rice with remote sensing data using support vector machines

Hyperspectral reflectance (350–2500 nm) measurements were made over two experimental rice fields containing two cultivars treated with three levels of nitrogen application. Four different transformations of the reflectance data were analyzed for their capability to predict rice biophysical parameters, comprising leaf area index (LAI; m² green leaf area m⁻² soil) and green leaf chlorophyll density (GLCD; mg chlorophyll m⁻² soil), using stepwise multiple regression (SMR) models and support vector machines (SVMs). Four transformations of the rice canopy data were made, comprising reflectances (R), first-order derivative reflectances (D1), second-order derivative reflectances (D2), and logarithm transformation of reflectances (LOG). The polynomial kernel (POLY) of the SVM using R was the best model to predict rice LAI, with a root mean square error (RMSE) of 1.0496 LAI units. The analysis of variance kernel of SVM using LOG was the best model to predict rice GLCD, with an RMSE of 523.0741 mg m⁻². The SVM approach was not only superior to SMR models for predicting the rice biophysical parameters, but also provided a useful exploratory and predictive tool for analyzing different transformations of reflectance data.     

PRODUCTIVITY AND COMPOSITION OF A BALDCYPRESS-WATER TUPELO SITE AND A BOTTOMLAND HARDWOOD SITE IN A LOUISIANA SWAMP

The productivity and composition of two study sites in a southern Louisiana freshwater swamp were studied from October 1973 to November 1974. Net productivity was determined from measurements of litter-fall, stem growth of woody species, and harvest samples of annual herbaceous understory. Annual stem growth was calculated from biomass estimates on two different dates. The annual increase in stem biomass was 800 g dry wt/m² for a bottomland hardwood site (BLH) and 500 g dry wt/m² for a baldcypress-water tupelo site (CT). Litter-fall was 574 g dry wt/m²/yr for BLH and 620 g dry wt/m²/yr for CT. Harvest samples within the two plots yielded 200 g dry wt/m² and 20 g dry wt/m² for BLH and CT, respectively. Minimum net primary production was calculated as the sum of the three: 1574 g dry wt/m²/ yr for BLH and 1140 g dry wt/m²/yr for CT. Maximum estimates of herbaceous production and insect consumption were made by using values from the literature. Estimated total net primary productivity was 1733 g dry wt/m²/yr for BLH and 1516 g dry wt/m²/yr for CT. Tree composition was determined by the point-centered quarter method. Relative frequency, relative density, absolute density, relative dominance, and importance value (IV) were calculated for the tree species along each transect. In the bottomland hardwood area many woody species exist with Acer rubrum var. drummondii (IV = 23.9) and Nyssa aquatica (IV = 18.4) the most dominant. In the baldcypress-water tupelo area, fewer woody species exist and Taxodium distichum (IV = 39.2) and N. aquatica (IV = 37.6) dominated. Comparison of productivity data from several southeastern swamps indicate that flowing water regimes tend to result in the highest swamp forest productivity.     

Reintroduction of rare arable plants by seed transfer. What are the optimal sowing rates?

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Population structure and spatial patterns for trees in a temperate old-growth evergreen broad-leaved forest in Japan

The population structure and spatial pattern of major tree species in a warm-temperate old-growth evergreen broad-leaved forest in the Tatera Forest Reserve of Japan were investigated. All stems 5 cm in diameter at breast height (DBH) were mapped on a 4 ha plot and analyses were made of population structure and the spatial distribution and spatial association of stems in different vertical layers for nine species. This was done in the context of scale dependency. The plot was located on a very gentle slope and 17.1% of its canopy layer was in gaps. It contained 45 woody plant species and 4570 living stems with a basal area of 63.9 m² ha^–1. Castanopsis cuspidata var. sieboldii, the most dominant species for the basal area, had the maximum DBH among the species present, fewer smaller stems and a lower coefficient of statistical skewness of the DBH distribution. The second most dominant species, Dystilium racemosum, had the highest stem density (410 ha^–1), more abundant smaller stems and a relatively higher coefficient of skewness. Most stems in different vertical layers showed a weakly aggregated distribution with loose colonies as basic units. Gap dependency for the occurrence of stems under the canopy layer was weak. Maximum slope degree of the plot also weakly affected the occurrence of stems. Spatial associations varied among intra- and interspecific cohorts in the different layers and spatial scales examined, and positive associations among cohorts were found more frequently as the scales examined became larger. This tendency suggests that key factors forming observed spatial associations might vary with the spatial scales.     

Possible involvement of the cytoskeleton in the regulation of barley caryopsis dormancy and germination

This study was conducted on barley cv. Ars. caryopses collected at full ripeness and divided into two batches. From one batch (dormant caryopses) polysomes were isolated from embryos immediately after harvesting and after two days of germination. From the other batch (non-dormant caryopses) the same was done after eight months storage in a dry state. A low ionic strength cytoskeleton-stabilizing buffer was used for the isolation of polysomes. Four different fractions of polysomes were examined: free polysomes (FP), membrane-bound polysomes (MBP), cytoskeleton-bound polysomes (CBP) and cytoskeleton-membrane-bound polysomes (CMBP). In germs grown from non-dormant caryopses, the first two fractions (FP + MBP) made up about 78 % of the total ribosomal material, whereas in embryos of dormant, imbibed caryopses, two last fractions (CBP + CMBP) made up about 71 %. The percentage of polysomes after 48 hours of imbibition of dormant caryopses in the FP, MBP and CBP was only about 13 % (i.e., 87 % monosomes), whereas a greater proportion (19.4 %) was found in the CMBP. The highest incorporation of ³H-uridine and ¹⁴C-amino acids (after 48 hours of germination and 0.5, 3 and 6 hrs incubation with precursors) took place in trhc CMBP both in dormant and non-dormant caryopses The major amount of the two polysome fractions associated with the cytoskeleton (CBP and CMBP) and the higher activity of CMBP in protein synthesis in embryos of dormant, imbibed triticale caryopses may indicate a significant role for polysomes associated with the cytoskeleton in the control of protein synthesis in dormant and germinating caryopses.     

Block-Surface Staining for Differentiation of Starch and Cell Walls in Wheat Endosperm

A staining technique for differentiating starch granules and cell walls was developed for computer-assisted studies of starch granule distribution in cells of wheat [Triticum aestivum L.] caryopses. Blocks of embedded caryopses were sectioned, exposing the endosperm tissue, and stained with iodine potassium iodide (IKI) and Calcofluor White. Excessive tissue hydration during staining was avoided by using stains prepared in 80% ethanol and using short staining times. The IKI quenched background fluorescence which facilitated the use of higher concentrations of Calcofluor White. Cell wall definition was improved with the IKI-Calcofluor staining combination compared to Calcofluor alone. The high contrast between darkly stained starch granules and fluorescent cell walls permitted computer assisted analysis of data from selected hard and soft wheat varieties. The ratio of starch granule area to cell area was similar for both wheat classes. The starch granule sizes ranged from 2.1 μm³ to 22,000 μm³ with approximately 90% of the granules measuring less than 752 μm³ (ca. 11 μm in diameter). Hard wheat samples had a greater number of small starch granules and a lower mean starch granule area compared to the soft wheat varieties tested. The starch size distribution curve was bimodal for both the hard and soft wheat varieties. Three-dimensional starch size distribution was measured for four cells near the central cheek region of a single caryopsis. The percentage of small granules was higher at the ends than at the mid-section of the cells.