Genotypic differences in root penetration ability of wheat through thin wax layers in contrasting water regimes and in the field

A significant proportion of arable land in south-western Australia is highly susceptible to subsoil compaction, which limits access of roots of wheat to water and nutrients at depth. Genotypic variation in the ability of roots to penetrate a hardpan has been reported for other cereals, using a pot technique, where a thin wax-layer of paraffin wax and petroleum jelly is placed in a soil column to simulate a hardpan. Previously we have modified and validated this technique for measuring root penetration ability of wheat seedlings under contrasting water regimes. Here we report on a series of five experiments (runs), two in well-watered and three in drought stress conditions, which evaluated seminal and nodal root penetration ability through thin wax layers among 24 Australian wheat cultivars and breeding lines (entries). These results were compared with observations on their rooting depths in two contrasting soil types in field trials, including a sandy duplex that contained a hardpan and a red clay that increased in soil strength with depth. Nodal roots ceased growth early under soil water deficit, and water uptake was instead dependant on seminal roots under conditions imposed in the pots. Plants were then reliant on the ability of seminal roots to penetrate the wax layer. Eight entries had superior root penetration ability in both well-watered and drought stressed conditions. Roots of three other entries, which failed to penetrate the wax layers, died under drought stress conditions. In field trials, there was a significant interaction between site and entry for maximum root depth. Our results from the pot studies and field trials indicate that there exists genotypic variation in root traits that are required to penetrate uniformly hard soil, dry soil or soil containing a hardpan. As four of the eight superior entries also showed superior root penetration ability at both sites in the field, there was an overall consistency, but there were exceptions at individual field sites. Factors likely to result in such exceptions were discussed, and topics for further research identified.     

Screening the ability of rice roots to overcome the mechanical impedance of wax layers: importance of test conditions and measurement criteria

The development of a wax layer method for screening the ability of rice (Oryza sativa L.) roots to overcome mechanical impedance is described. Wax layers (3 mm thick) made of mixtures of white soft paraffin and paraffin wax were installed 50 mm deep in tubes of sand. The sand was watered with nutrient solution and planted with 3-d old rice seedlings. The numbers of root axes per plant that had penetrated the wax layers 24 d after planting were counted. The ratio of penetrated to total root axes per plant gave a misleading measure of root penetration ability, as rice varieties differed in the ratio of penetrated to total axes in a low impedance (3% wax) control. In non-flooded conditions, a 60% wax layer decreased root penetration (number of roots penetrating the wax layer per plant) to a mean of 74% of the low impedance control, whereas an 80% wax layer decreased mean root penetration to 31% of the control. The best measure of root penetration in non-flooded conditions was the number of axes penetrating an 80% wax layer. Flooding decreased root penetration of a 60% wax layer to a mean of 26% of the low impedance control. The best measure of root penetration in flooded conditions was the number of axes penetrating a 60% wax layer.     

Field evaluation of laboratory techniques for predicting the ability of roots to penetrate strong soil and of the influence of roots on water sorptivity

The ability of two laboratory screening techniques to predict the abilities of roots of eight crop species to penetrate a compacted soil were evaluated and compared in a field experiment. A soil tilled to remove the effects of mechanical resistance was planted with the same species to serve as a control. Depth of root penetration, root density and the influence of the roots on the sorptivity of water were measured.Roots of all species penetrated deeper in the deep tilled than compacted soils. There were differences in the ability of roots of the species to penetrate the compacted soil. Generally dicotyledonous species had more roots penetrating to depth in both the compact and deep tilled soils. Within the main species classifications, lupin and safflower (dicotyledons) and oats and barley (monocotyledons) had the highest penetration into the compacted soil.Water sorptivities in the deep tilled soils were higher than those of the compact soil. Soil from planted treatments had higher sorptivities than soil which had not been planted. This is attributed to biopores left by the roots. Sorptivities of soils which had dicotyledonous species were generally higher than those of monocotyledons. The soil planted with safflower produced the highest sorptivity in the compacted layer (0.1–0.3 m).A comparison of the accuracy of the two laboratory screening methods in predicting the field penetration of roots suggest that the method involving mechanical stress was better than that involving osmotic stress. Relative root diameter was found to be a better indicator of the penetration ability of roots than relative root elongation.     

Genotypic variation in anther culture and effect of ovary coculture in durum wheat

The response of anthers to in vitro culture and the effect of coculture of ovaries on anther culturability have been studied in responsive and recalcitrant cultivars of durum wheat (Triticum turgidum ssp. durum) from Morocco and ICARDA. A large genotypic-dependence of anther culture has been shown in 18 cultivars. Their response in term of callus and embryo induction varied from 0 to 13%. Coculture of ovaries with anthers enhanced the response of the most responsive genotype (cv. Sarif) and removed the recalcitrance in Cocorit and Isly cultivars. However, there was no effect of anther-ovary coculture on green plant regeneration. The implication of the genome and the media conditioning by the ovaries on anther response is discussed. This revised version was published online in June 2006 with corrections to the Cover Date.     

The ability of non-classical meteorological parameters to penetrate into buildings

The penetration ability of four physical variables whose value in the undisturbed atmosphere is influenced by meteorological processes and which have repeatedly been mentioned in biometeorological studies has been investigated. Ion concentration, sferics, pressure fluctuations, and temperature fluctuations were simultaneously recorded outside and inside buildings. Measurements were made in a classical wood/brick house as well as in a fully airconditioned reinforced concrete/glass construction. It was found that pressure fluctuations with periods longer than one minute penetrate the buildings practically unchanged while sferics (measured at a frequency of 10 kHz) are attenuated. The attenuation depends strongly on the construction type. The concentration of ions and the temperature fluctuations inside buildings are independant of their respective values in the atmosphere. Their value is determined only by local activity such as smoking, open windows, heaters, or the presence of persons.     

Morphological plasticity of wheat and barley roots in response to spatial variation in soil strength

Root systems of individual crop plants may encounter large variations in mechanical impedance to root penetration. Split-root experiments were conducted to compare the effects of spatial variation in soil strength on the morphological plasticity of wheat and barley roots, and its relationship to shoot growth. Plants of spring barley (Hordeum vulgare cv Prisma) and spring wheat (Triticum aestivum cv Alexandria) were grown for 12 days with their seminal roots divided between two halves of a cylinder packed with sandy loam soil. Three treatment combinations were imposed: loose soil where both halves of the cylinder were packed to 1.1 g cm^–3 (penetrometer resistance 0.3 MPa), dense soil where both halves were packed to 1.4 g cm^–3 (penetrometer resistance 1 MPa), and a split-root treatment where one half was packed to 1.1 and the other to 1.4 g cm^–3. In barley, uniform high soil strength restricted the extension of main seminal root axes more than laterals. In the split-root treatment, the length of laterals and the dry weight of main axes and laterals were increased in the loose soil half and reduced in the dense soil half compared with their respective loose and dense-soil controls. No such compensatory adjustments between main axis and laterals and between individual seminal roots were found in wheat. Variation in soil strength had no effect on the density of lateral roots (number per unit main axis length) in either barley or wheat. The nature and extent of wheat root plasticity in response to variation in soil strength was very different from that in response to changes in N-supply in previous experiments. In spite of the compensatory adjustments in growth between individual seminal roots of barley, the growth of barley shoots, as in wheat, was reduced when part of the root system was in compacted soil.     

Cuticular wax of wheat

n-Alkanes, esters, aldehydes, free alcohols, -diketones and hydroxy--diketones were found to be the lipid components of the cuticular waxes of common wheat Chinese Spring (Triticum aestivum L.). The ditelosomic lines 7A-L and 7D-S showed a dramatic decrease in the amount of -diketones and hydroxy -diketones which are reduced to traces. The homologue composition within each class of compounds has also been determined for all three of the lines of wheat. The effects of chromosomal deficiencies have been demonstrated. Chromatographic techniques and mass spectrometry have been used for the separation and identification of the substances which compose the waxes. This study has provided further evidence of the role of genes situated on well defined chromosomes in determining the nature of classes of compounds composing wax and governing the homologous composition within each class of substances.     

Genotypic differences in phosphorus efficiency of wheat

In an attempt to evaluate whether breeding and selection for high yielding capacity did change the P requirements of modern wheat cultivars, the response of two wheat cultivars to different levels of P supply was investigated. A traditional cultivar (Peragis) and a modern cultivar (Cosir) were cultivated in a C-loess low in available P and high in CaCO₃ in 120 cm high PVC pots. Shoot and root growth at different developmental stages was compared. The grain yield of the modern cultivar Cosir was higher at limiting and non-limiting P supply and, therefore, this cultivar can be considered as more P-efficient than the traditional cultivar. From the results it can be concluded that the main factors contributing to the higher P efficiency of the modern cultivar are (i) efficient use of assimilates for root growth characteristics which enhance P acquisition: smaller root diameter, and longer root hairs, (ii) efficient remobilization of P from vegetative plant organs to the grains, and (iii) lower P requirement for grain yield formation because of lower ear number per plant but higher grain number per ear.     

Genotypic variation in tissue water relations of leaves and roots of black walnut (Juglans nigra) seedlings

Genetic variation in the drought response of leaf and root tissue water relations of seedlings of eight sources of black walnut ( Juglans nigra L.) was investigated using the pressure-volume technique. Tissue water relations were characterized at three stages of a drying cycle during which well-watered plants were allowed to desiccate and then were reirrigated.
Sources varied both in the capacity for and degree of leaf and root osmotic adjustment, and in the mechanism by which it was achieved. A decrease in osmotic potential at the turgor loss point (ψ_πp) of 0.4 MPa was attributable to increased leaf tissue elasticity in seedlings of four sources, while seedlings of an Ontario source exhibited a 0.7–0.8 MPa decline in ψ_πp as a result of both increased solute content and increased leaf tissue elasticity. Seedlings of a New York source showed no detectable osmotic adjustment.
In roots, decreased ψ_πp (osmotic potential at full hydration) and ψ_πp were observed under drought. Sources that exhibited significant leaf osmotic adjustment also generally showed a similar response in roots. Tissue elasticity and ψ_πp of roots were higher than those of shoots, whereas ψ_πp of the two organs was similar for most sources. Because of greater elasticity, roots exhibited a more gradual decline in turgor and total water potential than did leaves as tissue relative water content decreased.     

Contribution to the estimation of nucleic acids in wheat roots

Several methods of NA isolation and estimation were examined in order to determine RNA in the whole root systems of young wheat plants. It was found out that during the hydrolysis of roots with perchloric acid according toOgur andRosen (1950),Spirin (1958) andHeitefuss (1965) a considerable amount of orcine-positive compounds is released which cannot be adequate to the RNA content. Therefore the separate RNA determination in the presence of DNA was excluded even after the NA fractionation by hydrolysis at various temperature and perchloric acid concentration. Besides NA hydrolysates contained a high amount of other compounds absorbing in the UV-region. Compounds interfering with both these methods were present especially in the basal parts of roots. Using the method ofKern (1960) a nucleoprotein fraction was isolated containing about 75 per cent of the total NA content in the tissue. This fraction consisted of more than 90 per cent RNA combined with proteins, namely the ribosomal RNA. After the hydrolysis it could be estimated spectrophotometrically for the hydrolysate did not contain other compounds absorbing in the UV-region. In the root tips the content of this fraction was high in comparison with the basal parts of roots and it changed with the kind of nutrition.     

Genotypic variation of the responses to chromium toxicity in four oilseed rape cultivars

Heavy metal toxicity in soils has been considered as major constraints for oilseed rape (Brassica napus L.) production. In the present study, toxic effects of chromium (Cr) were studied in 6-d-old seedlings of four different cultivars of B. napus (ZS 758, Zheda 619, ZY 50, and Zheda 622). The elevated content of Cr inhibited seedling growth, decreased the content of photosynthetic pigments, and activities of antioxidant enzymes, as well as increased the content of malondialdehyde and reactive oxygen species in all the cultivars. The Cr content in different parts of plants was higher in Zheda 622 than in other cultivars. The electron microscopic study showed changes in ultrastructure of leaf mesophyll and root tip cells at 400 μmol Cr, and these changes were more prominent in Zheda 622. An increased size and number of starch grains and number of plastoglobuli, damaged thylakoid membranes, and immature nucleoli and mitochondria were observed in leaves. In roots, enlarged vacuoles, disrupted cell walls and cell membranes, an increased number of mitochondria and a size of nucleolus, as well as plasmolysis (in Zheda 622) were observed. On the basis of these findings, it can be concluded that cv. Zheda 622 was more sensitive to Cr as compared to other three cultivars.     

Differing sperm ability to penetrate the oocyte in vivo and in vitro as revealed using colloidal preparations

The penetration ability of boar (Sus scrofa domestica) spermatozoa exposed to viscous preparations under in vivo and in vitro fertilization conditions has been examined. Experiments involving induced ovulation in prepubertal animals and surgical insemination directly into the oviduct isthmus revealed an advantage of colloidal preparations. Based on within-animal comparisons, the incidence of penetration was 100% using both spermatozoa suspended in a viscous preparation of plant extracts and spermatozoa suspended in a control medium. However, percentages of monospermy were 22.2% in 54 oocytes inseminated with the control suspension compared with 62.5% in 48 oocytes inseminated with the colloidal preparation. An in vitro study involving 355 oocytes from slaughterhouse ovaries inseminated with in vitro-capacitated spermatozoa gave similar percentages of penetrated oocytes for both the control and colloidal suspensions. In this case, however, the percentage of monospermy was 32.7% in the control group compared with 10.6% for spermatozoa suspended in the colloidal preparation. Higher mean numbers of sperm inside the oocytes and higher numbers of sperm bound to the zona pellucida were also observed with the colloidal suspensions. In vitro motility and viability for spermatozoa in the colloidal suspensions were enhanced compared with that of the control group. Lower sperm membrane lipid disorder and reactive oxygen species generation were also observed in the viscous solution. These findings suggest that viscous fluids can enhance the ability of sperm to move, bind, and penetrate the oocyte in vitro, although this influence may be masked in vivo due to the already high viscosity in the oviductal fluid close to the time of ovulation.     

Genotypic variation in zinc efficiency and resistance to crown rot disease (Fusarium graminearum Schw. Group 1) in wheat

A crown rot disease in wheat caused by the fungusFusarium graminearum Schw. Group 1 is a widespread problem in chronically Zn-deficient Australian soils. A link between crown rot and Zn deficiency was established by Sparrow and Graham (1988). This paper reports a test of a further hypothesis, that wheat genotypes more efficient at extracting zinc from low-zinc soils are more resistant to infection by this pathogen. Three wheat cultivars (Excalibur, Songlen and Durati) of differential Zn efficiency were tested at three zinc levels (0.05, 0.5 and 2.0 mg Zn kg⁻¹ of soil) and three levels ofF. graminearum S. Group 1 inoculum (0.1 g and 0.3 g kg⁻¹ live chaff-inoculum and control having 0.1 g kg⁻¹ dead chaff inoculum). Six weeks after sowing dry matter production of shoots and roots was decreased byFusarium inoculation at 0.05 mg and 0.5 mg kg⁻¹ applied Zn.Fusarium inoculum at 0.1 g was as effective as 0.3 g kg⁻¹ for infection and decreasing dry matter. The infection at the basal part of culm decreased significantly by increasing the rate of Zn application. Excalibur, a Zn-efficient cultivar (tolerant to Zn deficiency) produced significantly more shoot and root dry matter, and showed less disease infection compared with Zn-inefficient cultivars (Durati and Songlen) at low (0.05 mg Zn kg⁻¹ soil) and medium (0.5 mg Zn kg⁻¹ soil) Zn fertilization rates. Higher rate of Zn fertilization (2.0 mg Zn kg⁻¹ soil) reduced the disease level in Durati to the level of Excalibur but the disease level of Songlen was still high, indicating its high Zn requirement and or sensitivity to crown rot. The data on Zn uptake show that Excalibur, being Zn-efficient, was able to scavenge enough Zn from Zn-deficient soil, we suggest that besides sustaining growth Excalibur was able to build and maintain resistance to the pathogen; inefficient cultivars needed extra Zn fertilization to achieve performance comparable to that of Excalibur. The present study indicates that growing Zn-efficient cultivars of wheat along with judicious use of Zn fertilizer in Zn-deficient areas where crown rot is a problem may sustain wheat production by reducing the severity of the disease as well as by increasing the plant vigour through improved Zn nutrition. ei]Section editor: R Rodriques-Kalana     

Relationship between heparin binding characteristics and ability of human spermatozoa to penetrate hamster ova

Heparin binding site affinity and density on human spermatozoa were compared between fertile and infertile men with normal or abnormal results in the zona-free hamster ova-sperm penetration assay (SPA). A portion of fresh semen from fertile donors and potentially infertile men was processed through the SPA while the remainder of the ejaculate was used to quantitate heparin binding on spermatozoa. Saturation binding assays with [3H]heparin (15-375 nM) were analysed for 3 groups of men: (1) infertile patients with abnormal SPA results, (2) infertile patients with normal SPA results and (3) fertile donors. The heparin binding site density was significantly higher in men who possessed normal SPA results (infertile men and fertile donors) than in infertile men with abnormal scores in the SPA. There was no difference in heparin binding affinity between the three groups. These findings suggest that the heparin binding-site density may be related to the ability of human spermatozoa to undergo successfully the acrosome reaction.     

Genotypic and environmental variation in autumn-sown onions

Summary Seven onion cultivars of Japanese and European origin were evaluated in autumn-sown trials, at six sites over two seasons. Within each season genotypic differences were detected for winter-kill, bolting, maturity time and yield. In general the Japanese cultivars showed consistently lower levels of bolting and winter-kill and earlier maturity relative to the European cultivars. Environmental effects were important with differences between seasons, sites and sowings recorded for most characters. It was concluded that the use of early and late August sowings would provide suitable screening environments for bolting and winter-kill respectively. There were also differences between genotypes in their linear response to environments as shown by joint regression analyses. Express Yellow O-X (hybrid) showed least response to environments for bolting and winter-kill and Senshyu semi-globe Yellow gave the most consistent time to maturity over environments.     

Application of Brachypodium to the genetic improvement of wheat roots

To meet the demands of a larger and more affluent global population, wheat yields must increase faster this century than last, with less irrigation, fertilizer, and land. Modelling and experiments consistently demonstrate a large potential for increasing wheat productivity by improving root systems; however, application of research to new varieties is slow because of the inherent difficulties associated with working underground. This review makes the case for the use of the model grass Brachypodium distachyon to simplify root research and accelerate the identification of genes underlying wheat root improvement. Brachypodium is a small temperate grass with many genomic, genetic, and experimental resources that make it a tractable model plant. Brachypodium and wheat have very similar root anatomies which are distinct from rice root anatomy that is specialized to help it overcome anaerobic conditions associated with submerged roots. As a dicotyledonous plant, Arabidopsis has an even more divergent root system that features a tap root system and cambia with secondary growth, both of which are lacking in the grasses. The major advantage of Brachypodium is its small stature that allows the adult grass root system to be readily phenotyped, unlike rice and maize. This will facilitate the identification of genes in adult roots that greatly influence yield by modulating water uptake during flowering and grain development. A summary of the advantages of Brachypodium for root studies is presented, including the adult root system architecture and root growth during grain development. Routes to translate discoveries from Brachypodium to wheat are also discussed.     

Genotypic variation in silicon concentration of barley grain

Soluble silicon (Si) in foods and drinks has been suggested to have a protective effect against neurotoxicity of Al. We investigated the genotypic variation in Si concentration of barley grain, which has many uses including in livestock feeds, malts for beer and whisky, and some foods for human consumption. Two collections of barley, grown in the same field, were subjected to analysis; 274 standard varieties selected at the Barley Germplasm Center of the Research Institute for Bioresources, Okayama University (SV), and 135 varieties from the Barley Core Collection of Americans (BCCAM). The Si concentration of barley grain showed large variation, ranging from 0 (under detection) to 3600 mg kg^–1 in SV and from 0 to 3800 mg kg^–1 in BCCAM barleys. The Si concentration was much lower in hull-less barley than in hulled barley. The Si concentration of two-row barley was similar to that of six-row barley, suggesting that Si concentration is not affected by the number of spike rows. Si concentration also did not differ with the origin of the barley variety. More than 80% of total Si was localized in the hull. The Si concentration of the hull was between 15343 and 27089 mg kg^–1 in the varieties tested. A close correlation was obtained between the Si concentrations of barley grains harvested in different years, suggesting that the variation in Si concentration of barley grain is controlled genetically. These results provide fundamental data for breeding Si-rich cultivars.     

Antioxidant response of wheat roots to drought acclimation

Wheat (Triticum aestivum L.) seedlings of a drought-resistant cv. C306 were subjected to severe water deficit directly or through stress cycles of increasing intensity with intermittent recovery periods. The antioxidant defense in terms of redox metabolites and enzymes in root cells and mitochondria was examined in relation to membrane damage. Acclimated seedlings exhibited higher relative water content and were able to limit the accumulation of H₂O₂ and membrane damage during subsequent severe water stress conditions. This was due to systematic up-regulation of superoxide dismutase, ascorbate peroxidase (APX), catalase, peroxidases, and ascorbate–glutathione cycle components at both the whole cell level as well as in mitochondria. In contrast, direct exposure of severe water stress to non-acclimated seedlings caused greater water loss, excessive accumulation of H₂O₂ followed by elevated lipid peroxidation due to the poor antioxidant enzyme response particularly of APX, monodehydroascorbate reductase, dehydroascorbate reductase, glutathione reductase, and ascorbate–glutathione redox balance. Mitochondrial antioxidant defense was found to be better than the cellular defense in non-acclimated roots. Termination of stress followed by rewatering leads to a rapid enhancement in all the antioxidant defense components in non-acclimated roots, which suggested that the excess levels of H₂O₂ during severe water stress conditions might have inhibited or down-regulated the antioxidant enzymes. Hence, drought acclimation conferred enhanced tolerance toward oxidative stress in the root tissue of wheat seedlings due to both reactive oxygen species restriction and well-coordinated induction of antioxidant defense.