Variation for stomatal characteristics and water use efficiency among diploid,tetraploid and hexaploid Iranian wheat landraces

Wheat is one of the most important crops in the world. Its study has involved anatomy, ecology, physiology, molecular biology, biotechnology, and cultivation. Knowledge of the relationship of diploid, tetraploid and hexaploid wheat with stomatal characteristics and water-use efficiency is spare and is subject to argument. The present study was aimed to examine different characteristics of stomata in 28 Iranian landraces belonging to the species Triticum monococcum (2n = 2x = 14), Triticum durum (2n = 4x = 28) and Triticum aestivum (2n = 6x = 42), respectively and investigated a possible relationship between these characteristics and water use efficiency by pot cultivation experiments. The results revealed large variation among landraces as well as between the different species for stomatal frequency and size. The diploid wheat species had the highest stomatal frequency and the lowest stomatal length and width. The hexaploid species had a lower stomatal frequency than the tetraploid species. A highly significant relationship was noted between the value of those traits on the adaxial and abaxial surfaces of the leaf. The ratio of adaxial to abaxial values appeared to be constant for a given species. Both stomatal frequency and size were associated with ploidy level, but also with the nature of the constituting genomes of the species. A highly significant negative association was found among accessions between stomatal frequency and size. As a consequence, the variation for stomatal area per unit leaf area was less than for the other traits. Diploid and hexaploid wheat have significantly higher water use efficiency, compared to tetraploid wheat. No clear association was found, however, between water use efficiency and stomatal characteristics.     

Allopatric distributions of tetraploid and hexaploid forms of Aegilops neglecta Req. ex Bertol.

The genus Aegilops L. comprises 22 annual wild species that are closely related to wheat (Hammer in Kulturpflanze 28: 33–180, 1980). Aegilops neglecta Req. ex Bertol. is a member of the Aegilops section of this genus and is distributed from Morocco and Spain in the west to Transcaucasia and western Iran in the east. This species includes tetraploid (2n = 28, genome UUMM) and hexaploid forms (2n = 42, UUMMNN). However, the geographical distributions of the two cytological forms remain unclear. Clarifying the distribution of the two cytological forms is essential for a better understanding of the diffusion of Ae. neglecta and its tetraploid and hexaploid forms. In the present study, chromosome numbers were determined for accessions of Ae. neglecta from a total of 137 populations, located in the western area of the species distribution from the Aegean Islands to Morocco. Taken together with previous studies, the present data reveal a difference in the geographical distribution of tetraploid and hexaploid forms: tetraploid form is distributed in the eastern part of the species area and hexaploid form predominantly occurs in the western part with their border on the western margin of the Aegean Sea. Near the border, tetraploid and mixed populations are sporadically found among hexaploid populations in the Balkan and Peloponnesus Peninsulas, while a few hexaploid and mixed populations are found among tetraploid populations in the East Aegean Islands and West Anatolia.     

Influence of beech litter fragmentation and glucose concentration on the microbial biomass in three different litter layers of a beechwood

In February 1993 samples of litter from three different litter layers (upper, intermediate, and lower) were taken from a beechwood growing on basalt soil. Using the substrate-induced respiration method, we investigated the influence of fragmentation and glucose concentration on the maximum initial respiratory response. Glucose concentrations ranged between 0 and 160000 g g^-1 dry weight. The initial respiratory response reached a maximum at 80000 g glucose g^-1 dry weight. The addition of higher concentrations of glucose resulted in negligible changes in respiration. Litter materials of four different size classes (intact leaves, fragmented <100 mm², <25 mm², and <5 mm²) were amended with 80000 g glucose g^-1 dry weight. Substrate-induced respiration was at a maximum in the size class <25 mm². The addition of glucose to intact litter did not result in microbial growth. It is concluded that C is not the primary limiting element for the microflora in litter layers of the study site. Fragmentation of beech litter enabled the microorganisms to grow. Presumably, nutrients that limited microbial growth in intact litter were mobilized by the fragmentation procedure and enabled microorganisms to grow in fragmented litter materials.     

The effects of feeding by Oniscus asellus on leaf litter sulfur constituents

Summary Feeding by the isopod, Oniscus asellus, produced changes in the sulfur constituents of leaf litter substrates (Acer negundo, A. saccharum, Fagus grandifolia, Picea rubens, and Tsuga canadensis). Isopod consumption of leaf litter generally accelerated the mineralization of carbon-bonded S and increased the formation of ester sulfate in all substrates. After the isopod egestion of A. negundo leaves, fecal decomposition over 6 weeks increased total S concentration from 68 to 120 mol S/g due to the catabolism of organic carbon. During the same period sulfate decreased from 34 to 20 mol S/g and carbon-bonded S increased from 34 to 100 mol S/g. Thus the total S pool in aged feces became enriched with organic S (83% of total S). Macroinvertebrate consumption accelerated the transformation of S constituents and the carbon-bonded S concentration approached that of the Oa organic horizon of a northern hardwood forest.Dedicated to the late Prof. Dr. M.S. Ghilarov     

Identification of genomic constitutions of <Emphasis Type="Italic">Oryza</Emphasis> species with the B and C genomes by the PCR-RFLP method

Oryza officinalis complex is the largest and the most complicated group in the genus Oryza L., consisting of about ten species with the B, C, BC, CD, and E genomes. Taxonomy and identification of the species, particularly those with the B, C and BC genomes, are difficult due to the similar morphology and overlapping distribution of some species. The difference in ploidy levels of some species adds more complexity. In the present study, we surveyed 64 accessions of rice germplasm in the O. officinalis complex using RFLP analysis of PCR-amplified Adh genes in addition to chromosome counting. The results confirmed that all O. rhizomatis accessions are diploids with the C genome, whereas all O. minuta accessions are tetraploids having the BC genome. However, both diploid and tetraploid forms were found for the accessions identified in the genebank as O. officinalis, O. punctata and O. eichingeri. The tetraploid form of O. officinalis with the BC genome is exclusively distributed in India and has been described as O. malampuzhaensis. The tetraploid form of O. punctata which has been called O. schweinfurthiana by some workers was found to be as widely distributed as its diploid form in Africa. It is noteworthy that two accessions that had been determined as tetraploid O. officinalis were actually belonging to a species with the CD genome (O. latifolia). Our results promote a better understanding of the genomic constitutions of many accessions in the O. officinalis complex and correct determination of the genebank material, which serves as an important basis of germplasm cataloguing for further research and utilization.     

Characterization of a world collection of <Emphasis Type="Italic">Agropyron cristatum</Emphasis> accessions

The genetic diversity was studied of 115 Agropyron cristatum accessions from 17 countries. Tetraploids were the most common (74.8%), followed by diploid (16.3%) and hexaploid (6.9%). We observed a relation between geographic distribution and ploidy level. The tetraploids, the most widespread, were found from Europe through Russia to East Asia. The diploids appeared over the same general range, except in Turkey, Iran and Georgia where no diploid accessions were found. Hexaploid accessions mainly came from a region comprising the east of Turkey, the north of Iran and Georgia. A selection of 71 accessions, including all three ploidy levels, were analyzed by capillary electrophoresis using six wheat simple sequence repeat (SSR) markers. All markers presented high levels of polymorphism, generating 166 different alleles ranging in size between 84 and 256 bp. Based on polymorphic information content values obtained (0.579–0.968), all the SSRs were classified as informative markers (values?>?0.5). According to the dendrogram generated, all the A. cristatum accessions were distinctly classified. Diploid, tetraploid and hexaploid accessions are not clearly differentiated from each other on the basis of SSR markers. A field experiment was conducted to morphologically characterize 18 accessions including the three ploidy levels. Significant differences were found between the accessions in spike length, spike width and number of spikelets per spike. All the cytological, molecular, and morphological data demonstrate the high genetic diversity present in A. cristatum, making it a valuable resource for future breeding programs.     

Identification of ergosterol in vesicular-arbuscular mycorrhizae

Summary Ergosta-5,7,22-tri-3-enol (ergosterol) was identified by gas chromatography-mass spectrometry in roots of berseem (Trifolium alexandrinum L., cv. Landsorte) and sweet corn (Zea mays L., cv. Honeycomb-F1) infected with the vesicular-arbuscular mycorrhizal (VAM) fungus Glomus intraradices. The fungal-derived compound ergosterol was determined quantitatively in root extracts using reverse-phase high performance liquid chromatography. The concentrations of ergosterol in VAM-infected roots reached 72 g^-1 dry material in berseem and 52 g^-1 in sweet corn after 80 days of growth, whereas concentrations in non-infected roots remained below 8 g^-1 dry weight. Additionally, phytosterols such as -sitosterol, campesterol, and stigmasterol were detected in both infected and non-infected roots. Ergosterol, as a characteristic fungal substance, is proposed as an indicator of fungal biomass in the early stages of VAM infection.     

Powdery mildew resistance in Aegilops tauschii coss. and synthetic hexaploid wheats

Summary A collection of 400 Ae. tauschii (syn. Ae. squarrosa) Coss. accessions were screened for powdery mildew resistance based on the response patterns of 13 wheat cultivars/lines possessing major resistance genes to nine differential mildew isolates. 106 accessions showed complete resistance to all isolates, and 174 accessions revealed isolate-specific resistance, among which were 40 accessions exhibiting an identical response pattern as wheat cultivar Ulka/^*8Cc which is known to possess resistance gene Pm2. Expression of both complete and isolate-specific resistance from Ae. tauschii was observed in some synthetic hexaploid wheats derived from four mildew susceptible T. durum Desf. parents, each crossed with five to 38 resistant diploid Ae. tauschii accessions. Synthetic amphiploids involving different combinations of T. durum and Ae. tauschii generally showed a decrease in resistance compared with that expressed by the Ae. tauschii parental lines.     

Estimation of Deposition Velocities for 85Sr, 131I, 137Cs on Spinach,Radish and Beans Leaves in a Tropical Region Under Simulated Fallout Conditions

Poly dispersive aerosols of SrCl₂, NaI, CsCl having a size distribution of 2.33 m (Activity Median Aerodynamic Diameter, [AMAD]) with a geometric standard deviation of 1.83 m are used for the determination of deposition velocity for ⁸⁵Sr, ¹³¹I and ¹³⁷Cs aerosols on tropical spinach (Spinicia olericia), radish (Raphanous sative) and beans (Phasolous valugeries) plant leaves. The experiments were carried out in a specially designed exposure chamber to simulate deposition under accidental releases of radioactivity in tropical environment. The rates of particles deposition are expressed as a function of plant surface area and of plant dry weight. The deposition velocities obtained are in the range of 10^–6 to 10^–7 m/s. These values differ significantly from those in temperate regions. The lower values for deposition velocities (v _g) for the tropical environment is attributed due to high humidity. Therefore, the deposition of suspended aerosols on leaf surfaces caused by impaction and Brownian diffusion becomes slower in tropical environment and resulted into lower value of deposition velocities.     

Intrinsic antibiotic resistance and competition in fast- and slow-growing soybean rhizobia on a hybrid of Asian and US cultivars

Summary Six fast-growing soybean rhizobia (Rhizobium fredii) and thirteen slow-growing soybean rhizobia (Bradyrhizobium japonicum) were examined for resistance to 10 antibiotics. Axenic studies were carried out to determine the competitiveness of dual-strain inocula consisting of fast- and slow-growing rhizobia isolated from subtropical-tropical soils for nodule occupancy on a hybrid of Asian and US soybean cultivars. Nodule occupancy was determined by intrinsic resistance to erythromycin and neomycin. The results showed wide variability in resistance to 10 antibiotics for fast- and slow-growing rhizobia. The intrinsic antibiotic resistance of fast- and slow-growing rhizobia was extremely high against nalidixic acid (400 g ml^–1) and penicillin (200 g ml^–1). The competitive ability of inoculant strains for nodule occupancy varied for different combination sets and with the plant growing media. Our results show that fast-growing rhizobia nodulate a hybrid of Asian and US soybean cultivars. Fast-growing soybean rhizobia did not completely exclude nodulation by the slow-growing strains, which formed 0–79% nodules, depending on the strain used in the inoculum.     

Genetic diversity of HMW glutenin subunits in diploid,tetraploid and hexaploid <Emphasis Type="Italic">Triticum</Emphasis> species

The genetic variations of high-molecular-weight (HMW) glutenin subunits in 1051 accessions of 13 Triticum subspecies were investigated using sodium dodecyl sulfate polyacrylamide-gel electrophoresis. A total of 37 alleles were detected, resulting in 117 different allele combinations, among which 20, 68 and 29 combinations were observed in diploid, tetraploid and hexaploid wheats, respectively. Abundance and frequency of allele and combinations in tetraploid wheats were higher than these in hexaploid wheats. Allele Glu-A1c was the most frequent subunit at Glu-A1 locus in tetraploid and hexaploid wheats. Consequently, the results also suggested that the higher variations occurred at Glu-B1 locus compared to Glu-A1 and Glu-D1. Therefore, carthlicum wheat possessing the allele 1Ay could be presumed a special evolutional approach distinguished from other tetraploid species. Furthermore, this provides a convenient approach of induction of the 1Ay to common wheat through direct cross with carthlicum wheat. Alleles Glu-B1c and Glu-B1i generally absent in tetraploid wheats were also found in tetraploid wheats. Our results implied that tetraploid and hexaploid wheats were distinguished in dendrogram, whereas carthlicum and spelta wheats and however displayed the unique performance. In addition, founder effect, no-randomness of diploidization, mutation and artificial selection could cause allele distribution of HMW-GS in Triticum. All alleles of HMW-GS in Triticum could be further utilized through hybrid in the quality improvement of common wheat.     

Size homoplasy and mutational behavior of chloroplast simple sequence repeats (cpSSRs) inferred from intra- and interspecific variations in four chloroplast regions of diploid and polyploid <Emphasis Type="Italic">Triticum</Emphasis> and <Emphasis Type="Italic">Aegilops</Emphasis> species

Chloroplast simple sequence repeats (cpSSRs) are widely distributed in the chloroplast genomes of all plant species, and are frequently employed for genotypic and phylogenetic analysis. However, information on intra- and interspecies variation in cpSSRs is lacking. In this study, we sequenced four intergenic (non-coding) chloroplast DNA regions in 57 accessions of 12 tetraploid, and 16 accessions of 4 hexaploid species of Triticum and Aegilops. These sequence data added to our previous data for diploid species in the same chloroplast regions. Intra- and interspecific genetic variation was analyzed for a total of 189 accessions of 13 diploid, 12 tetraploid, and 4 hexaploid species of Triticum and Aegilops, such that all species were represented by multiple accessions. The data were used to infer phylogenetic relationships within and among Triticum and Aegilops species. Based on this robust phylogenetic tree, seven of eight cpSSR loci clearly exhibited “size homoplasy,” referring to the fact that cpSSRs of identical size and DNA sequence can arise even if the alleles are not descended from a common ancestor. These data indicate that cpSSRs should be used with caution in phylogenetic analyzes. Interestingly, as observed from several previous studies, our data also suggest that observed mutation rates may increase significantly when mononucleotide (homopolymer) repeat numbers reach or exceed 9 bp. In the present report, using this sequence data set involving cpSSRs, 81 unique haplotypes among 189 accessions were detected, and five tetraploid Triticum and Aegilops species were successfully identified and genotyped. Our results indicate that combinations of nucleotide substitutions, indels and SSRs of chloroplast nucleotide sequences are available for genotyping at the species accession level.     

Uptake and toxicity of copper and zinc for the African earthworm,Eudrilus eugeniae (Oligochaeta)

Growth rate change in earthworms is considered to be a suitable endpoint when determining sublethal effects. In this study we evaluated growth and maturation in the vermicomposting earthworm speciesEudrilus eugeniae as marker of sublethal toxicity of copper and zinc. We also compared routes of uptake. Apart from exposing worms experimentally for 73 days to contaminated food, a series of contact filter paper tests was also performed to determine LD₅₀ for copper and zinc. Both copper and zinc at sublethal concentrations affected growth and maturation in worms exposed to contaminated food. These worms had a copper content of 34.5 g g^–1 after 73 days and a zinc content of 184.9 g g^–1, showing a differential uptake. Copper was more toxic than zinc. Also in the contact test worms did take up more zinc than copper and the LD₅₀ (48 h) for copper was 0.011 mg cm^–2 and for zinc 0.066 mg cm^–2, which translated to body burdens of 6 g g^–1 for copper and 131 g g^–1 for zinc. Indications were that a regulatory mechanism existed for both metals. Both metals were taken up through the body wall at a relatively fast rate. This study indicated that the skin was the major route of metal uptake. This study also showed a poor relation between the two types of tests for purposes of evaluating lethality of zinc and copper.     

Competition among strains of Bradyrhizobium and vesicular-arbuscular mycorrhizae for groundnut (Arachis hypogaea L.) root infection and their effect on plant growth and yield

Summary Strains of Bradyrhizobium influenced root colonization by a species of vesicular-arbuscular mycorrhizae (VAM), and species of VAM influenced root nodulation by strains of Bradyrhizobium in pot experiments. In a field experiment, the effects of VAM on competition amongst inoculated bradyrhizobia were less evident, but inoculation with Bradyrhizobium strains increased root colonization by VAM. Certain VAM/Bradyrhizobium inoculum strain combinations produced higher nodule numbers. Plants grown without Bradyrhizobium and VAM, but supplied with ammonium nitrate (300 g ml^–1) and potassium phosphate (16 g ml^–1), produced higher dry-matter yields than those inoculated with both symbionts in the pot experiment. Inoculation with either symbiont in the field did not result in higher pod and haulm yields at harvest.ICRISAT Journal Article No. 886     

Polymorphism of high molecular weight glutenin subunits in three species of Aegilops

A collection of 136 accessions of Aegilops umbellulata (39), Ae. comosa (75) and Ae. markgrafii (22) was analysed for high-molecular-weight (HMW) glutenin subunits composition. The homogeneity of the accessions was studied and 55.1% of the collection was homogeneous for HMW glutenin subunits (29 Ae. umbellulata, 33 Ae. comosa and 14 Ae. markgrafii). The HMW glutenin subunits of Ae. umbellulata are encoded by the Glu-U1 locus; in Ae. comosa results showed that this proteins are encoded at the 1M chromosome, and the locus was named Glu-M1. In Ae. markgrafii it was assumed that HMW glutenin subunits were encoded by an homoeologous locus and it was named Glu-C1. All the accessions of Ae. umbellulata and Ae. markgrafii expressed both, x-type and y-type subunits. Among the Ae. comosa accessions, only one expressed an x-type subunit alone. All the accessions of Ae. umbellulata and some of Ae. comosa had x-type glutenins of higher molecular weights than those commonly present in bread wheat. A total of 8 alleles were detected at the Glu-U1 locus, 11 at the Glu-M1 and 4 at the Glu-C1. The new HMW glutenin variation found in this work suggests their possible utilisation in breeding for wheat quality.     

Detection of adult-plant resistance to Puccinia triticina in a collection of wild Triticum species

Three hundred and fifty three Triticum accessions, several also classified as Aegilops and comprising 13 diploid, tetraploid or hexaploid species, were screened for seedling and adult-plant resistance to Puccinia triticina Eriks. using a mixture of pathotypes UVPrt2, 3, 9 and 13. Seedlings were spray-inoculated with a suspension of freshly collected urediospores in distilled water containing Tween 20^® seven days after planting. Infection types (ITs) were scored 10 days post-inoculation (d.p.i.). Fully expanded flag leaves were inoculated and ITs and leaf rust severity were scored 16 d.p.i. One hundred and eighty two of the accessions were resistant to moderately resistant in the adult stage, whereas 126 were resistant or moderately resistant as seedlings to the pathotype mixture. Hypersensitive adult-plant resistance was particularly apparent in lines of T. timopheevii, T. sharonense, T. longissimum, T. searsii and T. turgidum. In T. turgidum, which comprised 272 accessions, approximately 44% of the adult plants were resistant to moderately resistant compared to 28% of the seedlings. The expression of these adult-plant resistances varied between hypersensitive flecking of flag leaves, and small pustules commonly associated with chlorosis and/or necrosis of leaf tissue. Partial resistance, expressed by small pustules without any apparent chlorosis, was observed in species such as T. tauschii, T. turgidum ssp. durum and T. turgidum ssp. pyramidale.     

Physiological and symbiotic characteristics of Rhizobium fredii isolated from subtropical-tropical soils

Summary Physiological and symbiotic characteristics were identified in Rhizobium fredii isolated from subtropical-tropical soils. The generation times of R. fredii Taiwan isolated-SB 357 and -SB 682 were 1.7 and 2.5 h, respectively. These strains were associated with acid production in yeast-extract mannitol medium. They were able to use hexoses, pentose, sucrose, trehalose and raffinose. Strain SB 357 can resist a high concentration of kanamycin (100 g ml^–1 and penicillin (400 g ml^–1). It can tolerate up to 2.34% NaCl and 1031.3 mosmol kg^–1 (23.4 bars). The growth rate of R. fredii SB 357 under the concentration of approximately 450 mosmol kg^–1 (10.2 bars) was not affected by salinity, but responded to osmotic pressure. Both strains (SB 357 and SB 682) isolated from subtropical-tropical soils were able to form an effective N₂-fixing symbiosis with the US soybean cv Clark lanceolate leaflet.     

Analysis of Wheat Disease Resistance Data Originating from Screenings of Gatersleben Genebank Accessions during 1933 and 1992

Series of 10,348 accessions belonging to 21 species (hexaploid, tetraploid, diploid) of the genus Triticum and 489 accessions belonging to 20 species of the genus Aegilops were scored for disease resistance during a period of 60 years. Tests were performed at the seedling stage for powdery mildew (Erysiphe graminis DC. f. sp. tritici March.), leaf rust (Puccinia recondita Rob. ex Desm. f. sp. tritici Erikss.), stripe rust (Puccinia striiformis West. f. sp. tritici Erikss.) and eyespot (Pseudocercosporella herpotrichoides (Fron.) Deight.) but also at the adult plant stage considering powdery mildew, leaf rust, stripe rust, eyespot and glume blotch (Septoria nodorum Berk.). About 150,000 disease scores recorded on index cards using different scoring scales were transferred to the computer, converted into a 1–9 scale and used to summarise the results. Within the genus Triticum 20% of the material analysed was classified as heterogeneous. For the accessions without detectable segregation a large variability for resistance/susceptibility was detected. At the adult plant stage resistant accessions without visible infections were identified for all diseases. The percentages of resistant accessions at that growth stage were always higher than the ones found in the material tested at the seedling stage. The probability for finding resistant material was shown to be highest in the diploid species ( > 50%) but decreased with increasing ploidy level to about 10% in the hexaploids. For Aegilops it was shown that most of the accessions were homogeneous and highly resistant against powdery mildew (seedling and adult plant stage), leaf rust (adult plant stage) and eyespot (seedling and adult plant stage/natural infection). The data obtained for the individual accessions are available via Internet (http://www.ipk-gatersleben.de). An erratum to this article is available at .     

Resistance to wheat leaf rust in Aegilops tauschii Coss. and inheritance of resistance in hexaploid wheat

A collection of 164 Aegilops tauschii accessions, obtained from Gatersleben, Germany, was screened for reaction to leaf rust under controlled greenhouse conditions. We have also evaluated a selection of synthetic hexaploid wheats, produced by hybridizing Ae. tauschii with tetraploid durum wheats, as well as the first and second generation of hybrids between some of these resistant synthetic hexaploid wheats and susceptible Triticum aestivum cultivars. Eighteen (11%) accessions of Ae. tauschii were resistant to leaf rust among which 1 was immune, 13 were highly resistant and 4 were moderately resistant. Six of the synthetic hexaploid wheats expressed a high level of leaf rust resistance while four exhibited either a reduced or complete susceptibility compared to their corresponding diploid parent. This suppression of resistance at the hexaploid level suggests the presence of suppressor genes in the A and/or B genomes of the T. turgidum parent. Inheritance of leaf rust resistance from the intercrosses with susceptible bread wheats revealed that resistance was dominant over susceptibility. Leaf rust resistance from the three synthetics (syn 101, syn 701 and syn 901) was effectively transmitted as a single dominant gene and one synthetic (syn 301) possessed two different dominant genes for resistance.