Identifying global centers of genetic diversity for grain sorghum with the use of rice DNA markers

STS and InDel markers developed on the basis of DNA sequences of certain rice genes are used for a comparative study of grain sorghum accessions. The main sorghum genetic diversity centers are revealed, two in Africa and a third in regions of Central, East, and South Asia.     

Use of prolamine polymorphism in studying genetic resources of forage grasses

Summary Electrophoresis of single seed prolamines was used for the analysis ofLolium perenne L.,Festuca pratensis Huds., andDactylis glomerata L. populations. Identification and registration of populations was carried out according to the frequencies of occurrence of genotypes with corresponding types of prolamine banding patterns. The publication sums up the problems of applied use of molecular markers for identification and registration of world genetic resources of forage grasses, analysis of the dynamics of population composition and other problems of plant growing, genetics, breeding and seed control. The approaches mentioned in the article are promising for their use in genetic banks as well as at the institutions which store collections of genetic resources of given crops and at universities and breeding stations.     

Gliadin Electrophoretic Analysis of the Genetic Integrity of Wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) Accessions After Frequent Seed Reproductions

A standard electrophoretic method for wheat cultivar identification was used on single seeds to analyse the genetic integrity of 11 wheat (Triticum aestivum L.) accessions after up to 24 seed reproductions in the Gatersleben genebank. It was clearly demonstrated that the gliadin pattern of single seeds can be used to analyse the genotype composition of wheat accessions. Stability of electrophoretic banding patterns was detected in eight accessions. Very week genetic drift was observed in three accessions. Our investigations confirm experiences of the successful utilisation of protein markers for cultivar verification and genetic integrity testing and demonstrate the high standard of wheat accessions maintenance in the Gatersleben genebank.     

Interaction of insect digestive enzymes with plant protein inhibitors and host-parasite coevolution

Summary A complex of new approaches was used to study the insect-plant coevolution by using cereal pest digestive -amylases and proteinases and their proteinaceous inhibitors in cereals. During evolution, plants can weaken the destructive affects of insect hydrolases at the expense of inhibitors in various ways, including (1) increasing the inhibitor activity and heterogeneity, (2) increasing the complexity of an inhibitor set and (3) producing highly specific insect enzyme inhibitors and bifunctional amylase/proteinase inhibitors. Insects, in turn, can decrease the influence of inhibitors by (1) increasing digestive enzyme activities, (2) by modifying a set of related activities of various digestive hydrolases, (3) by decreasing enzyme sensitivity to inhibitors and (4) by destroying inhibitors in guts by proteinases.Abbreviations IEF isoelectric focusing - TI trypsin inhibitor - CI and C/SI chymotrypsin and chymotrypsin/subtilisin inhibitors - InsAI12 insect amylase inhibitor with molecular weight value of about 12 kDa     

Dynamics of the composition of gliadin biotypes during creation of the spring triticale cultivar Zolotoi Grebeshok

The dynamics of the composition of gliadin biotypes of a Mexican population of triticale Merino/Jlo//Zebra32 during selection for a set of economically valuable traits under northwestern Russia conditions is studied. The characteristics of the structure of the cultivar Zolotoi Grebeshok created on this basis are registered in the form of a “protein passport” that gives additional information for legal protection of the cultivar and allows checking its purity and authenticity at all stages of seed production.     

Characterization of a glutenin-specific serine proteinase of Sunn bug Eurygaster integricepts Put

Glutenin hydrolyzing proteinases (GHPs) have been purified, by affinity chromatography, from wheat seeds damaged by the Sunn bug Eurygaster integriceps (Hemiptera, Scutelleridae). A 28 kDa protein was partially sequenced by mass spectrometry and Edman degradation which showed homology to serine proteases from various insects. Three full length clones were obtained from cDNA isolated from Sunn bug salivary glands using degenerate PCR based on the sequences obtained. The cleavage site of the protease was determined using recombinant and synthetic peptides and shown to be between the consensus hexapeptide and nonapeptide repeat motifs present in the high molecular weight subunits of wheat glutenin (PGQGQQ∧GYYPTSLQQ). Homology models were generated for the three proteinases identified in this study using the high resolution X-ray structure of a crayfish (Pontastacus leptodactylus) trypsin complexed with a peptide inhibitor as template (PDB accession 2F91). The novel specificity of this protease may find applications in both fundamental and applied studies.     

Analysis of gluten-hydrolyzing proteinase polymorphism in wheat grains damaged by Sunn Pest Eurygaster integriceps Put. and related bugs

The heterogeneity and variability of gluten-hydrolyzing proteinases in accessions of wheat grains from Russia and Turkey damaged by Sunn pest and related bugs have been first estimated using isoelectric focusing (IEF) in combination with a new version of the glutenin replicas method. The variability revealed reflects peculiarities of bug populations and their adaptive reaction to wheat variety. Approaches proposed can be used for diagnostics of damage, studying of interaction of pest with food plant and development of resistant forms of wheat.     

Analysis of oil composition in cultivars and wild species of oat (Avena sp.)

Oil quality and content were analyzed in 33 accessions from 13 wild species and 10 accessions of cultivated oat. Wild oat species tended to have higher oil and 18:1 fatty acid (FA) contents and lower amounts of 18:2 and 18:3 FAs as compared to cultivated oats. In addition to common FAs, minor amounts of several hydroxy and epoxy FAs were also present in the oat oil and mainly confined to specific lipid classes. These unusual FAs included the previously reported 15-hydroxy 18:2 (Delta9,12) (avenoleic acid) mostly found among polar lipids and a novel 7-hydroxyhexadecanoic acid located to 1,2-diacylglycerol. The present study highlights the potential of making use of the existing germplasm, consisting of wild oat species, in breeding programs for achieving new oat varieties that produce a range of oils with different FA compositions as well as having high oil contents. However, in one matter, oats apparently lack genetic diversity and that is for oil qualities that are highly enriched in the omega 3 (omega-3) FA 18:3. Consequently, developing oat cultivars with highly unsaturated oils will need involvement of other techniques such as biotechnology.     

Proteinase inhibitor polymorphism in the genus Vigna subgenus Ceratotropis and its biosystematic implications

The diversity of components for fourproteinase inhibitors found in species ofthe genus Vigna subgenus Ceratotropis are described. Trypsin,chymotrypsin, subtilisin and cysteineproteinase inhibitors were analyzedby isoelectric focusing followed by thegelatin replica method. Of these proteinaseinhibitors, trypsin inhibitors showedmost polymorphism both within and betweenspecies. Many trypsin inhibitor componentswere also active to chymotrypsin. Severalaccessions had very low levels or absenceof some inhibitors, such as very low levelsof trypsin inhibitor in two accessions ofthe V. tenuicaulis and absence ofchymotrypsin inhibitors in V.grandiflora and V. subramaniana.Proteinase inhibitor polymorphism broadlyagreed with the taxonomic system for thesubgenus Ceratotropis. Based oninhibitor variation species analyzed couldbe divided into three groups whichcorresponding to sections Aconitifoliae, Angulares and Ceratotropis. Some species have verylittle variation in trypsin inhibitorsdespite wide distribution, such as, V.radiata and V. reflexo-pilosa.Accessions of other species showedconsiderable intraspecific variation fortrypsin inhibitors, such as, V.grandiflora, V. aconitifolia andV. stipulacea. Proteinase inhibitorpolymorphism provides an indication of thespecies that may have contributed a genometo the tetraploid species, V. reflexo-pilosa.     

Polymorphism and inheritance of methionine-rich 2S albumins in sunflower

2S albumin fractions were isolated by a modified acetone precipitationmethod (Kortt and Caldwell 1990) from seeds of 103 sunflower (Helianthus annuus L.) accessions and analysed by SDS-PAGE, IEF andRP-HPLC. Two methionine-rich albumins SFA7 and SFA8 showed nodifferences in mobility on SDS-PAGE gels but were readily separated byRP-HPLC. Their levels also varied widely between different genotypes, inrelation to each other and as proportions of the total albumin fraction. Avariant form of SFA8 was identified which differed from the normal SFA8in its pI (6.5 compared to 6.0) and mobility on SDS-PAGE. N-terminal sequences of both the variant form of SFA8 and the majorform of SFA7 were identical to that reported previously for the normalform of SFA8 from the cultivar Hysun (Kortt et al., 1991) indicating theirstructural relatedness. Analysis of segregation in the F₂ of the crossbetween lines VIR130 (variant SFA8) and VIR104 (normal SFA8) showedthat the normal and variant forms of SFA8 are encoded by alleles at asingle Mendelian locus. The levels of SFA7 and SFA8 in the seeds ofparental lines, F₁ hybrids and individual F₂ seeds classifiedfrom SDS-PAGE and IEF as homozygous for normal SFA8 (VIR104 type),homozygous for variant SFA8 (VIR130 type) and heterozygous (F₁type) were determined by RP-HPLC. Seeds of the parental line VIR130contained 3.7% SFA7 and 19.0% SFA8 whereas seeds of VIR104contained 9.9% SFA7 and 12.8% SFA8. The F₁ hybrid seedscontained a higher total amount of SFA7+8 proteins (32% comparingto 22% in each parent) which was largely accounted for by a highproportion of SFA7. The mean combined proportions of SFA7+8 in eachof the three phenotypic classes of F₂ seeds were about 18–19% ofthe total. However, the combined proportions of SFA7+8 varied in therange 10–20% among the individual seeds. The ratio of SFA7 to SFA8was highest in the VIR104-type and heterozygous seeds, with the amountof SFA7 exceeding that of SFA8 in six heterozygous seeds. Theproportions of SFA7 and SFA8 were inversely correlated among individualF₂ seeds. The results suggest that the amounts and proportions ofSFA7 and SFA8 are determined by genetic factors in addition toavailability of sulphur.