Nigrosine staining of wheat endosperm proteolipid patterns on starch gels

We have previously characterized a group of proteolipids from wheat endosperm, designated CM proteins, which are soluble in chloroformmethanol (2:1, v/v) and have a molecular weight lower than 25,000 daltons (1-3). These have been also studied by Redman and Ewart (4). The CM proteins are suitably fractionated into several components by starch gel electrophoresis at pH 3.2 (1). A sensitive staining procedure was required in connection with genetic studies of these proteins because phenotypes had to be ascertained in small endosperm fractions dissected without impairing normal germination and plant development. We report here on Nigrosine staining conditions for CM proteins under which high sensitivity and selectivity are achieve

HETEROGENEITY OF WHEAT ENDOSPERM PROTEOLIPIDS (CM PROTEINS)

Abstract-Proteins extracted with CHCl 3 -MeOH from wheat endosperm have been fractionated by Sephadex G-100 and the 15 000-20 000 MW range fraction, designated CM protein, has been examined by combined electrofocusing (pH range 5-8) and electrophoresis (pH 3-2) and the heterogeneity of the electrophoretic components has been ascertained. It has been shown by joint mapping and by sequential extraction that CM proteins are extracted by 70% EtOH but not by H 2 0, although they can be made water-soluble after dialysis against an acid buffer, pH 3-2, 3 M urea, without losing their solubility in CHCl 3 -MeOH mixtures. It isconcluded that CM proteins fit the definition of a Folch-Lees proteolipid. The Triticum aestivum (genomes ABD) map can be reconstructed by mixing T. durum (AB) and Aegilops squarrosa (D). The low intragenomic variability of CM protein is confirmed

Heterogeneity of wheat endosperm proteolipids (CM proteins)

Proteins extracted with CHCl3-MeOH from wheat endosperm have been fractionated by Sephadex G-100 and the 15 000–20 000 MW range fraction, designated CM protein, has been examined by combined electrofocusing (pH range 5–8) and electrophoresis (pH 3.2) and the heterogeneity of the electrophoretic components has been ascertained. It has been shown by joint mapping and by sequential extraction that CM proteins are extracted by 70% EtOH but not by H2O, although they can be made water-soluble after dialysis against an acid buffer, pH 3.2, 3 M urea, without losing their solubility in CHCl3-MeOH mixtures. It is concluded that CM proteins fit the definition of a Folch—Lees proteolipid. The Triticum aestivum (genomes ABD) map can be reconstructed by mixing T. durum (AB) and Aegilops squarrosa (D). The low intragenomic variability of CM protein is confirmed

Equivalent locations of sucrose synthase genes in chromosomes 7D of wheat, 7Ag of Agropyron elongatum, and 7H of barley

Wheat/Agropyron 7D/7Ag recombinant chromosomes were analysed by Southern blotting, using cDNA radioactive probes corresponding to the Ss1 and Ss2 sucrose synthase genes of wheat. The genes were located in the central segments of the short arms of both chromosomes 7D and 7Ag. A similar analysis of wheat/barley addition lines demonstrated that DNA sequences that cross-hybridized with the wheat probes were located in chromosome 7H of barley

Eyespot resistance gene Pch-1 from Aegilops ventricosa is associated with a different chromosome in wheat line H-93-70 than the resistance factor in "Roazon" wheat

The hexaploid wheat line H-93-70 carries a gene (Pch-1) that has been transferred from the wild grass Aegilops ventricosa and confers a high degree of resistance to eyespot diesease, caused by the fungus Pseudocercosporella herpotrichoides. Crosses of the resistant line H-93-70 with the susceptible wheat Pané 247 and with a 7D/7Ag wheat/Agropyron substitution line were carried out and F2 kernels were obtained. The kernels were cut transversally and the halves carrying the embryos were used for the resistance test, while the distal halves were used for genetic typing. Biochemical markers were used to discriminate whether the transferred Pch-1 gene was located in chromosome 7D, as is the case for a resistance factor present in Roazon wheat. In the crosses involving Pané 247, resistance was not associated with the 7D locus Pln, which determines sterol ester pattern (dominant allele in H-93-70). In the crosses with the 7D/7Ag substitution line, resistance was neither associated with protein NGE-11 (7D marker), nor alternatively inherited with respect to protein C-7 (7Ag marker). It is concluded that gene Pch-1 represents a different locus and is not an allele of the resistance factor in Roazon whea

Chromosomal control of non-gliadin proteins from the 70% ethanol extract of wheat endosperm

The non-gliadin fraction of the 70% ethanol extracts of compensated nulli-tetrasomics and ditelosomics of Triticum aestivum cv. Chinese Spring has been analyzed by combined electrofocusing and electrophoresis. Seventeen of the 21 protein map components of the euploid have been ascribed to eight chromosomes: 4A, 3BS, 6BS, 7BS, 3D, 4D, 5D and 7DS. The relationship of the different map components with other proteins previously associated with the same chromosomes is discusse

Ingeniería molecular en "Triticum"

Se resumen las investigaciones realizadas sobre genética bioquímica de la especie aloploide de Triticum aestivum L. (trigo común). Dichas investigaciones abarcan los siguientes aspectos: a) localización cromosómica de genes que controlan sistemas moleculares; b) formas de interacción entre los genomios integrados en dicha especie; c) transferencia genética extraespecífica y extragenérica desde Aegilops ventricosa, Sécale cereale y Agropyron elongatum a trigo. Se especula sobre las implicaciones evolutivas de los resultados obtenidos y se enumeran las aplicaciones prácticas

A tetrameric inhibitor of insect α-amylase from barley

A tetrameric inhibitor that is active against α-amylase from the larvae of the insect Tenebrio molitor, but inactive against the enzyme from human saliva and against the endogenous one, has been described in barley endosperm. The subunits of the inhibitor have been identified as the previously characterized proteins CMa, CMb and CMd, of which only CMa was inhibitory by itself

N-terminal amino acid sequences of chloroform/methanol-soluble proteins and albumins from endosperms of wheat, barley and related species: Homology with inhibitors of α-amylase and trypsin and with 2 S storage globulins

The N-terminal amino acid sequences of two chloroform/methanol soluble globulins from barley and one form wheat are reported. They are homologous with N-terminal sequences previously reported for α-amylase and trypsin inhibitors from cereals and 2 S storage proteins from castor bean and rape. Three albumins were also purified from Aegilops squarrosa and Triticum monococcum. These had N-terminal amino acid sequences most closely related to the α-amylase and trypsin inhibitors. The relationships of this superfamily of seed proteins are discussed

Genetics of CM-proteins (A-hordeins) in barley

The CM-proteins, which are the main components of the A-hordeins, include four previously described proteins (CMa-1, CMb-1, CMc-1, CMd-1), plus a new one, CMe-1, which has been tentatively included in this group on the basis of its solubility properties and electrophoretic mobility. The variability of the five proteins has been investigated among 38 Hordeum vulgare cultivars and 17 H. spontaneum accessions. Proteins CMa-1, CMc-1 and CMd-1 were invariant within the cultivated species; CMd was also invariant in the wild one. The inheritance of variants CMb-1/CMb-2 and CMe-1/CMe-2,2 was studied in a cross H. spontaneum x H. vulgare. The first two proteins were inherited as codominantly expressed allelic variations of a single mendelian gene. Components CMe-2,2 were jointly inherited and codominantly expressed with respect to CMe-1. Gene CMb and gene(s) CMe were found to be unlinked. The chromosomal locations of genes encoding CM-proteins were investigated using wheat-barley addition lines. Genes CMa and CMc were associated with chromosome 1, and genes CMb and CMd with chromosome 4. These gene locations further support the proposed homoeology of chromosomes 1 and 4 of barley with chromosomes groups 7 and 4 of wheat, respectively. Gene(s) CMe has been assigned to chromosome 3 of barley. The accumulation of protein CMe-1 is totally blocked in the high lysine mutant Riso 1508 and partially so in the high lysine barley Hiproly