Rapid identification of Triticeae genotypes from single seeds using the polymerase chain reaction

An easy and quick protocol has been developed for DNA analysis via PCR. Single cereal endosperm or small leaf pieces can be separately processed in several PCR reactions. The resultant PCR patterns are equivalents to those obtained with standard DNA extraction protocols using either specific or random primers. Intra-and inter-specific variability can be detected. This method allows the analysis of a large number of individuals in early stages prior to the plant sowing.     

Pollen germination and tube growth in rye (Secade cereale L.) homozygous and heterozygous for reciprocal translocations

Summary To contribute to the knowledge of the role of reciprocal translocations in rye, a component of fertility was estimated by comparing germination and pollen tube growth in homozygous and heterozygous plants for reciprocal translocations. The results obtained indicate that there are no differences in germination and pollen tube growth rate when homozygous and heterozygous plants as a whole are compared. However, there are significant differences in pollen tube growth between plants carrying different translocations. This suggests that the chromosome constitution of a plant is relevant to these fitness-estimating parameters together with its particular genetic background.     

mTOR activates the VPS34–UVRAG complex to regulate autolysosomal tubulation and cell survival

Lysosomes are essential organelles that function to degrade and recycle unwanted, damaged and toxic biological components. Lysosomes also act as signalling platforms in activating the nutrient‐sensing kinase mTOR. mTOR regulates cellular growth, but it also helps to maintain lysosome identity by initiating lysosomal tubulation through a process termed autophagosome‐lysosome reformation (ALR). Here we identify a lysosomal pool of phosphatidylinositol 3‐phosphate that, when depleted by specific inhibition of the class III phosphoinositide 3‐kinase VPS34, results in prolonged lysosomal tubulation. This tubulation requires mTOR activity, and we identified two direct mTOR phosphorylation sites on UVRAG (S550 and S571) that activate VPS34. Loss of these phosphorylation sites reduced VPS34 lipid kinase activity and resulted in an increase in number and length of lysosomal tubules. In cells in which phosphorylation at these UVRAG sites is disrupted, the result of impaired lysosomal tubulation alongside ALR activation is massive cell death. Our data imply that ALR is critical for cell survival under nutrient stress and that VPS34 is an essential regulatory element in this process.     

Biochemical evidence of homoeology between Triticum aestivum and Agropyron intermedium chromosomes

Summary The alcohol dehydrogenase (ADH), phosphoglucose mutase (PGM), glucosephosphate isomerase (GPI), glutamic oxaloacetic transaminase (GOT), malate dehydrogenase (MDH), leaf esterases (ESTL), leaf acid (ACPH) and endosperm alkaline (PHE) phosphatases, leaf peroxidases (PERL) zymogram phenotypes of Triticum aestivum, Agropyron intermedium, Triticum aestivum — Agropyron intermedium octoploids and six Agropyron intermedium chromosome additions to Triticum aestivum and two ditelocentric addition lines were determined. It was found that the six disomic chromosome addition lines and one ditelocentric chromosome addition line could be distinguished from one another and from the other possible lines on the basis of the zymogram phenotypes of these isozymes. The structural gene Acph-X1 was located on Agropyron chromosome L1, the genes Got-X3 and Mdh-X2 on chromosome L2, the gene Gpi-X1 on chromosome L3, the genes Adh-X1, Pgm-X1 and Phe-3 on chromosome L4, gene Perl-1 on chromosome L5 and the gene Estl-2 on chromosome L7 and chromosome arm L7d2. These gene locations provide evidence of homoeology between Agropyron chromosomes L1, L2, L3, L4, L5 and L7 and the Triticum aestivum chromosomes of homoeologous groups 7, 3, 1, 4, 2 and 6, respectively.     

Genetics of rye phosphatases: evidence of a duplication

Summary Genetic analyses were conducted on alkaline phosphatases of the endosperm of dry kernels and leaf acid phosphatases in four open pollinated and one inbred line of cultivated rye (Secale cereale L.). A total of seven alkaline phosphatase isozymes were observed occurring at variable frequencies in the different cultivars analyzed. We propose that at least five loci control the alkaline phosphatases of rye endosperm — Alph-1, Alph-2, Alph-3, Alph-4 and Alph-5 — all of which have monomeric behaviour. The leaf acid phosphatases are controlled by one locus and have a dimeric quaternary structure. All loci coding for alkaline phosphatase isozymes showed one active, dominant allele and one null, recessive allele, except for the locus Alph-3 which showed two active, dominant alleles and one null, recessive one. The linkage analyses suggest the existence of two linkage groups for alkaline phosphatases: one of them would contain Alph-2, Alph-4, Alph-5 and the locus/loci coding isozymes 6 and 7. This linkage group is located in the 7RS chromosome arm. The other group would include Alph-1 and Alph-3 loci, being located in the 1RL chromosome arm. Leaf acid phosphatases have been previously located in the 7RL chromosome arm. Our data also support an independent relationship between loci controlling the endosperm alkaline phosphatases and leaf acid phosphatases.     

On the consequences of aluminium stress in rye: repression of two mitochondrial malate dehydrogenase mRNAs

Plants have developed several external and internal aluminium (Al) tolerance mechanisms. The external mechanism best characterised is the exudation of organic acids induced by Al. Rye (Secale cereale L.), one of the most Al‐tolerant cereal crops, secretes both citrate and malate from its roots in response to Al. However, the role of malate dehydrogenase (MDH) genes in Al‐induced stress has not been studied in rye. We have isolated the ScMDH1 and ScMDH2 genes, encoding two different mitochondrial MDH isozymes, in three Al‐tolerant rye cultivars (Ailés, Imperial and Petkus) and one sensitive inbred rye line (Riodeva). These genes, which have seven exons and six introns, were located on the 1R (ScMDH1) and 3RL (ScMDH2) chromosomes. Exon 1 of ScMDH1 and exon 7 of ScMDH2 were the most variable among the different ryes. The hypothetical proteins encoded by these genes were classified as putative mitochondrial MDH isoforms. The phylogenetic relationships obtained using both cDNA and protein sequences indicated that the ScMDH1 and ScMDH2 proteins are orthologous to mitochondrial MDH1 and MDH2 proteins of different Poaceae species. The expression studies of the ScMDH1 and ScMDH2 genes indicate that it is more intense in roots than in leaves. Moreover, the amount of their corresponding mRNAs in roots from plants treated and not treated with Al was higher in the tolerant cultivar Petkus than in the sensitive inbred line Riodeva. In addition, ScMDH1 and ScMDH2 mRNA levels decreased in response to Al stress (repressive behaviour) in the roots of both the tolerant Petkus and the sensitive line Riodeva.     

Evolution of androgen-regulated mRNA expression in mouse kidney

To gain information on the evolution of mammalian gene expression patterns, we studied the androgen-inducible expression of three kidney mRNAs in several mouse species (genus Mus). The RP2, ornithine decarboxylase, and beta-glucuronidase mRNAs have each evolved independently, in that the pattern of variation among species is unique for each. This suggests a role for gene-specific, cis-acting genetic elements. Relationships between the regulatory phenotypes and the species phylogeny suggest that the variations in hormone-inducible mRNA expression were generated by a series of independent mutations that occurred in specific lineages, resulting in modifications of the progenitor phenotype. Alternatively, the variations may have preexisted within the progenitor population as polymorphisms that were fixed during establishment of individual lineages. Thus, significant alterations in the androgen-regulated mRNA phenotype have occurred either prior to or during speciation within the Mus genus. These alterations are presumed to be in regulatory sequences that control the expression of the corresponding genes and their response to testosterone; as such, they should be useful in further studying the genetic determinants of gene expression and its evolution.      

PCR derived molecular markers and phylogenetic relationships in theSecale genus

DNA from 22 different species, accessions, cultivars and lines included in theSecale genus were analyzed by the polymerase chain reaction (PCR), using as primers five pairs of oligonucleotides derived from specific sequences. A total of 42 amplified bands were considered, and some of them appeared to be potentially useful as molecular markers for some of the analyzed groups. These amplified bands were used to generate molecular phenograms inside theSecale genus.     

Red tide assemblage formation in an estuarine upwelling ecosystem: Ria de Vigo

Red tides are conspicuous in the upwelling system of Galicia(NW Iberian Peninsula). At present, there are conflicting hypothesesabout the generation site of these phytoplankton assemblages.It is interesting to know whether the rias can be sites of redtide formation or if they act only as accumulation sites ofpopulations advected from shelf waters. A study in the Ra deVigo, carried out during late September 1990, showed the developmentof a red tide assemblage, composed of Alexandrium affinis, Ceraiiumfusus and Gymnodinium catenaium, during a 2 week upwelling-downwellingcycle. Growth occurred at the bottom of the thermocline-topof the nutricline. Above this assemblage, a diatom assemblage(large diatoms) was blooming. Prior to the formation of thered tide, a subsurface chlorophyll maximum made up of smalldiatoms (Nilzschia f. seriaia, Chaeloceros socialis), smallflagellates (<30 µm) and small gymnodinid forms (<30µm) was observed. In the nutrient-depleted upper layer,several autotrophic and large heterotrophic dinoflagellatesdominated. It is suggested that the ratio between the velocityof upward water movement and the depth of the stratified upperlayer (flushing rate, day^–1) is the critical parameterwhich triggers active phytoplankton growth. It can be concludedthat upward water velocities of {small tilde}2.5 m day^–1and a stratified upper layer of 10 m depth (flushing rate 0.25day^–1) are the main physical constraints for red tidedevelopment.     

High mutability in rye (Secale cereale L.).

Analysis of the rye cultivar Ailés of several descents derived from crosses between plants carrying specific genotypes and/or chromosome constitutions resulted in the detection of high chromosome (2.05 x 10(-2)) and gene (9.3 x 10(-3)) mutation frequencies. The existence of a transposon system responsible for this instability is suggested.