Vol 50, No 2 (2016)

To exploit the potential benefits of mycorrhizas, we need to investigate the processes that occur in these symbiotic interactions, particularly in the arbuscular compartment where nutrients are exchanged between the plant and the fungus. Progress in this area is restricted due to the intricacy and complexity of this plant-fungus interface and many techniques that have been employed successfully in other plants and animal systems cannot be used. An effective approach to study processes in arbuscules is to examine transcript composition and dynamics. We applied laser capture microdissection (LCM) to isolate approximately 3000 arbuscules from Glomus intraradices colonised Medicago truncatula roots. Total RNA was extracted from microdissected arbuscules and subjected to T7 RNA polymerase-based linear amplification. Amplified RNA was then used for construction of a cDNA library. The presence and level of enrichment of mycorrhiza-specific transcripts was determined by quantitative Real-time and conventional PCR. To improve enrichment a cDNA library subtraction was performed. Complementation of yeast mutants deficient in the uptake of potassium, phosphate, sulphate, amino acids, ammonium and of a Mn²⁺ sensitive strain, demonstrates the functionality of our cDNA library.

The barley genes Rpg5, RGA1 and Adf3, which provide a strong resistance to many pathotypes of stem rust, were cloned a few years ago, but it was still unclear whether their homologues were represented in wheat and in related species. The paper describes the results of a bioinformatic research to determine the homologues of Rpg5, RGA1 and Adf3 in the genomes of Triticum aestivum and several wild grasses, which breeders usually use as sources of stem rust resistance, and which are available in the genome databases. It was found that the Th. elongatum sequence Q9FEC6 and T. aestivum sequence Q43655 were the highly identical homologues of the Adf3 sequence. T. urartu M8A999 sequence and T. aestivum W5FCU1 sequence were found to be the closest homologues of Rpg5 complete protein sequence, but the identity of their kinase domains was not as clear as that of the other domains. The separate Rpg5 kinase part analysis did not provide the strong evidences that its orthologs were present in our corn species. T. urartu M7ZZX9 sequence and T. aestivum W5FFP0 and W5FI33 sequences were shown to be the homologues of RGA1. The analysis of the predicted active sites allowed finding out the difference between sequences of Rpg5, RGA1, Adf3 protein and their homologues.

Samples of trees and seed embryos of five Pinus kochiana Klotzsch ex Koch populations in Crimea were comparatively studied at 12 allozymic loci for its genetic variability and mating system. It has been shown that the embryos of seeds reproduce the allelic diversity of maternal plants, but the level of their observed heterozygosity (H_o) is significantly lower, i.e., 0.286 vs. 0.189, respectively. Unlike maternal trees, embryos are characterized by a significant divergence between the observed distribution of genotypes and the theoretically expected one according to the Hardy–Weinberg law for the majority of analyzed loci. The frequency of cross pollination in populations varied from 0.384 to 0.673 according to single-locus (t_s) estimates and from 0.639 to 0.841 according to multilocus (t_m) estimates.

The intercellular translocation of chromatin material along with other cytoplasmic contents among the proximate meiocytes lying in close contact with each other commonly referred as cytomixis was reported during microsporogenesis in Phaseolus vulgaris L., a member of the family Fabaceae. The phenomenon of cytomixis was observed at three administered doses of gamma rays viz. 100, 200, and 300 Gy respectively in the diploid plants of Phaseolus vulgaris L. The gamma rays irradiated plants showed the characteristic feature of inter-meiocyte chromatin/chromosomes transmigration through various means such as channel formation, beak formation or by direct adhesion between the PMC’s (Pollen mother cells). The present study also reports the first instance of syncyte formation induced via cytomictic transmigration in Phaseolus vulgaris L. Though the frequency of syncyte formation was rather low yet these could play a significant role in plant evolution. It is speculated that syncyte enhances the ploidy level of plants by forming 2n gametes and may lead to the production of polyploid plants. The phenomenon of cytomixis shows a gradual inclination along with the increasing treatment doses of gamma rays. The preponderance of cytomixis was more frequent during meiosis I as compared to meiosis II. An interesting feature noticed during the present study was the channel formation among the microspores and fusion among the tetrads due to cell wall dissolution. The impact of this phenomenon is also visible on the development of post-meiotic products. The formation of heterosized pollen grains; a deviation from the normal pollen grains has also been reported. The production of gametes with unbalanced chromosomes is of utmost importance and should be given more attention in future studies as they possess the capability of inducing variations at the genomic level and can be further utilized in the improvement of germplasm.

Stringent response is a pleiotropic physiological response of cells caused by deficiency of aminoacetylated tRNAs and, correspondingly, by the arrest of protein synthesis. This response can experimentally be induced by amino acid deficiency in a culture medium and limitation of the aminoacylation capacity of tRNA molecules even in the presence of the respective amino acids in the cell. Many traits of this response indicate its dependence on the accumulation of ppGpp molecules. There are links between the growth rate of actinomycetes and the biosynthesis of secondary metabolites by the bacteria. In particular, it has been established that the introduction of additional relA gene copies of the ppGpp synthetase can affect the production of antibiotics in streptomycetes. The survey presents the authors’ own experimental data obtained in the studies on the effect of heterologous relA gene expression in Streptomyces nogalater, the nogalamycin producer.