Assessment of genetic diversity and relatedness in the Latvian potato genetic resources collection by DArT genotyping

Potato (Solanum tuberosumL.) has been cultivated in Latvia since the 17th century, and formal breeding programmes have been established since the start of the 20th century. The Latvian potato genetic resource collection consists of 83 accessions of Latvian origin, including landraces, old cultivars released starting from the 1930's, modern cultivars and breeding material. These are maintained in field andin vitrocollections. Pedigree information about the potato cultivars is often limited, and the use of hybrids of local cultivars as parents is common in the Latvian potato breeding programme. Ninety-four Latvian potato varieties and breeding lines and some commonly used foreign accessions w…

Global effects of ade8 deletion on budding yeast metabolism

AbstractPurine auxotrophy is a typical marker for many laboratory yeast strains. Supplementation of additional purine source (like adenine) is necessary to cultivate these strains. If not supplied in adequate amounts, purine starvation sets in. We tested purine starvation effects in budding yeast Saccharomyces cerevisiae ade8 knockout. We explored effects brought by purine starvation in cellular, central carbon metabolism and in the global transcriptome level.We observed that cells cultivated in purine depleted media became significantly more tolerant to severe thermal, oxidative and desiccation stresses when compared to the cells cultivated in media with all necessary supplements. When sta…

Purine Auxotrophic Starvation Evokes Phenotype Similar to Stationary Phase Cells in Budding Yeast

Purine auxotrophy is an abundant trait among eukaryotic parasites and a typical marker for many budding yeast strains. Supplementation with an additional purine source (such as adenine) is necessary to cultivate these strains. If not supplied in adequate amounts, purine starvation sets in. We explored purine starvation effects in a model organism, a budding yeast Saccharomyces cerevisiae ade8 knockout, at the level of cellular morphology, central carbon metabolism, and global transcriptome. We observed that purine-starved cells stopped their cycle in G1/G0 state and accumulated trehalose, and the intracellular concentration of AXP decreased, but adenylate charge remained stable. Cells becam…

Miežu slimības izturības gēnu ģenētiskais un funkcionālais raksturojums.

Miežiem viens no miltrasas izturību nosakošajiem gēniem ir Mla. Molekulāro marķieru izstrāde šim lokusam ievērojami atvieglinātu selekcionāru darbu. Tāpēc šī darba mērķis ir izpētīt Mla lokusu Latvijas miežu selekcijas materiālā, definēt iespējamos molekulāros marķierus Mla lokusam, kas pielietojami Latvijas materiālā. Paša Mla lokusa uzbūves kompleksums neļauj izveidot marķierus gēna sekvencē, tādēļ izmantoti šo lokusu flankējoši reģioni (ABC15612, 538P8, 206i20_T7). Lokusi sekvenēti, lokusu haplotipu analīze parāda, ka pastāv saistība starp fenotipiskajām miltrasas izturības alēlēm un haplotipiem, kā arī Latvijas miežiem ir augsta daudzveidība flankējošos lokusos. Darbā identificēti haplo…

Adenine auxotrophy--be aware: some effects of adenine auxotrophy in Saccharomyces cerevisiae strain W303-1A.

Adenine auxotrophy is a commonly used genetic marker in haploid yeast strains. Strain W303-1A, which carries the ade2-1 mutation, is widely used in physiological and genetic research. Yeast extract-based rich medium contains a low level of adenine, so that adenine is often depleted before glucose. This could affect the cell physiology of adenine auxotrophs grown in rich medium. The aim of our study was to assess the effects of adenine auxotrophy on cell morphology and stress physiology. Our results show that adenine depletion halts cell division, but that culture optical density continues to increase due to cell swelling. Accumulation of trehalose and a coincident 10-fold increase in desicc…

Purine auxotrophy: Possible applications beyond genetic marker

Exploring new drug candidates or drug targets against many illnesses is necessary as "traditional" treatments lose their effectivity. Cancer and sicknesses caused by protozoan parasites are among these diseases. Cell purine metabolism is an important drug target. Theoretically, inhibiting purine metabolism could stop the proliferation of unwanted cells. Purine metabolism is similar across all eukaryotes. However, some medically important organisms or cell lines rely on their host purine metabolism. Protozoans causing malaria, leishmaniasis, or toxoplasmosis are purine auxotrophs. Some cancer forms have also lost the ability to synthesize purines de novo. Budding yeast can serve as an effect…

Immobilisation of yeast cells on the surface of hydroxyapatite ceramics

Hydroxyapatite (HAP) ceramics was tested for the first time for the possibility of being used as a new carrier for the immobilisation of yeast cells that are both model organisms for eukaryotic cell investigations and producers, which is important in classical and modern biotechnological processes. It was shown that under typical immobilisation conditions yeast (Saccharomyces cerevisiae) has no affinity to HAP ceramics. A novel method for yeast immobilisation was developed. This new method includes the joint incubation of a carrier with the cells, the sedimentation and adhesion of cells on the carrier and the dehydration of obtained preparations. It was shown that the sedimentation and adhe…

Whey lactose bioconversion to valuable products by non-conventional yeasts Kluyveromyces marxianus

Model-based biotechnological potential analysis of Kluyveromyces marxianus central metabolism

Abstract The non-conventional yeast Kluyveromyces marxianus is an emerging industrial producer for many biotechnological processes. Here, we show the application of a biomass-linked stoichiometric model of central metabolism that is experimentally validated, and mass and charge balanced for assessing the carbon conversion efficiency of wild type and modified K. marxianus. Pairs of substrates (lactose, glucose, inulin, xylose) and products (ethanol, acetate, lactate, glycerol, ethyl acetate, succinate, glutamate, phenylethanol and phenylalanine) are examined by various modelling and optimisation methods. Our model reveals the organism’s potential for industrial application and metabolic engi…

Utilization of different carbohydrates by various Kluyveromyces marxianus strains