Dr. Emill Molle is an Associate Professor at the University of Forestry in Sofia, Bulgaria. His most recent areas of expertise comprise of Modeling in Ecology, Biostatistics, and Bioinformatics. His main focus in selecting the teaching materials and forming the curriculum for his classes is the use of the Open Source environment R. Most of Dr. Molle’s current research projects are conducted under the auspices of The Faculties of Forestry and Agriculture at the UoF.
The results described in these studies proved that the successful in vitro bioproduction of galan... more The results described in these studies proved that the successful in vitro bioproduction of galanthamine from L. aestivum shoot-clumps required mainly the selection of in vitro clones with a genetically determined high ability to produce the desired alkaloids, although the expression of this ability could be additionally influenced by diverse exterior factors, such as some components of the nutrient medium, or the cultivation conditions of the ambience. Tissue differentiation was also of great importance for the biosynthetic capacity of the cultures. The most suitable inocula for in vitro biosynthesis of galanthamine in liquid medium were the directly regenerated shoot-clumps, ensuring high alkaloid concentrations between 1 and 2 mg/g DW for the selected clones. We observed astonishing clone-specific dynamics of the biosynthetic activity of all of the studied in vitro clones. The dynamics were obviously related to the strong biological clock of the species, persisting even in several-year old cultures. These dynamics did not coincide with those usual for the plants growing in situ and under controlled field conditions. In our opinion, the clone specificity of the biosynthetic dynamics could be due to the disturbance of the plant regulation mechanism under the equal conditions of the ambience in the culture room. The sharp decrease of the alkaloid concentrations were transient, followed by an increase, so that cultures were retaining their biosynthetic capacity. The biosynthesis of the main alkaloids, galanthamine and lycorine, was influenced by diverse stimulants such as substances causing stress (JA), feeding with alkaloid precursors (the amino acids phenylalanine and tyrosine, and CH), and physical treatment (acoustic waves). However, the course of the biosynthetic dynamics during the period of the treatments was always the most important factor for the success of secondary metabolism stimulation. As far as scaling-up of the in vitro biosynthesis of valuable compounds, a stable and predictable yield is required, and additional investigations aimed at the annulment of the effect plant biological clock on alkaloid biosynthesis are needed. The elucidation of the relative influences of the diverse factors modulating alkaloid biosynthesis was of great importance. The high galanthamine concentrations of the selected in vitro clones are a promising basis for future studies.
Comptes rendus de l'Académie bulgare des sciences: sciences mathématiques et naturelles
Directly regenerated shoot-clumps of Leucojum aestivum manifested high biosynthetic capacity duri... more Directly regenerated shoot-clumps of Leucojum aestivum manifested high biosynthetic capacity during long-term cultivation conditions. However, the contents and the proportions of the main alkaloids galanthamine (Gal) and lycorine (Lyc) depended on the culture genotype and were distinguished for their fluctuations in the course of time. Addition of the precursors tyrosine and phenylalanine in the medium enhanced the biosynthetic activity: Gal of the best clone doubled its content reaching up to 1.94 mg/g DW, and Lyc was up to 5.68 mg/g DW which was several times more than the control. Most important for the success of the biosynthesis stimulation effect was the course of the alkaloid dynamics during the period of treatment.
In vitro clones of Leucojum aestivum cultured 4 years under equal conditions on agar MS medium su... more In vitro clones of Leucojum aestivum cultured 4 years under equal conditions on agar MS medium supplemented with BAP and NAA kept their inherent alkaloid profiles. However, the contents of the main alkaloids Gal and Lyc varied in the course of time.The dynamics were studied in 12 clones for at least 24 months (HPLC analysis every 3 months). Clones differed in the way of the changes of their alkaloid content. Different correlations were proved, using R, between the fluctuations of Gal, Lyc and cultures' dry matter of some clones and between clones.
ABSTRACT The purpose of this study is to obtain a comparative assessment of dihaploide lines and ... more ABSTRACT The purpose of this study is to obtain a comparative assessment of dihaploide lines and oriental tobacco varieties for number of genetic parameters applying the combinatory approach to diallel analysis. Two basic morphological traits “plant height” and “leaf number” are investigated. The conclusions valid for both the dihaploid varieties and dihaploid lines are that dominance is the basic inheritance type of the traits and the dihaploid lines arc more stable in some aspects.
The influence of variable concentrations of the nutritive components of Gauze 1 medium on the β-l... more The influence of variable concentrations of the nutritive components of Gauze 1 medium on the β-lactamase synthesis by Streptomyces galbus (F) subsp. achromogenes—p154–59 has been studied. Twelve carbon, ten nitrogen, and four phosphate sources have been examined and the data has been used for carrying out Complete Factor Experiment (CFE 23). As a result of our experiments a complete optimisation of the fermentation medium has been obtained which enhances ten-fold β-lactamase synthesis by Streptomyces galbus.
Comptes rendus de l'Académie bulgare des sciences: sciences mathématiques et naturelles
Directly regenerated shoot-clumps of Leucojum aestivum manifested high biosynthetic capacity duri... more Directly regenerated shoot-clumps of Leucojum aestivum manifested high biosynthetic capacity during long-term cultivation conditions. However, the contents and the proportions of the main alkaloids galanthamine (Gal) and lycorine (Lyc) depended on the culture genotype and were distinguished for their fluctuations in the course of time. Addition of the precursors tyrosine and phenylalanine in the medium enhanced the biosynthetic activity: Gal of the best clone doubled its content reaching up to 1.94 mg/g DW, and Lyc was up to 5.68 mg/g DW which was several times more than the control. Most important for the success of the biosynthesis stimulation effect was the course of the alkaloid dynamics during the period of treatment.
In vitro clones of Leucojum aestivum cultured 4 years under equal conditions on agar MS medium su... more In vitro clones of Leucojum aestivum cultured 4 years under equal conditions on agar MS medium supplemented with BAP and NAA kept their inherent alkaloid profiles. However, the contents of the main alkaloids Gal and Lyc varied in the course of time.The dynamics were studied in 12 clones for at least 24 months (HPLC analysis every 3 months). Clones differed in the way of the changes of their alkaloid content. Different correlations were proved, using R, between the fluctuations of Gal, Lyc and cultures' dry matter of some clones and between clones.
ABSTRACT The effect of three mutated genes affecting anthocyanin biosynthesis in tomato (anthocya... more ABSTRACT The effect of three mutated genes affecting anthocyanin biosynthesis in tomato (anthocyaninless of Hoffmann - ah, anthocyaninwithout - aw and baby lea syndrome - bls) on germination capacity of the brown seed (bs) mutant was investigated. Mutations brown seed ( bs, bs 2, bs 4) controlling darkness of the seed testa are known to have a negative impact on the velocity of seed germination. In contrast, the three anthocyaninless mutants, characterised by absence of condensed tannins in the testa inner epidermal layer were found to enhance germination under optimal and stress conditions. The study was performed on a series of isogenic/near isogenic lines (IL/NILs) as follows: Ailsa Craig (AC), AC bs, AC ah, AC aw, AC bls, AC ah bs, AC aw bs and AC bls bs. Under optimal conditions, low temperature and salt stress, the anthocyaninless IL/NILs possessing brown seeds germinated more rapidly than the seeds of the anthocyanin-containing bs line (AC bs), the differences in time to 50% germination being significant. The histochemical tests of the seed coat provided evidence that, as in ah, aw and bls mutants, no condensed tannins were detected in the testa of double-mutant seeds. We concluded that the absence of condensed tannins in the testa of the brown seeds contributed to their more rapid germination.
... Bulgarian Academy of Sciences, Department of Cytogenetics, Tzarigradsko chosse, 13 km., Sofia... more ... Bulgarian Academy of Sciences, Department of Cytogenetics, Tzarigradsko chosse, 13 km., Sofia 1113, Bulgaria 3 University of Sofia, St. Kl. ... that might contribute to their increased moisture uptake during storage, determined by Villers and Edgcumbe (1975) as a factor causing ...
in Vitro Cellular & Developmental Biology-plant, 2009
The investigation deals with in vitro clonal propagation of L. aestivum L. (summer snowflake), a ... more The investigation deals with in vitro clonal propagation of L. aestivum L. (summer snowflake), a threatened Amaryllidaceae plant species in Bulgaria used in the pharmaceutical industry as raw material for production of galanthamine-based medicines. Plants of known origin and with different alkaloid profile were taken from the living collection of the Institute of Botany, Sofia. Bulbs were used to initiate in vitro cultures and 24 clones were multiplied. The influence of the clone origin on the propagation coefficient, shoot and bulblet morphology, alkaloid profile and content of galanthamine, lycorine, and four related alkaloids was evaluated. Clones kept stable alkaloid profiles and for most of them, high regeneration rates were noted. Galanthamine content of some clones was commensurable with that of Bulgarian populations of L. aestivum of commercial importance. Five clones: four galanthamine-type and one lycorine-type were selected as promising for further investigation.
ABSTRACT Fragile X syndrome is a genetic neuro-developmental disorder, caused by the transcriptio... more ABSTRACT Fragile X syndrome is a genetic neuro-developmental disorder, caused by the transcriptional inactivation of the gene Fmr1(Fragile X mental retardation 1). The lack of its protein product (FMRP) is accompanied by defects in synaptic maturation and morphology and by synaptic dysfunction throughout the nervous system. FMRP is an RNA-binding protein, which interacts with specific neuronal mRNA-targets and with other proteins and is involved in mRNA-transport, stability and translation Drosophila model closely recapitulates the neuronal defects of the clinical disease phenotype and its FMRP homolog (dFMRP) shares similar biochemical features. Using this model and the GAL4/UAS system for targeted gene expression, we performed genetic interaction experiments. We over-expressed the gene dfmr1 (Drosophia fragile X mental retardation 1) in the developing wing imaginal discs and looked for possible suppressor/enhancer effects of the genes scrib(sribbled) and dlg1(discs large 1). We also looked for similar genetic interactions in the adult brain neurons. We found that scrib acted as an enhancer of the mutant wing phenotype, induced by the over-expression of dfmr1. scrib also enhanced the mutant synaptic phenotype in the adult brain neurons, caused by the overexpression or lack of expression of dfmr1. dlg1 had an opposite suppressor effect in both systems analyzed. It rescued the mutant wing phenotype and the mutant synaptic phenotype as well. Our results demonstrate that dfmr1 interacts genetically with the members of the Lgl-family of scaffolding proteins and functions with them in the synaptic architecture of Drosophila adult brain neurons.
The results described in these studies proved that the successful in vitro bioproduction of galan... more The results described in these studies proved that the successful in vitro bioproduction of galanthamine from L. aestivum shoot-clumps required mainly the selection of in vitro clones with a genetically determined high ability to produce the desired alkaloids, although the expression of this ability could be additionally influenced by diverse exterior factors, such as some components of the nutrient medium, or the cultivation conditions of the ambience. Tissue differentiation was also of great importance for the biosynthetic capacity of the cultures. The most suitable inocula for in vitro biosynthesis of galanthamine in liquid medium were the directly regenerated shoot-clumps, ensuring high alkaloid concentrations between 1 and 2 mg/g DW for the selected clones. We observed astonishing clone-specific dynamics of the biosynthetic activity of all of the studied in vitro clones. The dynamics were obviously related to the strong biological clock of the species, persisting even in several-year old cultures. These dynamics did not coincide with those usual for the plants growing in situ and under controlled field conditions. In our opinion, the clone specificity of the biosynthetic dynamics could be due to the disturbance of the plant regulation mechanism under the equal conditions of the ambience in the culture room. The sharp decrease of the alkaloid concentrations were transient, followed by an increase, so that cultures were retaining their biosynthetic capacity. The biosynthesis of the main alkaloids, galanthamine and lycorine, was influenced by diverse stimulants such as substances causing stress (JA), feeding with alkaloid precursors (the amino acids phenylalanine and tyrosine, and CH), and physical treatment (acoustic waves). However, the course of the biosynthetic dynamics during the period of the treatments was always the most important factor for the success of secondary metabolism stimulation. As far as scaling-up of the in vitro biosynthesis of valuable compounds, a stable and predictable yield is required, and additional investigations aimed at the annulment of the effect plant biological clock on alkaloid biosynthesis are needed. The elucidation of the relative influences of the diverse factors modulating alkaloid biosynthesis was of great importance. The high galanthamine concentrations of the selected in vitro clones are a promising basis for future studies.
Comptes rendus de l'Académie bulgare des sciences: sciences mathématiques et naturelles
Directly regenerated shoot-clumps of Leucojum aestivum manifested high biosynthetic capacity duri... more Directly regenerated shoot-clumps of Leucojum aestivum manifested high biosynthetic capacity during long-term cultivation conditions. However, the contents and the proportions of the main alkaloids galanthamine (Gal) and lycorine (Lyc) depended on the culture genotype and were distinguished for their fluctuations in the course of time. Addition of the precursors tyrosine and phenylalanine in the medium enhanced the biosynthetic activity: Gal of the best clone doubled its content reaching up to 1.94 mg/g DW, and Lyc was up to 5.68 mg/g DW which was several times more than the control. Most important for the success of the biosynthesis stimulation effect was the course of the alkaloid dynamics during the period of treatment.
In vitro clones of Leucojum aestivum cultured 4 years under equal conditions on agar MS medium su... more In vitro clones of Leucojum aestivum cultured 4 years under equal conditions on agar MS medium supplemented with BAP and NAA kept their inherent alkaloid profiles. However, the contents of the main alkaloids Gal and Lyc varied in the course of time.The dynamics were studied in 12 clones for at least 24 months (HPLC analysis every 3 months). Clones differed in the way of the changes of their alkaloid content. Different correlations were proved, using R, between the fluctuations of Gal, Lyc and cultures' dry matter of some clones and between clones.
ABSTRACT The purpose of this study is to obtain a comparative assessment of dihaploide lines and ... more ABSTRACT The purpose of this study is to obtain a comparative assessment of dihaploide lines and oriental tobacco varieties for number of genetic parameters applying the combinatory approach to diallel analysis. Two basic morphological traits “plant height” and “leaf number” are investigated. The conclusions valid for both the dihaploid varieties and dihaploid lines are that dominance is the basic inheritance type of the traits and the dihaploid lines arc more stable in some aspects.
The influence of variable concentrations of the nutritive components of Gauze 1 medium on the β-l... more The influence of variable concentrations of the nutritive components of Gauze 1 medium on the β-lactamase synthesis by Streptomyces galbus (F) subsp. achromogenes—p154–59 has been studied. Twelve carbon, ten nitrogen, and four phosphate sources have been examined and the data has been used for carrying out Complete Factor Experiment (CFE 23). As a result of our experiments a complete optimisation of the fermentation medium has been obtained which enhances ten-fold β-lactamase synthesis by Streptomyces galbus.
Comptes rendus de l'Académie bulgare des sciences: sciences mathématiques et naturelles
Directly regenerated shoot-clumps of Leucojum aestivum manifested high biosynthetic capacity duri... more Directly regenerated shoot-clumps of Leucojum aestivum manifested high biosynthetic capacity during long-term cultivation conditions. However, the contents and the proportions of the main alkaloids galanthamine (Gal) and lycorine (Lyc) depended on the culture genotype and were distinguished for their fluctuations in the course of time. Addition of the precursors tyrosine and phenylalanine in the medium enhanced the biosynthetic activity: Gal of the best clone doubled its content reaching up to 1.94 mg/g DW, and Lyc was up to 5.68 mg/g DW which was several times more than the control. Most important for the success of the biosynthesis stimulation effect was the course of the alkaloid dynamics during the period of treatment.
In vitro clones of Leucojum aestivum cultured 4 years under equal conditions on agar MS medium su... more In vitro clones of Leucojum aestivum cultured 4 years under equal conditions on agar MS medium supplemented with BAP and NAA kept their inherent alkaloid profiles. However, the contents of the main alkaloids Gal and Lyc varied in the course of time.The dynamics were studied in 12 clones for at least 24 months (HPLC analysis every 3 months). Clones differed in the way of the changes of their alkaloid content. Different correlations were proved, using R, between the fluctuations of Gal, Lyc and cultures' dry matter of some clones and between clones.
ABSTRACT The effect of three mutated genes affecting anthocyanin biosynthesis in tomato (anthocya... more ABSTRACT The effect of three mutated genes affecting anthocyanin biosynthesis in tomato (anthocyaninless of Hoffmann - ah, anthocyaninwithout - aw and baby lea syndrome - bls) on germination capacity of the brown seed (bs) mutant was investigated. Mutations brown seed ( bs, bs 2, bs 4) controlling darkness of the seed testa are known to have a negative impact on the velocity of seed germination. In contrast, the three anthocyaninless mutants, characterised by absence of condensed tannins in the testa inner epidermal layer were found to enhance germination under optimal and stress conditions. The study was performed on a series of isogenic/near isogenic lines (IL/NILs) as follows: Ailsa Craig (AC), AC bs, AC ah, AC aw, AC bls, AC ah bs, AC aw bs and AC bls bs. Under optimal conditions, low temperature and salt stress, the anthocyaninless IL/NILs possessing brown seeds germinated more rapidly than the seeds of the anthocyanin-containing bs line (AC bs), the differences in time to 50% germination being significant. The histochemical tests of the seed coat provided evidence that, as in ah, aw and bls mutants, no condensed tannins were detected in the testa of double-mutant seeds. We concluded that the absence of condensed tannins in the testa of the brown seeds contributed to their more rapid germination.
... Bulgarian Academy of Sciences, Department of Cytogenetics, Tzarigradsko chosse, 13 km., Sofia... more ... Bulgarian Academy of Sciences, Department of Cytogenetics, Tzarigradsko chosse, 13 km., Sofia 1113, Bulgaria 3 University of Sofia, St. Kl. ... that might contribute to their increased moisture uptake during storage, determined by Villers and Edgcumbe (1975) as a factor causing ...
in Vitro Cellular & Developmental Biology-plant, 2009
The investigation deals with in vitro clonal propagation of L. aestivum L. (summer snowflake), a ... more The investigation deals with in vitro clonal propagation of L. aestivum L. (summer snowflake), a threatened Amaryllidaceae plant species in Bulgaria used in the pharmaceutical industry as raw material for production of galanthamine-based medicines. Plants of known origin and with different alkaloid profile were taken from the living collection of the Institute of Botany, Sofia. Bulbs were used to initiate in vitro cultures and 24 clones were multiplied. The influence of the clone origin on the propagation coefficient, shoot and bulblet morphology, alkaloid profile and content of galanthamine, lycorine, and four related alkaloids was evaluated. Clones kept stable alkaloid profiles and for most of them, high regeneration rates were noted. Galanthamine content of some clones was commensurable with that of Bulgarian populations of L. aestivum of commercial importance. Five clones: four galanthamine-type and one lycorine-type were selected as promising for further investigation.
ABSTRACT Fragile X syndrome is a genetic neuro-developmental disorder, caused by the transcriptio... more ABSTRACT Fragile X syndrome is a genetic neuro-developmental disorder, caused by the transcriptional inactivation of the gene Fmr1(Fragile X mental retardation 1). The lack of its protein product (FMRP) is accompanied by defects in synaptic maturation and morphology and by synaptic dysfunction throughout the nervous system. FMRP is an RNA-binding protein, which interacts with specific neuronal mRNA-targets and with other proteins and is involved in mRNA-transport, stability and translation Drosophila model closely recapitulates the neuronal defects of the clinical disease phenotype and its FMRP homolog (dFMRP) shares similar biochemical features. Using this model and the GAL4/UAS system for targeted gene expression, we performed genetic interaction experiments. We over-expressed the gene dfmr1 (Drosophia fragile X mental retardation 1) in the developing wing imaginal discs and looked for possible suppressor/enhancer effects of the genes scrib(sribbled) and dlg1(discs large 1). We also looked for similar genetic interactions in the adult brain neurons. We found that scrib acted as an enhancer of the mutant wing phenotype, induced by the over-expression of dfmr1. scrib also enhanced the mutant synaptic phenotype in the adult brain neurons, caused by the overexpression or lack of expression of dfmr1. dlg1 had an opposite suppressor effect in both systems analyzed. It rescued the mutant wing phenotype and the mutant synaptic phenotype as well. Our results demonstrate that dfmr1 interacts genetically with the members of the Lgl-family of scaffolding proteins and functions with them in the synaptic architecture of Drosophila adult brain neurons.
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Papers by Emil D Molle