IN VITRO РЕГЕНЕРАЦИЯ ОТ ЕКСПЛАНТИ ОТ ЛИСТНИ ДРЪЖКИ НА PHASEOLUS VULGARIS L. ЧРЕЗ ОРГАНОГЕНЕЗИС

A system for somatic organogenesis in common bean (Phaseolus vulgaris L.) was developed. Precultivation of seeds on different media was investigated for the ability to influence the process of in vitro regeneration. Leave petioles excised from in vitro derived seedlings at different ages (7 and 14 days-old), were used as primary explants. Precultivation of the seedlings on medium MS-BAP 1 μM for 7 days and dark cultivation of their leave petioles on medium MSI (2 μM TDZ, 0.6 μM NAA and 2 μM paclobutrazol) benefit the process of shoot initiation in all investigated genotypes. Shoot elongation took place on MSE (22.2 μM BAP and 0.057 μM IAA) medium. Plant recovery was established on MSG3 (4.44 μM BAP and 0.58 μM GA3) medium.Разработена е система за соматичен органогенезис при фасул (Phaseolus vulgaris L.). Изследвано е предкултивирането на семена върху различни среди за способността да се повлияе върху процеса на in vitro регенерацията. Листни дръжки нарязани от in vitro развити прорастъци на различна възраст (7 и 14 дневни) са използвани като първични експланти. Предкултивирането на прорастъците върху среда MS-BAP 1 μM за 7 дни и култивиране на тъмно на експлантите от техните листни дръжки върху среда MSI (2 μM TDZ, 0.6 μM NAA и 2 μM паклобутразол) подобрява процеса на иницииране на прорастъци от тях при всички изследвани генотипи. Удължаването на прорастъците се проявява на среда MSE (22.2 μM BAP и 0.057 μM IAA). Растенийца се развиват на среда MSG3 (4.44 μM BAP и 0.58 μM GA3)

IN VITRO РЕГЕНЕРАЦИЯ ОТ ЕКСПЛАНТИ ОТ ЛИСТНИ ДРЪЖКИ НА PHASEOLUS VULGARIS L. ЧРЕЗ ОРГАНОГЕНЕЗИС

A system for somatic organogenesis in common bean (Phaseolus vulgaris L.) was developed. Precultivation of seeds on different media was investigated for the ability to influence the process of in vitro regeneration. Leave petioles excised from in vitro derived seedlings at different ages (7 and 14 days-old), were used as primary explants. Precultivation of the seedlings on medium MS-BAP 1 μM for 7 days and dark cultivation of their leave petioles on medium MSI (2 μM TDZ, 0.6 μM NAA and 2 μM paclobutrazol) benefit the process of shoot initiation in all investigated genotypes. Shoot elongation took place on MSE (22.2 μM BAP and 0.057 μM IAA) medium. Plant recovery was established on MSG3 (4.44 μM BAP and 0.58 μM GA3) medium.Разработена е система за соматичен органогенезис при фасул (Phaseolus vulgaris L.). Изследвано е предкултивирането на семена върху различни среди за способността да се повлияе върху процеса на in vitro регенерацията. Листни дръжки нарязани от in vitro развити прорастъци на различна възраст (7 и 14 дневни) са използвани като първични експланти. Предкултивирането на прорастъците върху среда MS-BAP 1 μM за 7 дни и култивиране на тъмно на експлантите от техните листни дръжки върху среда MSI (2 μM TDZ, 0.6 μM NAA и 2 μM паклобутразол) подобрява процеса на иницииране на прорастъци от тях при всички изследвани генотипи. Удължаването на прорастъците се проявява на среда MSE (22.2 μM BAP и 0.057 μM IAA). Растенийца се развиват на среда MSG3 (4.44 μM BAP и 0.58 μM GA3)

ВЛИЯНИЕ НА ЕТИЛМЕТАН СУЛФОНАТА (EMС) И N-НИТРОЗО-N´-ETИЛ КАРБАМИДА (НEК) ВЪРХУ РАСТЕЖА НА КАЛУС ОТ ФАСУЛ

Infl uence of ethyl methanesulfonate (EMS) and N-nitrose-N´-ethyl urea (ENU) mutagenic treatments was investigated on three time sub-cultured calli obtained from leaf petiole explants of 7-day old sterile plants. Calibrated sterile seeds of the common bean Bulgarian variety Plovdiv 11M were pre-cultivated on MS basal medium supplemented with 1 μmM BAP. Then, both mutagens EMS and ENU were applied for different times such as 15, 30, 60 and 90 min on the explants at the concentrations of: 2.5 . 10-2 M and 6.2 . 10-3 M, respectively. Times of the mutagenic treatments infl uenced callus growth, calli from 30-min treatment with both mutagens showing the highest weights. In both cases, the 90-min mutagen application caused a too relevant effect either on callus browning or growth inhibition. In general, ENU showed a stronger effect than EMS. The effect of subcultures on callus growth was higher than mutagenic treatments. Interactions between these factors checked by by correlation ratio (η%) were quite low.Изследвано е влиянието на третирането с етилметан сулфонат (EMС) и N-нитрозо-N´-етил карбамид (НEК) върху трикратно прехвърлен на свежа среда калус получен от експланти от листни дръжки на 7-дневни стерилни растения. Калибрирани стерилни семена от българския сорт фасул Пловдив 11М са предкултивирани на основна MS среда допълнена с 1 μmM BAP. След това, двата мутагена EMС и НEК, са приложени в концентрации съответно: 2.5 . 10-2 M и 6.2 . 10-3 M за различно време 15, 30, 60 и 90 min. Времето на мутагенното третиране влияе върху растежа на калуса като калусът получен след 30-min третиране с двата мутагена има най-високи тегла. 90-min третиране причинява подобен ефект от двата мутагена – покафеняване на калуса или инхибиране на калусния растеж. НЕК показва по-силен ефект от ЕМС. Ефектът от прехвърлянето на свежа среда върху растежа на калуса е по-силен от мутагенните третирания. Взаимодействията между тези фактори, отчетени чрез корелационното съотношение (η%), са сравнително ниски

INDUCTION OF CHLOROPHYLL MUTANTS IN COMMON BEAN UNDER THE ACTION OF CHEMICAL MUTAGENS ENU AND EMS

Effect of treatment with different concentrations of N-nitroso-N-ethyl urea (ENU) and etylmethan sulfonate (ЕМS) on seeds of Bulgarian common bean Dobroudjanski 7, Dobroudjanski 2, Plovdiv 10, Plovdiv 11М and snap bean Tcher Starozagorski varieties, for induction of chlorophyll mutants, was studied. It was established that investigated varieties manifested specifi c reactions to the treatment with ENU and EMS. Different mutation frequencies and width of mutation spectra were induced under the action of different concentrations of the two applied mutagens. ENU induced chlorophyll mutants with higher frequency in all studied varieties, in comparison to the action of EMS. Sixteen types of chlorophyll mutants were found, for all studied varieties, and mutagenic treatments. Mutant types chlorina (19,8%), xantha (19,3%), viridissima (15,4%) and chimerical leaves (9,1%) were with the highest frequency, comparing to the total number of observed mutants. Results were statistically elaborated by the Fisher’s method “ϕ”

DROUGHT TOLERANCE OF BULGARIAN COMMON BEAN GENOTYPES, CHARACTERISED BY SOME BIOCHEMICAL MARKERS FOR OXIDATIVE STRESS

The aim of this study was to characterize drought tolerance of 20 common bean genotypes using some biochemical markers for oxidative stress. 10 common bean cultivars (9 Bulgarian and a Mexican - BAT 477) and 10 mutant lines M(19–20), previously obtained by us after the treatment of seeds from Dobroudjanski 2 and Dobroudjanski 7 cultivars with ethyl methan sulphonate (EMS) and N-nithroso-N-ethyl urea (NEU) were used in this investigation. BAT 477 was chosen as a control and it was presented in unique cluster group. Three biochemical markers – malondialdehyde (MDA), hydrogen peroxide (H2O2) and proline were analyzed. The results were statistically elaborated by mono-, bifactorial ANOVA and cluster analyses. Our preliminary results demonstrated that to obtain more valuable information, concerning drought tolerance of both common bean cultivars and mutant lines, MDA, H2O2 and proline should be used as early warning markers. Genotypes studied could be of interest in future investigations being a geneplasme source of common bean drought tolerance

Influence of Diflufenican the Herbicide on Some Biological Traits of Bulgarian Common Bean Cultivar Plovdiv 15M

Three-year trials (2006-2008) were conducted in order to determine the influence of the herbicide diflufenican (trade name – Pelikan 50 CK) on certain biological traits of Bulgarian common bean cultivar Plovdiv 15 M. The herbicide was applied in the doses of 200, 250 and 300 ml/ha after the sowing and before the growth of the plants. It has been found that the traits related to the height of the plant, number of fruit-yielding branches, pods and seeds, mass of the seeds and average length of 10 pods per plant were influenced by different doses of the herbicide, by the year and the interaction between these two factors. A 300 ml/ha dose treatment differs most from other variants in the conducted clustering as a result of its strong influence on the traits: number of seeds, fruit-yielding branches and mass of the seeds per plant. The highest stability indices have been obtained for the traits related to the mass of the pods per plant, which showed greater stability regarding the treatment with the herbicide

INVESTIGATIONS ON THE RESISTANCE OF SOME BULGARIAN COMMON BEAN GENOTYPES TOWARDS BEAN WEEVIL (ACANTHOSCELIDES OBTECTUS SAY)

The most effective, environmentally sound and safety way to fight pests with biological means is the use of resistant varieties to them. In the present study were indicated the reactions of 30 Bulgarian common bean genotypes to the most economically important enemy – bean weevil (Acanthoscelidis obtectus Say). For this purpose, the following indicators were traced – seed damages and young adult insects, which largely characterized the response of different common bean genotypes to that biological pest enemy. The results of this investigation present a sensitive response to the sustainability of different genotypes to the bean weevil. The Bulgarian common bean varieties Plovdiv 11M, Abritus, Crystal and Bulgari can be used in breeding programs as donors of resistance to the bean weevil

Evaluation of the yield and the yield stability of perspective lines of barley

The study was carried out in the Experimental field of the Department of Genetics and Plant Breeding – Agricultural University, Plovdiv during the period 2009-2011. New breeding lines were investigated using block design with 4 replications and 20 m2 plot size. The aim of the investigation was to estimate barley lines by yield, stability and some important traits, using Obzor as a standard. Applying biometrical and cluster analysis and index of yield stability, it has been established that some  breeding lines: numbers 21411003, 21401409 and 21001008, respectively, had better traits in comparison with the standard cultivar “Obzor” as well as stable yields regardless of differences in the climatic conditions over the years. These lines can be certified as new cultivars according to the government’s requirements for cultivar testing

DROUGHT TOLERANCE OF BULGARIAN COMMON BEAN GENOTYPES, CHARACTERISED BY SOME BIOCHEMICAL MARKERS FOR OXIDATIVE STRESS

The aim of this study was to characterize drought tolerance of 20 common bean genotypes using some biochemical markers for oxidative stress. 10 common bean cultivars (9 Bulgarian and a Mexican - BAT 477) and 10 mutant lines M(19–20), previously obtained by us after the treatment of seeds from Dobroudjanski 2 and Dobroudjanski 7 cultivars with ethyl methan sulphonate (EMS) and N-nithroso-N-ethyl urea (NEU) were used in this investigation. BAT 477 was chosen as a control and it was presented in unique cluster group. Three biochemical markers – malondialdehyde (MDA), hydrogen peroxide (H2O2) and proline were analyzed. The results were statistically elaborated by mono-, bifactorial ANOVA and cluster analyses. Our preliminary results demonstrated that to obtain more valuable information, concerning drought tolerance of both common bean cultivars and mutant lines, MDA, H2O2 and proline should be used as early warning markers. Genotypes studied could be of interest in future investigations being a geneplasme source of common bean drought tolerance

Characteristics of Common Bean Mutant Lines and Cultivars Grown under Rainfed and Irrigated Conditions

The experiments were conducted in the field of Agricultural University in Plovdiv, Bulgaria. A standard method was applied for cultivation in 5 replicates. Biometric evaluation of common bean (Phaseolus vulgaris L.), 10 mutant lines and 10 varieties, grown under rainfed and irrigated conditions was conducted. Main traits, associated with productivity in common bean: plant height, mass of plants with pods, number of branches, height of the first pod, number of fruit branches, number of pods per plant, weight of pods with seeds, number of seeds per plant, weight of seeds and average length per 10 pods were characterised. Stronger degree of variation in studied traits was observed in genotypes grown under irrigated conditions. It was found that the studied Bulgarian varieties are promising in terms of germoplasm for their introduction in hybridisation breeding schemes as well as in application of mutagenesis and biotechnological practices. D2-0,0125 M EMS  mutant line (6) has the best manifestation of the studied traits among other mutant lines and it may be included in breeding schemes for evaluation as a new cultivar. BAT 477 cultivar (20) differs significantly by its traits from other genotypes, irrespective of the cultivation mode