Physical Mutagens Research Articles

Genetic Divergence and Heritability Study of Some NERICA Mutant Lines and Their Parents Using Microsatellites Marker and Morphological Traits in Rice

New Rice of Africa (NERICA) is drought tolerant and early maturing inter specific rice variety which was introduced in Bangladesh from Uganda in 2009. But the field record of NERICA was not very promising. In order to develop NERICA varieties suitable for agro-climatic conditions of Bangladesh different variations in yield contributing characters are required.In the experiment, physical mutagen treated (250, 300 and 350 Gy Gamma-rays) 18 NERICA mutant lines were selected from M4 to M5 generations along with 3 parents (NERICA-1, NERICA-4, NERICA-10) for morphological and molecular evaluation in orderto identify desired mutant linesusing 12 yield attributing characters and 3 simple sequence repeat (SSR) markers. We also estimated heritability, genetic advance and correlation among the studied morphological traits to identify besttraits based on which further selection could be made. Pearson’s correlation co-efficient of 12 morphological traits indicated that yield per plant had significant positive correlation with the number of tiller, number of effective tiller, panicle length, number of filled grainsper panicle and 1000 grain weight but negative correlation with Plant height and days to maturity. Broad sense heritability ( ) ranged from 71% to 99% while genetic advances in percent mean (GA%) varied from 10% to 60%. Yield per plant, unfilled grains per panicle, number of total tiller, number of effective tiller showed high heritability along with genetic advance as percent of the mean (GA%) value. A total of 24 alleles were detected by 3 SSR markers. The mean gene diversity and Polymorphism Information Content (PIC) values was 0.821 and 0.797, respectively. Dendrogram constructed based on SSR markers clustered the genotypes into six distinct clusters. Combining molecular and morphological evaluation data eight mutant lines, N10/300/P-2-3-5, N10/300/P-2-3-5, N10/300/P-2-3-5-2, N1/300/P-2-3-5, N1/300/P-2-3-5, N1/250/P-7-6-4-1, N10/300/P-2(1)-4-1 and N1/250/P-7-3-7-1were selected as desired mutant lines having good yield attributing characters and could be recommended for further evaluation in rice breeding program.

Physical and chemical mutagens improved Sporotrichum thermophile, strain ST20 for enhanced Phytase activity.

ObjectivePhosphorous is an essential micronutrient of plants and involved in critical biological functions. In nature, phosphorous is mostly present in immobilized inorganic mineral and in the fixed organic form including phytic acid and phosphoesteric compounds. However, the bioavailability of bound phosphorous could be enhanced by the use of phosphate solubilizing microorganisms such as bacteria and fungi. The phytases are widespread in an environment and have been isolated from different sources comprising bacteria and fungi.MethodologyIn current studies, we show the successful use of gamma rays and EMS (Ethyl Methane Sulphonate) mutagenesis for enhanced activity of phytases in a fungal strain Sporotrichum thermophile.ResultsWe report an improved strain ST2 that could produce a clear halo zone around the colony, up to 24 mm. The maximum enzymatic activity was found of 382 U/mL on pH 5.5. However, the phytase activity was improved to 387 U/ml at 45 °C. We also report that the mutants produced through EMS showed the greater potential for phytase production.ConclusionThe current study highlights the potential of EMS mutagenesis for strain improvement over physical mutagens.

Biological effects of three types of ionizing radiation on creeping bentgrass

Purpose: Gamma-rays and carbon ions are frequently used for mutation breeding in diverse plant species, whereas proton ions have been rarely used for this purpose. This study assessed the potential of proton ions for plant mutation breeding.Materials and methods: We compared the effects of radiation on creeping bentgrass seeds with γ-rays, proton ions, and carbon ions on seed germination, plant growth parameters, and DNA fragmentation.Results and conclusions: The lethal dose 50 (LD50) doses based on seed germinability were 115.9 Gy (γ-rays), 225.1 Gy (proton ions), and 57.7 Gy (carbon ions). Threshold doses for survival were 150 Gy (γ-rays), 150 Gy (proton ions), and 25 Gy (carbon ions). Suppression of plant growth was displayed at 100 Gy (γ-rays), 25 Gy (proton ions), and 25 Gy (carbon ions). Similar patterns of decreasing head DNA percentage were observed for γ-rays and proton ions. Carbon ions induced the lowest frequency of DNA fragmentation. The biological effects of the ionizing radiation types on creeping bentgrass are summarizable as follows: germination, carbon ions (C)>γ-rays (G)>proton ions (P); survival, C > P = G; growth, C ≥ P > G; DNA fragmentation, G ≥ P > C. These results indicate that proton ions are useful as a physical mutagen in plant mutation breeding.

IMPROVING THE CHITINOLYTIC ACTIVITY OF STREPTOMYCES GRISEORUBENS E44G BY MUTAGENESIS

Genetic improvement trials of the chitinolytic activity of Streptomyces griseorubens E44G were made by using physical, chemical and site-directed mutagenesis. Although the UV radiation, as a physical mutagen, was shed on the tested bacteria for different durations (5, 10, and 15 min), no change in the chitinolytic activity was observed when compared with the wild type. To induce the chemical mutagenesis, S. griseorubens E44G was treated with ethylmethane sulfonate for varied durations (20, 40, and 60 min). The chitinolytic activity decreased with the increment in the exposure period. Four different sets of primers were designed based on the DNA sequence of the wild type of S. griseorubens E44G.Overexpressionof chitinase-encoding genes was observed as three of the amplified mutated genes comparing with the wild-type gene. The chitinolytic activity of the recombinant gene P2 increased by 1.39-fold comparing with the wild-type gene. The molecular weight of the chitinase protein produced by the mutated gene was determined by SDS-PAGE. In conclusion, these results demonstrated that the recombinant gene of S. griseorubens E44G possess a higher level of chitinolytic activity than that of the wild-type. Genetic improvement of the chitinolytic activity of S. griseorubens E44G may enhance their biocontrol potential against phytopathogenic fungi.

Мутаційна мінливість пшениці озимої (Triticum aestivum L.) при хімічному мутагенезі

Мутаційна мінливість пшениці озимої (Triticum aestivum L.) при хімічному мутагенезі

Banana diversity (Musa spp.) as a result of in vitro gamma ray irradiation based on morphological markers.

Abstract. Due MS, Yunus A, Susilowati A. 2019. Banana diversity (Musa spp.) as a result of in vitro gamma ray irradiation based on morpological markers. Pros Sem Nas Masy Biodiv Indon 5: 347-352. Banana is a horticultural commodity that has a high enough production so that it makes it a superior product in the country. Banana has long been domesticated by the community because it has many benefits. There are more than 200 banana cultivars growing in various regions, one of which is feather plantain. Banana plants are propagated vegetatively so that they have limitations in obtaining genetic variation and require a long generation time in their vegetative cycle. Improvements in the nature of banana plants can be done by increasing its genetic diversity through mutation induction using gamma ray physical mutagens combined with in vitro culture techniques.This study aims to find out the diversity of feather plantain as a result of in vitro gamma ray radiation based on morphological markers. This research is a type of experimental research using completely randomized design (CRD) method. The factors tested were 4 radiation doses (0 Gy, 10 Gy, 20 Gy and 30 Gy). Morphological observations are carried out based on quantitative and qualitative characters. The data obtained were analyzed using the SPSS ver. 10 and the NTSYS (Numerical Taxonomy and Multivariate Analysis System) program ver. 16. The result of this study shows that gamma ray radiation in various doses causes the emergence of variations in plants, especially in the stems and leaves. Gamma ray radiation at a dose of 30 Gy is the most optimal dose to produce diversity.

Determination of Effective Mutagen Dose for Carrot (Daucus carota ssp. sativus var. atrorubens alef and D. carota) Callus Cultures

Recep Tayyip Erdogan Universitesi, Doga Bilimleri Fakultesi, Bahce Bitkileri Bolumu, TAEK, RHTDB / AR-GE Projeleri Subesi Uludag Universitesi Ziraat Fakultesi Bahce Bitkileri Bolumu, Ankara Universitesi Ziraat Fakultesi BAhce Bitkileri Bolumu

Determination of lethal dose and effect of EMS and gamma ray on germination percentage and seedling parameters in barnyard millet variety Co(Kv)2

An experiment was conducted to estimate the lethal dose of the chemical mutagen EMS and physical mutagen gamma ray in barnyard millet variety Co(Kv) 2. Genetically pure seeds were treated with different doses of EMS viz. 30mM, 40mM, 50mM, 60mM, 70mM, 80mM, 90mM and 100mM and gamma rays viz., 500 Gy, 600 Gy, 700 Gy, 800 Gy, 900Gy and 1000 Gy. The LD50 values were observed based on growth reduction of seedlings after EMS and gamma ray treatment. The LD50 dose for EMS and gamma ray under in vitro and in vivo condition were fixed at 70mM and 800 Gy respectively, based on probit analysis. As the doses of applied EMS and gamma ray increased, there was a decrease in germination, survival rate of seedlings, root length, shoot length, seedling height, vigour index under in vitro conditions and emergence and survival under field (in vivo) conditions in M1 generation as compared to the control.

Radiation effect on germination and seedling traits in rice (Oryza sativa L.)

The biological effect of new source of physical mutagen viz., electron beam over gamma rays were studied with different dosages in the M1 generation of rice during 2017-18. The seeds of rice variety Anna (R) 4 rice were treated with 100 to 400 Gy of electron beam and gamma rays. At higher dosage level of 400 Gy both in gamma rays and electron beam, the germination percentage was reduced to 28 and 24 per cent respectively. The LD50 value was fixed as 273.27 Gy for electron beam, which is much lower dose than gamma rays (376.57Gy). During germination, root length was more affected than shoot length in both radiations. The electron beam irradiation affected maximum reduction in pollen fertility (61.87%) and in spikelet fertility (72.75%) at 400 Gy, whereas at same dose lesser reduction was observed in gamma rays. The relationship between pollen and spikelet fertility showed positive significant. The above information obtained from this study is more useful to understand the effect of electron beam on rice over gamma rays and also help to decide the dose for future mutation breeding programme in rice.

Research of photosynthesis and genomewide resequencing on a yellow‐leaf Lotus japonicus mutant induced by carbon ion beam irradiation

AbstractCarbon ion beam is an advanced physical mutagen that is used for mutation breeding and functional genomics research. Using carbon ion beams, a stable Lotus japonicus mutant C16 which showed yellow leaves and reduced height compared with the laboratory wild type was obtained in our previous study. Yellow‐leaf mutation is a desirable characteristic for studying photosynthesis. The photosynthetic‐related physiological indexes, including the content of chlorophylls and the ratio of variable fluorescence (Fv) over the maximum fluorescence value (Fm) (maximal photochemical efficiency of photosystem II in the dark) of leaves between laboratory wild type and C16, were investigated. C16 showed lower amount of chlorophylls than that of laboratory wild type, as well as a relatively photosynthetic capacity. Meanwhile, to comprehend the molecular mechanism underlying the mutant, genomewide resequencing was performed to identify the mutational site responsible for the yellow‐leaf phenotype causing the weaker photosynthetic capacity. Ninety‐five single‐base substitutions and 17 small insertions and deletions were found in C16 mutant. Among those, by association analysis of functional annotation and mutant phenotype, T>G SBS in chromosome 5 located in Lj5g3v1669360.1 which encodes arabinogalactan protein was supposed as candidate mutation sites. Quantitative real‐time polymerase chain reaction analysis indicated that the expression level of Lj5g3v1669360.1 in C16 was significantly lower than that of laboratory wild type. Decreased expression of the Lj5g3v1669360.1 may be responsible for the smaller amount of chlorophylls causing yellow leaves, the weaker photosynthetic capacity of C16, and resulting in slow growth.

Biogenic Synthesis of Silver Nanoparticle from Mushroom Exopolysaccharides and its Potentials in Water Purification

Objective:This study reports a novel eco-friendly biosynthesis of Silver Nanoparticles (AgNPs) from Exopolysaccharides (EPS) ofLentinus edodesafter an attempt to optimise the production of EPS through mutagenesis. It further describes some potential application of silver nanoparticles in water treatment.Methods:A wild strain ofL. edodeswas subjected to UV irradiation, a physical mutagen, at 254 nm. The wild and resultant irradiated strains were then assessed for the production of EPS and subsequent application of the crude EPSs for biosynthesis of AgNPs. The particles were characterised by colour pattern and UV-visible spectroscopy. Based on superior EPS production and nanoparticle attributes, nanoparticles obtained from UV irradiated process were further subjected to Scanning Electron Microscopy (SEM). EPS produced was quantified by the phenol-sulphuric acid method and studied by GC-MS.Results:Results obtained for EPS productivity indicated the presence of monomer sugars such as arabinose (50.65%), mannose (19.20%), mannitol (15.58%), fructose (7.96%), trehalose (6.49%), and glucuronic acid, xylose, galactose and glucose with low percentages of ≤ 0.11. EPS productivity of wild and mutant strains was obtained as 1.044 and 2.783 mg/ml, respectively, after 7 days of fermentation. The result of EPS production for UV irradiated strain corresponds to a yield improvement of 2.7 fold of the wild-type. UV Spectroscopy and SEM analysis studies on EPS nanoparticle product of the improved (UV irradiated) strain indicated the formation of AgNPs at the absorption band of 421 nm with a size range of 50-100 nm.Conclusion:This study, which aimed at eco-friendly synthesis of myco-nanoparticle has established the novel ability ofL. edodes’polysaccharide in silver nanoparticles biosynthesis. It expounded potential frontiers of silver nanoparticles application in the water industry. To the best of the authors’ knowledge, this result represents the first report on the biosynthesis of AgNPs usingL. edode’sEPS.

Studies on the Effects of Ultraviolet Irradiation on Pea (Pisum sativum L.)

1. Abstract Pea ( Pisum sativum L.) is one of the earliest domesticated cool season annual legume crop produced worldwide, mainly in temperate regions. In common with other grain legumes, pea plays an important role in food and nutritional security of humans as well as livestock. Pea is an annual plant which exhibits mainly self-pollination, although cross pollination through insects also occurs in nature. The improvement of pea through plant breeding requires considerable genetic variations in the key quantitative traits and the expression of those polygenic traits also depends on the environmental interactions. Therefore, UV irradiations have been employed in the present study to assess the genotypic sensitivity of the pea cultivar for possible application in mutation breeding of pea. Seed germination and seedling growth at different duration of exposures were calculated for estimating the effect of UV irradiations stress on pea. The findings of the present study conclude that the UV irradiation can be useful as non-ionizing physical mutagen for induction of selectable macromutations in local pea cultivars and the exposure to the increasing UV stress in the nature will be detrimental for the crop productions.

Molecular diversity and phenotypic responses of twoin vitro Solanum tuberosumvarieties by physical mutagen

Induction of mutation in potato (Solanum tuberosum L.) plants by gamma radiation is considered an effective method for plant improvement. Solanum tuberosum L. is a vegetative plant, its tuber is st...

Improvement of Secondary Metabolites from Phyllanthus odontadenius against Malaria by Mutagenesis

Aims: Majority of deaths in children aged under 5 years are due to Plasmodium falciparum malaria. Malaria deaths in children decreased but malaria remains a major killer of children, taking the life of a child every 2 minutes. This study aims to investigate the increasing of the in vitro antiplasmodial activities by mutagenesis techniques using gamma-rays (Cs-137) or sodium azide (NaN3) as utagens. It will allow the importance of mutagenesis use as tools for improvement of secondary metabolites against malaria parasites using chemical or physical mutagens. Study Design: Laboratory experiment tests : identification of plant material, immersion of seeds in SA (sodium azide) solutions or irradiation by Gamma-rays (Cs-137) of P. odontadenius seeds for improvement of secondary metabolites against malaria parasites, in vitro culture of seeds followed by the in situ culturing of plantlets for obtaining material of study, phytochemical screening of Phyllanthus odontadenius aerial parts to determine the change of compounds in comparison to controls, in vitro antiplasmodial tests for the determination of SA concentrations or those of gammarays doses which killing 50% of malaria parasite populations (IC50). Place and Duration of Study: Department of Biotechnology and Molecular Biology, Department of Biochemistry: General Atomic Energy Commission, Regional Nuclear Studies Center of Kinshasa, P.O. Box. 868 Kinshasa XI, Democratic Republic of the Congo (DRC). National Institute of Biomedical Research (NIBR) at Kinshasa/Gombe (DRC). Laboratory of UMR-MD3, Aix-Marseille University, Faculty of Pharmacy/Marseille, France. The experiments were conducted from Junuary - December 2010; August and December 2011 and during May and September 2012 until July 2014. Methodology: Aerial parts of plants M1 and M2 from Gamma-rays irradiation of P. odontadenius seeds or from immersion of P. odontadenius seeds in sodium azide solutions used as biological material for the in vitro antiplasmodial analysis. The in vitro antiplasmodial activities assays on clinical isolates of P. falciparum and on chloroquine-resistante P. falciparum strain K1 was determined using microscopic method, the isotopic micro-test method and using HRP2-based ELISA assay. Results: Gamma-rays (Cs-137) increased (multiplied) the in vitro antiplasmodial activities from 2.48 up to 7.6 in comparison to control. Thus, the in vitro antiplasmodial activities were improved or exceeded from 147.57% up to 660% than those of control plant. SA had increased (multiplied) the in vitro antiplasmodial activities from 1.24 up to 10.15 comparing to the control plants. The antiplasmodial activities in vitro were exceeded compared to the control plants from 24.43% up to 915%. Conclusion: Treatment of P. odontadenius seeds by Gamma-rays or by SA give plants high in vitro antiplasmodial activities. Values of in vitro antiplasmodial activities varried from 1.24 (147.57%) to 10.15 (915%). 125, 150 and 225 Gy of Gamma-rays (Cs-137) for physical mutagenesis and 10.15 and 17.5 M of SA solutions for the chemical mutagenesis could be used for improving in vitro antiplasmodial activities against P. falciparum (clinical isolates of P. falciparum or chloroquine-resistant P. falciparum strain K1). Thus, plants extracts from treated seeds have justified the usefulness of mutagens in plant breeding particularly in the increasing production of secondary metabolite against malaria parasite.

Strategies for identification of mutations induced by carbon-ion beam irradiation in Arabidopsis thaliana by whole genome re-sequencing

Heavy-ion beam irradiation is a powerful physical mutagen that has been used to create numerous mutant materials in plants. These materials are an essential resource for functional genomics research in the post-genome era. The advent of Next-Generation Sequencing (NGS) technology has promoted the study of functional genomics and molecular breeding. A wealth of information can be gathered from whole genome re-sequencing; however, understanding the molecular mutation profile at genome wide, as well as identifying causal genes for a given phenotype are big challenging issues for researchers. The huge outputs created by NGS make it difficult to capture key information. It is worthy to explore an effective and efficient data-sieving strategy for mutation scanning at whole genome scale. Re-sequencing data from one laboratory wild type (Columbia) and eleven M3Arabidopsis thaliana lines derived from carbon-ion beam irradiation were used in present study. Both the number and different combinations of samples used for analysis affected the sieving results. The result indicated that using six samples was sufficient to filter out the shared mutation (background interference) sites as well as to identify the true mutation sites in the whole genome. The final number of candidate mutation sites could be further narrowed down by combining traditional rough map-based cloning. Our results demonstrated the feasibility of a parallel sequencing analysis as an efficient tool for the identification of mutations induced by carbon-ion beam irradiation. For the first time, we presented different analysis strategies for handling massive parallel sequencing data sets to detect the mutations induced by carbon-ion beam irradiation in Arabidopsis thaliana with low false-positive rate, as well as to identify the causative nucleotide changes responsible for a mutant phenotype.

Lethal Dose (LD50) Fixation and Sensitivity of Fenugreek (Trigonellafoenum-graecumL.) to Gamma Radiation for Induction of Mutation

Fenugreek (Trigonella foenum-graecumL.) is an annual crop, mainly used as a spiceand leafy vegetable crop in many parts of the world. Classical breeding in fenugreek is restricted due to its low genetic variability and small flower size which hamper manual emasculation and pollination. Mutation breeding is an effective way to enrich genetic variability in crop plants. An experiment was conducted to determine the lethal dose of the physical mutagen gamma rays in fenugreek. The dry seeds of fenugreek were exposed to different doses of gamma rays i.e. 150Gy, 200Gy, 250Gy, 300Gy and 350Gy. These irradiated seeds were sown in the Petri plates with non-irradiated seeds (control). As the dose of gamma rays increased, there was a decrease in germination percentage, seedling survival, root length, shoot length and vigour index. Among five doses of gamma rays, the maximum seed germination was observed at lowest dose 150Gy (93%), followed by 200Gy (83%), 250Gy (76%), 300Gy (76%) and 350Gy (64%). The seedling survival was decreased from 90% (in control) to 56% in 350Gy dose of gamma rays. The gamma rays dose of 150Gy gave stimulatory effect on seedlings growth. The growth parameters were dose dependent, as the dose of gamma rays increased from 200Gy to 350Gy. The gamma rays dose of 350Gy showed 64% seeds germination and 56% of seedlings survival. Therefore, it is concluded that the LD50 dose for fenugreek is close to 350Gy. This information would be highly useful for initiating mutation breeding programme in fenugreek

Antimutagenic activity of vitamin B1 against damages induced by chemical and physical mutagens in Salmonella typhimurium and Escherichia coli

Thiamine (vitamin B1) is an essential nutrient acting mainly as an enzymatic cofactor on diverse cell processes. It has been reported that vitamin B1 has a significant role in the signaling pathways related to the response to adverse environmental conditions (chemical and physical). The objectives of this study were to evaluate the antimutagenic potential of vitamin B1 in front of DNA-alkylating agents in the presence/absence of ogt and ada repairing genes in Salmonella typhimurium strains and against damage induced by ultraviolet light type C in Escherichia coli strains mutated at the uvrABC system and recBCD enzymes. For S. typhimurium, an antimutagenesis test (Ames test) was performed using strains deficient in one or both genes (YG7100 ada−/ogt+, YG7104 ada+/ogt−, YG7108 ada−/ogt−). For E. coli, mutated strains (K-12 derived strains Hfr H180 uvrB+/recA+, W3110 uvrB+/recA− and ATCC®8739 uvrB−/recA+) were exposed to UV-C light at different time intervals, with and without vitamin B1. Our results showed that thiamine is an antimutagen against methyl-N-nitro-N-nitrosoguanidine or ethyl-N-nitro-N-nitrosoguanidine only when the ogt gene is present. While for E. coli, the presence of vitamin B1 increased the survival rate, implying an antimutagenesis independent of uvrABC repairing system and recBCD enzymes.

Gamma rays and EMS induced Flower Color Mutation in Grasspea (Lathyrus sativus Linn.)

To induce flower color mutants in Grasspea ( Lathyrus sativus Linn.) variety “Pusa-24“, the fresh seeds were exposed to 4 different doses of gamma radiation as physical mutagens (100, 200, 300 and 400 Gy) and treated with 4 different concentration of Ethyl Methanesulphonate as chemical mutagen (5, 10, 15 and 20 mM ) and also combination of both mutagens. Due to mutagenesis broad range of flower color mutants in M 2 , M 3 and M 4 were obtained. Various unique and different flower color mutations like pink, purple, white and yellow detected against blue which is natural color in control progenies. Maximum flower color frequency was recorded in EMS treated seeds (20 mM) in 4th generation and lowest mutation frequency in 2nd generation was obtained in gamma rays (100 Gy) treated seeds. The results showed more flower color mutations happened in more dose /concentration of mutagens in every generation

Rates and spectra of chromosome aberrations in winter wheat cells after dimethylsulfate action

In the article we report about our investigation of cytogenetic parameters in chromosomal complex of the new Ukrainian winter wheat varieties and some relations amid value of cytological indexes and different concentrations of DMS (dimethylsulfate). We used the anaphases method for the investigation of chromosomal rearrangements and determination the mutagen action on plants and identification the nature of mutagen. We studied combined the sensitivity of genotype by the cytological analysis of mutagen treated population of wheat and revealed the reaction of different varieties depending upon the way of its origin. This was done for determining its connections and differences on cytogenetics level together with specificity of mutagen action on cell level. Dry seeds of seven varieties and one line of winter wheat were subjected to DMS in 0.0125, 0.025, and 0.05 % concentrations, which are the most useful for winter wheat in terms of genetic investigation programs. We investigated the rates and spectra of chromosomal aberrations in winter wheat (primary roots of tips cells) during the mitosis. We identified the following types of chromosomal rearrangements: chromatid and chromosome bridges, single and double fragments, micronuclei, and lagging chromosomes. Investigation of DMS action confirmed reliability of fragments-bridges ratio (prevalence of fragments under the bridges for chemical mutagens and vice versa for physical mutagens) for the mutagen nature identification.

Mutagenic Effectiveness and Efficiency of Gamma Rays and HZ with Phenotyping of Induced Mutations in Lentil Cultivars

In mutation breeding, mutagenic effectiveness and efficiency are the base parameters to predict the mutagenic potency of any mutagen. Studies on mutagenic effectiveness and mutagenic efficiency of physical mutagen (gamma rays) and chemical mutagen (hydrazine hydrates; HZ) on two cultivars of lentil (Lens culinaris Medik.), viz. DPL 62 (macrosperma) and Pant L 406 (microsperma) have been reported. Dry and healthy seeds were treated with four doses of each gamma rays (100-400 Gy), HZ (0.1-0.4 %) and their combinations. Frequencies of the induced agro-morphological variations into different phenotypic categories were estimated in M2 population that resulted into identification and isolation of wide range of mutants with altered phenotypes. Data on effectiveness and efficiency of various mutagenic treatments calculated on the basis of the frequency of chlorophyll mutations, which showed effectiveness and efficiency were higher at the moderate doses of gamma rays and HZ, while in case of combination treatments; lower doses were most effective and efficient with few inter-varietal exceptions. Phenotyping of the mutants revealed that growth habits was the most sensitive category to which most of the mutant belongs, followed by leaf and flower/pod/seed in both the cultivars studied. Overall, the screened and isolated mutants with economically important agronomic traits can be further propagated in the subsequent generation for development of elite lentil mutant cultivars.