Concentration Of Ethyl Methane Sulphonate Research Articles

Effect of ethyl methane sulphonate on growth, yield and some physiological parameters of Bambara groundnut (Vigna subterranea (L.) verdc) in non - water stressed and water stressed conditions

Bambara groundnut (Vigna subterranea (L.) Verdc.) is a legume cultivatd majorly for it’s seeds and commonly grown by subsistence farmers in sub-Saharan Africa. In order to determine the effect of ethyl methane sulphonate (EMS) mutagen on the growth, yield and some physiological parameters of non-water stressed (NWS) and water stressed (WS) bambara groundnut. It’s seeds were treated with five concentrations (0.00 (control), 0.25, 0.50, 0.75 and 1.00% v/v) Ethyl methane sulphonate and were grown under two conditions: non-water stressed and water stressed. Analysis of variance (ANOVA) revealed significant differences (P ≤ 0.50) in the emergence percentage (6 DAS), germination percentage (7 DAS), plant height (PH), number of leaves per plant (NL), leaf length (LL), leaf width, (LW), relative water content (RWC), chlorophyll a, b and total chlorophyll contents in the non-water stressed and water stressed plants. After rewatering at 8 WAS, highest number of leaves in NWS and WS conditions were observed in 0.75% EMS concentration (229, 189), highest leaf length in NWS and WS plants were observed in 0.50% EMS concentration (9.47cm, 9.13cm). Highest leaf width in NWS and WS plants was observed in 0.25% EMS concentration (4cm, 3.63cm). Highest number of pods per plant (NP) and weight of pods per plant (WP) were observed in 0.75% EMS concentration only in NWS plants. Highest chlorophyll a, b and total chlorophyll content was observed in 1.00% EMS concentration. The result form this study showed that Ethyl methane sulphonate (EMS) mutagen concentration increased the growth, yield and physiological parameters of Bambara groundnut in non-water stressed condition at different doses, however there was no positive effect of EMS on yield in the water stressed plants.

Mutagenic effectiveness and efficiency of ethyl methane sulphonate (EMS) mutagen in linseed (Linum usitatissimum L.)

Induced mutation study was carried out to study the mutagenic effectiveness and efficiency ofthe mutagen ethyl methane sulphonate (EMS) in two genotypes of linseed, Him Alsi-2 and Kangra Local. On the basis of survivalpercentage in M1 generation under lab conditionsthe LD50 dose for both genotypes was estimated. Only three doses (one higher and one lower dose to LD50) were selected for raising M2 generation. Effects of these concentrations were studied on various morphological characteristics. Mutagen EMS was effective and efficient in producing chlorophyll and viable macromutations in M2 generation. Three types of chlorophyll mutants i.e. albina, xantha and radina were observed. Different types of alterations in seed color, seed texture, flower shape and plant type were observed. The macromutations although are not very useful for plant breeding but help in determining the effective mutagen doses and have also known to play an important role in evolution. EMS concentrations showing maximum effectiveness and efficiency in both varieties were different indicating varietal sensitivity towards EMS.

Mutagenic effect of gamma rays and EMS on agronomical traits of niger (Guizotia abyssinica Cass.)

This study was conducted to augment genetic variability in niger plant (Guizotia abyssinica Cass.) using gamma rays and ethyl methane sulphonate (EMS) Niger (G. abyssinica) seeds of cultivar N-71 were exposed to various doses of gamma rays which ranged from 24 to 28 KR and different concentrations of EMS which ranged from 0.2 to 0.4% and in combination. Morphogenic and agronomic traits in the M1 and M2 generations that analyzed included: percentage of germination and survival, plant height, number of branches, capitulum size, number of ray florets per capitulum, number of disc florets per capitulum, head size, number of capitula per plant, number of seeds per capitulum, seed yield per plant and 1000-seed weight. The results of the present study showed that the three gamma irradiation doses used (24, 26 and 28 KR) had decreasing effect in most of the agronomic and morphological traits evaluated in M1. However, combination treatments like 24 KR+0.2% EMS and 26KR+0.4% EMS showed a noteworthy increase of grain yield and yield related components. In general, for the three irradiation doses in M1 population, there were no significant changes observed in morphological traits, however, in progenies of collective mutagenic treated seeds some potential high-yielding mutants were identified.

Mutagenic impact of ethyl methane sulphonate on morphological and cytological effects in berseem (Trifolium alexandrinum L.)

Induced mutagenesis has been effectively demonstrated as one of the best possible methodology for the genetic improvement of crop species. Egyptian clover (Trifolium alexandrinum L.), an important green fodder plant species of the family Fabaceae with immense agricultural value, was selected for the present study to improve its genetics by using the strategy of induced mutagenesis. Dry and healthy seeds of Egyptian clover cv. ‘Wardan’ were exposed to different doses (0.1, 0.3 and 0.5%) of Ethyl methane sulphonate (EMS) for variable time (3 and 5 h). The germination and survival percentages were recorded in M1 generation and 0.3% EMS at 5 h was determined as LD50 dose; whereas frequency of chlorophyll mutants and viable mutants along with cytological observations were screened in M2 generation. A broad spectrum of chlorophyll and viable mutants with different frequencies were observed. Cytological abnormalities increased along with rising EMS concentrations and mutagen exposure time. The findings are encouraging and show that significant genetic variability had been induced by EMS. Thus, the comprehensive selection of the desirable mutants may bring about advancement of improved and high yielding mutants of T. alexandrinum in upcoming generation.

PERBAIKAN SIFAT LADA (Piper nigrum L.) MENGGUNAKAN MUTAGEN ETHYL METHANE SULPHONATE (EMS)

The study aimed to determine the LC50 using EMS on pepper and obtain a better mutant than the parents. The material used was Indian variety pepper, and the chemical mutagen was Ethyl Methane Sulphonate (EMS). Two hundred Indian Cultivar pepper seeds obtained from community gardens were treated with EMS mutagen at a concentration of 0; 0.2; 0.4; 0.6; 0.8, and 1.0% with 3 hours of immersion. Observations consisted of germination, mutagen damage and agronomic characters. The LC50 calculation was obtained from the regression relationship of dead sprouts with several doses of EMS application which had the best fitting curve with the highest determination value (R2). The results showed that pepper germination was relatively low (62%). The impact of using EMS mutagens causes growth inhibition or even death of sprouts. The higher the concentration, the higher the mortality rate. The relationship between the concentration of mutagens and the mortality rate follows the polynomial equation y = 188.45x – 97.21x2 + 2.71 with a determination value of 99%. Based on the regression model, the LC50 value is 0.3%. In this study, the EMS concentration of 0.2% succeeded obtaining a better mutant pepper than the parental. indicated by increasing the length and width of the cotyledons, respectively, 20 and 24% greater than the control.

Evaluation of physical and chemical mutagens on various genotypes of brinjal (Solanum melongena L.) in M2 generation

Mutation breeding in crop plants such as brinjal (Solanum melongena L.) is a successful approach in change of product having narrow genetic base and to explore it a field experiment was conducted toevaluate the effect of physical mutagen such as gamma rays and chemical mutagens such as Ethyl methane sulphonate (EMS) were used on two brinjal genotypes namely Banarasi Gol and Kashi Taru. The seeds were treated with different concentrations of gamma irradiation (5, 10, 15 and 20 kR) and EMS (0.05, 0.10, 0.15 and 0.20%).The increasing concentration of mutagens was recorded with a negative response on all growth and yield attributes of both the genotypes which ultimately reduce the yield. In this context the 5kR gamma rays and 0.05% EMS concentration were recorded with superior values of all characters in M2 generation. Early flowering and early edible fruit maturity was observed under 5kR gamma rays over all other treatments. The fruit yield per plant increased by 19.87% and 20.30% of Banarasi Gol and Kashi Taru, respectively under 5kR gamma rays over control. Lowest value of all characters was recorded under 0.20% EMS as compared to all other treatments including control. Thus it can be concluded on present study basis that the increasing concentration of mutagens were showed an adverse effect for all characters.

Emergence of micronuclei as polymorphism induced genomic instability in Fennel (Foeniculum vulgare Mill.).

Micronuclei (MN) are extra-nuclear bodies containing damaged chromosomal fragments as well as complete chromosomes that are not otherwise incorporated into the nucleus after cell division. The present experiment was conducted to assess mutagenic effects of ethyl methane sulphonate (EMS) on the meiotic behavior of pollen mother cells of fennel (Foeniculum vulgare Mill.). Cytological analysis revealed that micronuclei were incited at all the three concentrations of EMS. A peculiar correlation of chromosomal abnormalities, mainly bridge and laggard at anaphase I and II with this nuclear polymorphism were experienced for genomic instability. Pollen viability was additionally inspected through pollen fertility. Thus, the MN formations bring about incomplete elimination of the genome, which can be proficiently use as the light of hope inbreeding programmers to produce addition and substitution lines.

Mutagenic potential of cadmium nitrate [Cd(NO3)2] and ethyl-methane sulphonate [EMS] in quantitative and cyto-physiological characters of Capsicum annum L. cultivars

Mutation breeding sustained food demand of the world through the introduction of desirable changes in the plants at the phenotypic as well as genotypic level. The study was primarily based on the aspects of beneficial and harmful effects of cadmium nitrate [Cd(NO 3 ) 2 ] and ethyl-methane sulphonate (EMS) as mutagenic agents in cyto-physiological and quantitative characters in Capsicum annum L. cultivars. The chilli seeds were treated with different concentrations of Cd(NO 3 ) 2 (0.01%, 0.02%, 0.03%, 0.04%, and 0.05%) and different concentrations of EMS (0.10%, 0.25%, 0.50%,0.75%, and 1.00%) alone to induce the cyto-physiological and quantitative changes and also to assess the mutagenicity of both chemicals. The results were revealed that Cd(NO 3 ) 2 has more toxic effects even in low concentrations than EMS with high concentrations. A high frequency of distortive chromosomes was found under treatment of Cd(NO 3 ) 2 while EMS revealed the least chromosomal effects. The physiological parameters were also recorded and results shown a decreasing trend in all the characters. The yield and yield contributing characters were recorded to moderate with increasing concentrations of Cd(NO 3 ) 2 while in EMS treated it decreased with increasing concentrations and showed some positive effects. The stomata aperture is reduced to close at high concentration of Cd(NO 3 ) 2 while EMS has little effect on stomata movement. The proline content and superoxide dismutase (SOD) enzyme activity increased with the increasing concentration of Cd(NO 3 ) 2 , whereas a small increment in proline content and SOD activity also was reported in EMS at higher concentrations. The mutagenic efficiency of a chemical mutagen can be calculated by determining the effectiveness of inducing the trait of interest in chilli crop plants having a narrow genetic base. The following changes in chilli cultivars can be further used to improve the quantitative and physiological characters through the plant breeding programs.

Protocol for ethyl methanesulphonate (EMS) mutagenesis application in rice.

Background: Non-transgenic chemical mutagen application, particularly ethyl methanesulfonate (EMS), is an important tool to create mutations and gain a new genetic makeup for plants. It is useful to obtain a sufficient number of mutant plants instead of working with a severe mutation in a few plants. EMS dose and exposure period have been previously studied in several crops; however, EMS used to create point mutations in presoaked rice seeds has not been sufficiently studied and there is no standard protocol for such treatment. The aim of this study is to establish a standard protocol for EMS mutagenesis application in rice. Methods: Two studies were conducted to evaluate the effect of four durations of rice seed presoaking (0, 6, 12, and 24 hours), four EMS concentration doses (0.0%, 0.5%, 1.0%, and 2.0%), and four EMS exposure periods (6, 12, 24, and 48 hours). Germination rate, plumula and radicle length, seedling survival,LD 50(Lethal Dose) determination, shoot length, root length and fresh seedling weight were evaluated. Results: Results showed that a 12-hour presoaking duration, 0.5% EMS dose, and six hours of EMS exposure were the best practices for the optimum number of mutant plants. Conclusions: In light of both this study and the literature, a standard application protocol was established. This application protocol, detailed in this article, contains the following guidelines: (1) Presoaking: 12 hours, (2) EMS application: 0.5% dose EMS and six hours, (3) Final washing: six hours, (4) Drying: 72 hours at 38°C. A user-friendly protocol has been presented for utilization by researchers.

Protocol for ethyl methanesulphonate (EMS) mutagenesis application in rice

Background: Non-transgenic chemical mutagen application, particularly ethyl methanesulfonate (EMS), is an important tool to create mutations and gain a new genetic makeup for plants. It is useful to obtain a sufficient number of mutant plants instead of working with a severe mutation in a few plants. EMS dose and exposure period have been previously studied in several crops; however, EMS used to create point mutations in presoaked rice seeds has not been sufficiently studied and there is no standard protocol for such treatment. The aim of this study is to establish a standard protocol for EMS mutagenesis application in rice. Methods: Two studies were conducted to evaluate the effect of four durations of rice seed presoaking (0, 6, 12, and 24 hours), four EMS concentration doses (0.0%, 0.5%, 1.0%, and 2.0%), and four EMS exposure periods (6, 12, 24, and 48 hours). Germination rate, plumula and radicle length, seedling survival, LD50 (Lethal Dose) determination, shoot length, root length and fresh seedling weight were evaluated. Results: Results showed that a 12-hour presoaking duration, 0.5% EMS dose, and six hours of EMS exposure were the best practices for the optimum number of mutant plants. Conclusions: In light of both this study and the literature, a standard application protocol was established. This application protocol, detailed in this article, contains the following guidelines: (1) Presoaking: 12 hours, (2) EMS application: 0.5% dose EMS and six hours, (3) Final washing: six hours, (4) Drying: 72 hours at 38°C. A user-friendly protocol has been presented for utilization by researchers.

Protocol for ethyl methanesulphonate (EMS) mutagenesis application in rice

Background: Non-transgenic chemical mutagen application, particularly ethyl methanesulfonate (EMS), is an important tool to create mutations and gain a new genetic makeup for plants. It is useful to obtain a sufficient number of mutant plants instead of working with a severe mutation in a few plants. EMS dose and exposure period have been previously studied in several crops; however, EMS used to create point mutations in presoaked rice seeds has not been sufficiently studied and there is no standard protocol for such treatment. The aim of this study is to establish a standard protocol for EMS mutagenesis application in rice. Methods: Two studies were conducted to evaluate the effect of four durations of rice seed presoaking (0, 6, 12, and 24 hours), four EMS concentration doses (0.0%, 0.5%, 1.0%, and 2.0%), and four EMS exposure periods (6, 12, 24, and 48 hours). Germination rate, plumula and radicle length, seedling survival, shoot length, root length and fresh seedling weight were evaluated. Results: Results showed that a 12-hour presoaking duration, 0.5% EMS dose, and six hours of EMS exposure were the best practices for the optimum number of mutant plants. Conclusions: In light of both this study and the literature, a standard application protocol was established. This application protocol, detailed in this article, contains the following guidelines: (1) Presoaking: 12 hours, (2) EMS application: 0.5% dose EMS and six hours, (3) Final washing: six hours, (4) Drying: 72 hours at 38°C. A user-friendly protocol has been presented for utilization by researchers.

Effect of EMS on seed germination and seedling vigour in mungbean [Vigna radiata (L.)

The pure, healthy and dry seeds of four mungbean varieties i.e., Meha, K 851, GM 3 and GM 4 were treated with different concentrations of Ethyl Methane Sulphonate (0.00 M, 0.02 M, 0.03 M and 0.04 M) for study the effect on seed germination and seedling vigour such as germination per cent, shoot length (cm), root length (cm), fresh weight (g) and dry weight (g). For LD50 recorded near to the 0.04 M EMS as per survival per cent in the all four mungbean varieties. The present results clearly indicated that different concentrations of Ethyl Methane Sulphonate can be effectively utilized to create variability for different quantitative characters.

Selection and Analysis of Polymorphisms in Somaclonal Variants of Agave americana Resistant to Fusarium oxysporum via an Ethyl Methanesulphonate Treatment

Agave americana L. callus were exposed to different concentrations of ethyl methanesulphonate (EMS) 0, 15, 30, 45 and 60 mM and to different times of exposure (2 and 4 h). The viability and capacity of shoot formation were shown to be affected when the callus were exposed to high concentrations (30–60 mM). Only the callus exposed to 15 mM EMS presented shoot formation; the exposure time of two hours produced the largest quantity of shoots regenerated per callus (21 shoots/callus). In order to generate somaclonal variants resistant to Fusarium oxysporum, a selection pressure was applied through of a culture filtrate (CF) of 100 ppm of the fungus. This was made in callus obtained in the treatment with 15 mM EMS during 2 h of exposure. The CF caused oxidation and necrosis in 71.25% of the callus; however, they were capable of generating shoots (3.5 shoots/callus). Molecular markers type RAPD, ISSR and DAMD were used to evaluate the genetic variation arising from the mutations caused by EMS on control plants and 16-month-old somaclonal variants. The polymorphic information content (PIC) for each one of the initiating groups was: 0.28 (DAMD), 0.09 (ISSR) and 0.14 (RAPD). DAMD revealed a greater percentage of polymorphism than RAPD and ISSR. Polymorphic bands were detected in the somaclonal variants. This indicated that the EMS caused genetic variation in the regenerated plants conferring resistance to them against Fusarium oxysporum.

Determination of optimum dose of chemical mutagen for large scale seed treatment of white seeded sesame (Sesamum indicum L.) varieties

In the present study two white seeded sesame varieties VRI 3 and SVPR 1 were treated with seven different concentration viz., 0.2 %, 0.4 %, 0.6 %, 0.8 %, 1 %, 1.2 % and 1.4 % of chemical mutagen Ethyl Methane Sulphonate (EMS). Germination percentage and germination index were tested using two different germination methods. Seed germination percentage and germination index was decreased with increased doses of EMS in both the varieties. The higher concentration of EMS completely arrested the seed germination. The optimum dose of the mutagen should be tested to maintain the require plant population in the M1 generation and increasing the effectiveness and efficiency of the mutagen. LD 50 value was determined using probit analysis. For SVPR 1 and VRI 3, LD 50 value was found to be 0.36% and 0.44 % respectively. For fixing the lethal dose for the chemical mutagen, Pro tray method of germination test was found to be reliable than germination paper method.

Effect of EMS on seed germination in white seeded sesame varieties (Sesamum indicum L.)

In the present investigation, two white seeded sesame varieties VRI 3 and SVPR 1 were treated with sevendifferent concentrations of EMS. Germination percentage and germination index were tested using two differentgermination methods. Seed germination decreased with increased concentration of Ethyl Methane Sulphonate (EMS) in both the varieties. For fixing the lethal dose for the chemical mutagen Pro tray method of germination test was found reliable than germination paper method.

Mutagenic components and dosage effects of ethyl methanesulphonate on <i>Arachis hypogea</i> (Samnut 24 VR.)

Determination of the optimum concentrations of a specific mutagen for crop improvement has always been a challenge. The present study assessed the mutagenic effect of Ethyl methanesulphonate (EMS) concentrations on growth, yield and nutrient composition of Arachis hypogea (Samnut 24 vr.). Seeds treated with varying concentrations (0.00, 0.25, 0.50, 0.75, 1.00 and 1.25 % v/v) of EMS for 12 hours were sown in randomized complete block design with three replicates. Alkylating effects of the EMS was studied on growth and yield parameters while nutritional quality was determined by proximate analysis at M and M generations. 1 2 Concentrations of 0.75% and 1.00% increased growth, biological yield and seed related parameters significantly (at P< 0.05). Maturity time was significantly shortened by EMS application across the generations. Higher percentage seed crude fat and protein were induced by the treatments with the optimal performance at 0.75- 1.00% concentrations. Mineral content indicated by the percentage ash was marginally increased. The trend in response to the EMS was similar across the generations, however, decreased vigour, reduced growth, less nut and biological yields were observed at M . In most traits evaluated, performance of 0.00, 0.25 and 0.50% 2 treatments were similar, 0.75-1.00% treatments were optimal while 1.25% v/v of EMS was detrimental. Correlation analysis revealed poor linkage of characters in the groundnut; nevertheless, positive significant associations exist between number of leaves and branches, pod length and seed number, seed dimension (length and circumference) and seed weight. The study concluded that since groundnut is basically cultivated for biomass yield a well as source of plant vegetable oil and fat, EMS application at optimal concentration could be used for the crop improvement. Keywords : Alkylating effect, Ethyl methanesulphonate, Mutagenesis, Proximate analysis, Samnut 24.

EMS Induced Desynaptic Male Sterile Lines in Buckwheat (Fagopyrum esculentum Moench)

Cytological analysis of EMS (Ethyl methane sulphonate) treated population of Fagopyrum esculentum revealed an abnormal behaviour of microsporogenesis that affected the meiotic events resulting in the formation of abnormal meiotic products that prevent the gamete formation and impair pollen fertility. The two desynaptic mutants were recorded, showing distinctive variations in the morphology as compared to the control plants. The seeds of Fagopyrum esculentum were treated with EMS solution in different concentrations such as 0.1, 0.3 and 0.5% using potassium phosphate buffer (pH 7) for 5 h. During cytological investigation, 0.5% concentration of EMS enhanced the univalent frequency per cell at diakinesis/Metaphase I, respectively along with unequal segregation at anaphase I which is pronounced to be higher in contrast to bivalents. Because of higher frequency of univalents desynapsis has been categorized as medium-strong type. EMS induced desynaptic plants showed abnormal meiotic behaviour leading to pollen sterility and no seed setting was recorded. The chemical mutagen has acted on recombination genes system which is accountable for synapsis and chiasma formation and further disrupted the complete chiasma assembly. The desynaptic mutant is a potential tool that provides genetic information on the maintenance of chiasma and the study offers the possibility for formation of aneuploids production which may be exploited successfully through chemical mutagenesis in breeding programmes.

Determination of absorption dose in chemical mutagenesis in plants.

Chemical mutagenesis is a useful tool for inducing mutations in plants. Seeds are often used as the material for chemical mutagenesis. The biological effect of a chemical mutagen on seeds is determined by absorption dose (the product of mutagen concentration and acting time, which starts after the mutagen is absorbed by the seeds). In practice, however, the concept of exposure dose (the product of mutagen concentration and treating time) is usually used instead because the time for absorbing mutagen is unknown. In this study, we conducted an experiment using ethyl methane sulphonate (EMS) to treat cauliflower seeds, in which five EMS concentrations (0%, 0.5%, 1.0%, 1.5% and 2.0%), three treating time lengths (4 h, 6 h and 8 h) and two pretreatments (non-presoaking and presoaking of seeds for 2 h) were set. We obtained a well-fitted nonlinear regression model for the relationship between seedling survival rate and the EMS treatment, and its marginal models for the two pretreatments. Based on the models, we determined the EMS absorption doses under the two different pretreatments and identified their 50% lethality dose (LD50). We found that presoaking could delay EMS absorption and therefore reduce the injury caused by EMS within a given treating time, but could hardly change the biological effect of EMS after it is absorbed. The conclusions about absorption dose and presoaking effect obtained in this study might be generally applicable to plant chemical mutagenesis in principle.

Development and evaluation of diverse promising rapeseed (Brassica napus L.) mutants using physical and chemical mutagens

Genetic variability is a prerequisite for any plant breeding program, and mutagenesis is a proven way of creating new variation within a crop germplasm. Novel genetic variability in rapeseed was induced by gamma rays, Ethyl Methane Sulphonate (EMS) and combined mutagen treatment, using various doses and concentrations. The objective was to evaluate and compare the obtained M2 mutants for important quantitative traits in two contrasted environments. Data on phenological, morphological and agronomic parameters were recorded. A large variability was observed and mutagenic treatments had a significant effect on all traits studied. Compared to control plants, mutant genotypes derived from seeds treated with low EMS concentrations during moderate time were earlier and characterized by a higher number of pods per plant. For high concentration of EMS during long time and for combinations of physical and chemical mutagens, a significant decrease in plant height and stature was noticed, as compared to control. Besides, plants derived from gamma rays-treated seeds exhibited the highest 1000-seed weight. The novel induced variability may be integrated in rapeseed breeding program as a new germplasm with improved agronomic traits. Particularly, EMS1-7-stable mutant may be exploited to develop efficiently and quickly a new rapeseed cultivar with some desirable traits. The present study highlights once more the possibility to bring novel genetic diversity for rapeseed desirable traits improvement through mutation breeding.

Enigmatic Phenomenon of Secondary Association among Bivalents in Guar (Cyamopsis tetragonoloba (L.) Taub.)

Cytogenetical assessment of pollen mother cells of Guar (Cyamopsis tetragonoloba (L.) Taub.) in response to treatment of chemical mutagen was done. Ethyl Methane Sulphonate (EMS) revealed possession of an enigmatic behaviour among chromosomal bivalents i.e. formation of secondary association. Chromosomes exhibited a unique tendency of getting associated in close proximity and form discrete pairs. The phenomenon of secondary chromosomal association has been ascribed to the existence of similar cohesive homology or malfunctioning in chiasma formation among the associating bivalents. The association of bivalents was witnessed to be executed in either end to end fashion or sideways association. Various other chromosomal abnormalities were found, but secondary association was the preponderant abnormality which exhibited a continuous increase in its magnitude with increase in EMS concentration. Along with secondary associations, stickiness of chromosomes at anaphase was observed in considerable frequency. A significant influence of chromosomal association on the post meiotic product was also experienced, which in turn, led to influence the meiotic index. Abnormal sporads in the form of triads, dyads, monads and polyads, alongwith normal tetrad were seen. Pollen fertility percent was seen to decline from control to the highest concentration of EMS due to anomalies of preceding stages.