Volume 8, Issue 6, December Issue - 2020, Pages:774-788 |
Authors: Parthasarathi G, Sugitha Thankappan, M. Arumugam Pillai, R. Kannan, S MerinaPrem Kumari, Asish K. Binodh |
Abstract: The present study envisaged the effects of two mutagens, gamma rays and EMS on the phenotypes of two sesame varieties viz., TMV7 and SVPR1. A known quantity of dry, uniform, and healthy seeds of TMV7 and SVPR 1 were irradiated using Co60 (Cobalt 60) with different doses (250, 300, 350, 400, 450 Gy) of gamma rays. For chemical mutagenesis, different concentrations of EMS @ 0.20%, 0.40% and 0.60% was used and treated for 8 h. The dose-response curve of the probit analysis showed that the optimal lethal dose for SVPR1was lower than TMV7. The expected LD50 values of gamma radiation for TMV 7 and SVPR1 were 40.3.91Gy and 343.84Gy, respectively. For EMS, the expected LD50 values are 0.525 % and 0.276% for TMV7 and SVPR1 respectively. Germination and pollen fertility declined linearly with an increase in dose or concentration of the mutagens. Three classes of chlorophyll mutants viz., xantha, chlorine, and viridis in M2 generation reveals a dose dependent relationship between mutagens and frequency of chlorophyll mutants. Mutagenic effectiveness was higher at lower doses whereas mutagenic efficiency was observed higher at extremity doses in both the varieties. The overall considerations on M1 generation effects showed that SVPR1was highly sensitive to gamma rays and TMV7 produced more viable mutationsthanSVPR1. The current studies suggest gamma rays as an efficient mutagen to induce essential mutations in TMV7 for the further crop improvement program. |
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Full Text: Sesame (Sesamum indicum L.) the Indian olive oil is one of the world’s important ancient oilseed crops known as “Queen of Oil seeds”. Seeds are highly protein-rich and the essential amino acids particularly methionine are considered as rejuvenating and anti-aging factors (Namiki, 2007). Oil and protein content of sesame ranges from 48 to 55 % and 20 to 28 % respectively (Pathak et al., 2014). Fats in sesame oil are relatively stable and resist oxidative rancidity as compared to other oils. Sesame contains a high percentage of antioxidants such as sesamol, sesamin, sesamolin, and sesaminol; fatty acids like palmitic, stearic, oleic, and linoleic, and rich in minerals like vitamin E, calcium, magnesium, and phosphorus (Pusadkar et al., 2015; Myint et al., 2020). The sesame oil aids in reducing blood cholesterol, high blood pressure and prevents atherosclerosis, heart diseases, and cancers (Kumar & Singh, 2015). It is a self-pollinated annual shrub widely grown in tropical, subtropical, and temperate areas of the world, exceptionally in India, China, South America, and Africa (Tufail et al., 2020). The architecture of sesame is poorly adapted to modern farming owing to its indeterminate growth habit, non-uniform ripening of capsules, sensitivity to wilting under intensive management, and absence of non-shattering cultivars suited for mechanical harvest (Cagirgan, 2001). Lack of elite cultivars, with inherent resistance to biotic and abiotic stresses, leads to low productivity of sesame (0.33 tonnes ha-1) (Rao et al., 2017). Introgression of genes from wild relatives to cultivated varieties is largely unsuccessful due to the operation of high crossability barriers (Rajeswari et al., 2010; Chaudhary et al., 2019). To generate variability, attempts have been made by crossing to some extent, but desirable success could not have been attained. Mutation breeding serves as an inevitable tool to rectify specific defects by creating desirable genetic variation with the help of physical or chemical mutagens (Chaudhary et al., 2019). Ionizing radiations include X-rays, gamma rays, protons, neutrons, alpha, and beta particles. Gamma rays are widely employed for mutation studies due to their shorter wavelength and more penetration ability (Khin, 2006). Furthermore, chemical mutagens such as sodium azide, ethyl methane sulphonate (EMS), and N-ethyl-N-nitrosourea (ENU), shows distinct negative and positive impacts on the mutated populations (Mohd-Yusoff et al., 2015). EMS is a common mutagen for mutation breeding, that pairs with T instead of C. The mispairing results in G/C to A/T transitions, apart from causing deletions and rearrangements (Rafi et al., 2016). Induced mutations with ionizing radiations and chemical mutagens offer desired phenotypic as well as genotypic effects, which can be further harnessed through recombination breeding or as such (Flibotte et al., 2010). Limited mutant population is advantageous in achieving genome-wide saturation mutagenesis (Henikoff & Comai, 2003). Mutagenic effectiveness and efficiency are two distinct properties of mutagens in plant breeding (Shah et al., 2008; Girija & Dhanavel,2009). The product of maximum desirable changes associated with the least possible undesirable changes is termed efficient mutagenesis. These two criterions, viz., mutagenic efficiency and effectiveness of any mutagen, determines the success of a breeding scheme. Mutagenic efficiency is the rate of mutation produced to biological damage or injury by any mutagen. However, the efficacy of a mutagen is the rate of mutation produced relative to the dose of the mutagen (Anbarasan et al., 2015). Thus the choice of effective mutagen and dose to be used (preferably one that induces enormous variability in any crop) is highly crucial for untargeted induced mutagenesis. In general, the frequency of chlorophyll mutants is used for assessing the potency of any mutagen such as Albina, Striata, Chlorina, Xantha and Viridis. Cultivated species of sesame is under the major threat of phyllody, caused by mycoplasma like organism. Recent studies highlighted that the area and production of sesame crop is declining in the traditional areas. Lack of resistant cultivars is one of the major constraints in sesame growing regions of India (Gupta et al., 2018). With an attempt to develop an improved sesame variety via induced mutagenesis, the present study deals with the mutagenic effectiveness and efficiency of two mutagens gamma ray (Physical mutagen) and ethyl methane sulphonate (chemical mutagen) in M2 generations of two commercial sesame varieties of Tamil Nadu, TMV7 and SVPR1. 2 Materials and Methods The investigation was carried out at the experimental farm of plant breeding and Genetics, Agricultural College and Research Institute, Killikulam, Vallanad Tuticorin Dt, Tamil Nadu (8 ° 46 'N latitude and 77 ° 42' E longitude) in two seasons Kharif (June –Sept) and Rabi season (October – March) of 2019-20. The elevation of the site is 40 m above the mean sea level with relative humidity of 60 to 80%., sub-tropical monsoon climate, and red lateritic soil (pH : 6.8; EC: 0.05 dSm-1). 2.1. Plant materials Seeds of two popular sesame cultivars TMV7 and SVPR1 were obtained from Oil seeds Research Station, Tindivanam, and Cotton Research Station, Srivilliputtur, Tamil Nadu Agricultural University (Table 1). 2.2 Gamma irradiation A known quantity (2 g per dosage) of dry, uniform, and healthy seeds of two genotypes of sesame (TMV7 and SVPR 1) were irradiated using Co60 (Cobalt 60) gamma source with different doses (250, 300, 350, 400, 450 Gy) of gamma rays at Indira Gandhi Centre for Atomic Research (IGCAR), Kalpakkam, Chennai, Tamil Nadu. The irradiated seeds were carefully transferred to sterile containers and sown in the experimental plot within 24 h. 2.3. Chemical Mutagenesis For EMS treatment, 2 g of dry, uniform, healthy, matured and viable seeds of the two varieties viz., TMV7 and SVPR1 were pre-soaked in distilled water for 12 h to enhance imbibing capacity. Pre-soaked seeds were treated in different concentrations of EMS, an alkylating chemical mutagen solution @ 0.20%, 0.40%, and 0.60% for 8 h under controlled conditions at room temperature with intermittent shaking (Kim et al., 2006). After soaking, the treated seeds were washed in running water for an hour to remove the chemical residues. Sodium thiosulphate (100mM) was used to clean beakers and tips that can inactivate residual EMS and enables safe disposal. 2.4. Lethal dose of gamma and EMS The LD50 values for gamma rays and EMS for both the genotypes were calculated according to the probit analysis (Finney, 1978). The probit function represents the inverse cumulative distribution function or quantile function associated with the standard normal distribution. The steps for probit analysis are (i) transformation of the dose concentration of mutagens into log10 values (ii) determination of the mortality % due to treatment doses. The proportions were corrected for control mortality if it is more than 10% using Abbot Formula (Ramchander et al., 2015). |
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