Analyzing frequency and spectrum of chlorophyll mutation induced through Gamma ray and Combination treatment (Gamma + EMS) on genetic paradigm of Artemisia annua L.
Keywords:Artemisia annua L. (Linnaeus), chlorophyll mutants, cytological anomalies, gamma rays, EMS
For the development of genetic programs with novel characteristics induced mutagenesis has been used extensively. Chlorophyll (chl) mutations are considered as the most dependable indices for assessing the efficiency of different mutagens in inducing the genetic variability in crop plants and are also used as genetic markers in basic and applied research. In the present scenario of high health susceptibility, the global demand for natural medicine derived from plant species has increased enormously. Sweet wormwood (Artemisia annua Linnaeus) – an important medicinal plant species with immense remedial values, was selected for the present study and exposed to gamma rays at 100 Gy, 200 Gy, 300 Gy and combination treatments with 100 Gy + 0.1%EMS, 200 Gy + 0.1% EMS, 300 Gy + 0.1%EMS. Meiotic study was also done and various cytological aberrations were observed in M2 generation like stickiness, precocious, scattering, laggard and bridge etc. The frequency of induced chl mutation varied in different mutagen treatments. Eight different types of chl mutants namely albina, chlorina, xantha, aurea, viridis, yellow viridis and tigrina etc. were recorded in M2 generation on plant population basis. The frequency of xantha mutants was quite high in both the treatments but in gamma exposed set it was followed by albina whereas in combination treatments viridis was second highest mutant. In different mutants quantitative analysis of chl pigments was also done and content was highest in viridis i.e. 3.86µg/ml FW and lowest in albina i.e. 0 µg/ml FW . Although chlorophyll mutations thought to be lethal in nature, but present study has proven to be a milestone in identifying the threshold dose of a mutagen that would increase the genetic variability and induces new trait in Artemisia annua Linnaeus.
Arisha MH, Liang BK, Shah SM, Gong ZH, Li DW. 2014. Kill curve analysis and response of first generation Capsicum annuum L. B12 cultivar to ethyl methane sulfonate. Genet Mol Res. 13:49–61.
Arisha MH, Shah SN, Gong ZH, Jing H, Li C, Zhang HX. 2015. Ethyl methane sulfonate induced mutations in M2 generation and physiological variations in M1 generation of peppers (Capsicum annuum L.). Front Plant Sci. 6:399.
Avijeet C, Shukla S, Rastogi A, Mishra BK, Ohri D, Singh SP. 2011. Impact of mutagenesis on cytological behavior in relation to specific alkaloids in Opium Poppy (Papaver somniferum L.). Caryologia. 64:14–24.
Blixt S. 1961. Quantitative studies of induced mutations in peas. V. Chlorophyll mutations. Agri Hort Genet. 1:19.
Bolbhat SN, Dhumal KN. 2009. Induced macromutations in horsegram [Macrotyloma uniflorum (Lam.) Verdc]. Legum Res. 32:278–281.
Bosland PW. 2002. Inheritance of a novel flaccid mutant in Capsicum annuum. J Hered. 93:380–392.
Botticella E, Sestili F, Hernandez-Lopez A, Phillips A, Lafiandra D. 2011. High resolution melting analysis for the detection of EMS induced mutations in wheat Sbella genes. BMC Plant Biol. 11:156.
Dhumal KN, Bolbhat SN. 2012. Induction of genetic variability with gamma radiation and its applications in improvement of horsegram. Gamma Radiation. 21:207–228.
Eckhardt U, Grimm B, Hörtensteiner S. 2004. Recent advances in chlorophyll biosynthesis and breakdown in higher plants. Plant Mol Biol. 56:1–4.
Flood PJ, Harbinson J, Aarts MG. 2011. Natural genetic variation in plant photosynthesis. Trends Plant Sci. 16:327–335.
Gandhi S, Umavathi S, Mullainathan L. 2014. Studies on induced chlorophyll mutants in green gram (Vigna radiata (L.) Wilczek), Int J of Adv Res. 2:00–04.
Gaur LB, Singh SP, Srivastava K. 2013. Frequency and spectrum of chlorophyll mutation in chilli (Capsicum annuum L.) Int J Plant Sci. 8:70–74.
Gustafsson Å. 1940. The mutation system of the chlorophyll apparatus. Kungliga Fysiografiska Sallskapets i Lund Handlingar. 51.
Hanafiah DS, Trikoesoemoningtyas T, Yahya S, Wirnas D. 2010. Induced mutations by gamma ray irradiation to Argomulyo soybean (Glycine max) variety. Nusantara Bioscience. 2.
Haq MA. Genetic and physiologic studies on induced mutants of chickpea (Doctoral dissertation, Ph. D. Thesis. University of the Punjab, Lahore, Pakistan), 1990.
Hasbullah NA, Taha RM, Saleh A, Mahmad N. 2012. Irradiation effect on in vitro organogenesis, callus growth and plantlet development of Gerbera jamesonii. Hortic Bras. 30:252–257.
International Atomic Energy Agency (IAEA)., Mutant variety database. https://mvd.iaea.org, 2017.
Jabee F, Ansari MY, Shahab D. 2008. Studies on the effect of maleic hydrazide on root tip cells and pollen fertility in Trigonella foenum-graecum L. Turk J Bot. 32:337–344.
Jafri IF, Khan AH, Gulfishan M. 2011. Genotoxic effects of 5-bromouracil on cytomorphological characters of Cichorium intybus L. Afr J Biotechnol. 10:10595–10599.
Khan S, Wani MR, Bhat M, Parveen K. 2005. Induced chlorophyll mutations in chickpea (Cicer arietinum L.). Int J Agric Biol. 7:764–767.
Kharkwal MC. 1998. Induced mutations in chickpea (Cicer arietinum L.) II. Frequency and spectrum of chlorophyll mutations. Indian J Genet plant Breed. 58:465–474.
Kirchhoff WR, Hall AE, Thomson WW. 1989. Gas exchange, carbon isotope discrimination, and chloroplast ultrastructure of a chlorophyll‐deficient mutant of cowpea. Crop sci. 29:109–115.
Kolar F, Pawar N, Dixit G. 2011. Induced chlorophyll mutations in Delphinium malabaricum (Huth) Munz. J Appl Hortic. 13:18–24.
Konzak CF, Woo SC, Dickey J. 1969. Induced dominant semi-dwarf plant height mutation in spring wheat. Wheat Inform Serv.
Koobkokkruad T, Chochai A, Kirdmanee C, De-Eknamkul W. 2008. Effects of low-dose gamma irradiation on artemisinin content and amorpha-4, 11-diene synthase activity in Artemisia annua L. Int J Radiat Biol. 84:878–884.
Kulkarni GB, Mogle UP. 2013. Effects of mutagen on chlorophyll mutation in horse gram [Macrotyloma uniflorum (Lam) Verdcourt]. Bio Disc. 4:214–229.
Kumar A, Parmhansh P, Prasad R. 2009. Induced chlorophyll and morphological mutations in mungbean (Vigna radiata L. Wilczek). Legum Res. 32:41–45.
Kumar AP, Boualem A, Bhattacharya A, Parikh S, Desai N, Zambelli A, Leon A, Chatterjee M, Bendahmane A. 2013. SMART–sunflower mutant population and reverse genetic tool for crop improvement. BMC Plant Biol. 13:38.
Kumar G, Gupta P. 2009. Induced karyomorphological variations in three phenodeviants of Capsicum annuum L. Turk J Biol. 33:123–128.
Lal GM, Toms B, Smith S. 2009. Induced chlorophyll mutations in black gram. Asian J Agric Sci. 1:1–3.
Lichtenthaler HK, Wellburn AR. 1983. Determinations of total carotenoids and chlorophylls a and b of leaf extracts in different solvents. Biochem Soc Trans. 11:591–592.
Meherchandani M. 1975. Effect of gamma radiation on dormant seeds of Avena sativa L. Rad Bot. 15:439–445.
Mitra PK. 1996. Studies in the frequency and segregation of induced chlorophyll mutations in Nigella sativa L. Adv Plant Sci. 12:125–139.
Nielsen NC, Smillie RM, Henningsen KW, Von Wettstein D, French CS. 1979. Composition and function of thylakoid membranes from grana-rich and grana-deficient chloroplast mutants of barley. Plant Physiol. 63:174–182.
Østergaard L, Yanofsky MF. 2004. Establishing gene function by mutagenesis in Arabidopsis thaliana. Plant J. 39:682–696.
Page DR, Grossniklaus U. 2002. The art and design of genetic screens: Arabidopsis thaliana. Nat Rev Genet. 3:124–136.
Panigrahi KK, Mohanty A and Jyotshnarani BB. 2015. Mutagenic efficiency and effectiveness of gamma rays, ethyl methane sulphonate (ems), nitrosoguanidine (ng) and their synergistic effect for different polygenic traits in black gram (Vigna mungo (l.) hepper) through induced mutagenesis. Int J Plant Anim Environ Sci. 5:292.
Parry MA, Madgwick PJ, Bayon C, Tearall K, Hernandez-Lopez A, Baudo M, Rakszegi M, Hamada W, Al-Yassin A, Ouabbou H, Labhilii M, Phillips AL. 2009. Mutation discovery for crop improvement. J Exp Bot. 60:2817–2825.
Prasad AB, Das AK. 1980. Studies of induced chlorophyll mutations in Lathyrus sativus L. Cytologia. 45:335–341.
Qin R, Qiu Y, Cheng Z, Shan X, Guo X, Zhai H, Wan J. 2008. Genetic analysis of a novel dominant rice dwarf mutant 986083D. Euphytica. 160:379–387.
Reddy CS, Smith JD. 1981. Mutagenic effects of combination treatments of hadrazine, ethyl methanesulphonate and gamma rays in Sorghum bicolor (L.) Moench. Indian J Bot. 4:5–14.
Reddy PM, Munirajappa M. 2012. Gamma ray induced meiotic abnormalities IN S13 Mulberry. Int J Sci Nat. 3:170–172.
Reddy VR, Gupta PK. 1989. Induced mutations in triticale: Frequency and spectrum of chlorophyll mutations. Indian J Genet plant Breed. 49:183–190.
Roychowdhury R, Tah J. 2013. Mutagenesis—A potential approach for crop improvement. In Crop Improvement pp. 149–187. Springer, Boston, MA.
Sato M, Gaul H. 1967. Effect of ethyl methanesulfonate on the fertility of barley. Rad Bot. 7:7–15.
Serrat X, Esteban R, Guibourt N, Moysset L, Nogués S, Lalanne E. 2014. EMS mutagenesis in mature seed-derived rice calli as a new method for rapidly obtaining TILLING mutant populations. Plant Methods. 10:1–4.
Shah TM, Mirza JI, Haq MA, Atta BM. 2006. Induced genetic variability in chickpea (Cicer arietinum L.). I. Frequency and spectrum of chlorophyll mutations. Pak J Bot. 38:1217.
Shahwar D, Ansari MY, Choudhary S, Aslam R. 2017. Evaluation of yield attributing variants developed through ethyl methane sulphonate in an important proteinaceous crop-Vicia faba. Asian J Crop Sci. 9:20–7.
Shahwar D, Khan Z, Ansari MY. 2020. Evaluation of mutagenized lentil populations by caffeine and EMS for exploration of agronomic traits and mutant phenotyping. Ecol Genet Genom. 14:100049.
Shakoor A, Sadiq MS, Hasan MU, Saleem M. 1978. Selection for useful semidwarf mutants through induced mutations in bread wheat. In Proc 5th Int Wheat Genet Symp, New Delhi (Vol. 1, pp. 540–546).
Sharma RP. 1970. Increased mutation frequency and wider mutation spectrum in barley induced by combining gamma-rays with ethyl methane sulphonate. Indian J Genet plant Breed. 30:180–196.
Sharma S, Sharma P, Datta SP, Gupta P. 2010. Morphological and Biochemical Response of Cicer arietinum L. var. pusa-256 towards an Excess of Zinc Concentration. Life Sci. 7:95–98.
Sharma SK, Sharma B. 1981. Induced chlorophyll mutations in lentil.Indian J Genet plant Breed.41:328–333.
Shukla nee Tripathi R, Kumar G. 2010. Comparative effect of ageing and gamma irradiation on the somatic cells of Lathyrus sativus L. J Cent Eur Agric. 11:437–442.
Singh GR, Sareen PK, Saharan RP. 1999. Clastogenic effect of gamma rays, EMS and ECH in Vigna radiata (L.) Wilczek. J Cytol Genet. 34:21–23.
Singh O. 1988. Induced mutations and cytogenetic studies in chickpea (Cicer arietinum L.). Meerut University, Meerut.
Sinuraya M, Rosmayati H, Hanafiah DS. 2017. The effect of gamma rays iradiation to morphological and agronomical character of local Samosir shallot. International network for natural sciences. 10:126–134
Solanki IS, Phogat DS, Waldia RS. 2004. Frequency and spectrum of morphological mutations and effectiveness and efficiency of chemical mutagens in Macrosperma lentil. National Journal of Plant Improvement. 6:22–25.
Solanki IS, Sharma B. 1994. Mutagenic effectiveness and efficiency of gamma rays, ethylene imine and N-nitroso-N-ethyl urea in macrosperma lentil (Lens culinaris Medik.). Indian J Genet plant Breed. 54:72–76.
Solanki IS. 2005. Isolation of macromutations and mutagenic effectiveness and efficiency in lentil (Lens culinaris Medik.). Indian J Genet plant Breed. 65:264–278
Swaminathan MS, Chopra VL, Bhaskaran S. 1962. Chromosome aberrations and the frequency and spectrum of mutations induced by ethylmethane sulphonate in barley and wheat. Indian J Genet. 22:192–207.
Talebi AB, Talebi AB, Shahrokhifar B. 2012. Ethyl methane sulphonate (EMS) induced mutagenesis in Malaysian rice (cv. MR219) for lethal dose determination. Am J of PlantSci. 3:1661–1665.
Thilagavathi C, Mullainathan L. 2011. Influence of physical and chemical mutagens on quantitative characters of Vigna mungo (L. Hepper). IntMultidiscip Res J. 1:6–8.
Van Harten AM. 1998. Mutation breeding: theory and practical applications. Cambridge University Press.
Vardhan PV, Shukla LI. 2017. Gamma irradiation of medicinally important plants and the enhancement of secondary metabolite production. Int J Radiat Biol. 93:967–79.
Vaughn KC, Wilson KG, Stewart KD. 1978. Light-harvesting pigment-protein complex deficiency in Hosta (Liliaceae). Planta. 143:275–278.
Waghmare VN, Mehra RB. 2000. Induced genetic variability for quantitative characters in grasspea (Lathyrus sativus L.). Indian J Genet. 60:81–87.
Walles B. Plastid structures and mutations. In: Structure and Function of Chloroplasts, Gibba M (ed), Springer Verlag New York,1973;51.
Wang ZK, Huang YX, Miao ZD, Hu ZY, Song XZ, Liu L. 2013. Identification and characterization of BGL11 (t), a novel gene regulating leaf-color mutation in rice (Oryza sativa L.) Genes Genomics. 35:491.
Wani AA, Anis M. 2004. Spectrum and frequency of chlorophyll mutations induced by gamma rays and EMS in Cicer arietinum L. J Cytol Genet. 5:143–147.
Williams JG, Kubelik AR, Livak KJ, Rafalski JA, Tingey SV. 1990. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res. 18:6531–6535.
Wu Z, Zhang X, He B, Diao L, Sheng S, Wang J, Guo X, Su N, Wang L, Jiang L, Wang C. 2007. A chlorophyll-deficient rice mutant with impaired chlorophyllide esterification in chlorophyll biosynthesis. Plant physiol. 145:29–40.
How to Cite
Copyright (c) 2022 RAJANI SINGH, Girjesh Kumar
This work is licensed under a Creative Commons Attribution 4.0 International License.
- Copyright on any open access article in a journal published byCaryologia is retained by the author(s).
- Authors grant Caryologia a license to publish the article and identify itself as the original publisher.
- Authors also grant any third party the right to use the article freely as long as its integrity is maintained and its original authors, citation details and publisher are identified.
- The Creative Commons Attribution License 4.0 formalizes these and other terms and conditions of publishing articles.
- In accordance with our Open Data policy, the Creative Commons CC0 1.0 Public Domain Dedication waiver applies to all published data in Caryologia open access articles.