Isolation of a NaCl-tolerant mutant of Chrysanthemum morifolium by gamma radiation: in vitro mutagenesis and selection by salt stress
Zahed Hossain A , Abul Kalam Azad Mandal A , Subodh Kumar Datta A C and Amal K. Biswas BA Botanic Garden and Floriculture, National Botanical Research Institute, Lucknow 226 001, Uttar Pradesh, India.
B Cytogenetics and Plant Breeding Laboratory, Botany Department, University of Kalyani, Kalyani 741 235, West Bengal, India.
C Corresponding author. Email: subodhdatta@usa.net
Functional Plant Biology 33(1) 91-101 https://doi.org/10.1071/FP05149
Submitted: 20 June 2005 Accepted: 16 September 2005 Published: 3 January 2006
Abstract
A stable NaCl-tolerant mutant (R1) of Chrysanthemum morifolium Ramat has been developed by in vitro mutagenesis with gamma radiation (5 gray; Gy). Salt tolerance was evaluated by the capacity of the plant to maintain both flower quality and yield under NaCl stress. Enhanced salt tolerance of the R1 mutant was attributed to increased activities of reactive oxygen species (ROS)-scavenging enzymes, namely superoxide dismutase (SOD), monodehydroascorbate reductase (MDAR), dehydroascorbate reductase (DHAR) and glutathione reductase (GR), and to reduced membrane damage, higher relative water content (RWC), chlorophyll and carotenoids contents. RAPD analysis revealed two polymorphic bands (956 and 1093 bp) for the R1 mutant that might be considered as specific RAPD markers associated with salt tolerance. Better performance of the R1 progeny under identical salinity stress conditions, even in the second year, confirmed the genetic stability of the induced salt tolerance character. The R1 mutant developed by gamma ray treatment can be considered a salt-tolerant mutant showing all the positive characteristics of tolerance to NaCl stress.
Keywords: antioxidant enzymes, Chrysanthemum, gamma ray, in vitro mutagenesis, NaCl-tolerant, reactive oxygen species.
Acknowledgments
The authors thank the Director, National Botanical Research Institute, Lucknow for providing facilities. Zahed Hossain thankfully acknowledges the financial support provided by Council of Scientific and Industrial Research (CSIR), New Delhi as S.R.F.
Acar O,
Türkan I, Ozdemir F
(2001) Superoxide dismutase and peroxidase activities in drought sensitive and resistant barley (Hordeum vulgare L.) varieties. Acta Physiologiae Plantarum 3, 351–356.
Bajaj, YPS ,
and
Gosal, SS (Eds) (1982).
Banerji BK, Datta SK
(1992) Gamma ray induced flower shape mutation in Chrysanthemum cv. ‘Jaya’. Journal of Nuclear Agriculture and Biology 21, 73–79.
Barakat MN, Abdel-Latif TH
(1996)
In vitro selection of wheat callus tolerant to high levels of salt and plant regeneration. Euphytica 91, 127–140.
Bartoli CG,
Simontacchi M,
Guiamet JJ, Montaldi E
(1995) Antioxidant enzymes and lipid peroxidation during aging of Chrysanthemum morifolium RAM petals. Plant Science 104, 161–168.
| Crossref | GoogleScholarGoogle Scholar |
Bates LS,
Waldren RP, Teare ID
(1973) Rapid determination of free proline for water stress studies. Plant and Soil 39, 205–207.
| Crossref | GoogleScholarGoogle Scholar |
Beyer WF, Fridovich I
(1987) Assaying for superoxide dismutase activity: some large consequences of minor changes in conditions. Analytical Biochemistry 161, 559–566.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Binh DQ,
Heszky LE,
Gyulai G, Csillag A
(1992) Plant regeneration of NaCl-pretreated cells from long-term suspension culture of rice (Oryza sativa L.) in high saline conditions. Plant Cell, Tissue and Organ Culture 29, 75–82.
| Crossref | GoogleScholarGoogle Scholar |
Blum A, Ebercon A
(1981) Cell membrane stability as measure of drought and heat tolerance in wheat. Crop Science 21, 43–47.
Bor M,
Ozdemir F, Turkan I
(2003) The effect of salt stress on lipid peroxidation and antioxidants in leaves of sugar beet Beta vulgaris L. and wild beet Beta maritima L. Plant Science 164, 77–84.
| Crossref | GoogleScholarGoogle Scholar |
Bowler C,
Montagu TV, Inze D
(1992) Superoxide dismutase and stress tolerance. Annual Review of Plant Physiology and Plant Molecular Biology 43, 83–116.
| Crossref | GoogleScholarGoogle Scholar |
Bradford MM
(1976) A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. Analytical Biochemistry 72, 248–254.
| Crossref |
PubMed |
Brennan T, Frenkel C
(1977) Involvement of hydrogen peroxide in regulation of senescence in pear. Plant Physiology 59, 411–416.
Carlberg I, Mannervik B
(1985) Glutathione reductase. In ‘Methods in enzymology’. (Ed. Alton Meister)
pp. 484–490. (Academic Press: San Diego)
Chandler SF, Thorpe TA
(1987) Characterization of growth, water relations, and proline accumulation in sodium sulfate tolerant callus of Brassica napus L. cv. Westar (Canola). Plant Physiology 84, 106–111.
Crute I, Benyon J, Dangl J, Holub E, Mauchi-Mani B, Slusarenko A, Staskawicz B, Ausubel F
(1994) Microbial pathogenesis in Arabidopsis. In ‘’. (Eds EM Meyerowitz, CR Somerville)
pp. 705–747. (Cold Spring Harbor Laboratory Press: Cold Spring Harbor)
Datta SK, Gupta MN
(1981) Effects of gamma irradiation on rooted cuttings of Korean type Chrysanthemum cv. ‘Nimrod’. Bangladesh Journal of Botany 10, 124–131.
Demiral T, Türkan I
(2005) Comparative lipid peroxidation, antioxidant defense systems and proline content in roots of two rice cultivars differing in salt tolerance. Environmental and Experimental Botany 53, 247–257.
| Crossref | GoogleScholarGoogle Scholar |
Desikan R,
Neill SJ, Hancock JT
(2000) Hydrogen peroxide-induced gene expression in Arabidopsis thaliana. Free Radical Biology and Medicine 28, 773–778.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Desikan R,
Reynolds A,
Hancock JT, Neill SJ
(1998) Harpin and hydrogen peroxide both initiate programmed cell death but have differential effects on gene expression in Arabidopsis suspension cultures. The Biochemical Journal 330, 115–120.
| PubMed |
Dionisio-Sese M, Tobita S
(1998) Antioxidant responses of rice seedlings to salinity stress. Plant Science 135, 1–9.
| Crossref | GoogleScholarGoogle Scholar |
Dix PJ, Street HE
(1975) Sodium chloride-resistant cultured cell lines from Nicotiana sylvestris and Capsicum annuum. Plant Science Letters 5, 231–237.
| Crossref | GoogleScholarGoogle Scholar |
Fridovich I
(1986) Biological effects of superoxide radical. Archives of Biochemistry and Biophysics 247, 1–11.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Fukutaku Y, Yamada Y
(1984) Sources of proline nitrogen in water stressed soybean (Glycine max) II, fate of 15N-labeled protein. Physiologia Plantarum 61, 622–628.
Gosal SS, Bajaj YP
(1984) Isolation of sodium chloride cell lines in some grain-legumes. Indian Journal of Experimental Biology 22, 209–214.
Gossett DR,
Millhollon EP, Lucas MC
(1994) Antioxidant response to NaCl stress in salt-tolerant and salt-sensitive cultivars of cotton. Crop Science 34, 706–714.
Gossett DR,
Banks SW,
Millhollon EP, Lucas MC
(1996) Antioxidant response to NaCl stress in a control and an NaCl-tolerant cotton cell line grown in the presence of paraquat, buthionine sulfoximine and exogenous glutathione. Plant Physiology 112, 803–809.
| PubMed |
Greenway H, Munns R
(1980) Mechanism of salt tolerance in non-halophytes. Annual Review of Plant Physiology 31, 149–190.
| Crossref | GoogleScholarGoogle Scholar |
Gupta MN, Datta SK
(1978) Effects of gamma rays on growth, flowering behaviour and induction of somatic mutations in Chrysanthemum. SABRAO Journal 10, 149–161.
Halliwell, B ,
and
Gutteridge, JMC (1989).
Hanson AD,
Nelsen CE, Everson EH
(1977) Evaluation of free proline accumulation as an index of drought resistance using two contrasting barley cultivars. Crop Science 17, 720–726.
Hanson AD,
Nelsen CE,
Pederson AR, Everson EH
(1979) Capacity for proline accumulation during water stress in barley and its implication for breeding for drought resistance. Crop Science 19, 489–493.
Hernandez JA,
Olmos E,
Corpas FJ,
Sevilla F, Rio LA
(1995) Salt induced oxidative stress in chloroplast of pea plants. Plant Science 105, 151–167.
| Crossref | GoogleScholarGoogle Scholar |
Jain RK,
Dhawan RS,
Sharma DR, Chowdhury JB
(1987) Salt-tolerance and proline accumulation: a comparative study in salt-tolerant and wild type cultured cells of eggplant. Plant Cell Reports 6, 382–384.
| Crossref | GoogleScholarGoogle Scholar |
Jain, SM ,
Brar, DS ,
and
Ahloowalia, BS (Eds) (1998).
Kalir A,
Omri G, Poljakoff-Mayber A
(1984) Peroxidase and catalase activity in leaves of Halimione portulacoides exposed to salinity. Physiologia Plantarum 62, 238–244.
Karadimova M, Djambova G
(1993) Increased NaCl-tolerance in wheat (Triticum aestivum L. and durum Desf.) through in vitro selection. In Vitro Cellular and Developmental Biology 29, 180–182.
Kellogg EW, Fridovich I
(1975) Superoxide, hydrogen peroxide and singlet oxygen in lipid peroxidation by a xanthine oxidase system. The Journal of Biological Chemistry 250, 8812–8817.
| PubMed |
Lichtenthaler HK
(1987) Chlorophylls and carotenoids: pigments of photosynthetic bio-membranes. In ‘Methods in enzymology’. (Eds L Packer, R Douce)
pp. 350–382. (Academic Press: San Diego)
Lin CC, Kao CH
(1996) Proline accumulation is associated with inhibition of rice seedling root growth caused by NaCl. Plant Science 114, 121–128.
| Crossref | GoogleScholarGoogle Scholar |
Martinez CA,
Maestri M, Lani EG
(1996)
In vitro salt tolerance and proline accumulation in Andean potato (Solanum spp.) differing in frost resistance. Plant Science 116, 177–184.
| Crossref | GoogleScholarGoogle Scholar |
Mittal R, Dubey RS
(1991) Behaviour of peroxidases in rice: changes in enzyme activity and isoforms in relation to salt tolerance. Plant Physiology and Biochemistry 29, 31–40.
Miyake C, Asada K
(1992) Thylakoid-bound ascorbate peroxidase in spinach chloroplasts and photoreduction of its primary oxidation product monodehydroascorbate radicals in thylakoids. Plant and Cell Physiology 33, 541–553.
Moftah AE, Michel BE
(1987) The effect of sodium chloride on solute potential and proline accumulation in soybean leaves. Plant Physiology 83, 238–240.
Mullineaux P,
Ball L,
Escobar C,
Karpinska B,
Creissen G, Karpinski S
(2000) Are diverse signaling pathways integrated in the regulation of Arabidopsis antioxidant defense gene expression in response to excess excitation energy? Philosophical Transactions of the Royal Society of London. 355, 1531–1540.
| Crossref | GoogleScholarGoogle Scholar |
Murashige T, Skoog F
(1962) A revised medium for rapid growth and bioassay with tobacco tissue culture. Physiologia Plantarum 15, 473–497.
Nakano Y, Asada K
(1981) Hydrogen peroxide is scavenged by ascorbate specific peroxidase in spinach chloroplast. Plant and Cell Physiology 22, 867–880.
Neill S,
Desikan R, Hancock J
(2002) Hydrogen peroxide signaling. Current Opinion in Plant Biology 5, 388–395.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Nguyen NT,
Moghaieb REA,
Saneoka H, Fujita K
(2004) RAPD markers associated with salt tolerance in Acacia auriculiformis and Acacia mangium. Plant Science 167, 797–805.
| Crossref | GoogleScholarGoogle Scholar |
Noctor G, Foyer C
(1998) Ascorbate and glutathione: keeping active oxygen under control. Annual Review of Plant Physiology and Plant Molecular Biology 49, 249–279.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Ohkawa H,
Ohishi N, Yagi K
(1979) Assay for lipid peroxidation in animal tissues by thiobarbituric acid reaction. Analytical Biochemistry 95, 351–358.
| Crossref | GoogleScholarGoogle Scholar |
Olmos E,
Hernandez JA,
Sevilla F, Hellin E
(1994) Induction of several antioxidant enzymes in the selection of a salt tolerant cell line of Pisum sativum. Journal of Plant Physiology 144, 594–598.
Patnaik J, Debata BK
(1997a)
In vitro selection of NaCl tolerant callus lines of Cymbopogon martinii (Roxb.) Wats. Plant Science 124, 203–210.
| Crossref | GoogleScholarGoogle Scholar |
Patnaik J, Debata BK
(1997b) Regeneration of plantlets from NaCl tolerant callus lines of Cymbopogon martinii (Roxb.) Wats. Plant Science 128, 67–74.
| Crossref | GoogleScholarGoogle Scholar |
Perez-Alfocea F,
Santa-Cruz A,
Guerrier G, Bolarin MC
(1994) NaCl stress-induced organic solute changes on levels and calli of Lycopersicon esculentum, L. pennellii and their interspecific hybrid. Journal of Plant Physiology 143, 106–111.
Premachandra GS,
Soneoka H,
Kanaya M, Ogata S
(1991) Cell membrane stability and leaf surface wax content as affected by increasing water deficits in maize. Journal of Experimental Botany 42, 167–171.
Saghai-Maroof MA,
Soliman KM,
Jorgensen RA, Allard RW
(1984) Ribosomal DNA spacer-length polymorphisms in barley: Mendelian inheritance, chromosomal location, and population dynamics. Proceedings of the National Academy of Sciences USA 81, 8014–8018.
Sairam RK, Srivastava GC
(2002) Changes in antioxidant activity in sub-cellular fractions of tolerant and susceptible wheat genotypes in response to long term salt stress. Plant Science 162, 897–904.
| Crossref | GoogleScholarGoogle Scholar |
Salin ML
(1988) Toxic oxygen species and protective systems of the chloroplasts. Physiologia Plantarum 72, 681–689.
Shalata A, Tal M
(1998) The effect of salt stress on lipid peroxidation and antioxidants in the leaf of the cultivated tomato and its wild salt-tolerant relative Lycopersicon pennellii. Physiologia Plantarum 104, 169–174.
| Crossref | GoogleScholarGoogle Scholar |
Sheoran IS, Garg OP
(1979) Quantitative and qualitative changes in peroxidase during germination of mung bean under salt stress. Physiologia Plantarum 46, 147–150.
Siefermann-Harms D
(1987) The light harvesting and protective functions of carotenoids in photosynthetic membrane. Physiologia Plantarum 68, 561–568.
Stewart GR, Lee JA
(1974) The role of proline accumulation in halophytes. Planta 120, 279–289.
| Crossref | GoogleScholarGoogle Scholar |
Vaidyanathan H,
Sivakumar P,
Chakrabarty R, Thomas G
(2003) Scavenging of reactive oxygen species in NaCl-stressed rice (Oryza sativa L.) — differential response in salt-tolerant and sensitive varieties. Plant Science 165, 1411–1418.
| Crossref | GoogleScholarGoogle Scholar |
Vajrabhaya M,
Thanapaisal T, Vajrabhaya T
(1989) Development of salt tolerant lines of KDML and LPT rice cultivars through tissue culture. Plant Cell Reports 8, 411–414.
| Crossref | GoogleScholarGoogle Scholar |
Whetherley PE
(1950) Studies in the water relations of cotton plants. I. The field measurement of water deficit in leaves. New Phytologist 49, 81–87.
Williams JGK,
Kubelik AR,
Livak KJ,
Rafalski JA, Tigey SV
(1990) DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Research 22, 6531–6535.
Winicov I
(1996) Characterization of rice (Oryza sativa L.) plants regenerated from salt-tolerant cell lines. Plant Science 113, 105–111.
| Crossref | GoogleScholarGoogle Scholar |
Young AJ
(1991) The protective role of carotenoids in higher plants. Physiologia Plantarum 83, 702–708.
| Crossref | GoogleScholarGoogle Scholar |
Zheng Z-L,
Yang Z,
Jang J-C, Metzger JD
(2001) Modification of plant architecture in chrysanthemum by ectopic expression of the Tobacco Phytochrome B1 gene. Journal of the American Society for Horticultural Science 126, 19–26.
