Functional Plant Biology Functional Plant Biology Society
Plant function and evolutionary biology
RESEARCH ARTICLE

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 B

A 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 http://dx.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.


References

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 Gosal SS 1982 ‘Tissue culture in economically important plants.’ Singapore

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
DOI

Bates LS Waldren RP Teare ID 1973 Rapid determination of free proline for water stress studies. Plant and Soil 39 205 207 DOI

Beyer WF Fridovich I 1987 Assaying for superoxide dismutase activity: some large consequences of minor changes in conditions. Analytical Biochemistry 161 559 566 DOI

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 DOI

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
DOI

Bowler C Montagu TV Inze D 1992 Superoxide dismutase and stress tolerance. Annual Review of Plant Physiology and Plant Molecular Biology 43 83 116 DOI

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

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’. Alton Meister 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 ‘Arabidopsis’. Meyerowitz EM Somerville CR 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
DOI

Desikan R Neill SJ Hancock JT 2000 Hydrogen peroxide-induced gene expression in Arabidopsis thaliana. Free Radical Biology and Medicine 28 773 778 DOI

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

Dionisio-Sese M Tobita S 1998 Antioxidant responses of rice seedlings to salinity stress. Plant Science 135 1 9
DOI

Dix PJ Street HE 1975 Sodium chloride-resistant cultured cell lines from Nicotiana sylvestris and Capsicum annuum. Plant Science Letters 5 231 237 DOI

Fridovich I 1986 Biological effects of superoxide radical. Archives of Biochemistry and Biophysics 247 1 11 DOI

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


Greenway H Munns R 1980 Mechanism of salt tolerance in non-halophytes. Annual Review of Plant Physiology 31 149 190
DOI

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 Gutteridge JMC 1989 ‘Free radicals in biology and medicine.’ Clarendon Press Oxford

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
DOI

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 DOI

Jain SM Brar DS Ahloowalia BS 1998 ‘Somaclonal variation and induced mutation in crop improvement.’ Kluwer Academic publishers Dordrecht

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


Lichtenthaler HK 1987 Chlorophylls and carotenoids: pigments of photosynthetic bio-membranes. In ‘Methods in enzymology’. Packer L Douce R 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
DOI

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 DOI

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
DOI

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
DOI

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 DOI

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 DOI

Ohkawa H Ohishi N Yagi K 1979 Assay for lipid peroxidation in animal tissues by thiobarbituric acid reaction. Analytical Biochemistry 95 351 358 DOI

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 1997 a In vitro selection of NaCl tolerant callus lines of Cymbopogon martinii (Roxb.) Wats. Plant Science 124 203 210
DOI

Patnaik J Debata BK 1997 b Regeneration of plantlets from NaCl tolerant callus lines of Cymbopogon martinii (Roxb.) Wats. Plant Science 128 67 74 DOI

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
DOI

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
DOI

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
DOI

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 DOI

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 DOI

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
DOI

Young AJ 1991 The protective role of carotenoids in higher plants. Physiologia Plantarum 83 702 708 DOI

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



Export Citation