Polyploidy increases tolerance to salt stress in Anise hyssop (Agastache foeniculum [Pursh.] Kuntze)

Polyploidy increases tolerance to salt stress in Anise hyssop


  • Yavar Sharafi Shahed university
  • Seyyedeh Farahnaz Talebi
  • Mohammad jamal saharkhiz
  • Maryam Jafarkhani Kermani




Anise hyssop. Essential oil. Polyploidy. Salt tolerance


Salinization is one of the most serious environmental problems in agriculture. Polyploid induction could increase abiotic stress tolerance in plants. In this study, the effect of different NaCl concentrations (0, 50, 100 and 150 mM) was studied on diploid (2x) and tetraploid (4x) plants of anise hyssop (Agastache foeniculum) in vitro. The results indicated that salt stress reduced survival percent, stem length, leaf and shoot number in both tetraploid and diploid plants. However, tetraploid plants had better survival and growth rates compared to diploids. The highest antioxidant enzymes activity was observed in the plants treated with 100 mM NaCl, while increasing the salinity to 150 mM NaCl lowered the activity of antioxidant enzymes significantly. Essential oil content in diploid and tetraploid plants decreased as the concentration of NaCl was elevated. Also, salinity stress affected the chemical composition of essential oil in both diploid and tetraploid plants. In conclusion, the results indicated that tetraploids showed greater tolerance to salt stress compared to diploids, and polyploidy might be a useful breeding method in anise hyssop to amplify its tolerance to salt stress under soil salinity.


Ahmadi T, Jafarkhani Kermani M, Mashayekhi K, Hasanloo T, Shariatpanahi ME .2013. Comparing plant morphology, fertility and secondary metabolites in Rosa hybrida CV Iceberg and its chromosome-doubled progenies. Int Res J of App and Bas Sci 4:3840-3849.
Apse MP, Blumwald E. 2002. Engineering salt tolerance in plants. Plant biotech. 13: 146–150.
Ben Taarit MK, Msaada K, Hosni M, Hammami M, Kchouk E, Marzouk B .2009. Plant growth, essential oil yield and composition of sage (salvia officinalis L.) fruits cultivated under salt stress conditions. Ind Crops Prod 30:333-7
Breusegem FV, Vranova E, Dat JF, Inzé D .2001. The role of active oxygen species in plant signal transduction. Plant Sci 16:405–414.
Campos JMS, Davide LC, Salgado CC, Santos FC, Costa PN, Silva PS, Alves CCS, Viccini LF, Pereira AV. 2009. In vitro induction of hexaploid plants from triploid hybrids of Pennisetum purpureum and Pennisetum glaucum. Plant Breed 128:101-104.
Castro M, (2018) Evolutionary ecology of polyploids: understanding species coexistence at the contact zones. PhD thesis Department of Life Sciences, University of Coimbra, Coimbra.
Chance B, Maehly AC. 1955. Assay of catalases and peroxidase. Methods in Enzymology 2:764–775.
Chao DY, Dilkes B, Luo H, Douglas A, Yakubova E, Lahner B, Salt DE .2013. Polyploids Exhibit Higher Potassium Uptake and Salinity Tolerance in Arabidopsis. Science 341:658–659.
Comai L .2005. The advantages and disadvantages of being polyploid. Nature Genetics 6:836-846
Costich DE, Ortiz R, Meagher TR, Bruederle LP, Vorsa N. 1993. Determination of ploidy level and nuclear DNA content in blueberry by flow cytometry. Theo. App Gen 86:1001–1006
Deng B, Du W, Liu C, Sun W, Tian S, Dong H. 2012. Antioxidant response to drought, cold and nutrient stress in two ploidy levels of tobacco plants: low resource requirement confers polytolerance in polyploids?. Plant Gro Regu 66:37-47.
Ezz El-Din AA, Aziz EE, Hendawy SF, Omer EA. 2009. Response of thymus vulgaris L. to salt stress and alar (B9) in newly reclaimed soil. J Appl Sci Res 5:2165-70.
Ghani A, Neamati SH, Azizi M, Saharkhiz MJ, Farsi M. 2014. Artificial Autotetraploidy Induction Possibility of Two Iranian Endemic Mint (Mentha mozaffarianii) Ecotypes. Not Sci Biol 6:185-191.
Hansen NJP, Andersen SB. 1996. In vitro chromosome doubling potential of colchicine,oryzalin, trifluralin and APM in Brassica napus microspore culture. Euphytica 88:159–164.
Kermani MJ. 2001. Chromosome doubling and the breeding of disease resistant roses. PhD Thesis University of East London, London, UK.
Kermani MJ, Sarasan V, Roberts AV, Yokoya K, Wentworth J, Sieber VK. 2003. Oryzalin-induced chromosome doubling in Rosa and its eVect on plant morphology and pollen viability. Theo Appl Gen 107:1195-1200.
Khosravi P, Jafarkhani Kermani M, Nematzade GA, Bihamta MR. 2008. Role of mitotic inhibitors and genotype on chromosome doubling of Rosa. Euphytica 160:267-275.
Kolar F, Dortová M, Lepš J, Pouzar M, Krejčovám A, Štech M. 2014. Serpentine ecotypic differentiation in a polyploid plant complex: shared tolerance to Mg and Ni stress among di- and tetraploid serpentine populations of Knautia arvensis (Dipsacaceae). Plant Soil 374:435–447.
Li WD, Biswas, D.K., Xu, H., Xu, CQ, Wang X Z, Liu JK, Jiang GM. 2009. Photosynthetic responses to chromosome doubling in relation to leaf anatomy in Lonicera japonica subjected to water stress. Plant Biol 36:783–792.
Liu S, Chen S, Chen Y, Guan Z, Yin D, Chen F. 2011. In vitro induced tetraploid of Dendranthema nankingense (Nakai) Txvel. Shows an improved level of abiotic stress tolerance. Scien Horti 127:411-419
Ma Y, Byrne DH, Chen J (1997) Amphidiploid induction from diploid rose interspeciWc hybrids. Hort Sci 32:292–295.
Meng F, Luo Q, Wang Q, Zhang X, Qi Z, Lei X, Cao Y, Chow WS, Sun G, Xu F. 2016. Physiological and proteomic responses to salt stress in chloroplasts of diploid and tetraploid black locust (Robinia pseudoacacia L.). Scien Rep 1-15.
Meng HB, Jiang SS, Hua SJ, Lin XY, Li YL, Guo WL, Jiang LX. 2011. Comparison between a tetraploid turnip and its diploid progenitor (Brassica rapa L.): The adaptation to salinity stress. Agri Sci. China 10:363-375.
Moghbel N, Khalili Borujeni M, Bernard F. 2015. Colchicine effect on the DNA content and stomata size of Glycyrrhiza glabra var.glandulifera and Carthamus tinctorius L. cultured in vitro. Gen Eng. Biot 13:1–6
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497.
Najafian S, Khoshkhui M, Tavallali V, Saharkhiz MJ. 2009. Effect of salicylic acid and salinity in thyme (thymus vulgaris L.): investigation on changes in gas exchange, water relations, and membrane stabilization and biomass accumulation. Aust J Basic Appl Sci 3:2620-6.
Ojiewo CO, Murakam KI, Masinde PW, Agong SG. 2007. Polyploidy breeding of African nightshade (Solanum section Solanum). Plant Breed 1:10–21.
Omami EN, Hammes PS, Robbertse PJ. 2006. Differences in salinity tolerance for growth and water-use efficiency in some amaranth (Amaranthus spp.) genotypes. New Zeal J Crop Hort Sci 34:11-22.
Ozturk A, Unlukara A, Ipekl A, Gurbuz B. 2004. Effect of salt stress and water deficit on plant growth and essential oil content of lemon balm (melissa officinalis L.). Pak J Bot 36:787-92.
Pillay M, Ogundiwin E, Tenkouano A, Dolezel J. 2006. Ploidy and genome composition of Musa germplasm at the International Institute of Tropical Agriculture (IITA). Afr J of Biotech 5:1224-1232.
Podda A, Checcucci G, Mouhaya W, Centeno D, Rofidal V, Del Carratore R, Luro F, Morillon R, Ollitrault P, Maserti BE. 2013. Salt-stress induced changes in the leaf proteome of diploid and tetraploid mandarins with contrasting Na+ and Cl-accumulation behaviour. Plant Physiol 170:1101–1112.
Quan K, Guolu L, Qigao G, Xiaolin L .2004. Polyploid induc-tion of Arctium lappa by colchicine. Plant Physiol Communi 40:157-158
Queslati S, Karray-Bouraoui N, Attia H, Rabhi M, Ksouri R, Lachaal M. 2010. Physiological and antioxidant responses of mentha pulegium (Pennyroyal) to salt stress. Acta Physiol Plant 32:289-96.
Riddle NC, Jiang H, An L, Doerge RW, Birchler JA. 2010. Gene expression analysis at the intersection of ploidy and hybridity in maize. Theor Appl Gen 120:341–353.
Saharkhiz MJ. 2006. The effects of some environmental factors and ploidy level on morphological and physiological characteristics of feverfew (Tanacetum parthenium L.) medicinal ornamental plant. PhD Thesis of Tarbiat Modares University, Tahran, Iran.
Said-Al Ahl HAH, Mahmoud AA. 2010. Effect of zinc and/or iron foliar application on growth and essential oil of sweet basil (Ocimum basilicum L.) under salt stress. Ozean J Appl Sci 3:97-111.
Sakhanokho HF, Rajasekaran K, Kelley RV, Farjdj NI. 2009. Induced polyploidy in diploid ornamental ginger (Hedychiummuluense R. M. Smith) using colchicine and oryzalin. Hort Sci 44:1809–1814.
Sartor ME, Quarin CL, Urbani MH, Espinoza F. 2011. Ploidy levels and reproductive behaviour in natural populations of five Paspalum species. Plant Sys and Evo 293:31-41.
Shafieizargar A, Awang Y, Juraimz AS, Othman R. 2013. Comparative studies between diploid and tetraploid Dez Orange (Citrus sinensis (L.) Osb.) under salinity stress. Aus J. Crop Sci 7:1436-1441.
Shalan MN, Abdel-Latif TAT, Ghadban EA. 2006. Effect of water salinity and some nutritional compounds of the growth and production of sweet marjoram plants (majorana hortensis L.). Egypt J Agric Res 84:959.
Stanys V, Weckman A, Staniene G, Duchovskis P. 2005. In vitro induction of polyploidy in japanese quince (Chaenomeles japonica). Plant Cell Tiss Org Cult 84:263–268.
Taira T, Shao ZZ, Hamawaki H, Larter EN. 1991. The effect of colchicine as a chromosome doubling agent for wheatrye hybrids as influenced by pH, method of application, and post-treatment environment. Plant Breed 109:329- 333.
Talebi SF, Saharkhiz MJ, Jafarkhani Kermani M, Sharafi Y, Raouf Fard F. 2017. Effect of different antimitotic agents on polyploidy induction of anise hyssop (Agastache foeniculum L.). Caryologia http://dx.doi.org/10.1080/00087114.2017.1318502.
Tavan M, Mirjalili MH, Karimzadeh G. 2015. In vitro polyploidy induction: changes in morphological, anatomical and phytochemical characteristics of Thymus persicus (Lamiaceae). Plant Cell Tiss Organ Cult 22:573–583.
Temel A, Gozukirmizi N. 2015. Cytotoxic effects of metaphase-arresting methods in barley. Cytol Gen 49: 382
Thao N, Ureshino K, Miyajima I, Ozaki Y, Okubo H (2003) Induction of tetraploids in ornamental Alocasia through colchicine and oryzalin treatments. Plant Cell Tiss Organ Cult 72:19–25.
Tilney-Bassett RAE (1986) Plant cimeras. Edward Arnold, London, UK.
Timbó ALDO, Pereira RC, Sobrinho FS, Davide LC. 2014. Nuclear DNA content and chromosome number in Brachiaria spp. Genotypes. Revis Ciênc Agron 45:62-67.
Tseng MJ, Liu CW, Yiu JC. 2007. Enhanced tolerance to sulfur dioxide and salt stress of transgenic Chinese cabbage plants expressing both superoxide dismutase and catalase in chloroplasts. Plant Physiol Bioch 45:822–833.
Urwin NAR, Horsnell J, Moon T. 2007. Generation and characterization of colchicine-induced autotetraploid Lavandula angustifolia. Euphytica 156:257-266.
Van Laere K, França S, Vansteenkiste H, Van Huylenbroeck J, Steppe K, Van Labeke MC. 2010. Influence of ploidy level on morphology, growth and drought susceptibility in Spathiphyllum wallisii. Acta Phy Plant 33:1149-1156.
Verhoeven HA, Ramulu KS, Dijkhuis P. 1990. A comparison of the effects of various spindle toxins on metaphase arrest and formation of micronuclei in cellsuspension culture of Nicotiana plumbaginifolia. Planta 182:408-414.
Wang Z, Wang M, Liu L, Meng F. 2013. Physiological and proteomic responses of diploid and tetraploid black locust (Robinia pseudoacacia L.) subjected to salt stress. Mol Sci 14:20299–20325.
Wannakrairoj S, Wondyifraw T. 2013. In vitro chromosome doubling in korarima [Aframomum corrorima (braun) p.c.m. jansen] using colchicine and oryzalin. Kaset J Nat Sci 47:684 – 694
Warner DA, Edwards GE. 1989. Effects of polyploidy on photosynthetic rates, photosynthetic enzymes, contents of DNA, chlorophyll and sizes and numbers of photosynthetic cells in the C 4 dicot Atriplex confertifolia. Plant Physio 91:1143–1151.
Wu HZ, Zheng S, He Y, Yan G, Bi Y, Zhu Y. 2007. Diploid female gametes induced by colchicine in oriental Lilies. Sci Hort 114:50-53.
Yang XM, Cao ZY, An LZ, Wang YM, Fang XW. 2006. In vitro tetraploid induction via colchicine treatment from diploid somatic embryos in grapevine (Vitis vinifera L.). Euphytica 152:217-224
Yokoya K, Roberts AV, Mottley J, Lewis R, Brandham PE. 2000. Nuclear DNA amounts in roses. Ann Bot 85:557– 56
Zhang XY, Hu CG, Yao JL .2010. Tetraploidization of diploid Dioscorea results in activation of the antioxidant defense system and increased heat tolerance. Plant Physio 167:88–94.




How to Cite

Sharafi, Y., Talebi, S. F., saharkhiz, M. jamal, & Jafarkhani Kermani , M. (2021). Polyploidy increases tolerance to salt stress in Anise hyssop (Agastache foeniculum [Pursh.] Kuntze): Polyploidy increases tolerance to salt stress in Anise hyssop . Caryologia. International Journal of Cytology, Cytosystematics and Cytogenetics. https://doi.org/10.36253/caryologia-783