Abstract
Abnormal physiological responses of plant cultures such as shoot tip necrosis, callus, and hyperhydricity are some of the most difficult challenges in shoot micropropagation, and their causes are not well understood. Five Murashige and Skoog mineral salt factors, which influence the growth of pear shoot cultures, were tested in a five-dimensional surface response experimental design. Pyrus communis ‘Old Home × Farmingdale 87,’ ‘Horner 51,’ and ‘Winter Nelis’; Pyrus dimorphophylla; and Pyrus ussuriensis ‘Hang Pa Li’ shoot cultures were grown on 43 computer-designed treatments to represent the design space of all possible treatment combinations. Analysis of shoot response to these treatments identified the factors that both contributed to physiological disorders and remedied them. Undesirable callus formation was common for pear shoots cultured on standard medium and decreased on formulations with increased NH4NO3, Fe, and mesos (CaCl2, KH2PO4, and MgSO4) for most genotypes. Shoot tip necrosis varied with the genotype, but low mesos or low nitrogen concentrations contributed to the necrosis. Hyperhydricity was more prominent with low mesos or low NH4NO3. Hooked and upwardly curled new leaves were seen in most genotypes and resulted from use of low mesos in P. communis and low nitrogen for ‘Hang Pa Li’ and P. dimorphophylla. Fasciation and hypertrophy were seen infrequently and resulted from wide imbalances in several nutrients simultaneously. In general, standard concentrations of Murashige and Skoog iron and micros combined with high mesos and moderate nitrogen compounds produced normal shoots without physiological disorders.
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References
Abousalim A.; Mantell S. H. A practical method for alleviating shoot-tip necrosis symptoms in in vitro shoot cultures of Pistacia vera cv. Mateur. J Hortic Sci 69: 357–365; 1994.
Bairu M. W.; Stirk W. A.; Van Staden J. Factors contributing to in vitro shoot-tip necrosis and their physiological interactions. Plant Cell Tiss Organ Cult 98: 239–248; 2009.
Bosela M. J.; Michler C. H. Media effects on black walnut (Juglans nigra L.) shoot culture growth in vitro: evaluation of multiple nutrient formulations and cytokinin types. In Vitro Cell Dev Biol - Plant 44: 316–329; 2008.
Bottcher I.; Zohlauer K.; Goring H. Induction and reversion of vitrification of plants cultured in vitro. Physiol Plant 72: 560–564; 1988.
Chauhan M.; Kothari S. L. Optimization of nutrient levels in the medium increases the efficiency of callus induction and plant regeneration in recalcitrant Indian barley (Hordeum vulgare L.) in vitro. In Vitro Cell Dev Biol - Plant 40: 520–527; 2004.
Debergh P.; Aitken-Christie J.; Cohen D.; Grout B.; Von Arnold S.; Zimmerman R.; Ziv M. Reconsideration of the term ‘vitrification’ as used in micropropagation. Plant Cell Tiss Organ Cult 30: 135–140; 1992.
Design-Expert. Stat-Ease, Inc., Minneapolis, MN; 2010
Epstein E.; Bloom A. (eds). Mineral nutrition of plants: principles and perspectives. Sinauer Associates, Sunderland, pp 1–380; 2005.
Evens T. J.; Niedz R. P. ARS-Media: ion solution calculator. U.S. Horticultural Research Laboratory, Ft. Pierce; 2008.
Grigoriadou K.; Leventakis N.; Vasilakakis M. Effects of various culture conditions on proliferation and shoot tip necrosis in the pear cultivars ‘Williams’ and ‘Highland’ grown in vitro. Acta Hortic 520: 03–108; 2000.
Hamant O.; Traas J. The mechanics behind plant development. New Phytol 185: 369–385; 2010.
Hazarika B. N. Morpho-physiological disorders in in vitro culture. Scientia Hortic 108: 105–120; 2006.
Iliev I.; Kitin P. Origin, morphology, and anatomy of fasciation in plants cultured in vitro and in vivo. Plant Growth Regul 63: 115–129; 2011.
Ishida T.; Anno T.; Matsukawa S.; Nagano T. Hydraulic conductivity and diffusion coefficient in gels for plant tissue culture. Environ Cont Biol 38: 165–171; 2000.
Ivanova M.; Van Staden J. Nitrogen source, concentration, and NH4 +:NO3 - ratio influence shoot regeneration and hyperhydricity in tissue cultured Aloe polyphylla. Plant Cell Tiss Organ Cult 99: 167–174; 2009.
Kintzios S.; Drossopoulos J. B.; Lymperopoulos C. Effect of vitamins and inorganic micronutrients on callus growth and somatic embryogenesis from leaves of chilli pepper. Plant Cell Tiss Organ Cult 67: 55–62; 2001.
Kintzios S.; Stavropoulou E.; Skamneli S. Accumulation of selected macronutrients and carbohydrates in melon tissue cultures: association with pathways of in vitro dedifferentiation and differentiation (organogenesis, somatic embryogenesis). Plant Sci 167: 655–664; 2004.
Leyser H.; Ottoline M.; Furner I. J. Characterisation of three shoot apical meristem mutants of Arabidopsis thaliana. Development 116: 397–403; 1992.
Lloyd G.; McCown B. Commercially feasible micropropagation of mountain laurel, Kalmia latifolia, by use of shoot-tip culture. Comb Proceed Int Plant Propag Soc 30: 421–427; 1980.
Martin K. P.; Zhang C.-L.; Slater A.; Madassery J. Control of shoot necrosis and plant death during micropropagation of banana and plantains (Musa spp.). Plant Cell Tiss Organ Cult 88: 51–59; 2007.
Murashige T.; Skoog F. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15: 473–497; 1962.
Niedz R. P.; Evens T. J. A solution to the problem of ion confounding in experimental biology. Nat Methods 3: 417; 2006.
Niedz R. P.; Evens T. J. Regulating plant tissue growth by mineral nutrition. In Vitro Cell Dev Biol - Plant 43: 370–381; 2007.
Paques M. Vitrification and micropropagation: causes, remedies and prospects. Acta Hortic 289: 283–290; 1991.
Piagnani C.; Eccher T. Factors affecting the proliferation and rooting of chestnut in vitro. Acta Hortic 227: 384–386; 1988.
Podwyszynska M.; Goszczynska D. M. Effect of inhibitors of ethylene biosynthesis and action, as well as calcium and magnesium on rose shoot rooting, shoot-tip necrosis and leaf senescence in vitro. Acta Physiol Plant 20: 91–89; 1998.
Reed B. M. Screening Pyrus germplasm for in vitro rooting response. HortScience 30: 1292–1294; 1995.
Reed B. M.; DeNoma J.; Luo J.; Chang Y.; Towill L. Cryopreservation and long-term storage of pear germplasm. In Vitro Cell Dev Biol - Plant 34: 256–260; 1998.
Reed B. M.; Wada S.; DeNoma J.; Niedz R. P. Improving in vitro mineral nutrition for diverse pear germplasm. In Vitro Cell Dev Biol - Plant 49: 343–355; 2013.
Singha S.; Townsend E.; Oberly G. Relationship between calcium and agar on vitrification and shoot-tip necrosis of quince (Cydonia oblonga Mill.) shoots in vitro. Plant Cell Tiss Organ Cult 23: 135–142; 1990.
Thakur A.; Kanwara J. S. Effect of phase of medium, growth regulators and nutrient supplementations on in vitro shoot-tip necrosis in pear. New Zealand J Crop Hort Sci 39: 131–140; 2011.
Wada S.; Niedz R. P.; DeNoma J.; Reed B. M. Mesos components (CaCl2, MgSO4, KH2PO4) are critical for improving pear micropropagation. In Vitro Cell Dev Biol - Plant 49: 356–365; 2013.
Yadav M. K.; Gaur A. K.; Garg G. K. Development of suitable protocol to overcome hyperhydricity in carnation during micropropagation. Plant Cell Tiss Organ Cult 72: 153–156; 2003.
Ziv M. Quality of micropropagated plants—vitrification. In Vitro Cell Dev Biol - Plant 27: 64–69; 1991.
Acknowledgments
We thank the NCGR lab personnel for assistance with the collection of data for this study. This project was funded by a grant from the Oregon Association of Nurseries and the Oregon Department of Agriculture, and by the United States Department of Agriculture-Agricultural Research Service, CRIS project 5358-21000-0-38-00D.
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Reed, B.M., Wada, S., DeNoma, J. et al. Mineral nutrition influences physiological responses of pear in vitro . In Vitro Cell.Dev.Biol.-Plant 49, 699–709 (2013). https://doi.org/10.1007/s11627-013-9556-2
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DOI: https://doi.org/10.1007/s11627-013-9556-2