Selecting plants with increased total polyphenol oxidases in the genus Trifolium

https://doi.org/10.17221/107/2015-CJGPBCitation:Jakešová H., Řepková J., Nedělník J., Hampel D., Dluhošová J., Soldánová M., Ošťádalová M. (2015): Selecting plants with increased total polyphenol oxidases in the genus Trifolium. Czech J. Genet. Plant Breed., 51: 155-161.
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One of the aims in red clover (Trifolium pratense) breeding is to increase the polyphenol oxidase (PPO) activity, which may effectively reduce protein breakdown in silage and when cattle are fed by fresh clover. We analysed total PPO activity spectrophotometrically and on the level of gene expression using real-time quantitative PCR in single plants derived from an interspecific T. pratense × T. medium hybrid. Experiments were performed for two years and evaluated according to the general linear model with three factors (family, year, and cut). The analysis revealed considerable variability in total PPO activity between individuals and between families. Four families and two individuals with significantly higher PPO activity were selected. Their PPO activity ranged from 3.411 to 3.547 mkatal/min/g and from 4.041 to 5.731 mkatal/min/g, respectively, in comparison with the control variety Amos (2.370 mkatal/min/g). The majority of PPO transcripts were expressed by the two genes PPO1/5 and PPO2. In some genotypes, the PPO5 gene was expressed. Quantitative PCR confirmed the highest activity of PPO genes in seven hybrid plants with higher DNA contents corresponding to 30 chromosomes with 815 013 copies per plant. Our results indicate the suitability of combining two methods for improved selection: initial expression analysis to assess the PPO transcript level indicating gene activity and subsequent enzymatic assay.
References:
Gadeyne F., Van Ranst G., Vlaeminck B., Vossen E., Van der Meeren P., Fievez V. (2015): Protection of polyunsaturated oils against ruminal biohydrogenation and oxidation during storage using a polyphenol oxidase containing extract from red clover. Food Chemistry, 171, 241-250 https://doi.org/10.1016/j.foodchem.2014.08.109
 
Gregory RPF, Bendall DS (): The purification and some properties of the polyphenol oxidase from tea ( Camellia sinensis L.). Biochemical Journal, 101, 569-581 https://doi.org/10.1042/bj1010569
 
Jakešová H., Řepková J., Hampel D., Čechová L., Hofbauer J. (2011): Variation of morphological and agronomic traits in hybrids of Trifolium pratense × T. medium and a comparison with the parental species. Czech Journal of Genetics and Plant Breeding, 47: 28–36.
 
Jones Beth A, Hatfield Ronald D, Muck Richard E (1995): Screening legume forages for soluble phenols, polyphenol oxidase and extract browning. Journal of the Science of Food and Agriculture, 67, 109-112 https://doi.org/10.1002/jsfa.2740670117
 
Mehdi Khanlou Khosro, Van Bockstaele Erik (2012): A critique of widely used normalization software tools and an alternative method to identify reliable reference genes in red clover (Trifolium pratense L.). Planta, 236, 1381-1393 https://doi.org/10.1007/s00425-012-1682-2
 
Kyung No J., Yu Soung D., Jung Kim Y., Hee Shim K., Soo Jun Y., Hee Rhee S., Yokozawa T., Young Chung H. (1999): Inhibition of tyrosinase by green tea components. Life Sciences, 65: 241–246.
 
Lee Michael R. F. (2014): Forage polyphenol oxidase and ruminant livestock nutrition. Frontiers in Plant Science, 5, - https://doi.org/10.3389/fpls.2014.00694
 
Lee Michael R.F., Tweed John K.S., Minchin Frank R., Winters Ana L. (2009): Red clover polyphenol oxidase: Activation, activity and efficacy under grazing. Animal Feed Science and Technology, 149, 250-264 https://doi.org/10.1016/j.anifeedsci.2008.06.013
 
Li Li, Steffens John (2002): Overexpression of polyphenol oxidase in transgenic tomato plants results in enhanced bacterial disease resistance. Planta, 215, 239-247 https://doi.org/10.1007/s00425-002-0750-4
 
Mayer Alfred M., Harel Eitan (1979): Polyphenol oxidases in plants. Phytochemistry, 18, 193-215 https://doi.org/10.1016/0031-9422(79)80057-6
 
Řepková J., Jungmannová B., Jakešová H. (2003): Interspecific hybridisation prospects in the genus Trifolium. Czech Journal of Genetics and Plant Breeding, 39 (Special Issue): 306–308.
 
Repkova Jana, Jungmannova Barbara, Jakesova Hana (2006): Identification of barriers to interspecific crosses in the genus Trifolium. Euphytica, 151, 39-48 https://doi.org/10.1007/s10681-006-9126-3
 
Steinite Ineta, Gailite Agnese, Ievinsh Gederts (2004): Reactive oxygen and ethylene are involved in the regulation of regurgitant-induced responses in bean plants. Journal of Plant Physiology, 161, 191-196 https://doi.org/10.1078/0176-1617-01098
 
Stewart R.J., Sawyer B.J.B., Bucheli C.S., Robinson S.P. (2001): Polyphenol oxidase is induced by chilling and wounding in pineapple. Australian Journal of Plant Physiology, 28: 181–191.
 
Sullivan Michael L., Hatfield Ronald D. (2006): Polyphenol Oxidase and -Diphenols Inhibit Postharvest Proteolysis in Red Clover and Alfalfa. Crop Science, 46, 662- https://doi.org/10.2135/cropsci2005.06-0132
 
Sullivan M. L. (2004): Cloning and Characterization of Red Clover Polyphenol Oxidase cDNAs and Expression of Active Protein in Escherichia coli and Transgenic Alfalfa. PLANT PHYSIOLOGY, 136, 3234-3244 https://doi.org/10.1104/pp.104.047449
 
Taylor N.L., Quesenberry K.H. (1996): Red Clover Science. Dordrecht, Kluwer Academy Publishing: 170–187.
 
Thipyapong P., Joel D.M., Steffens J.C. (1997): Differential expression and turnover of the tomato polyphenol oxidase gene family during vegetative and reproductive development. Plant Physiology, 113: 707–718.
 
Vámos‐Vigyázó Lilly, Haard Norman F. (): Polyphenol oxidases and peroxidases in fruits and vegetables. C R C Critical Reviews in Food Science and Nutrition, 15, 49-127 https://doi.org/10.1080/10408398109527312
 
Winters Ana, Heywood Sue, Farrar Kerrie, Donnison Iain, Thomas Ann, Webb K Judith (2009): Identification of an extensive gene cluster among a family of PPOs in Trifolium pratense L. (red clover) using a large insert BAC library. BMC Plant Biology, 9, 94- https://doi.org/10.1186/1471-2229-9-94
 
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