Effects of Culture Media on Phytophthora palmivora Growth, α-elicitin Production and Toxicity to Dendrobium

Apinya KHAIRUM, Oythip POOLSAWAT, Paniti PORNBUNGKERD, Akkawat THARAPREUKSAPONG, Sopone WONGKAEW, Piyada A. TANTASAWAT

Abstract


Four culture media were evaluated for their ability to induce Phytophthora palmivora growth and produce culture filtrate (CF), and to determine a CF concentration and culture period effective for in vitro screening of black rot resistance in Dendrobium cv. ‘Earsakul’. Mycelial fresh weights of P. palmivora cultured in potato dextrose broth (PDB; the most commonly used medium for fungi), pea sucrose broth (PSB; a medium frequently used for Phytophthora spp.), and Murashige and Skoog broth (MSB; the most popular plant tissue culture medium) were found to be significantly higher than that in the newly developed modified oat meal broth (MOMB). When the total proteins of CFs were analysed with SDS-PAGE, a protein band of 10.5 kDa MW was found in CFs from all media with the highest level in PSB. LC-MS/MS analysis identified this protein as α-elicitin that had an identical amino acid sequence to the α-elicitin hibernalin of P. hibernalis and syringicin from P. syringae. The optimum conditions for in vitro selection of Dendrobium for black rot resistance using α-elicitin-containing CFs were also determined by evaluating the CF toxicity on Dendrobium protocorm-like bodies (PLBs) when cultured in all media supplemented with 0, 30, 50 and 100% CFs for seven, 14 and 21 d. The levels of PLB necrosis varied according to medium types, CF concentrations and culture periods. The maximum percentage of PLB necrosis (100%) was obtained in PSB supplemented with 50 and 100% CFs, and the severity of PLB necrosis was highest when treated with 100% CF for 14 and 21 d.


Keywords


black rot; in vitro; selection; mycelial growth; orchid; pea sucrose broth; toxin

Full Text:

PDF

References


Amusa NA (2006). Microbially produced phytotoxins and plant disease management. African Journal of Biotechnology 5:405-414.

Awuah RT, Frimpong M (2002). Cocoa-based media for culturing Phytophthora palmivora (Butl.) Butl., causal agent of black pod disease of cocoa. Mycopathologia 155:143-147.

Baillieul F, de Ruffray P, Kauffmann S (2003). Molecular cloning and biological activity of ?, ?, and ?-megaspermin, three elicitins secreted by Phytophthora megasperma H20. Plant Physiology 131:155-166.

Beever RE, Bollard EG (1970). The nature of the stimulation of fungal growth by potato extract. Journal of General Microbiology 60:273-279.

Behnke M (1979). Selection of potato callus for resistance to culture filtrates of Phytophthora infestans and regeneration of resistant plants. Theoretical and Applied Genetic 55:69-71.

Capasso R, Cristinziob G, Di Maroc A, Ferrantid P, Parentee A (2001). Syringicin, a new alpha-elicitin from an isolate of Phytophthora syringae, pathogenic to citrus fruit. Phytochemistry 58:257-262.

Capasso R, Di Maro A, Cristinzio G, De Martino A, Chambery A, Daniele A, … Parente A (2008). Isolation, characterization and structure–elicitor activity relationships of hibernalin and its two oxidized forms from Phytophthora hibernalis Carne 1925. Journal of Biochemistry 143:131-141.

Cating RA, Palmateer AJ, Stiles CM, Rayside PA (2010). Black rot of orchids caused by Phytopthora cactorum and Phytophthora palmivora in Florida. Plant Health Progress doi: 10.1094/PHP-2010-0614-01-DG.

Chee C-H, Newhook FJ (1965a). Improved methods for use in studies on Phytophthora cinnamomi Rands and other Phytophthora species. New Zealand Journal of Agricultural Research 8:88-95.

Chee C-H, Newhook FJ (1965b). Nutritional studies with Phytophthora cinnamomi Rands. New Zealand Journal of Agricultural Research 8:523-529.

Churngchow N, Rattarasarn M (2000). The elicitin secreted by Phytophthora palmivora, a rubber tree pathogen. Phytochemistry 54:33-38.

Dahlin P, Srivastava V, Ekengren S, McKee LS, Bulone V (2017). Comparative analysis of sterol acquisition in the oomycetes Saprolegnia parasitica and Phytophthora infestans. PLoS One 12(2), e0170873.

Dehgahi R, Zakaria L, Joniyas A, Subramaniam S (2014). Fusarium proliferatum culture filtrate sensitivity of Dendrobium sonia-28’s PLBs derived regenerated plantlets. Malaysian Journal of Microbiology 10:241-248.

Derakhshan-Honaroarvar M, Hamedi MM, Pirouzifard MKh (2010). Rice bran phytosterols of three widespread Iranian cultivars. Journal of Agricultural Science and Technology 12:167-172.

Derevnina L, Dagdas FY, De la Concepcion JC, Bialas A, Kellner R, Petre B, … Kamoun S (2016). Nine things to know about elicitins. New Phytologist 212:888-895.

Dokladal L, Oboril M, Stejskal K, Zdrahal Z, Ptackova N, Chaloupkova R, … Lochman J (2012). Physiological and proteomic approaches to evaluate the role of sterol binding in elicitin-induced resistance. Journal of Experimental Botany 63:2203-2215.

Dong S, Kong G, Qutob D, Yu X, Tang J, Kang J, … Wang Y (2012). The NLP toxin family in Phytophthora sojae includes rapidly evolving groups that lack necrosis-inducing activity. Molecular Plant-Microbe Interactions 25:896-909.

Du J, Verzaux E, Chaparro-Garcia A, Bijsterbosch G, Keizer LCP, Zhou J, ... Vleeshouwers VG (2015). Elicitin recognition confers enhanced resistance to Phytophthora infestans in potato. Nature Plants 1:15034.

Elliott CG (1994). Reproduction in fungi: genetical and physiological aspects. Chapman and Hall, London.

Erwin DC, Ribeiro OK (1996). Phytophthora disease worldwide. American Phytopathological Society, Minnesota.

Esmaiel NM, Al-Doss AA, Barakat MN (2012). In vitro selection for resistance to Fusarium oxysporum f. sp. dianthi and detection of genetic polymorphism via RAPD analysis in carnation. Journal of Medicinal Plants Research 6:3997-4004.

Fernanda AP, Cristiana CB, Sílvia LRM, Jaqueline GB, Eliana BF (2013). Activity of rice bran protein extracts against Fusarium graminearum. African Journal of Agricultural Research 8:6283-6290.

Gangopadhyay N, Hossain MB, Rai DK, Brunton NP (2015). A review of extraction and analysis of bioactives in oat and barley and scope for use of novel food processing technologies. Molecules 20:10884-10909.

Hendrix JW (1964). Sterol induction of reproduction and stimulation of growth of Pythium and Phytophthora. Science 144:1028-1029.

Ho HH (1970). A study of growth and sporulation of Phytophthora megasperma var. sojae. Sydowia 24:51-58.

Jadon KS, Shah R, Gour HN, Sharma P (2015). Effect of environmental factors on toxin production of Drechslera bicolor, a causal agent of leaf blight in bell pepper. African Journal of Microbiology Research 9:521-526.

Jiang RHY, Tyler BM, Whisson SC, Hardham AR, Govers F (2006). Ancient origin of elicitin gene clusters in Phytophthora genomes. Molecular Biology and Evolution 23:338-351.

Kamoun S, Young M, Glascock C, Tyler BM (1993). Extracellular protein elicitors from Phytophthora: host-specificity and induction of resistance to fungal and bacterial phytopathogens. Molecular Plant-Microbe Interactions 6:15-25.

Khairum A, Tharapreuksapong A, Wongkaew S, Tantasawat PA (2016). Cultural characteristics and pathogenicity analysis of Phytophthora palmivora, causal pathogen of black rot in orchids. In: Rahman MA, Dasic P (Eds). Proceedings of the International Conference on Agricultural, Food, Biological and Health Sciences (AFBHS-16). Kuala Lumpur, Malaysia pp 101-106.

Laemmli UK (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680-685.

Lagarda ML, García-Llatas G, Farré R (2006). Analysis of phytosterols in foods. Journal of Pharmaceutical and Biomedical Analysis 41:1486-1496.

Lecomte M, Hamama L, Voisine L, Gatto J, Hélesbeux J-J, Séraphin D, … Berruyer R (2014). Partial resistance of carrot to Alternaria dauci correlates with in vitro cultured carrot cell resistance to fungal exudates. PLoS One 9(7), e101008.

Levesque R, SPSS Inc. (2006). SPSS programming and data management. SPSS Institute (3rd ed), New York.

McMillan RT, Jr, Palmateer A, Cating RA (2009). Problems in controlling Phytopthora cactorum on Cattleya orchids. Proceedings of the Florida State Horticultural Society 122:426-428.

Moreau RA, Whitaker BD, Hicks KB (2002). Phytosterols, phytostanols, and their conjugates in foods: structural diversity, quantitative analysis, and health-promoting uses. Progress in Lipid Research 41:457-500.

Murashige T, Skoog F (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum 15:473-496.

Orlikowski LB, Szkuta G (2006). Phytophthora rot of some orchids - new disease in Poland. Phytopathologia Polonica 40:57-61.

Osman H, Vauthrin S, Mikes V, Milat M-L, Panabières F, Marais A, … Blein J-P (2001). Mediation of elicitin activity on tobacco is assumed by elicitin-sterol complexes. Molecular Biology of the Cell 12:2825-2834.

Özdestan Ö, Erol T, Acar A (2014). Phytosterols in rice bran and usage of rice bran in food industry. In: Straumite E (Ed). Proceedings of 9th Baltic Conference on Food Science and Technology “Food for Consumer Well-Being” FoodBalt 2014, Jelgava pp 24-27.

Oßwald W, Fleischmann F, Rigling D, Coelho AC, Cravador A, Diez J, … Werres S (2014). Strategies of attack and defence in woody plant – Phytophthora interactions. Forest Pathology 44:169-190.

Phungamngoen A, Sungsri-in M (2011). Antifungal activity of rice bran extracts against Colletotrichum gloeosporioides isolated from fresh and fresh-cut mango. Agricultural Science Journal 42:323-325.

Piironen V, Toivo J, Lampi A-M (2002). Plant sterols in cereals and cereal products. Cereal Chemistry 79:148-154.

Piironen V, Toivo J, Puupponen-Pimiä R, Lampi A-M (2003). Plant sterols in vegetables, fruits and berries. Journal of the Science of Food and Agriculture 83:330-337.

Ponchet M, Panabières F, Milat M-L, Mikes V, Montillet J-L, Suty L, … Blein J-P (1999). Are elicitins cryptograms in plant-oomycete communications?. Cellular and Molecular Life Sciences 56:1020-1047.

Qi F, Gong Z-H, Huang W (2006). Study on the selection of cell mutant lines resistant to Phytophthora capsici in pepper. Journal of Northwest Sci-Tech University of Agriculture and Forestry (Natural Science Edition) 34:83-88.

Rao S, Ramgoapl S (2010). Effect of Alternaria helianthi culture filtrate on callus and regeneration of plantlets from tolerant callus in sunflower (Helianthus annuus L.). Indian Journal of Biotechnology 9:187-191.

Rockefeller University (2016). Protocol for silver staining. Proteomics Resource Center. Retrieved 2016 Jan 10 from http://www.proteomics.rockefeller.edu/ms_ silverStaining.

Rodpai R, Intapan PM, Thanchomnang T, Sanpool O, Janwan P, Laummaunwai P, … Maleewong W (2016). Strongyloides stercoralis diagnostic polypeptides for human strongyloidiasis and their proteomic analysis. Parasitology Research 115:4007-4012.

Ryan E, Galvin K, O’Connor TP, Maguire AR, O’Brien NM (2007). Phytosterol, squalene, tocopherol content and fatty acid pro?le of selected seeds, grains, and legumes. Plant Foods for Human Nutrition 62:85-91.

Saubeau G, Gaillard F, Legentil L, Nugier-Chauvin C, Ferrières V, Andrivon D, Val F (2014). Identification of three elicitins and a galactan-based complex polysaccharide from a concentrated culture filtrate of Phytophthora infestans efficient against Pectobacterium atrosepticum. Molecules 19:15374-15390.

Savita, Virk GS, Nagpal A (2011). In vitro selection of calli of Citrus jambhiri Lush. for tolerance to culture filtrate of Phytophthora parasitica and their regeneration. Physiology and Molecular Biology of Plants 17:41-47.

Sharma A, Wongkham C, Prasongwattana V, Boonnate P, Thanan R, Reungjui S, Cha’on U (2014). Proteomic analysis of kidney in rats chronically exposed to monosodium glutamate. PLoS One 9(12), e116233.

Sopee J, Sangchote S, Stevenson WR (2012). Modified agar-based media for culturing Phytophthora infestans. Phytoparasitica 40:269-278.

Souza MM, Oliveira MS, Rocha M, Furlong EB (2010). Antifungal activity evaluation in phenolic extracts from onion, rice bran, and Chlorella phyrenoidosa. Ciência e Tecnologia de Alimentos [Food Science and Technology] (Campinas) 30:680-685.

Stong RA, Kolodny E, Kelsey RG, González-Hernández MP, Vivanco JM, Manter DK (2013). Effect of plant sterols and tannins on Phytophthora ramorum growth and sporulation. Journal of Chemical Ecology 39:733-743.

Sumbali G (2010). The fungi. Alpha Science International Ltd., Oxford.

Uhlíková H, Oboril M, Klempová J, Šedo O, Zdráhal Z, Kašparovský T, … Lochman J (2016). Elicitin-induced distal systemic resistance in plants is mediated through the protein–protein interactions influenced by selected lysine residues. Frontiers in Plant Science 7:59.

Valencia LDC, Pascual CB, Delfin EF (2014). In-vitro selection of pineapple cv. 'Queen' with resistance to culture filtrate of Phytophthora cinnamomi Rands. Philippine Journal of Crop Science 39:58-66.

Widmer TL (2014). Phytophthora palmivora. Forest Phytophthoras. Retrieved 2017 August 12 from http://dx.doi.org/10.5399/ osu/fp.4.1.3557.

Yoder OC (1980). Toxins in pathogenesis. Annual Review of Phytopathology 18:103-129.

Zipfel C, Robatzek S (2010). Pathogen-associated molecular pattern-triggered immunity: veni, vidi…?. Plant Physiology 154:551-554.




DOI: http://dx.doi.org/10.15835/nbha46211076

June 1, 2017: Notulae Botanicae Horti Agrobotanici Cluj-Napoca in CiteScore rank (Scopus – Elsevier) 28/66 in Horticulture
 
http://not-bot-horti-agrobo.blogspot.com/
https://www.facebook.com/NotBotHA
https://twitter.com/NotBotHA