Sequential infections by 32 isolates of Phoma medicaginis increase production of phytoestrogens in Medicago polymorpha var. brevispina
Mahtab Omidvari A , Gavin R. Flematti B , Ming Pei You A , Payman Abbaszadeh-Dahaji C and Martin J. Barbetti
A *
+ Author Affiliations
- Author Affiliations
A School of Agriculture and Environment and the UWA Institute of Agriculture, University of Western Australia, 35 Stirling Highway, Crawley, WA 6009, Australia.
B School of Molecular Sciences, University of Western Australia, 35 Stirling Highway, Crawley, WA 6009, Australia.
C Department of Soil Sciences, Faculty of Agricultural Science, Vali-e-Asr University of Rafsanjan, Rafsanjan, Iran.
Crop & Pasture Science 73(12) 1367-1384 https://doi.org/10.1071/CP22098
Submitted: 22 March 2022 Accepted: 29 May 2022 Published: 13 July 2022
© 2022 The Author(s) (or their employer(s)). Published by CSIRO Publishing. This is an open access article distributed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (CC BY-NC-ND)
Abstract
Context: Studies of Phoma black stem and leaf spot disease (caused by Phoma medicaginis) in annual medics (Medicago spp.) normally involve a ‘once-only’ inoculation not reflecting multiple pathogen infection and phytoestrogen production cycles in the field. Phytoestrogen production by plants can result in lower ovulation rates in grazing animals.
Aims: We aimed to determine whether sequential infections by P. medicaginis increase production of phytoestrogens in annual medics, and to measure the genetic diversity of isolates.
Methods: In a greenhouse experiment, pathogenicity and virulence were investigated across 32 isolates of P. medicaginis following one, two or three rounds of inoculation of M. polymorpha var. brevispina. Production of the phytoestrogens coumestrol and 4′-O-methyl coumestrol was measured, and correlation with disease parameters assessed. DNA sequencing using ITS, β-tubulin, calmodulin and P. medicaginis-specific EFNI-1α was applied for phylogenetic analysis of isolates from Western Australia and elsewhere.
Key results: Across isolates, highest leaf disease incidence was 76%, petiole disease incidence 61%, leaf disease severity 52% and petiole disease severity 53%. Stem coumestrol content range was 45–1247 mg kg−1, and 4′-O-methyl coumestrol 0–344 mg kg−1. All measures were highest after three rounds of inoculation. Overall, there was a positive correlation of leaf disease incidence with coumestrol content (P < 0.05) and of both leaf and petiole disease incidence with 4′-O-methylcoumestrol content (P < 0.01, P < 0.05, respectively). Phylogenetic analysis revealed a high degree of genetic similarity among Western Australian isolates, generally grouping into a single separate cluster across the four markers, and genetically distinct from isolates sourced outside Australia.
Conclusions: Leaf disease incidence was the best discriminating disease parameter for coumestrol and 4′-O-methylcoumestrol content. Western Australian isolates of P. medicaginis were genetically similar and unique, possibly due to geographic separation.
Implications: The study emphasised the importance of sequential inoculations when screening annual Medicago genotypes towards developing cultivars with superior disease resistance and enhanced animal reproductive outcomes.
Keywords: 4′-O-methylcoumestrol, annual medic, annual Medicago spp, coumestrol, genetic variation, phoma black stem and leaf spot, Phoma medicaginis, phytoestrogen.
References
Barbetti MJ (1983) Fungal foliage diseases of pasture legumes. Journal of the Department of Agriculture, Western Australia 24, 10–12.Barbetti MJ (1984) Subterranean clover foliage fungi as root pathogens. Australasian Plant Pathology 13, 38–40.
| Subterranean clover foliage fungi as root pathogens.Crossref | GoogleScholarGoogle Scholar |
Barbetti MJ (1987) Effects of temperature and humidity on disease caused by Phoma medicaginis, resistance in some Medicago cultivars and the incidence of seed-borne inoculum. Australian Journal of Experimental Agriculture 27, 851–856.
| Effects of temperature and humidity on disease caused by Phoma medicaginis, resistance in some Medicago cultivars and the incidence of seed-borne inoculum.Crossref | GoogleScholarGoogle Scholar |
Barbetti MJ (1989a) Response of Medicago cultivars to fungal root pathogens associated with Trifolium subterraneum. Plant Protection Quarterly 4, 1–3.
Barbetti MJ (1989b) Strategies for control of Phoma black stem in annual Medicago species. Australian Journal of Experimental Agriculture 29, 635–640.
| Strategies for control of Phoma black stem in annual Medicago species.Crossref | GoogleScholarGoogle Scholar |
Barbetti MJ (1990) Resistance in annual Medicago species to Phoma medicaginis under controlled environment and field conditions. Australian Journal of Experimental Agriculture 30, 209–214.
| Resistance in annual Medicago species to Phoma medicaginis under controlled environment and field conditions.Crossref | GoogleScholarGoogle Scholar |
Barbetti MJ (1991) Effects of temperature and humidity on diseases caused by Phoma medicaginis and Leptosphaerulina trifolixs in lucerne (Medicago sativa). Plant Pathology 40, 296–301.
| Effects of temperature and humidity on diseases caused by Phoma medicaginis and Leptosphaerulina trifolixs in lucerne (Medicago sativa).Crossref | GoogleScholarGoogle Scholar |
Barbetti MJ (1995a) Relative resistance, associated yield losses and phyto-oestrogen production from fungal foliar diseases in new and old annual Medicago cultivars. Australian Journal of Agricultural Research 46, 441–450.
| Relative resistance, associated yield losses and phyto-oestrogen production from fungal foliar diseases in new and old annual Medicago cultivars.Crossref | GoogleScholarGoogle Scholar |
Barbetti MJ (1995b) Resistance in annual Medicago species to Phoma medicaginis and Leptosphaerulina trifolii under field conditions. Australian Journal of Experimental Agriculture 35, 209–214.
| Resistance in annual Medicago species to Phoma medicaginis and Leptosphaerulina trifolii under field conditions.Crossref | GoogleScholarGoogle Scholar |
Barbetti MJ (2007) Resistance in annual Medicago spp. to Phoma medicaginis and Leptosphaerulina trifolii and its relationship to induced production of a phyto-oestrogens. Plant Disease 91, 239–244.
| Resistance in annual Medicago spp. to Phoma medicaginis and Leptosphaerulina trifolii and its relationship to induced production of a phyto-oestrogens.Crossref | GoogleScholarGoogle Scholar | 30780554PubMed |
Barbetti MJ, Fang CS (1991) Relationship between Phoma black stem severity and herbage and seed yield and coumestrol content in three Medicago polymorpha var. brevispina cultivars. Australian Journal of Agricultural Research 42, 409–415.
| Relationship between Phoma black stem severity and herbage and seed yield and coumestrol content in three Medicago polymorpha var. brevispina cultivars.Crossref | GoogleScholarGoogle Scholar |
Barbetti MJ, Khan TN (1987) Cross-pathogenicity of Phoma medicaginis isolates from the genera Medicago, Pisum and Trifolium. Phytophylactica 19, 517–519.
Barbetti MJ, Nichols PGH (1991) Effect of Phoma medicaginis and Leptosphaerulina trifolii on herbage and seed yield and coumestrol content of annual Medicago species. Phytophylactica 23, 223–228.
Barbetti MJ, Riley IT, You MP, Li H, Sivasithamparam K (2006) The association of necrotrophic fungal pathogens and plant parasitic nematodes with the loss of productivity of annual medic-based pastures in Australia and options for their management. Australasian Plant Pathology 35, 691–706.
| The association of necrotrophic fungal pathogens and plant parasitic nematodes with the loss of productivity of annual medic-based pastures in Australia and options for their management.Crossref | GoogleScholarGoogle Scholar |
Barbetti MJ, You M, Jones RAC (2020) Medicago truncatula and other annual Medicago spp.: interactions with root and foliar fungal, oomycete, and viral pathogens. In ‘The model legume Medicago truncatula’. Chapter 5.2.1.1. (Eds FJ de Bruijn, DY Liu) pp. 293–306. (Wiley: Chichester, UK)
Boerema GH, Bollen GJ (1975) Conidiogenesis and conidial septation as differentiating criteria between Phoma and Ascochyta. Persoonia 8, 111–144.
Castell-Miller CV, Zeyen RJ, Samac DA (2007) Infection and development of Phoma medicaginis on moderately resistant and susceptible alfalfa genotypes. Canadian Journal of Plant Pathology 29, 290–298.
| Infection and development of Phoma medicaginis on moderately resistant and susceptible alfalfa genotypes.Crossref | GoogleScholarGoogle Scholar |
Cenis JL (1992) Rapid extraction of fungal DNA for PCR amplification. Nucleic Acids Research 20, 2380
| Rapid extraction of fungal DNA for PCR amplification.Crossref | GoogleScholarGoogle Scholar | 1594460PubMed |
Chatterton L, Chatterton B (1996) ‘Sustainable dryland farming: combining farmer innovation and medic pasture in a Mediterranean climate.’ (Cambridge University Press: Cambridge, UK)
Chihaoui S-A, Djébali N, Mrabet M, Barhoumi F, Mhamdi R, Mhadhbi H (2015) Phoma medicaginis colonizes Medicago truncatula root nodules and affects nitrogen fixation capacity. European Journal of Plant Pathology 141, 375–383.
| Phoma medicaginis colonizes Medicago truncatula root nodules and affects nitrogen fixation capacity.Crossref | GoogleScholarGoogle Scholar |
Croker KP, Barbetti MJ, Nichols PGH (1994a) Incidence of coumestrol in medic pastures in Western Australia. In ‘Proceedings of the Australian Society of Animal Production’. Vol. 20, p. 416. (Australian Society of Animal Production)
Croker K, Nichols PGH, Barbetti MJ, Adams N (1994b) Sheep infertility from pasture legumes. Farmnote No. 6/94. Department of Agriculture Western Australia, Perth, WA, Australia.
Croker K, Nichols PGH, Barbetti MJ, Adams N (1999) Sheep infertility from pasture legumes. Farmnote No. 79/99. Department of Agriculture Western Australia, Perth, WA, Australia.
Croker K, Nichols PGH, Barbetti MJ, Adams N (2005) Sheep infertility from pasture legumes. Farmnote No. 41/2005. Department of Agriculture Western Australia, Perth, WA, Australia.
Deavours BE, Dixon RA (2005) Metabolic engineering of isoflavonoid biosynthesis in alfalfa. Plant Physiology 138, 2245–2259.
| Metabolic engineering of isoflavonoid biosynthesis in alfalfa.Crossref | GoogleScholarGoogle Scholar | 16006598PubMed |
Deavours BE, Liu C-J, Naoumkina MA, Tang Y, Farag MA, Sumner LW, Noel JP, Dixon RA (2006) Functional analysis of members of the isoflavone and isoflavanone O-methyltransferase enzyme families from the model legume Medicago truncatula. Plant Molecular Biology 62, 715–733.
| Functional analysis of members of the isoflavone and isoflavanone O-methyltransferase enzyme families from the model legume Medicago truncatula.Crossref | GoogleScholarGoogle Scholar | 17001495PubMed |
Dhingra OD, Sinclair JB (1995) ‘Basic plant pathology methods.’ 2nd edn. p. 359. (CRC Press: Boca Raton, FL, USA)
Dixon RA (2001) Natural products and plant disease resistance. Nature 411, 843–847.
| Natural products and plant disease resistance.Crossref | GoogleScholarGoogle Scholar | 11459067PubMed |
Dixon RA, Achnine L, Kota P, Liu C-J, Reddy MSS, Wang L (2002) The phenylpropanoid pathway and plant defence: a genomics perspective. Molecular Plant Pathology 3, 371–390.
| The phenylpropanoid pathway and plant defence: a genomics perspective.Crossref | GoogleScholarGoogle Scholar | 20569344PubMed |
Djebali N (2013) Aggressiveness and host range of Phoma medicaginis isolated from Medicago species growing in Tunisia. Phytopathologia Mediterranea 52, 3–15.
Ellwood SR, Kamphuis LG, Oliver RP (2006a) Identification of sources of resistance to Phoma medicaginis isolates in Medicago truncatula SARDI core collection accessions, and multigene differentiation of isolates. Phytopathology 96, 1330–1336.
| Identification of sources of resistance to Phoma medicaginis isolates in Medicago truncatula SARDI core collection accessions, and multigene differentiation of isolates.Crossref | GoogleScholarGoogle Scholar | 18943665PubMed |
Ellwood SR, D’Souza NK, Kamphuis LG, Burgess TI, Nair RM, Oliver RP (2006b) SSR analysis of the Medicago truncatula SARDI core collection reveals substantial diversity and unusual genotype dispersal throughout the Mediterranean basin. Theoretical and Applied Genetics 112, 977–983.
| SSR analysis of the Medicago truncatula SARDI core collection reveals substantial diversity and unusual genotype dispersal throughout the Mediterranean basin.Crossref | GoogleScholarGoogle Scholar | 16402186PubMed |
Ellwood S, Kamphuis LG, Pfaff T, Oliver RP, Samac DA, Foster-Hartnett D, Tivoli B, Onfroy C, Moussart A, Villegas AM, Sillero JC, Rubiales D (2007) Inoculation and growth with foliar pathogenic fungi. In ‘The Medicago truncatula handbook’. (Eds U Mathesius, EP Journet, LW Sumner) pp. 1–14. (The Noble Foundation: Ardmore, OK USA)
Fatehi J, Bridge PD, Punithalingam E (2003) Molecular relatedness within the ‘Ascochyta pinodes–complex’. Mycopathologia 156, 317–327.
| Molecular relatedness within the ‘Ascochyta pinodes–complex’.Crossref | GoogleScholarGoogle Scholar | 14682458PubMed |
Fields RL, Barrell GK, Gash A, Zhao J, Moot DJ (2018) Alfalfa coumestrol content in response to development stage, fungi, aphids, and cultivar. Agronomy Journal 110, 910–921.
Francis CM, Millington AJ (1965) Wether bioassay of annual pasture legumes. IV. The oestrogenic activity of annual medic pastures. Australian Journal of Agricultural Research 16, 927–35.
| Wether bioassay of annual pasture legumes. IV. The oestrogenic activity of annual medic pastures.Crossref | GoogleScholarGoogle Scholar |
Francis CM, Millington AJ (1971) The presence of methylated coumestans in annual Medicago species: response to a fungal pathogen. Australian Journal of Agricultural Research 22, 75–80.
| The presence of methylated coumestans in annual Medicago species: response to a fungal pathogen.Crossref | GoogleScholarGoogle Scholar |
Fuchs A, Davidse LC, De Waard MA, De Wit PJGM (1983) Contemplations and speculations on novel approaches in the control of fungal plant diseases. Pesticide Science 14, 272–293.
| Contemplations and speculations on novel approaches in the control of fungal plant diseases.Crossref | GoogleScholarGoogle Scholar |
Hammond-Kosack KE, Parker JE (2003) Deciphering plant–pathogen communication: fresh perspectives for molecular resistance breeding. Current Opinion in Biotechnology 14, 177–193.
| Deciphering plant–pathogen communication: fresh perspectives for molecular resistance breeding.Crossref | GoogleScholarGoogle Scholar | 12732319PubMed |
He XZ, Dixon RA (2000) Genetic manipulation of isoflavone 7-O-methyltransferase enhances biosynthesis of 4′-O-methylated isoflavonoid phytoalexins and disease resistance in alfalfa. Plant Cell 12, 1689–1702.
| Genetic manipulation of isoflavone 7-O-methyltransferase enhances biosynthesis of 4′-O-methylated isoflavonoid phytoalexins and disease resistance in alfalfa.Crossref | GoogleScholarGoogle Scholar | 11006341PubMed |
Jayasiri SC, Hyde KD, Jones EBG, Jeewon R, Ariyawansa HA, Bhat JD, Camporesi E, Kang JC (2017) Taxonomy and multigene phylogenetic evaluation of novel species Boeremia and Epicoccum with new records of Ascochyta and Didymella in Didymellaceae. Mycosphere 8, 1080–1101.
| Taxonomy and multigene phylogenetic evaluation of novel species Boeremia and Epicoccum with new records of Ascochyta and Didymella in Didymellaceae.Crossref | GoogleScholarGoogle Scholar |
Kamphuis LG, Lichtenzveig J, Oliver RP, Ellwood SR (2008) Two alternative recessive quantitative trait loci influence resistance to spring black stem and leaf spot in Medicago truncatula. BMC Plant Biology 8, 30
| Two alternative recessive quantitative trait loci influence resistance to spring black stem and leaf spot in Medicago truncatula.Crossref | GoogleScholarGoogle Scholar | 18366746PubMed |
Kamphuis LG, Williams AH, Kuster H, Trengove RD, Singh KB, Oliver RP, Ellwood SR (2012) Phoma medicaginis stimulates the induction of the octadecanoid and phenylpropanoid pathways in Medicago truncatula. Molecular Plant Pathology 13, 593–603.
| Phoma medicaginis stimulates the induction of the octadecanoid and phenylpropanoid pathways in Medicago truncatula.Crossref | GoogleScholarGoogle Scholar | 22212347PubMed |
Kernkamp MF, Hemerick GA (1953) The relation of Ascochyta imperfecta to alfalfa seed production in Minnesota. Phytopathology 43, 378–383.
Lamprecht SC, Knox-Davies PS (1984) Preliminary survey of foliage diseases of annual Medicago spp. in South Africa. Phytophylactica 16, 177–183.
Leath KT (1990) Spring black stem and leaf spot. In ‘Compendium of alfalfa diseases’. (Eds DL Stuteville, DC Erwin) pp. 16–17. (American Phytopathological Society: St. Paul, MN, USA)
Lüscher A, Mueller-Harvey I, Soussana JF, Rees RM, Peyraud JL (2014) Potential of legume-based grassland–livestock systems in Europe: a review. Grass and Forage Science 69, 206–228.
| Potential of legume-based grassland–livestock systems in Europe: a review.Crossref | GoogleScholarGoogle Scholar | 26300574PubMed |
Marshall T, Parkin RJ (1970) Phosphate applications affect the coumestrol level of medics. Journal of the Department of Agriculture Western Australia, Series 4 11,
Mhadhbi H, Jebara M, Limam F, Huguet T, Elarbi Aouania M (2005) Interaction between Medicago truncatula lines and Sinorhizobium meliloti strains for symbiotic efficiency and nodule antioxidant activities. Physiologia Plantarum 124, 4–11.
| Interaction between Medicago truncatula lines and Sinorhizobium meliloti strains for symbiotic efficiency and nodule antioxidant activities.Crossref | GoogleScholarGoogle Scholar |
Omidvari M, Flematti G, You MP, Abbaszadeh-Dahaji P, Barbetti MJ (2021) Phoma medicaginis isolate differences determine disease severity and phytoestrogen production in annual Medicago spp. Plant Disease 105, 2851–2860.
| Phoma medicaginis isolate differences determine disease severity and phytoestrogen production in annual Medicago spp.Crossref | GoogleScholarGoogle Scholar | 33851866PubMed |
Omidvari M, Flematti G, You MP, Abbaszadeh-Dahaji P, Barbetti MJ (2022) Phoma black stem severity and phytoestrogen production in annual Medicago spp. is primarily determined by interaction of cultivar and pathogen isolate. Plant Pathology 71, 860–872.
| Phoma black stem severity and phytoestrogen production in annual Medicago spp. is primarily determined by interaction of cultivar and pathogen isolate.Crossref | GoogleScholarGoogle Scholar |
Patisaul HB, Jefferson W (2010) The pros and cons of phytoestrogens. Frontiers in Neuroendocrinology 31, 400–419.
| The pros and cons of phytoestrogens.Crossref | GoogleScholarGoogle Scholar | 20347861PubMed |
Pierson LM, Ferkin MH (2015) The impact of phytoestrogens on sexual behavior and cognition in rodents. Mammalian Biology 80, 148–154.
| The impact of phytoestrogens on sexual behavior and cognition in rodents.Crossref | GoogleScholarGoogle Scholar |
Reed KFM (2016) Fertility of herbivores consuming phytoestrogen-containing Medicago and Trifolium species. Agriculture 6, 35
| Fertility of herbivores consuming phytoestrogen-containing Medicago and Trifolium species.Crossref | GoogleScholarGoogle Scholar |
Reid RL (1990) ‘Manual of Australian agriculture.’ 5th edn. (Australian Institute of Agricultural Science: Sydney, Australia)
Rodriguez RdP (2005) Histological studies on the development of Phoma root rot of alfalfa. Journal of Agriculture 89, 251–262.
Rodriguez RDP, Leath KT (1992) Pathogenicity of Phoma medicaginis var. medicaginis to crowns of alfalfa. Plant Disease 76, 1237–1240.
| Pathogenicity of Phoma medicaginis var. medicaginis to crowns of alfalfa.Crossref | GoogleScholarGoogle Scholar |
Smith JF, Jagusch KT, Brunswick LFC, Kelly RW (1979) Coumestans in lucerne and ovulation in ewes. New Zealand Journal of Agricultural Research 22, 411–416.
| Coumestans in lucerne and ovulation in ewes.Crossref | GoogleScholarGoogle Scholar |
Tivoli B, Baranger A, Sivasithamparam K, Barbetti MJ (2006) Annual Medicago: from a model crop challenged by a spectrum of necrotrophic pathogens to a model plant to explore the nature of disease resistance. Annals of Botany 98, 1117–1128.
| Annual Medicago: from a model crop challenged by a spectrum of necrotrophic pathogens to a model plant to explore the nature of disease resistance.Crossref | GoogleScholarGoogle Scholar | 16803846PubMed |
Wasserman MD, Milton K, Chapman CA (2013) The roles of phytoestrogens in primate ecology and evolution. International Journal of Primatology 34, 861–878.
| The roles of phytoestrogens in primate ecology and evolution.Crossref | GoogleScholarGoogle Scholar |
You MP, Sivasithamparam K, Riley IT, Barbetti MJ (2000) The occurrence of root-infecting fungi and parasitic nematodes in annual Medicago spp. in Western Australian pastures. Australian Journal of Agricultural Research 51, 435–444.
| The occurrence of root-infecting fungi and parasitic nematodes in annual Medicago spp. in Western Australian pastures.Crossref | GoogleScholarGoogle Scholar |
