Free Standard AU & NZ Shipping For All Book Orders Over $80!
Register      Login
Crop and Pasture Science Crop and Pasture Science Society
Plant sciences, sustainable farming systems and food quality
RESEARCH ARTICLE

Progress towards developing bloat-safe legumes for the farming industry

Kerry Hancock A D , Vern Collette A , Elisabeth Chapman B , Katherine Hanson B , Stephen Temple B , Roger Moraga A and John Caradus C
+ Author Affiliations
- Author Affiliations

A AgResearch, Grasslands Research Centre, Tennent Drive, Palmerston North 4442, New Zealand.

B Forage Genetics International, Gills Coulee Road, West Salem, WI 54669, USA.

C Grasslanz Technology Limited, Tennent Drive, Palmerston North 4474, New Zealand.

D Corresponding author. Email: kerry.hancock@agresearch.co.nz

Crop and Pasture Science 65(11) 1107-1113 https://doi.org/10.1071/CP13308
Submitted: 4 September 2013  Accepted: 19 November 2013   Published: 24 March 2014

Abstract

Proanthocyanidins (PAs) are polymeric flavonoids derived from the phenylpropanoid pathway, and they bind reversibly to forage proteins within the rumen, providing protection from bloat while enhancing protein utilisation and animal production. The occurrence of PAs varies greatly within forage legume species. Foliar PAs are present in Lotus corniculatus (birdsfoot trefoil) and Onobrychis viciifolia (sainfoin), but such species often show poor persistence under grazing. By contrast, Trifolium repens (white clover) and Medicago sativa (lucerne, or alfalfa) have good persistence but negligible amounts of foliar PAs. We altered the accumulation patterns present in lucerne and white clover by the overexpression of an R2R3-MYB transcription factor (TaMYB14) isolated from T. arvense (rabbit’s foot clover), a species with significant levels of leaf PA. Such plants effectively produced PAs of high degree of polymerisation (DP) in leaf tissue by upregulating genes of the PA pathway. By comparing transcriptome pools, we identified additional MYB transcription factors with putative involvement in PA synthesis in white clover and T. arvense, indicating that these species share a complex PA regulation system. Progress towards producing commercial cultivars of lucerne and white clover containing effective levels of PAs has begun. Such plants will provide a viable option for mitigating bloat in pastoral agriculture-based farming systems.

Additional keywords: MYB transcription factors, phenylpropanoid pathway, proanthocyanidins, transgenic alfalfa and clover.


References

Aerts RJ, Barry TN, McNabb WC (1999) Polyphenols and agriculture: beneficial effects of proanthocyanidins in forages. Agriculture, Ecosystems & Environment 75, 1–12.
Polyphenols and agriculture: beneficial effects of proanthocyanidins in forages.Crossref | GoogleScholarGoogle Scholar |

Barry TN, McNabb WC (1999) The implications of condensed tannins on the nutritive value of temperate forages fed to ruminants. The British Journal of Nutrition 81, 263–272.

Berg P, Majak W, McAllister TA, Hall JW, McCartney D, Coulman BE, Goplen BP, Acharya SN, Tait RM, Cheng KJ (2000) Bloat in cattle grazing alfalfa cultivars selected for a low initial rate of digestion: A review. Canadian Journal of Plant Science 80, 493–502.
Bloat in cattle grazing alfalfa cultivars selected for a low initial rate of digestion: A review.Crossref | GoogleScholarGoogle Scholar |

Bogs J, Jaffe FW, Takos AM, Walker AR, Robinson SP (2007) The grapevine transcription factor VvMYBPA1 regulates proanthocyanidin synthesis during fruit development. Plant Physiology 143, 1347–1361.
The grapevine transcription factor VvMYBPA1 regulates proanthocyanidin synthesis during fruit development.Crossref | GoogleScholarGoogle Scholar | 17208963PubMed |

Burggraaf VT, Woodward SL, Woodfield DR, Thom ER, Waghorn GC, Kemp PD (2006) Morphology and agronomic performance of white clover with increased flowering and condensed tannin concentration. New Zealand Journal of Agricultural Research 49, 147–155.
Morphology and agronomic performance of white clover with increased flowering and condensed tannin concentration.Crossref | GoogleScholarGoogle Scholar |

Carlsen SCK, Fomsgaard IS (2008) Biologically active secondary metabolites in white clover (Trifolium repens L.)—a review focusing on contents in the plant, plant–pest interactions and transformation. Chemoecology 18, 129–170.
Biologically active secondary metabolites in white clover (Trifolium repens L.)—a review focusing on contents in the plant, plant–pest interactions and transformation.Crossref | GoogleScholarGoogle Scholar |

Czemmel S, Stracke R, Weisshaar B, Cordon N, Harris NN, Walker AR, Robinson SP, Bogs J (2009) The grapevine R2R3-MYB transcription factor VvMYBF1 regulates flavonol synthesis in developing grape berries. Plant Physiology 151, 1513–1530.
The grapevine R2R3-MYB transcription factor VvMYBF1 regulates flavonol synthesis in developing grape berries.Crossref | GoogleScholarGoogle Scholar | 19741049PubMed |

Deluc L, Barrieu F, Marchive C, Lauvergeat V, Decendit A, Richard T, Carde JP, Merillon JM, Hamdi S (2006) Characterization of a grapevine R2R3-MYB transcription factor that regulates the phenylpropanoid pathway. Plant Physiology 140, 499–511.
Characterization of a grapevine R2R3-MYB transcription factor that regulates the phenylpropanoid pathway.Crossref | GoogleScholarGoogle Scholar | 16384897PubMed |

Dixon RA, Liu C, Jun JH (2013) Metabolic engineering of anthocyanins and condensed tannins in plants. Current Opinion in Biotechnology 24, 329–335.
Metabolic engineering of anthocyanins and condensed tannins in plants.Crossref | GoogleScholarGoogle Scholar | 22901316PubMed |

FAO (2011) ‘FAOSTAT.’ (Food and Agriculture Organization of the United Nations: Rome) available at: http://faostat3.fao.org

Hancock KR, Collette V, Fraser K, Greig M, Xue H, Richardson K, Jones C, Rasmussen S (2012) Expression of the R2R3-MYB transcription factor TaMYB14 from Trifolium arvense activates proanthocyanidin biosynthesis in the legumes Trifolium repens and Medicago sativa. Plant Physiology 159, 1204–1220.
Expression of the R2R3-MYB transcription factor TaMYB14 from Trifolium arvense activates proanthocyanidin biosynthesis in the legumes Trifolium repens and Medicago sativa.Crossref | GoogleScholarGoogle Scholar | 22566493PubMed |

Li W, Godzik A (2006) Cd-hit: a fast program for clustering and comparing large sets of protein or nucleotide sequences. Bioinformatics 22, 1658–1659.
Cd-hit: a fast program for clustering and comparing large sets of protein or nucleotide sequences.Crossref | GoogleScholarGoogle Scholar | 16731699PubMed |

Li YG, Tanner G, Larkin P (1996) The DMACA-HCl. Protocol and the threshold proanthocyanidin content for bloat safety in forage legumes. Journal of the Science of Food and Agriculture 70, 89–101.
The DMACA-HCl. Protocol and the threshold proanthocyanidin content for bloat safety in forage legumes.Crossref | GoogleScholarGoogle Scholar |

McMahon LR, McAllister TA, Berg BP, Majak W, Acharya SN, Popp JD, Coulman BE, Wang Y, Cheng K-J (2000) A review of the effects of forage condensed tannins on ruminal fermentation and bloat in grazing cattle. Canadian Journal of Plant Science 80, 469–485.
A review of the effects of forage condensed tannins on ruminal fermentation and bloat in grazing cattle.Crossref | GoogleScholarGoogle Scholar |

McNabb WC, Waghorn GC, Peters JS, Barry TN (1996) The effect of condensed tannins in Lotus pedunculatus on the solubilization and degradation of ribulose-1,5-bisphosphate carboxylase (EC 4.1.1.39; Rubisco) protein in the rumen and the sites of Rubisco digestion. The British Journal of Nutrition 76, 535–549.
The effect of condensed tannins in Lotus pedunculatus on the solubilization and degradation of ribulose-1,5-bisphosphate carboxylase (EC 4.1.1.39; Rubisco) protein in the rumen and the sites of Rubisco digestion.Crossref | GoogleScholarGoogle Scholar | 8942361PubMed |

Mellway RD, Tran LT, Prouse MB, Campbell MM, Constabel CP (2009) The wound-, pathogen-, and ultraviolet B-responsive MYB134 gene encodes an R2R3 MYB transcription factor that regulates proanthocyanidin synthesis in poplar. Plant Physiology 150, 924–941.
The wound-, pathogen-, and ultraviolet B-responsive MYB134 gene encodes an R2R3 MYB transcription factor that regulates proanthocyanidin synthesis in poplar.Crossref | GoogleScholarGoogle Scholar | 19395405PubMed |

Miyake K, Ito T, Senda M, Ishikawa R, Harada T, Niizeki M, Akada S (2003) Isolation of a subfamily of genes for R2R3-MYB transcription factors showing up-regulated expression under nitrogen nutrient-limited conditions. Plant Molecular Biology 53, 237–245.
Isolation of a subfamily of genes for R2R3-MYB transcription factors showing up-regulated expression under nitrogen nutrient-limited conditions.Crossref | GoogleScholarGoogle Scholar | 14756320PubMed |

Oliveros JC (2007) ‘VENNY. An interactive tool for comparing lists with Venn Diagrams.’ (Computational Genomics Service, Centro Nacional de Biotecnología: Madrid) Available at: http://bioinfogp.cnb.csic.es/tools/venny/index.html.

Patra AK, Saxena J (2011) Exploitation of dietary tannins to improve rumen metabolism and ruminant nutrition. Journal of the Science of Food and Agriculture 91, 24–37.
Exploitation of dietary tannins to improve rumen metabolism and ruminant nutrition.Crossref | GoogleScholarGoogle Scholar | 20815041PubMed |

Peel GJ, Pang Y, Modolo LV, Dixon RA (2009) The LAP1 MYB transcription factor orchestrates anthocyanidin biosynthesis and glycosylation in Medicago. The Plant Journal 59, 136–149.
The LAP1 MYB transcription factor orchestrates anthocyanidin biosynthesis and glycosylation in Medicago.Crossref | GoogleScholarGoogle Scholar | 19368693PubMed |

Pinares-Patiño CS, Waghorn GC, Hegarty RS, Hoskin SO (2009) Effects of intensification of pastoral farming on greenhouse gas emissions in New Zealand. New Zealand Veterinary Journal 57, 252–261.
Effects of intensification of pastoral farming on greenhouse gas emissions in New Zealand.Crossref | GoogleScholarGoogle Scholar | 19802038PubMed |

Radovic J, Sokolovic D, Markovic J (2009) Alfalfa—most important perennial forage legume in animal husbandry. Biotechnology in Animal Husbandry 25, 465–475.
Alfalfa—most important perennial forage legume in animal husbandry.Crossref | GoogleScholarGoogle Scholar |

Shelton D, Stranne M, Mikkelsen L, Pakseresht N, Welham T, Hiraka H, Tabata S, Sato S, Paquette S, Wang TL, Martin C, Bailey P (2012) Transcription factors of Lotus: regulation of isoflavonoid biosynthesis requires coordinated changes in transcription factor activity. Plant Physiology 159, 531–547.
Transcription factors of Lotus: regulation of isoflavonoid biosynthesis requires coordinated changes in transcription factor activity.Crossref | GoogleScholarGoogle Scholar | 22529285PubMed |

Stockdale CR (1994) Incidence of bloat in lactating dairy cows fed clover dominant herbage and maize silage. Australian Society of Animal Production 20, 15–17.

Tanner GJ 2004. Condensed tannins. In ‘Plant pigments and their manipulation’. Annual Plant Reviews Vol. 14 (Ed. KM Davies) pp. 150–184. (Blackwell Publishing-CRC Press: Boca Raton, FL)

Tanner GJ, Franki KT, Abrahams S, Watson JM, Larkin PJ, Ashton AR (2003) Purification of legume leucoanthocyanidin reductase and molecular cloning of its cDNA. The Journal of Biological Chemistry 278, 31 647–31 656.
Purification of legume leucoanthocyanidin reductase and molecular cloning of its cDNA.Crossref | GoogleScholarGoogle Scholar |

Terrier N, Torregrosa L, Ageorges A, Vialet S, Verriès C, Cheynier V, Romieu C (2009) Ectopic expression of VvMybPA2 promotes proanthocyanidin biosynthesis in grapevine and suggests additional targets in the pathway. Plant Physiology 149, 1028–1041.
Ectopic expression of VvMybPA2 promotes proanthocyanidin biosynthesis in grapevine and suggests additional targets in the pathway.Crossref | GoogleScholarGoogle Scholar | 19098092PubMed |

Verdier J, Zhao J, Torres-Jerez I, Ge SJ, Liu CG, He XZ, Mysore KS, Dixon RA, Udvardi MK (2012) MtPAR MYB transcription factor acts as an on switch for proanthocyanidin biosynthesis in Medicago truncatula. Proceedings of the National Academy of Sciences of the United States of America 109, 1766–1771.
MtPAR MYB transcription factor acts as an on switch for proanthocyanidin biosynthesis in Medicago truncatula.Crossref | GoogleScholarGoogle Scholar | 22307644PubMed |

Waghorn GC, Jones WT (1989) Bloat in cattle. 46. Potential of dock (Rumex obtusifolius) as an antibloat agent for cattle. New Zealand Journal of Agricultural Research 32, 227–235.
Bloat in cattle. 46. Potential of dock (Rumex obtusifolius) as an antibloat agent for cattle.Crossref | GoogleScholarGoogle Scholar |

Wang Y, Majak W, McAllister TA (2012) Frothy bloat in ruminants: Cause, occurrence, and mitigation strategies. Animal Feed Science and Technology 172, 103–114.
Frothy bloat in ruminants: Cause, occurrence, and mitigation strategies.Crossref | GoogleScholarGoogle Scholar |

Woodfield DR, McNabb W, Kennedy L, Cousins GR, Caradus JR (1998) Floral and foliar tannin content in white clover. Proceedings of the Trifolium Conference 15, 19.