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Evidence in support of a role for plant-mediated proteolysis in the rumens of grazing animals

Published online by Cambridge University Press:  08 March 2007

A. H. Kingston-Smith ,
R. J. Merry ,
D. K. Leemans ,
H. Thomas  and
M. K. Theodorou
A. H. Kingston-Smith*
Affiliation:
Department of Plant, Animal and Microbial Science, Institute of Grassland and Environmental Research, Plas Gogerddan, Aberystwyth SY23 3EB, UK
R. J. Merry
Affiliation:
Department of Plant, Animal and Microbial Science, Institute of Grassland and Environmental Research, Plas Gogerddan, Aberystwyth SY23 3EB, UK
D. K. Leemans
Affiliation:
Department of Plant, Animal and Microbial Science, Institute of Grassland and Environmental Research, Plas Gogerddan, Aberystwyth SY23 3EB, UK
H. Thomas
Affiliation:
Department of Plant, Animal and Microbial Science, Institute of Grassland and Environmental Research, Plas Gogerddan, Aberystwyth SY23 3EB, UK
M. K. Theodorou
Affiliation:
Department of Plant, Animal and Microbial Science, Institute of Grassland and Environmental Research, Plas Gogerddan, Aberystwyth SY23 3EB, UK
*
*Corresponding author: Dr A. H. Kingston-Smith, fax +44 (0) 1970 828357, email alison.kingston-smith@bbsrc.ac.uk
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Abstract

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The present work aimed to differentiate between proteolytic activities of plants and micro-organisms during the incubation of grass in cattle rumens. Freshly cut ryegrass was placed in bags of varying permeability and incubated for 16 h in the rumens of dairy cows that had previously grazed a ryegrass sward, supplemented with 4 kg dairy concentrate daily. Woven polyester bags (50 μm pore size) permitted direct access of the micro-organisms and rumen fluid enzymes to the plant material. The polythene was impermeable even to small molecules such as NH3. Dialysis tubing excluded micro-organisms and rumen enzymes/metabolites larger than 10 kDa. DM loss was 46·3 % in polyester, 36·2 % in polythene and 38·1 % in dialysis treatments. It is possible that the DM loss within polythene bags occurred due to a solubilisation of plant constituents (e.g. water-soluble carbohydrates) rather than microbial attachment/degradation processes. The final protein content of the herbage residues was not significantly different between treatments. Regardless of bag permeability, over 97 % of the initial protein content was lost during incubations in situ. Electrophoretic separation showed that Rubisco was extensively degraded in herbage residues whereas the membrane-associated, light-harvesting protein remained relatively undegraded. Protease activity was detected in herbage residues and bathing liquids after all incubation in situ treatments. Although rumen fluid contains proteases (possibly of plant and microbial origin), our results suggest that, owing to cell compartmentation, their activity against the proteins of intact plant cells is limited, supporting the view that plant proteases are involved in the degradation of proteins in freshly ingested herbage.

Keywords

RuminantGrazingFresh forageProteolysisRubisco
Type
Research Article
Information
British Journal of Nutrition , Volume 93 , Issue 1 , January 2005 , pp. 73 - 79
Copyright
Copyright © The Nutrition Society 2005

References

Anastassiou, R, Argyroudi-Akoyunglou, JH (1995) Thylakoid-bound activity against LHCII apoprotein in bean. Photosynth Res 43, 241–250.CrossRefGoogle Scholar
Baird, DB, Harding, SA, Lane, PW, Murray, DA, Payne, RW & Soutar, DM (2002) Genstat for Windows 6th ed. Oxford: VSN International.Google Scholar
Beever, DE & Siddons, RC (1986) Digestion and metabolism in the grazing ruminant Control of Digestion and Metabolism in Ruminants 479 – 497 Milligan LP Grovum WL Dobson A Englewood Cliffs, NJ Prentice-Hall Proceedings of the Sixth International Symposium on Ruminant Physiology held at Banff, Canada, September 10th–14th, 1984.Google Scholar
Beever, DE, Terry, RA, Cammell, SB & Wallace, AS (1978) The digestibility of spring and autumn harvested perennial ryegrass by sheep. J Agric Sci 90, 463–470.CrossRefGoogle Scholar
Beha, EM, Theodorou, MK, Kingston-Smith, AH (2002) Grass cells ingested by ruminants undergo autolysis which differs from senescence: implications for grass breeding targets and livestock production. Plant Cell Environ 25, 1299–1312.CrossRefGoogle Scholar
Bennett, J (1981) Biosynthesis of the light-harvesting chlorophyll a / b binding protein. Peptide turnover in darkness. Eur J Biochem 118, 61–70.CrossRefGoogle Scholar
Bradford, MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analyt Biochem 72, 248–254.CrossRefGoogle ScholarPubMed
Bushnell, TP, Bushnell, D & Jagendorf, AT (1993) A purified zinc-protease of pea chloroplasts, EP1, that degrades the large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase. Plant Physiol 103, 585–591.CrossRefGoogle ScholarPubMed
Buxton, DR, O'Kiely, P (2003) Preharvest plant factors affecting ensiling. In Silage Science and Technology, pp. 199–250 [Buxton, DR, Muck, RE, Harrison, JH, editors]. Agronomy No. 42 Madison, WI: American Society of Agronomy Inc, Crop Science Society of America, Inc, Soil Science Society of America Inc.CrossRefGoogle Scholar
Cochran, WG & Cox, GM (1957) Experimental Designs 2nd ed. New York: John Wiley & Sons.Google Scholar
Coruzzi, G & Last, R (2000) Amino acids. In Biochemistry and Molecular Biology of Plants, pp. 358–410 [Buchanan, B, Gruissem, W, Jones, R, editors]. Rockville MD: ASPB.Google Scholar
Dewhurst, RJ, Mitton, AM, Offer, NW & Thomas, C (1996) Effects of the composition of grass silages on milk production and nitrogen utilization by dairy cows. An Sci 62, 25–43.Google Scholar
Feller, U (1986) Plant proteolytic enzymes in relation to leaf senescence. In Plant Proteolytic Enzymes, pp. 49–68 [Dalling, MJ, editors]. Boca Raton, FL: CRC Press.Google Scholar
Kingston-Smith, AH & Theodorou, MK (2000) Post-ingestion metabolism of fresh forage. New Phytol 148, 37–55.CrossRefGoogle ScholarPubMed
Kingston-Smith, AH, Bollard, A, Armstead, IP, Thomas, BJ & Theodorou, MK (2003a) Proteolysis and cell death in clover leaves is induced by grazing. Protoplasma 220, 119–129.CrossRefGoogle ScholarPubMed
Kingston-Smith, AH, Bollard, A, Thomas, BJ, Brooks, AE & Theodorou, MK (2003b) Nutrient availability during the early stages of colonization of fresh forage by rumen micro-organisms. New Phytol 158, 119–130.CrossRefGoogle Scholar
Laemmli, UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.CrossRefGoogle ScholarPubMed
Lee, MRF, Jones, EL, Moorby, JM, Humphreys, MO, Theodorou, MK, MacRae, JC & Scollan, ND (2001) Production responses from lambs grazed on Lolium perenne selected for an elevated water-soluble carbohydrate concentration. Anim Res 50, 441–449.CrossRefGoogle Scholar
Lindahl, M, Yang, DH & Andersson, B (1995) Regulatory proteolysis of the major light-harvesting chlorophyll a/b protein of photosystem II by a light-induced membrane associated enzymic system. Eur J Biochem 231, 503–509.Google ScholarPubMed
McDougall, EI (1948) Studies on ruminant saliva. 1. The composition and output of sheep's saliva. Biochem J 43, 99–109.CrossRefGoogle ScholarPubMed
Mae, T, Thomas, H, Gay, AP, Makino, A & Hidema, J (1993) Leaf development in Lolium temulentum: photosynthesis and photosynthetic proteins in leaves senescing under different irradiances. Plant Cell Physiol 34, 391–399.Google Scholar
Matile, P (1997) The vacuole and cell senescence. In The Plant Vacuole, vol. 25. pp. 87–112 [Leigh, RA and Sanders, D, editors]. Advances in Botanical Research. London: Academic Press.CrossRefGoogle Scholar
Mehrez, AZ, Ørskov, ER (1977) A study of the artificial fibre bag technique for determining the digestibility of feeds in the rumen. J Agric Sci 88, 645–650.CrossRefGoogle Scholar
Meyer, JHF & Mackie, RI (1986) Microbiological evaluation of the intraruminal in sacculus digestion technique. Appl Environ Microbiol 51, 622–629.CrossRefGoogle ScholarPubMed
Morris, K, Thomas, H & Rogers, L (1996) Endopeptidases during the development and senescence of Lolium temulentum leaves. Phytochem 41, 377–384.CrossRefGoogle Scholar
Nakazono, M, Tsuji, H, Li, Y, Saisho, D, Arimura, S-I, Tsutsumi, N & Hirai, A (2000) Expression of a gene encoding mitochondrial aldehyde dehydrogenase in rice increases under submerged conditions. Plant Physiol 124, 587–598.CrossRefGoogle ScholarPubMed
Nugent, JHA, Jones, WT, Jordan, DJ & Mangan, JL (1983) Rates of proteolysis in the rumen of the soluble proteins casein, fraction 1 (18S) leaf protein, bovine serum albumin and bovine submaxillary mucoprotein. Br J Nutr 50, 357–388.CrossRefGoogle Scholar
Nugent, JHA & Mangan, JL (1981) Characterisation of the rumen proteolysis of fraction 1 (18S) leaf protein from lucerne ( Medicago sativa L.). Br J Nutr 46, 39–58.CrossRefGoogle Scholar
Pahlow, G, Muck, RE, Driehuis, F, Oude, Elferink, SJWH Speolstra, SF (2003) Biochemistry of ensiling. In Silage Science and Technology, pp. 31–93 [Buxton, DR, Muck, RE and Harrison, JH, editors]. Agronomy No. 42, Madison, WI: Americann Society of Agronomy Inc, Crop Science Society of America Inc, Soil Science Society of America Inc.Google Scholar
Papadopoulos, YA & McKersie, BD (1983) A comparison of protein degradation during wilting and ensiling of six forage species. Can J Plant Sci 63, 903–912.CrossRefGoogle Scholar
Roughan, PG (1995) Acetate concentrations in leaves are sufficient to drive in vivo fatty acid synthesis at maximum rates. Plant Sci 107, 49–55.CrossRefGoogle Scholar
Roulin, S & Feller, U (1998) Light-independent degradation of stromal proteins in intact chloroplasts isolated from Pisum sativum L. leaves: requirement for divalent cations. Planta 205, 297–304.CrossRefGoogle Scholar
Siddons, RC, Paradine, J, Gale, DL & Evans, RT (1985) Estimation of the degradability of dietary-protein in the sheep rumen by in vivo and in vitro procedures. Br J Nutr 54, 545–561.CrossRefGoogle ScholarPubMed
Spencer, D, Higgins, JTV, Freer, M, Dove, H & Coombe, JB (1981) Monitoring the fate of dietary proteins in rumen fluid using gel electrophoresis. Br J Nutr 60, 241–247.CrossRefGoogle Scholar
Stewart, CS, Flint, HJ & Bryant, MP (1997) The rumen bacteria. In The Rumen Microbial Ecosystem, pp. 10–72 [Hobson, PN, Stewart, CS, editors]. London: Blackie Academic & Professional.CrossRefGoogle Scholar
Theodorou, MK, Merry, RJ & Thomas, H (1996) Is proteolysis in the rumen of grazing animals mediated by plant enzymes?. Br J Nutr 75, 507–510.CrossRefGoogle ScholarPubMed
Thomas, H (1978) Enzymes of nitrogen metabolism in detached leaves of Lolium temulentum during senescence. Planta 142, 161–169.CrossRefGoogle ScholarPubMed
Thomas, H (1997) Chlorophyll: a symptom and a regulator of plastid development. New Phytol 136, 163–181.CrossRefGoogle Scholar
Wallace, RJ & Cotta, MA (1988) Metabolism of nitrogen containing compounds. In The Rumen Microbial Ecosystem, pp. 217–249 [Hobson, PN, editors]. Amsterdam: Elsevier.Google Scholar
Wallace, RJ, Atasoglu, C & Newbold, CJ (1999) Role of peptides in rumen microbial metabolism-review. Asian-Austral J Anim Sci 12, 139–147.CrossRefGoogle Scholar
Wallace, RJ, Newbold, CJ, Bequette, BJ, MacRae, JC & Lobley, GE (2001) Increasing the flow of protein from ruminal fermentation – review. Asian Austral J Anim Sci 14, 885–893.CrossRefGoogle Scholar
Zhu, W-Y, Kingston-Smith, AH, Troncoso, D, Merry, RJ, Davies, DR, Pichard, G, Thomas, H & Theodorou, MK (1999) Evidence of a role for plant proteases in the degradation of herbage proteins in the rumen of grazing cattle. J Dairy Sci 82, 2651–2658.CrossRefGoogle ScholarPubMed