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CCRSPI Feb 2011
Plant theme



Nutrient supply, below ground
processes and elevated CO2 change
the nutritional quality of cyanogenic
clover Trifolium repens
Ros Gleadow

Siobhan Isherwood, Tim Cavagnaro, Saman
Seneweera, Rebecca Miller,
2/20




Factors affecting food security

                Production
                    •Yield
                    •Area
                    •Agronomy

                                             Change in precipitation




 Availability                   Nutrition
    •Distribution                  •Safety
    •Cost                          •Value
    •Wealth

                                              Change in temperature
                                                                 IPCC WG1
3/20




How will climate change affect nutritional value?




1. Yield 
2. Nutritional value 
3. Availability 
4/20




  Why clover? 1. Interesting roots



                                                          VAM




  Legume – N fixing                                     Mycorrhizae – P uptake
Access to nutrients may facilitate positive response to CO2
5/20




Why clover? 2. Cyanogenic glycosides
   HOCH2
             O            CH3
                  O          CH3

  HO
                          CN
                                                  Linamarin (valine)
                                                   Lotaustralin (isoleucine)
             OH
                  Chewing
                  releases
                  -glucosidase
  glucose         (detonator)


                      
            HCN                Binds to cytochrome oxidase
           + ketone
                                                 Gleadow& Woodrow 2002, J Chem Ecol, 28, 1297.
6/20


Why clover?
3. Model for other important cyanogenic crops
       Forage sorghum                                          Cassava/Manioc




   Photo: Peter Stuart, Pac Seeds, Qld                     Photo: Julie Cliff, UEM, Moz




  5-10% of all plants cyanogenic         Gleadow, Conn et al, 2008 Phytochem, 69: 1870
  60% crop plants (some parts)           Jones 1998 Phytochem 47: 155; Miller et al. 2006 Ann Bot 97: 1017.
7/20



IF Less protein required for photosynthesis at high CO2
THEN resources may be diverted to defence

                          120
                                                 Is “excess” N used to
 Rubisco (% of control)




                          100
                                                 synthesise cyanide?
                          80

                          60
                                                                        Rubisco
                          40
                                                                        Other
                                                                        Cyanide
                          20

                           0
                                  350   700
                                  CO2 ppm

                                                 Gleadow and Woodrow 1999, Plants in Action –
                            Leaf protein, Rice   available www.plantsci.org.au/publications/
8/20




More Nitrogen allocated to cyanide at high-CO2

                                                      Sugar gums
                                                      Natimuk Road, Horsham
                  20       ~400 ppm CO2

                           ~ 800 ppm CO2
   CN-N per N %




                                              E
                                          A
                  10
                              E
                       A

                  0
                       Low Nitrogen   High Nitrogen




Gleadow et al. 1998, Plant Cell Environ, 12:21
9/20




CO2 effects on cyanogenesis in clover
                                    Siobhan Isherwood




• FACE
• Pot trials (ANU)
                                        CassavaFACE
• Nutrient trials –N, P, microbes
10/20




        Clover Biomass 6 months after sowing (Dec)

               400
Biomass g/m2




               300


               200


               100



                     380 ppm   550 ppm



                                         October 2010, 3 months
11/20




Cyanide in clover trending higher at 550 ppm


                      1000
Cyanide mg/g dry wt




                      800

                      600

                      400

                      200


                             380 ppm      550 ppm

                      • Is leaf protein lower?
                         • Nodulation rates: 6% higher significant??
                         • Mycorrhizae colonisation: 5% higher ns?
12/20



ANU study: More cyanide relative protein at
elevated CO2…ONLY with +P
                          370 ppm
                     12   700 ppm

                     10
 Cyanide : protein




                     8

                     6

                     4

                     2

                     0
                          -N-P      -N+P           +N+P
Gleadow et al. 2009 J Chem. Ecol 35, 476
Edwards & Evans 2005 Global Change Biol.11, 1968
13/20




   Nutrient cyanogenesis trials
                                      1.2
                                      1.0
                              CN-N/N% 0.8
                                      0.6
                                      0.4
                                      0.2
                                      0.0
                                            HP:HN   HP:LN       LP:HN      LP:LN
                                                    Nutrient Treatment

    • N15/N14 isotopes
         – P improved N fixation
                                                            High P: 0.125g/kg
         – Fixed N used to make HCN                         Low P: 0.025g/kg

                                                            High N: 8mM
                                                            Low N: 2mM


Isherwood, 2010 HONS THESIS
14/20




Conclusion: nutritional value IS affected by eCO2
   • Leaf composition
      – Less leaf protein
      – Increased cyanide, relative to protein
      – Increase tannins, phenolics


   • Soil environment is crucial

   • Microbes important in nutrient acquisition
      …How will elevated CO2 affect these symbioses???
     VIEWPOINT:
     Cavagnaro, Gleadow, Miller (2011) Functional Plant Biology 38, 87–96
15/20



                                          Monash Cyanogenesis Group
Collaborators                                 Cecilia Blomstedt
                                              Tim Cavagnaro
University of Copenhagen
                                              Rebecca Miller
      Birger Møller
      Kirsten Jørgensen, Morten Møldrup       Alan Neale
                                              John Hamill
RSB (ANU)                                     Anna Burns
     John Evans, Stephanie McCaffrey          Natalie O’Donnell
     Howard Bradbury, Bill Foley              Cara Griffiths
                                              Sam Fromhold
Pacific Seeds
                                              Melissa Bain
       Peter Stuart, Wayne Chesser
                                              Siobhan Isherwood
Mozambique/South Africa                       Kiara O’Gorman
     Julie Cliff UEM
     Anabela Zacarias, IIAM               FUNDING
     Simon Adams (Monash)                     ARC Linkage
                                              AusAID
University of Melbourne
                                              Finkel Foundation
      Saman Sereweena (AGFACE)
      Ian Woodrow, Jennifer Fox               Monash University
                                              Casstech
DPI: AGFACE                                   Pacific Seeds

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Nutrient supply, below ground processes and elevated CO2 change the nutritional quality of cyanogenic clover Trifolium repens - Ros Gleadow

  • 1. CCRSPI Feb 2011 Plant theme Nutrient supply, below ground processes and elevated CO2 change the nutritional quality of cyanogenic clover Trifolium repens Ros Gleadow Siobhan Isherwood, Tim Cavagnaro, Saman Seneweera, Rebecca Miller,
  • 2. 2/20 Factors affecting food security Production •Yield •Area •Agronomy Change in precipitation Availability Nutrition •Distribution •Safety •Cost •Value •Wealth Change in temperature IPCC WG1
  • 3. 3/20 How will climate change affect nutritional value? 1. Yield  2. Nutritional value  3. Availability 
  • 4. 4/20 Why clover? 1. Interesting roots VAM Legume – N fixing Mycorrhizae – P uptake Access to nutrients may facilitate positive response to CO2
  • 5. 5/20 Why clover? 2. Cyanogenic glycosides HOCH2 O CH3 O CH3 HO CN  Linamarin (valine) Lotaustralin (isoleucine) OH Chewing releases -glucosidase glucose (detonator)  HCN Binds to cytochrome oxidase + ketone Gleadow& Woodrow 2002, J Chem Ecol, 28, 1297.
  • 6. 6/20 Why clover? 3. Model for other important cyanogenic crops Forage sorghum Cassava/Manioc Photo: Peter Stuart, Pac Seeds, Qld Photo: Julie Cliff, UEM, Moz 5-10% of all plants cyanogenic Gleadow, Conn et al, 2008 Phytochem, 69: 1870 60% crop plants (some parts) Jones 1998 Phytochem 47: 155; Miller et al. 2006 Ann Bot 97: 1017.
  • 7. 7/20 IF Less protein required for photosynthesis at high CO2 THEN resources may be diverted to defence 120 Is “excess” N used to Rubisco (% of control) 100 synthesise cyanide? 80 60 Rubisco 40 Other Cyanide 20 0 350 700 CO2 ppm Gleadow and Woodrow 1999, Plants in Action – Leaf protein, Rice available www.plantsci.org.au/publications/
  • 8. 8/20 More Nitrogen allocated to cyanide at high-CO2 Sugar gums Natimuk Road, Horsham 20 ~400 ppm CO2 ~ 800 ppm CO2 CN-N per N % E A 10 E A 0 Low Nitrogen High Nitrogen Gleadow et al. 1998, Plant Cell Environ, 12:21
  • 9. 9/20 CO2 effects on cyanogenesis in clover Siobhan Isherwood • FACE • Pot trials (ANU) CassavaFACE • Nutrient trials –N, P, microbes
  • 10. 10/20 Clover Biomass 6 months after sowing (Dec) 400 Biomass g/m2 300 200 100 380 ppm 550 ppm October 2010, 3 months
  • 11. 11/20 Cyanide in clover trending higher at 550 ppm 1000 Cyanide mg/g dry wt 800 600 400 200 380 ppm 550 ppm • Is leaf protein lower? • Nodulation rates: 6% higher significant?? • Mycorrhizae colonisation: 5% higher ns?
  • 12. 12/20 ANU study: More cyanide relative protein at elevated CO2…ONLY with +P 370 ppm 12 700 ppm 10 Cyanide : protein 8 6 4 2 0 -N-P -N+P +N+P Gleadow et al. 2009 J Chem. Ecol 35, 476 Edwards & Evans 2005 Global Change Biol.11, 1968
  • 13. 13/20 Nutrient cyanogenesis trials 1.2 1.0 CN-N/N% 0.8 0.6 0.4 0.2 0.0 HP:HN HP:LN LP:HN LP:LN Nutrient Treatment • N15/N14 isotopes – P improved N fixation High P: 0.125g/kg – Fixed N used to make HCN Low P: 0.025g/kg High N: 8mM Low N: 2mM Isherwood, 2010 HONS THESIS
  • 14. 14/20 Conclusion: nutritional value IS affected by eCO2 • Leaf composition – Less leaf protein – Increased cyanide, relative to protein – Increase tannins, phenolics • Soil environment is crucial • Microbes important in nutrient acquisition …How will elevated CO2 affect these symbioses??? VIEWPOINT: Cavagnaro, Gleadow, Miller (2011) Functional Plant Biology 38, 87–96
  • 15. 15/20 Monash Cyanogenesis Group Collaborators Cecilia Blomstedt Tim Cavagnaro University of Copenhagen Rebecca Miller Birger Møller Kirsten Jørgensen, Morten Møldrup Alan Neale John Hamill RSB (ANU) Anna Burns John Evans, Stephanie McCaffrey Natalie O’Donnell Howard Bradbury, Bill Foley Cara Griffiths Sam Fromhold Pacific Seeds Melissa Bain Peter Stuart, Wayne Chesser Siobhan Isherwood Mozambique/South Africa Kiara O’Gorman Julie Cliff UEM Anabela Zacarias, IIAM FUNDING Simon Adams (Monash) ARC Linkage AusAID University of Melbourne Finkel Foundation Saman Sereweena (AGFACE) Ian Woodrow, Jennifer Fox Monash University Casstech DPI: AGFACE Pacific Seeds

Notas del editor

  1. Yield Nutritional value Protein contentPlant defence: cyanogenic glycosideAvailability 
  2. Below ground, : soil microbial associations MODEL FOR OTHER CYANOGENIC PLANTSsubterranean clover is the most important annual pasture legume sown in southern Australia. It originated in the Mediterranean region, parts of western Europe, southern England and Ireland.Subterranean clover (sub clover) was accidentally brought to Australia in hay, straw and pasture seed, probably as early as the 1830s, and was spread along transport routes, in stock camps and on town commons throughout southern Australia.The potential of sub clover to provide nutritious feed for livestock and improve soil fertility
  3. Ever since I did this work I’ve been wondering what the impact would be on crop plants.
  4. To investigate relationships between cyanogenesis in T. repens and nutrient supplyIs allocation of N to chemical defence affected by relative availability N and P?How this that affected by microbial associationsIs there a trade off between defence & growth?How is cyanogen concentration likely to change under future climate changes & eCO2? AgFACE elevated CO2 study, Horsham
  5. Does this translate into higher HCN levels?
  6. 27 vs. 36% (+/- 3) Mycorrhizae colonisation: 54 vs. 59 % (+/- 5) ns?
  7. Haven’t yet measured total N and other nutrients – expect CN per protein will be higher as in this study given sufficient nutrientsConsistent with this pot study – higher CN: protein at eCO2. This study also shows that importance of nutrient supply. These plants has rhizobium but were not inoculated mycrorrhizae.Effect only detected when P was added/sufficient.So here N – no difference with soil N because able to fix N, take up N to compensate for low soil N.
  8. IN order to understand importance of N and phosphorus and whether the source of N and P (i.e. whether from soil microbes) impacts on HCN content,Q…..N from rhizobium compensates for less soil N and plants can continue to synthesis cyanogenic glycosidesAs before under low N , rhizobium compensated i.e. low N plants actually had higher plant NBUT low P plants had lower PO4- mycorrhizae were unable to compensate.N treatment had no effect on HCN, but P treatment did.
  9. Available nitrogen is reallocated to cyanide in plants grown at elevated CO2ANDNitrogen (and Cyanogenesis) is dependent on soil microbesTHEN