Microbe Plant Mutalisms- Rhizobia/Legumes

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Microbe Plant Mutalisms- Rhizobia/Legumes

  1. 1. Microbe – Plant Mutualisms<br />So You Think You Are Alone<br />Fall 2009<br />
  2. 2. Mutualisms<br />Repetitive Intro<br />Mutualisms are interactions between two species in which both species gain a benefit<br />+ + interactions<br />
  3. 3. Natural Selection <br />Natural selection should favor organisms that behave in a way that maximizes their fitness (lifetime reproductive success)<br />Natural selection should favor traits that maximizes Benefit – Cost<br />
  4. 4. Mutualisms<br />Thus, both species involved in mutualistic relationships should receive benefits that are greater than the costs<br />
  5. 5. Origin of Mutualisms<br />How did these mutualistic relationships originate?<br />John Thompson (1984) proposed that most mutualistisms arose out of relationships that were originally antagonistic<br />Pollinators were originally pollen predators<br />Fruit dispersers were originally seed predators<br />Over time coevolution caused antagonistic relationships to become mutualistic<br />
  6. 6. Let’s Think About Mutualisms<br />You might have a mutualistic relationship with Dairy Queen<br />You benefit by getting Blizzards from DQ<br />DQ benefits by getting money from you<br />Everyone Benefits!!!<br />
  7. 7. Costs of Mutualisms<br />But there are costs associated with your relationship with Dairy Queen<br />Costs you money to buy a blizzard<br />Costs DQ money to make and sell a Blizzard<br />
  8. 8. Let’s Think About Units<br />The benefits and costs must be measured in some type of unit<br />If I told you that you would gain a benefit of 200 if you washed my car would you do it?<br />It might depend on whether the units of benefit were cents or dollars<br />DQ example<br />Your costs, DQ’s costs, & DQ’s benefit measured in dollars<br />Your benefit measured in enjoyment<br />
  9. 9. Different Units<br />It is often difficult to start making cost/benefit decisions when costs and benefits are measured in different units<br />How do you convert from one unit to another<br />E.g. how much money is the benefit that you receive from eating a blizzard really worth?<br />
  10. 10. DQ and You<br />In order for you to buy a Blizzard from DQ <br />the benefit you gain from eating the blizzard must be greater than the cost of buying the Blizzard<br />In order for DQ to sell the Blizzard to you <br />the benefit they gain must be greater than the cost for them to make and sell it to you<br />Both parties benefit<br />
  11. 11. DQ and You<br />DQ is a business designed to maximize their profits<br />Maximize money they make – money they spend<br />How should DQ price the Blizzard?<br />Imagine that it costs DQ $1.00 to make and sell the Blizzard.<br />Thus, the higher they price the Blizzard the more profit they make<br />$500 – $1 &gt;&gt;&gt;&gt;&gt; $2.99 - $1<br />
  12. 12. DQ and You<br />Thus, DQ would appear to make more profit by charging you $500 for a Blizzard than by charging you $2.99<br />Why doesn’t DQ charge so much for a Blizzard?<br />
  13. 13. DQ and You<br />In order for DQ to maximize their profit then they have to be aware that if they charge too much for a Blizzard that you won’t buy them<br />Your Benefit &lt; the cost<br />Thus, DQ can’t just gouge their customers because you could choose to shop somewhere else for your desert treats (e.g., Sonic Blast) or get by with no ice cream<br />
  14. 14. You and DQ<br />You want to maximize your benefit – cost<br />The enjoyment you get from eating a Blizzard is independent of the cost so you should want to pay as little as possible <br />However, if you are only willing to pay $0.50 then DQ will go out of business and you won’t be able to buy any more Blizzards.<br />
  15. 15. DQ and You & You and DQ<br />Thus, you and Dairy Queen are both trying to maximize your own benefit – cost while keeping the relationship going<br />However it is possible for the relationship to break down when either party is no longer receiving more benefit than cost<br />
  16. 16. Selfishness in Mutualisms<br />Both species in mutualisms are selected the maximize the benefit – cost<br />They shouldn’t worry about the benefit – cost of their partner as long as they can make sure that their partner benefits enough to remain in the partnership<br />Thus, organisms might be pretty selfish in these relationships!!!!!!<br />
  17. 17. Nitrogen<br />80 % of atmosphere is N2<br />Living organisms can not directly incorporate N2 into biological molecules<br />Living organisms need nitrogen<br />Proteins<br />DNA<br />RNA<br />Other molecules<br />
  18. 18. Nitrogen Fixation<br />N2 + 3H2 2NH3<br />The reduction of nitrogen gas to ammonia is energy intensive<br />
  19. 19. Nitrogen Fixation<br />Nitrogen can be fixed in three ways<br />Atmospheric fixation<br />this occurs spontaneously due to lightning<br />a small amount only is fixed this way.<br />Industrial fixation –<br />the Haber process, which is very energy inefficient<br />is used to make nitrogen fertilizers.<br />
  20. 20. Nitrogen Fixation<br />Biological fixation <br />nitrogen-fixing bacteria <br />16 molecules of ATP <br />a complex set of enzymes to break the nitrogen bonds so that it can combine with hydrogen<br />
  21. 21. Nitrogen Fixing Bacteria<br />Free living<br />Aerobic - Azotobacter<br />Anaerobic – Clostridium<br />Symbiotic association with plants<br />Legumes<br />e.g. peas, beans<br />Rhizobium<br />Non-legumes<br />e.g. alder tree<br />Frankia<br />
  22. 22. Rhizobium<br />These bacteria can infect the roots of leguminous plants, leading to the formation of lumps or nodules where the nitrogen fixation takes place. <br />About 90% of legume species can become nodulated.<br />
  23. 23. LegumesFamily Fabaceae<br />Alfalfa<br />
  24. 24. Rhizobia in Root Nodules<br />In root nodules the nitrogen-fixing form exists as irregular cells called bacteroids which are often club and Y-shaped.<br />
  25. 25. Free Living Rhizobium<br />In the soil the bacteria are free living and motile, feeding on the remains of dead organisms. <br />Free living Rhizobiumcannot (or rarely) fix nitrogen<br />
  26. 26. Origin of Rhizobium/Legume Mutualism<br />Don’t really know<br />An ancestral bacteria might have infected plant roots<br />Because the plant benefited from the fixed nitrogen, rather than excluding the bacteria the plant evolved to facilitate the relationship<br />Because the bacteria benefited from the sugar supplied by the plant they might have evolved to facilitate the relationship<br />
  27. 27. Genetics of Nodule Formation<br />Specificity genes determine which Rhizobiumstrain infects which legume. <br />the pea is the host plant to Rhizobiumleguminosarumbiovarviciae<br /> clover acts as host to R. leguminosarumbiovartrifolii.<br />
  28. 28. Genetics of Nodule Formation<br />Even if a strain is able to infect a legume, the nodules formed may not be able to fix nitrogen.<br />ineffective. <br />Effective strains induce nitrogen-fixing nodules.<br />Effectiveness is governed by a different set of genes in the bacteria from the specificity genes. <br />Nod genes direct the various stages of nodulation.<br />
  29. 29. Root Nodules<br />nodules<br />
  30. 30. Nodule Formation<br /><ul><li>Root cells release chemicals into the soil.
  31. 31. encourage the growth of the bacterial population in the area around the roots (the rhizosphere).
  32. 32. Bacterial cell wall and the root surface interactions are responsible for the rhizobia recognizing their correct host plant and attaching to the root hairs.
  33. 33. Flavonoids secreted by the root cells activate the nod genes in the bacteria which then induce nodule formation. </li></li></ul><li>Root Hairs<br />Root hair<br />Root hairs are surrounded by free<br />Living rhizobium<br />Root tip<br />
  34. 34. Nodule Formation<br />Once bound to the root hair, the bacteria excrete nod factors. These stimulate the hair to curl. <br />Rhizobia then invade the root through the hair tip where they induce the formation of an infection thread. This thread is constructed by the root cells and not the bacteria and is formed only in response to infection. <br />
  35. 35. Formation of Infection Thread<br />Root hair starts to curl<br />formation of an infection thread through which rhizobiaenter root cells<br />
  36. 36. Nodule Formation<br />The infection thread grows through the root hair cells and penetrates other root cells nearby often with branching of the thread. <br />The bacteria multiply within the expanding network of tubes, continuing to produce nod factors which stimulate the root cells to proliferate, eventually forming a root nodule. <br />
  37. 37. Formation of Bacteroids<br />infection thread spreads<br />into adjacent cells<br />bacteroids are released from<br />the infection thread<br />
  38. 38. Nodule Formation<br />Within a week of infection small nodules are visible to the naked eye. Each root nodule is packed with thousands of living Rhizobiumbacteria, most of which are in the misshapen form known as bacteroids.<br />
  39. 39. Bacteroids<br />
  40. 40. Nodules<br />
  41. 41. Nitrogenase<br />Nitrogenase is the enzyme that catalyses the conversion of nitrogen gas to ammonia in nitrogen-fixing organisms. <br />In legumes it only occurs within the bacteroids. <br />The reaction requires hydrogen as well as energy from ATP. <br />
  42. 42. Nitrogenase is Oxygen Sensitive<br />The nitrogenase complex is sensitive to oxygen, becoming inactivated when exposed to it. <br />Free Living Bacteria<br />have a variety of different mechanisms for protecting the nitrogenase complex, including high rates of metabolism and physical barriers.<br />E.g., Azotobacter overcomes this problem by having the highest rate of respiration of any organism, thus maintaining a low level of oxygen in its cells.<br />
  43. 43. Rhizobia in Nodules<br />Rhizobiacontrols oxygen levels in the nodule with leghaemoglobin. <br />This red, iron-containing protein has a similar function to that of haemoglobin; binding to oxygen. <br />provides sufficient oxygen for the bacteroidsbut prevents the accumulation of free oxygen that would destroy the activity of nitrogenase.<br />Leghaemoglobinis formed through the interaction of the plant and the rhizobia<br />neither can produce it alone.<br />Insides of nodules are red because of<br />leghaemoglobin<br />
  44. 44. Back to DQ and You<br />DQ wants you to buy a Blizzard every day.<br />However, your need for a Blizzard should vary seasonally<br />You gain a great benefit from a Blizzard in the summer when it is warm<br />You gain less benefit from a Blizzard in the winter when it is cold<br />
  45. 45. DQ and You<br />Who has control?<br />DQ advertises like crazy<br />‘that’s what I like about TEXAS” <br />You control your purse<br />Thus, you might like to only be able to buy Blizzards when it is warm.<br />
  46. 46. Rhizobia and Legumes<br />Who has control in this relationship?<br />Do the rhizobia determine how much sugar the plant send to the nodules? or<br />Does the plant control how much Nitrogen fixation takes place by controlling how much sugar it sends to the plant?<br />Some early data suggested that the plants had control.<br />
  47. 47. From the Plant’s Perspective<br />When does the plant benefit most from having a relationship with rhizobia?<br />When they need nitrogen<br />When do they need more nitrogen?<br />When they live in soil that has a low nitrogen content<br />Therefore we predict, that if plants have control of the system that plants should be more likely to have more nodules in low nitrogen soil.<br />
  48. 48. How would you test this prediction?<br />Natural Experiments<br />Examine the roots of the same species of plants living in soil with different nitrogen contents<br />Manipulative Experiments<br />Experimentally manipulate the soil nitrogen content <br />Field or lab<br />
  49. 49. Tests of Prediction<br />Many laboratory and field studies in crop plants and some undomesticated legumes have shown Lower nodulation rates with increasing soil nitrogen availability <br />This observation suggests that the relative benefits of nodulation decline with increasing abundance of reduced nitrogen, which plants can obtain directly from the soil.<br />
  50. 50. From the Plant’s Perspective<br />Plants should benefit less from increasing their access to nitrogen when they are limited by another resources (e.g. phosphorous)<br />Thus, predict that if the level of nitrogen in the soil is constant that plants should allow more nodules when the level of other limiting soil nutrients is high<br />
  51. 51. From the Plant’s Perspective<br />Legumes also restrict nodulation when inadequate supplies of other nutrients, especially phosphorus, limit plant growth<br />
  52. 52. Conclusions<br />The fact that plants reduce nodule formation under conditions of high nitrogen or low phosphorus availability illustrates that plants do not permit unlimited infection by compatible rhizobia.<br />Plants have some control over whether or not they are “infected” by rhizobia<br />
  53. 53. From the Plant’s Perspective<br />The value of nitrogen to plants varies over time<br />Once a plant is mature and has the appropriate concentration of nitrogen in its leaves, increasing the nitrogen content in the plant doesn’t help very much<br />When do mature legume plants require more nitrogen?<br />When they are filling their seeds.<br />
  54. 54. From the Plant’s Perspective<br />Legume seeds are rich in nitrogen<br />If the plant has control of the system then the plant would like for the rhizobia to provide more nitrogen during the period of seed filling<br />Data shows that to be the case.<br />It appears that plants can “turn on” nitrogen fixation by rhizobia<br />Maybe by increasing the amount of sugar that they send to the nodules<br />
  55. 55. Phloem<br />Xylem moves water and nutrients from roots to the leaves<br />Phloem moves sugar around the plant<br />Plant can control when and where sugar is moved<br />Phloem loading and unloading<br />
  56. 56. But As We Learn More Things Get More Complex<br />All Rhizobium are not created equally<br />Mutualisticrhizobiaprovide their legume hosts with nitrogen. <br />Form nodules & fix nitrogen<br />Parasitic rhizobia infect legumes, but fix little or no nitrogen. <br />Form nodules<br />Nonsymbioticstrains are unable to infect legumes at all. <br />Don’t form nodules<br />
  57. 57. Why have Rhizobiumstrains with one of these three strategies not displaced the others?<br />We need to think about the costs and benefits associated with the different strategies<br />
  58. 58. Things we need to think about<br />Competition<br />All rhizobia spend some time in the soil, where they compete for resources. <br />Symbiotic rhizobia, both mutualisticN2-fixers and parasitic nonfixers, compete for host plants to colonize<br />
  59. 59. Things we need to think about<br />Nitrogen fixation is an energetically expensive process<br />uses resources that rhizobia could otherwise use for their own growth and reproduction. <br />Therefore a good strategy might be to be a nonfixing bacteria that benefits from the resources provided by the plant<br />Get the benefit without paying either of the costs<br />Bacterial strain benefits but the plant does not.<br />“cheater”<br />
  60. 60. Benefits of Symbiosis<br />Founding a nodule can dramatically enhance the reproductive success of rhizobia.<br />A single rhizobium cell that infects a soybean root may produce up to 1010 descendants inside a large nodule<br />Thus, a rhizobium could produce many more descendants in the soil by founding a nodule than by remaining in the soil.<br />
  61. 61. Stuff Scientists Are Starting to Learn<br />First, the chances of infecting a legume may be quite low. <br />A soil in which a compatible host was last grown five years before can still contain2.5 x 104rhizobia per g of soil. <br />roughly 2 x 109g/ha of soil in the plow layer would then contain5 x 1013rhizobia. <br />A soybean field with 4 x 105 plants, each forming 100 nodules, would offer only 4 · 107 opportunities to found a nodule. <br />Thus, the chances that a given symbiotic rhizobium cell would successfully found a nodule would be about one in a million.<br />
  62. 62. Stuff Scientists Are Starting to Learn<br />Aggregations of rhizobia around a root might attract high populations of predatory protozoa increasing predation risk relative to living in “bulk soil”<br />Many rhizobiaproduce various antibiotics active against other rhizobia<br />exposure to these antibiotics could be greater for rhizobia attempting to infect a legume root for those that live in the bulk soil.<br />
  63. 63. Partner Choice<br />Plants might benefit from being able to choose which strain of rhizobia infects them or by being able to preferentially allocate sugars to nodules that contain mutualisticrhizobium.<br />It is well known that crop legumes nodulate non-fixing rhizobia, but allocate few resources to those nodules. <br />
  64. 64. Simms et al. 2006<br />Examined wild legume, Lupinusarboreusin experiments where they infected them with 3 strains of rhizobia (low, medium, and high nitrogen fixers)<br />Greenhouse experiments showed that plants frequently hosted less cooperative strains<br />Appears plants can’t control who infects them<br />However, the nodules occupied by low fixing strains were smaller. <br />Suggests they may be able to reduce resources sent to these nodules<br />
  65. 65. Simms et al. 2006<br />Survey of wild-grown plants showed that larger nodules house more high fixing strains<br />plants may prevent the spread of exploitation by favoring better cooperators.<br />
  66. 66. Mathematical modelling<br />
  67. 67. Mathematical Modelling<br />A mechanistic molecular test of the plant-sanction hypothesis in legume–rhizobia mutualism <br />Diana E. Marco et al. 2009<br />
  68. 68. Questions<br />Can plants tell whether rhizobia are fixing or non-fixing strains?<br />Do they preferentially form nodules with fixing strains?<br />After nodulation can plants determine which nodules contain fixing and non-fixing strains?<br />Do they “sanction” non-fixing strains by sending them less energy?<br />
  69. 69. Studied Soybeans in the Greenhouse<br />A mechanistic molecular test of the plant-sanction hypothesis in legume–rhizobia mutualism <br />Diana E. Marcoet al. 2009<br />
  70. 70. Split Root Experimental Design<br />
  71. 71. Can Plants Determine Whether or Not a Bacteria is a Fixer or Not Before Nodulation?<br />If they can then we predict to see more nodules on roots that have been grown in soil that contains fixing bacteria than on the roots of plants grown in soil that contains non-fixing bacteria<br />
  72. 72. Results- from plants infected by both strains<br />Closed circles- fixing strain and open circles- non fixing strain<br />
  73. 73. Conclusion<br />Because there were equal numbers of nodules on roots grown in soil with fixing and non-fixing bacteria we conclude either<br />plants are unable to determine whether or not bacteria are fixing or non-fixing strain or<br />If they can tell, they are unable to stop non-fixing strains from forming nodules<br />
  74. 74. Can Plants Determine Which Nodules Are Not Fixing Nitrogen and “Sanction” Them?<br />If so, we predict that over time nodules formed by non-fixing bacteria should have fewer bacteria living in them<br />
  75. 75. Split Root Study<br />Plants received one type of rhizobia<br />Predict more rhizobia in nodules of plants infected with fixing strain than in nodules infected with non-fixing strain<br />Plants that received both types of rhizobia<br />On the same plant, nodules that were infected with fixing strain should have more rhizobia than nodules infected by non-fixing strain<br />
  76. 76. A & B are two different strains of <br />soybeans<br />T1 plants- ½ fixing, ½ non-fixing<br />T2 plants- both halves fixing<br />T3 plants- both halves non-fixing<br />
  77. 77. Conclusions<br />Plants infected with non-fixing bacteria could tell and stop sending resources to the nodules so the bacteria in the nodules died<br />Plants infected by both fixing and non-fixing bacteria could not selective stop sending resources to the non-fixing nodules<br />Very Interesting Result!!!!!<br />
  78. 78. References<br />ELLEN L. SIMMS AND D. LEE TAYLOR. 2002. Partner Choice in Nitrogen-Fixation Mutualisms of Legumes and Rhizobia. INTEG. AND COMP. BIOL., 42:369–380<br />E. Toby Kiers, Robert A. Rousseau, Stuart A. Wes & R. Ford Denison. 2003. Host sanctions and the legume–rhizobium mutualism. NATURE 425, 78-81<br />R. Ford Denison & E. Toby Kiers. 2004. Lifestyle alternatives for rhizobia: mutualism, parasitism,andforgoing symbiosis. FEMS Microbiology Letters 237 187–193<br />Ellen L. Simms, D. Lee Taylor, Joshua Povich, Richard P. Shefferson,J. L. Sachs, M. Urbina and Y. Tausczik. 2006. An empirical test of partner choice mechanisms in a wild legume–rhizobium interaction. Proc. R. Soc. B. 273, 77–81<br />Katy D. Heath and Peter Tiffin. 2007. Context dependence in the coevolution of plant and rhizobialmutualists. Proc. R. Soc. B. 274, 1905–1912<br />E. Toby Kiers, Robert A. Rousseau and R. Ford Denison. 2006. Measured sanctions: legume hosts detect quantitative variation in rhizobium cooperation and punish accordingly. Evolutionary Ecology Research, 8: 1077–1086<br />Diana E. Marco, RebecaPérez-Arnedo, ÁngelesHidalgo-Perea, José Olivares, José E. Ruiz-Sainzand Juan Sanjuán. 2009. A mechanistic molecular test of the plant-sanction hypothesis in legume–rhizobiamutualism. ActaOecologica35: 664-667<br />

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