1. Course Project = Algal Lipid Production 1)Decide which algae to study http://www.cbs.umn.edu/lab/wackett/links/oil 2)Learn more about cell walls and lipid synthesis 3)Design some experiments 4)See where they lead us

2. Course Project = Algal Lipid Production 1)Decide which algae to study http://www.cbs.umn.edu/lab/wackett/links/oil Next assignment: each pick an alga and product and convince the group in 5-10 minutes why your choice is best. Next Wed?

3. Potential experiments 1.Effects of environment on lipid production pCO 2 Temperature Light quantity Light quality = color(s) Nutrition Adding acetate? 2. Effects of environment on cell walls

4. Potential experiments 1.Effects of environment on lipid production pCO 2 Temperature Light quantity Light quality = color(s) Nutrition 2. Effects of environment on cell walls 3. Effects of inhibitors on cell walls

5. Lipid metabolism Unique aspects in plants 1)Make fatty acids in plastids 2)large amounts of galactolipids 3)Oleosomes: oil-storing organelles with only outer leaflet

6. Plant Growth Size & shape depends on cell # & cell size

7. Plant Growth Size & shape depends on cell # & cell size Decide when,where and which way to divide

8. Plant Growth Size & shape depends on cell # & cell size Decide which way to divide & which way to elongate Periclinal = perpendicular to surface

9. Plant Growth Size & shape depends on cell # & cell size Decide which way to divide & which way to elongate Periclinal = perpendicular to surface: get longer

10. Plant Growth Size & shape depends on cell # & cell size Decide which way to divide & which way to elongate Periclinal = perpendicular to surface: get longer Anticlinal = parallel to surface

11. Plant Growth Size & shape depends on cell # & cell size Decide which way to divide & which way to elongate Periclinal = perpendicular to surface: get longer Anticlinal = parallel to surface: add more layers

12. Plant Growth Decide which way to divide & which way to elongate Periclinal = perpendicular to surface: get longer Anticlinal = parallel to surface: add more layers Now must decide which way to elongate

13. Plant Growth Decide which way to divide & which way to elongate Periclinal = perpendicular to surface: get longer Anticlinal = parallel to surface: add more layers Now must decide which way to elongate: which walls to stretch

14. Plant Cell Walls and Growth Carbohydrate barrier surrounding cell Protects & gives cell shape

15. Plant Cell Walls and Growth Carbohydrate barrier surrounding cell Protects & gives cell shape 1˚ wall made first mainly cellulose Can stretch!

16. Plant Cell Walls and Growth Carbohydrate barrier surrounding cell Protects & gives cell shape 1˚ wall made first mainly cellulose Can stretch! 2˚ wall made after growth stops Lignins make it tough

17. Plant Cell Walls and Growth 1˚ wall made first mainly cellulose Can stretch! Control elongation by controlling orientation of cell wall fibers as wall is made

18. Plant Cell Walls and Growth 1˚ wall made first mainly cellulose Can stretch! Control elongation by controlling orientation of cell wall fibers as wall is made 1˚ walls = 25% cellulose, 25% hemicellulose, 35% pectin, 5% protein (but highly variable)

19. Plant Cell Walls and Growth 1˚ walls = 25% cellulose, 25% hemicellulose, 35% pectin, 5% protein (but highly variable) Cellulose: ordered chains made of glucose linked  1-4

20. Plant Cell Walls and Growth 1˚ walls = 25% cellulose, 25% hemicellulose, 35% pectin, 5% protein (but highly variable) Cellulose: ordered chains made of glucose linked  1-4 Cross-link with neighbors to form strong, stable fibers

21. Plant Cell Walls and Growth Cellulose: ordered chains made of glucose linked  1-4 Cross-link with neighbors to form strong, stable fibers Made by enzyme embedded in the plasma membrane Guided by cytoskeleton Other wall chemicals are made in Golgi & secreted Only cellulose pattern is tightly controlled

22. Plant Cell Walls and Growth Cellulose pattern is tightly controlled 6 CES enzymes form a “ rosette ” : each makes 6 chains -> 36/fiber

23. Plant Cell Walls and Growth Cellulose pattern is tightly controlled 6 CES enzymes form a “ rosette ” : each makes 6 chains -> 36/fiber Rosettes are guided by microtubules

24. Plant Cell Walls and Growth Cellulose pattern is tightly controlled 6 CES enzymes form a “ rosette ” : each makes 6 chains Rosettes are guided by microtubules Deposition pattern determines direction of elongation

25. Plant Cell Walls and Growth Cellulose pattern is tightly controlled Deposition pattern determines direction of elongation New fibers are perpendicular to growth direction, yet fibers form a mesh

26. Plant Cell Walls and Growth New fibers are perpendicular to growth direction, yet fibers form a mesh Multinet hypothesis: fibers reorient as cell elongates Old fibers are anchored so gradually shift as cell grows

27. Plant Cell Walls and Growth New fibers are perpendicular to growth direction, yet fibers form a mesh Multinet hypothesis: fibers reorient as cell elongates Old fibers are anchored so gradually shift as cell grows Result = mesh

28. Plant Cell Walls and Growth 1˚ walls = 25% cellulose, 25% hemicellulose, 35% pectin, 5% protein (but highly variable) Hemicelluloses AKA cross-linking glycans: bind cellulose

29. Plant Cell Walls and Growth Hemicelluloses AKA cross-linking glycans: bind cellulose Coat cellulose & bind neighbor

30. Plant Cell Walls and Growth Hemicelluloses AKA cross-linking glycans Coat cellulose & bind neighbor Diverse group of glucans: also linked  1-4, but may have other sugars and components attached to C6

31. Hemicelluloses Diverse group of glucans: also linked  1-4, but may have other sugars and components attached to C6 makes digestion more difficult

32. Hemicelluloses Diverse group of glucans: also linked  1-4, but may have other sugars and components attached to C6 makes digestion more difficult Assembled in Golgi

33. Plant Cell Walls and Growth Hemicelluloses AKA cross-linking glycans A diverse group of glucans also linked  1-4, but may have other sugars and components attached to C6 makes digestion more difficult Assembled in Golgi Secreted cf woven

34. Plant Cell Walls and Growth 1˚ walls = 25% cellulose, 25% hemicellulose, 35% pectin, 5% protein (but highly variable) Pectins: fill space between cellulose-hemicellulose fibers

35. Pectins Pectins: fill space between cellulose-hemicellulose fibers Form gel that determines cell wall porosity(& makes jam)

36. Pectins Pectins: fill space between cellulose-hemicellulose fibers Form gel that determines cell wall porosity (& makes jam) Acidic, so also modulate pH & bind polars

37. Pectins Pectins: fill space between cellulose-hemicellulose fibers Form gel that determines cell wall porosity (& makes jam) Acidic, so also modulate pH & bind polars Backbone is  1-4 linked galacturonic acid

38. Pectins Backbone is  1-4 linked galacturonic acid Have complex sugar side-chains, vary by spp.

39. Pectins Backbone is  1-4 linked galacturonic acid Have complex sugar side-chains, vary by spp.

40. Plant Cell Walls and Growth Also 4 main multigenic families of structural proteins

41. Plant Cell Walls and Growth Also 4 main multigenic families of structural proteins Amounts vary between cell types & conditions

42. Plant Cell Walls and Growth Also 4 main multigenic families of structural proteins Amounts vary between cell types & conditions 1.HRGP: hydroxyproline-rich glycoproteins (eg extensin) Proline changed to hydroxyproline in Golgi

43. Plant Cell Wall Proteins 1.HRGP: hydroxyproline-rich glycoproteins (eg extensin) Proline changed to hydroxyproline in Golgi Highly glycosylated: helps bind CH 2 O

44. Plant Cell Wall Proteins 1.HRGP: hydroxyproline-rich glycoproteins (eg extensin) Proline changed to hydroxyproline in Golgi Highly glycosylated: helps bind CH 2 O Common in cambium, phloem

45. Plant Cell Wall Proteins 1.HRGP: hydroxyproline-rich glycoproteins (eg extensin) Proline changed to hydroxyproline in Golgi Highly glycosylated: helps bind CH 2 O Common in cambium, phloem Help lock the wall after growth ceases

46. Plant Cell Wall Proteins 1.HRGP: hydroxyproline-rich glycoproteins (eg extensin) Proline changed to hydroxyproline in Golgi Highly glycosylated: helps bind CH 2 O Common in cambium, phloem Help lock the wall after growth ceases Induced by wounding 2. PRP: proline-rich proteins

47. Plant Cell Wall Proteins 1.HRGP: hydroxyproline-rich glycoproteins (eg extensin) 2.PRP: proline-rich proteins Low glycosylation = little interaction with CH 2 O

48. Plant Cell Wall Proteins 1.HRGP: hydroxyproline-rich glycoproteins (eg extensin) 2.PRP: proline-rich proteins Low glycosylation = little interaction with CH 2 O Common in xylem, fibers, cortex

49. Plant Cell Wall Proteins 1.HRGP: hydroxyproline-rich glycoproteins (eg extensin) 2.PRP: proline-rich proteins Low glycosylation = little interaction with CH 2 O Common in xylem, fibers, cortex May help lock HRGPs together

50. Plant Cell Wall Proteins 1.HRGP: hydroxyproline-rich glycoproteins (eg extensin) 2.PRP: proline-rich proteins Low glycosylation = little interaction with CH 2 O Common in xylem, fibers, cortex May help lock HRGPs together 3.GRP: Glycine-rich proteins No glycosylation = little interaction with CH 2 O

51. Plant Cell Wall Proteins 1.HRGP: hydroxyproline-rich glycoproteins (eg extensin) 2.PRP: proline-rich proteins Low glycosylation = little interaction with CH 2 O Common in xylem, fibers, cortex May help lock HRGPs together 3.GRP: Glycine-rich proteins No glycosylation = little interaction with CH 2 O Common in xylem

52. Plant Cell Wall Proteins 1.HRGP: hydroxyproline-rich glycoproteins (eg extensin) 2.PRP: proline-rich proteins Low glycosylation = little interaction with CH 2 O Common in xylem, fibers, cortex May help lock HRGPs together 3.GRP: Glycine-rich proteins No glycosylation = little interaction with CH 2 O Common in xylem May help lock HRGPs & PRPs together

53. Plant Cell Wall Proteins 1.HRGP: hydroxyproline-rich glycoproteins (eg extensin) 2.PRP: proline-rich proteins 3.GRP: Glycine-rich proteins No glycosylation = little interaction with CH 2 O Common in xylem May help lock HRGPs & PRPs together 4. Arabinogalactan proteins

54. Plant Cell Wall Proteins 1.HRGP: hydroxyproline-rich glycoproteins (eg extensin) 2.PRP: proline-rich proteins 3.GRP: Glycine-rich proteins 4.Arabinogalactan proteins Highly glycosylated: helps bind CH 2 O

55. Plant Cell Wall Proteins 1.HRGP: hydroxyproline-rich glycoproteins (eg extensin) 2.PRP: proline-rich proteins 3.GRP: Glycine-rich proteins 4.Arabinogalactan proteins Highly glycosylated: helps bind CH 2 O Anchored to PM by GPI

56. Plant Cell Wall Proteins 1.HRGP: hydroxyproline-rich glycoproteins (eg extensin) 2.PRP: proline-rich proteins 3.GRP: Glycine-rich proteins 4.Arabinogalactan proteins Highly glycosylated: helps bind CH 2 O Anchored to PM by GPI Help cell adhesion and cell signaling

57. Plant Cell Wall Proteins 1.HRGP: hydroxyproline-rich glycoproteins (eg extensin) 2.PRP: proline-rich proteins 3.GRP: Glycine-rich proteins 4.Arabinogalactan proteins Highly glycosylated: helps bind CH 2 O Anchored to PM by GPI Help cell adhesion and cell signaling 5. Also many enzymes involved in cell wall synthesis and loosening

58. Plant Cell Walls and Growth Also many enzymes involved in cell wall synthesis and loosening As growth stops, start making lignins & linking HGRP

59. Plant Cell Walls and Growth As growth stops, start depositing lignins & linking HGRP Lignins = polyphenolic macromolecules: 2 nd most abundant on earth (after cellulose)

60. Plant Cell Walls and Growth Lignins = polyphenolic macromolecules: 2 nd most abundant on earth (after cellulose) Bond hemicellulose: solidify & protect cell wall (nature ’ s cement): very difficult to digest

61. Plant Cell Walls and Growth Lignins = polyphenolic macromolecules: 2 nd most abundant on earth (after cellulose) Bond hemicellulose: solidify & protect cell wall (nature ’ s cement): very difficult to digest Monomers are made in cytoplasm & secreted

62. Plant Cell Walls and Growth Monomers are made in cytoplasm & secreted Peroxidase & laccase in cell wall create radicals that polymerise non-enzymatically

63. Plant Cell Walls and Growth Monomers are made in cytoplasm & secreted Peroxidase & laccase in cell wall create radicals that polymerise non-enzymatically

64. Plant Cell Walls and Growth Peroxidase & laccase in cell wall create radicals that polymerise non-enzymatically Very difficult to digest, yet major plant component!

65. Plant Cell Walls and Growth As growth stops, start depositing lignins & linking HGRP Solidify & protect cell wall: very difficult to digest Elongation precedes lignification

66. Plant Cell Walls and Growth As growth stops, start depositing lignins & linking HGRP Solidify & protect cell wall: very difficult to digest Elongation precedes lignification Requires loosening the bonds joining the cell wall

67. Plant Cell Walls and Growth Elongation precedes lignification Requires loosening the bonds joining the cell wall Can ’ t loosen too much or cell will burst

68. Plant Cell Walls and Growth Elongation precedes lignification Requires loosening the bonds joining the cell wall Can ’ t loosen too much or cell will burst Must coordinate with cell wall synthesis so wall stays same

69. Plant Cell Walls and Growth Elongation: loosening the bonds joining the cell wall Can ’ t loosen too much or cell will burst Must coordinate with cell wall synthesis so wall stays same Must weaken crosslinks joining cellulose fibers

70. Plant Cell Walls and Growth Must weaken crosslinks joining cellulose fibers Turgor pressure then makes cells expand

71. Plant Cell Walls and Growth Must weaken crosslinks joining cellulose fibers Turgor pressure then makes cells expand Lower pH: many studies show that lower pH is sufficient for cell elongation

72. Plant Cell Walls and Growth Must weaken crosslinks joining cellulose fibers Lower pH: many studies show that lower pH is sufficient for cell elongation Acid growth hypothesis: Growth regulators cause elongation by activating H + pump

73. Plant Cell Walls and Growth Acid growth hypothesis: Growth regulators cause elongation by activating H + pump Inhibitors of H + pump stop elongation But: Cosgrove isolated proteins that loosen cell wall Test protein extracts to see if wall loosens

74. Plant Cell Walls and Growth Acid growth hypothesis: Growth regulators cause elongation by activating H + pump But: Cosgrove isolated proteins that loosen cell wall Test protein extracts to see if wall loosens Identified expansin proteins that enhance acid growth

75. Plant Cell Walls and Growth Acid growth hypothesis: Growth regulators cause elongation by activating H + pump But: Cosgrove isolated proteins that loosen cell wall Test protein extracts to see if wall loosens Identified expansin proteins that enhance acid growth Still don ’ t know how they work!

76. Plant Cell Walls and Growth Identified expansin proteins that enhance acid growth Still don ’ t know how they work! Best bet, loosen Hemicellulose/cellulose bonds

77. Plant Cell Walls and Growth Also have endoglucanases and transglucanases that cut & reorganize hemicellulose & pectin

78. Plant Cell Walls and Growth Also have endoglucanases and transglucanases that cut & reorganize hemicellulose & pectin XET (xyloglucan endotransglucosylase) is best-known

79. Plant Cell Walls and Growth Also have endoglucanases and transglucanases that cut & reorganize hemicellulose & pectin XET (xyloglucan endotransglucosylase) is best-known Cuts & rejoins hemicellulose in new ways

80. Plant Cell Walls and Growth XET is best-known Cuts & rejoins hemicellulose in new ways Expansins & XET catalyse cell wall creepage

81. Plant Cell Walls and Growth XET is best-known Cuts & rejoins hemicellulose in new ways Expansins & XET catalyse cell wall creepage Updated acid growth hypothesis: main function of lowering pH is activating expansins and glucanases

82. Plant Cell Walls and Growth Updated acid growth hypothesis: main function of lowering pH is activating expansins and glucanases Coordinated with synthesis of new cell wall to keep thickness constant

83. Plant Cell Walls and Signaling Pathogens must digest cell wall to enter plant

84. Plant Cell Walls and Signaling Pathogens must digest cell wall to enter plant Release cell wall fragments

85. Plant Cell Walls and Signaling Pathogens must digest cell wall to enter plant Release cell wall fragments Many oligosaccharides signal ” HELP! ”

86. Plant Cell Walls and Signaling Pathogens must digest cell wall to enter plant Release cell wall fragments Many oligosaccharides signal ” HELP! ” Elicit plant defense responses