QuickTime and a TIFF (Uncompressed) decompressor are needed to see this picture. Clay Carter Department of Biology QuickTime and a TIFF (LZW) decompressor are needed to see this picture.
Ornamental tobacco Nicotiana langsdorffii x Nicotiana sanderae
Are there antimicrobial proteins in nectar? kda 65 40 35 28 IV III II NecI SDS- PAGE proteins accumulate to ~240 μg/ml in tobacco nectar
HO OH OO - O NEC2/3 metal ions O OH monodehydroascorbate OH -. e - OH free radical trap HO OH OO O HO OH Ascorbate (930 μm) NEC5 glucose H 2 O 2 (4 mm) Antimicrobial NEC1 O 2 - NOX? X DPI O 2 The tobacco nectar redox cycle
Nectar Nectar + catalase
sugar (w/w) 49% suc/ (gluc+ fruc) 0.78 glucose / fructose 2.9 Primarily visited by Bombus sp. L. Galetto, Plant Syst Evol 196: 99-121
OH O OH PPO O R diphenol 1/2 O 2 H 2 O R o-benzoquinone Non-enzymatic polymerization melanins Antimicrobial and antiviral activities
QuickTime and a pellet pellet 84 kda 55 kda 45 kda 36 kda QuickTime and a TIFF (LZW) decompressor are needed to see this picture. TIFF (LZW) decompressor are needed to see this picture. QuickTime and a TIFF (LZW) decompressor are needed to see this picture. 29 kda SDS PAGE PPO Assay
QuickTime TIFF (LZW) dec are needed to see SDS PAGE PPO Assay
TDTDWLDAEFLFYDENAELVR QuickTime TIFF (LZW) dec are needed to see VDIPWINSPK SDS PAGE PPO Assay
pellet 84 kda 55 kda 45 kda 36 kda QuickTime and a TIFF (LZW) decompressor are needed to see this picture. QuickTime and a TIFF (LZW) decompressor are needed to see this picture. N-KPQVPCFFIFGDSLLDNG-C 29 kda Degenerate RT-PCR SDS PAGE 5 /3 RACE
K P Q V P C F F I F G D S L V D N G AAR CCN CAR GTN CCN TGY TTY TTC ATA TTT GGC GAT TCA TTG GTG GAC AAT GGT N N N N I Q S L A R A N Y L P Y G V AAC AAT AAT AAT ATC CAA TCC TTA GCC CGG GCT AAT TAC TTG CCT TAC GGT GTT D F P D G P T G R F S N G K T T V D GAT TTT CCT GAC GGG CCG ACC GGG AGG TTC TCC AAT GGT AAA ACC ACA GTT GAT V I A E L L G F D D Y I P P Y A S A GTC ATT GCT GAG CTG CTA GGT TTT GAT GAT TAT ATT CCC CCA TAC GCC AGT GCC S G D Q I L R G V N Y A S A A A G I AGT GGC GAT CAA ATA CTC AGG GGA GTA AAC TAT GCA TCT GCT GCT GCT GGA ATT R S E T G Q Q L G A R I D F T G Q V AGA TCA GAA ACT GGC CAG CAA TTG GGT GCT AGA ATT GAT TTT ACT GGG CAA GTA N N Y K N T V A Q V V D I L G D E D AAT AAC TAC AAG AAC ACA GTT GCA CAA GTG GTG GAC ATA TTA GGT GAT GAG GAT S A A N Y L S K C I Y S V G V G S N TCC GCG GCA AAT TAC TTG AGC AAA TGC ATA TAC TCA GTT GGA GTC GGC AGC AAC D Y L N N Y F M P L Y Y S S G R Q Y GAC TAT CTC AAC AAC TAT TTC ATG CCT CTC TAC TAC TCA AGT GGC CGC CAA TAC S P E Q Y S D L L I Q Q Y S E Q I R TCC CCT GAA CAA TAC TCC GAT CTC CTC ATT CAA CAA TAC TCT GAG CAA ATA AGG T L Y N Y G A R K F S L I G V G Q I ACT TTG TAC AAC TAT GGA GCT AGA AAG TTT TCT TTG ATT GGA GTG GGG CAA ATC JAC1 Length: 339 amino acids Predicted MW: 40 kda Function: GDSL-like lipase G C S P N A L A Q N S P D G S T C I GGG TGC AGC CCG AAT GCA TTG GCG CAA AAT AGC CCG GAT GGA TCT ACA TGT ATA R R I N D A N Q M F N N K L R A L V CGG AGA ATT AAC GAC GCG AAT CAG ATG TTC AAC AAC AAA CTG AGA GCA TTA GTT D E L N N G A Q D A K F I Y I N A Y GAT GAG TTG AAC AAC GGT GCT CAG GAT GCA AAG TTT ATC TAC ATT AAT GCA TAT G I F Q D L I D N P S A F G F R V T GGG ATT TTC CAG GAT TTG ATT GAC AAC CCT TCA GCT TTT GGT TTT AGA GTG ACA N A G C C G V G R N N G Q I T C L P AAT GCT GGA TGC TGT GGA GTG GGG AGG AAC AAT GGG CAA ATA ACA TGT CTC CCT M Q T P C Q N R D E Y L F W D A F H ATG CAG ACT CCA TGC CAG AAC AGA GAT GAA TAT TTG TTT TGG GAT GCA TTC CAT P T E A A N V V V G R R S Y R A E K CCC ACA GAA GCT GCA AAT GTT GTG GTC GGG AGG AGA TCG TAT CGT GCT GAA AAG A S D A Y P F D I Q R L A Q L GCA TCA GAT GCA TAT CCT TTT GAT ATT CAG CGC TTG GCT CAA CTC TAA
R O O C Lipase/ esterase H 2 O O ROH + HO C
Roles of lipases in plants growth development membrane composition disease resistance signaling direct antimicrobial activity
Lipase Activity in Crude Jacaranda Nectar Nonboiled nectar y = 0.0006x + 0.0007 R 2 = 1 y = 5E-05x + 0.0007 R 2 = 1 Boiled nectar Linear (Nonboiled nectar) Linear (Boiled nectar)
Lipase Activity in Centrifuged Jacaranda Nectar y = 0.0003x - 4E-05 R 2 = 0.9987 y = 4E-05x + 0.0003 R 2 = 0.1051 Supernatant Fraction Pelleted Fraction Linear (Supernatant Fraction) Linear (Pelleted Fraction)
T7 JAC1 His 6 pet22b-jac1 Vector only No induction
QuickTime and a TIFF (Uncompressed) decompressor are needed to see this picture. Leaky expression of Jac1 Full expression of Jac1
What is a lipase doing in nectar? Antimicrobial activity? or Modifying nectar?
QuickTime and a TIFF (Uncompressed) decompressor are needed to see this picture. QuickTime and a TIFF (Uncompressed) decompressor are needed to see this picture. 50 μm 10 μm QuickTime and a TIFF (Uncompressed) decompressor are needed to see this picture. 50 μm
H 2 C HC H 2 C O O O O C O C O C? JAC1 O H 2 C HC OH OH HO HO C O C O H 2 C OH HO C
QuickTime and a TIFF (LZW) decompressor are needed to see this picture. QuickTime and a TIFF (LZW) decompressor are needed to see this picture.
Two secretory processes may contribute to nectar production Aqueous portion from nectary? lipid portion from secretory trichomes?
Future Directions verify antimicrobial nature of nectar components determine lipid composition of nectar role(s) of lipids in pollinator attraction do similar systems occur in other plants? Summary Jacaranda nectar contains proteins known to be involved in plant defenses; may act to limit microbial growth polyphenol oxidase lipase Jacaranda nectar may be produced via two distinct mechanisms nectary - appears to produce aqueous components secretory trichomes - appears to produce lipids
Thanks to: Brian Kram QuickTime and a TIFF (LZW) decompressor are needed to see this picture. Natasha Raikhel Songqin Pan