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FLOWER HEATING
     FOLLOWING ANTHESIS
              &
 THE EVOLUTION OF GALL MIDGE
POLLINATION IN SCHISANDRACEAE



   AMERICAN JOURNAL OF BOTANY
           -JULY 2010-
Shi Xiao Luo & Dianxiang Zhan
     South China Botanical Garden,
          Guangzhou, China



       Shu-Miaw Chaw
      Biodiversity Research Center
            Taipei, Taiwan



     Susanne S. Renner
         Department of Biology,
          University of Munich
          Munich, Germany
KEY TERMS
• Agamospermy -the asexual formation of
  embryos and seeds without the occurrence of
  fertilization.
• Anthesis - the period during which a flower is
  fully open and functional.
• Oviposition - to deposit or lay eggs.
• Saprophytic - organisms that feed on dead or
  decaying matter.
•Background Information
• Objectives
• Materials
• Methods
• Results
• Discussion
BACKGROUND INFORMATION
STAR VINE FAMILY
                       ~90 SPECIES


Schisandraceae


                        Kadsura (22 species) - Endemic to Tropical Asia




              Schisandra (25 species) - Tropical Asia- (Exception - S. glabra)




Illicium (42 species) - SE Asia, SE U.S., Mexico, Greater Antilles
WHY IS THIS PAPER INTERESTING?


   • First Data Ever Published On
     Thermogenesis & Pollination
     In Any Asian Illicium Species



   • TR-71U THERMO RECORDER
PREVIOUS STUDIES
    POLLINATION BIOLOGY
       (5 of 90 Species)

 North American Schisandraceae
               &
      Asian Schisandraceae

            Thien et al., 1983;
          White and Thien, 1985;
           Deringer et al.,1999;
             Liu et al., 2007;
          Yuan et al., 2007, 2008
PREVIOUS EXPERIMENTS SUPPORT:


                          FLOWER HEATING:

               1.) is a direct energy reward for ectothermic pollinators


          2.) increases the volatilization of chemicals directed at pollinators


3.) helps the plant mimic mammalian feces or carrion attracting saprophytic pollinators


 4.) enhances the respiratory release of CO₂, which in combination with other volatile
                        chemicals may stimulate fly oviposition
NORTH AMERICAN
      SPECIES
               (New World Illicium & Schisandra)



-Pollinated predominantly by flies with beetles as co-
                    pollinators

                       Thien et al., 1983;
                     White and Thien, 1985;
                      Deringer et al.,1999;
                         Liu et al., 2007
Illicium floridanum




      Jack Scheper - www.Floridata.com
Illicium parviflorum




        Photo By:
Schisandra glabra
ASIAN SPECIES
                  (Old World Kadsura & Schisandra)




-Exclusively Pollinated by pollen-eating, Megomatta gall midges



                         Yuen et al., 2007, 2008
Kadsura longipedunculata




                           www.plant.ac.cn
Schisandra henryii
                     Photo By:
Schisandra henryii   PHOTO BY:
FLOWER HEATING
     FOLLOWING ANTHESIS
             &
 THE EVOLUTION OF GALL MIDGE
POLLINATION IN SCHISANDRACEAE
POST ANTHESIS FLOWER HEATING



Schisandraceae        Old World Illicium



   EVOLUTION OF GALL MIDGE POLLINATION
OBJECTIVES
QUESTIONS AUTHORS ARE TRYING TO ANSWER

1.) Given that some Schisandraceae exhibit flower heating, do Asian
Illicium species also possess this trait?


2.) Does any flower heating continue after a flower’s sexual function
is over, and if so, what is the adaptive significance of postanthetic
flower heating?


3.) Are Asian Illicium species pollinated by gall midges, or do they
show “generalized” fly and/or beetle pollination similar to New World
Illicium?


4.) Are flower heating and midge pollination functionally correlated?
MATERIALS
STUDY SPECIES 1: Illicium dunnianum
DESCRIPTION: Small shrub (0.5 – 2m high)
HABITAT: Riverbanks in wooded ravines (Elevations 300-750m)
LOCATION: Shiheqiguan - Nan Kun Shan National Forest Park, China
OBSERVED: Mid March to late April – 2008 & 2009
# OBSERVED: 53 individuals total: (38 at site A, 8 at site B, 5 at site C)




STUDY SPECIES 2: Illicium tsangii
DESCRIPTION: Shrub or small tree (up to 10m)
HABITAT: Mixed forests or thickets (Elevations 500-800m)
LOCATION: Along road - Nan Kun Shan National Forest Park,
China
OBSERVED: April to late May – 2008 & 2009
# OBSERVED: 56 individuals total
METHODS
FLORAL DEVELOPMENT, FUNCTION, & TEMPERATURE

             Open Pollinated,                         Male Phase,
             Cross Pollinated,                        Nursing Phase,

             Bagged Pre-Anthesis                      Female Phase

  •
      Relative position & color of tepals, stamens, and styles.
  •
      Presence or absence of secretion.
  •
      Timing of; style movements, anther dehiscence, and floral organ
      wilting/abscission.
  •
      Ambient and floral temperatures in open-pollinated flowers
      recorded for 50 h) using TR-71U thermo recorder (AWESOME!)
      Readings taken every 5 sec. (male phase, nursing phase, female phase)
  •
      One-way ANOVA F-tests and t-tests – using statistical package SPSS
  •
      G-test- using Microsoft Excel Poptools 3.0
Plant Reproductive Biology
                   -Mating Systems-


1.) Randomly selected flowers and marked as controls


2.) Flowers self pollinated from male-phase flower then bagged


3.) Flowers bagged to test for agamospermy


4.) Flowers cross pollinated then bagged
VISITORS
&
POLLINATORS


Diurnal & nocturnal observations made over 150
                     hours:
              -Recorded Kinds & Numbers of Visitors
                         -Duration of Visits
                          -Insect Behavior



    To investigate flowers functional phases
     •
         Visitor behavior was monitored inside flowers
     •
         Flowers were bagged at end of female phase
     •
         Flowers were bagged at end of male phase
     •
         Tepal tips were trimmed in male phase flowers
     •
         Tepal tips were trimmed at beginning of nursing phase
Molecular Phylogenetics
                                      &
            Ancestral Trait Reconstruction
•
    Sequenced complete internal transcribed spacer (ITS) of ribosomal DNA


•
    Sequenced part of the chloroplast trnL region


•
    Data set comprised 43 species – 2197 aligned nucleotides


•
    Performed Maximum Likelihood analysis under GTR + G model of
    substitution


•
    Bootstrap support values were estimated w/ 100 replicate heuristic searches
RESULTS
♦   1st night = Female flowers w/ fully receptive stigmas


♦   1st day = interim phase (female to male)


♦   2nd night = flowers entered male phase –last 2-3 nights (anther dehiscence)


♦   Day 3-7= Nursing phase begins
          Inner tepals close, forming a chamber,
          Adaxial filament surfaces and inner tepal bases produce a secretion,
          Filaments increased in diameter in control from female to nursing
           phase
          Filaments thickened from female to male and then wilted by nursing.
FEMALE PHASE
                 (Illicium dunnianum)
   Pollination occurred mostly at night


   Single midges entered same flower numerous times


   Only one midge at a time – if occupied other midges would fly away


   Midge eggs became visible & pollen grains appeared on stigmas after visits


   Single midges deposited 26 (+/- 3) pollen grains


   Captured midges carried numerous pollen grains on body


   New species of Clinodiplosis was likely discovered
MALE PHASE
                   (Illicium dunnianum)
   When anthers dehisced midges again visited to oviposit


   20 eggs were found per flower in the male phase, but flowers with trimmed tepals
    caused eggs to die


   The highest number of larvae/ flower were found in the bagged male-phase.


   Highest number of eggs was found in male phase flowers


   High temperatures beginning of female phase then dropped to ambient during male


   Produced little heat during day and throughout male stages
NURSING PHASE
                       (Illicium dunnianum)

   Onset
of
nursing
phase

chamber
temp.
increased
to
higher
than
in
female
phase



   Midges
did
not
try
to
enter
nursing
phase
flowers


   Larvae
in
nursing
phase
flowers
were
seen
feeding
on
filament
and
tepal
secre=on.


OBJECTIVES
1.   Asian
Illicium
species
do
possess
postanthe:c
flower
hea:ng.


2.   The
key
stage
of
thermal
warming
was
well
aEer
the
sexual
func:on
was
over
in
the
two

     studied
species.

The
only
observable
&
supported
benefit
of
post‐anthe:c
hea:ng
is

     exclusive
to
the
larvae
of
the
pollinator.

The
increase
in
temperature
provides
warm
enough

     temperatures
for
larvae
to
survive
and
the
heat
also
causes
filament
and
tepal
secre:on

     which
the
larvae
feed
on.

The
hea:ng
seems
to
be
a
reward
for
the
pollinator
in
a
plant‐
     pollinator
mutualis:c
rela:onship.



3.   In
contrast
to
the
New
World
Illicium,
the
two
study
species
Illicium
dunnianum
and
I.
tsangii

                           were
pollinated
only
by
Clinodiplosis
gall
midges.



4.   Despite
these
findings,
comparison
of
the
ancestral
state
reconstruc:ons
for
two
traits

     showed
that
flower
hea:ng
and
gall
midge
pollina:on
are
not
strictly
correlated.


DISCUSSION
‐Flower
hea:ng
evolved
during
the
evolu:on
of
Schisandraceae
and
became
co
opted
in
flowers

pollinated
by
flower
breeding
midges?


‐Floral
hea:ng
helps
odor
emission??


‐Flower
hea:ng
aids
in
pollen
tube
growth??


‐Obligate
outcrossing
may
be
enforced
by
self
incompatability??


‐Selec:ve
factors
behind
the
evolu:on
of
flower
hea:ng??


‐Are
midges
simply
co‐pollinators??
Thank You
For Your Patience
SOURCES

•   http://www.efloras.org/florataxon.aspx?flora_id=610&taxon_id=242326638

•

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Schisandraceae

  • 1. FLOWER HEATING FOLLOWING ANTHESIS & THE EVOLUTION OF GALL MIDGE POLLINATION IN SCHISANDRACEAE AMERICAN JOURNAL OF BOTANY -JULY 2010-
  • 2. Shi Xiao Luo & Dianxiang Zhan South China Botanical Garden, Guangzhou, China Shu-Miaw Chaw Biodiversity Research Center Taipei, Taiwan Susanne S. Renner Department of Biology, University of Munich Munich, Germany
  • 3. KEY TERMS • Agamospermy -the asexual formation of embryos and seeds without the occurrence of fertilization. • Anthesis - the period during which a flower is fully open and functional. • Oviposition - to deposit or lay eggs. • Saprophytic - organisms that feed on dead or decaying matter.
  • 4. •Background Information • Objectives • Materials • Methods • Results • Discussion
  • 6. STAR VINE FAMILY ~90 SPECIES Schisandraceae Kadsura (22 species) - Endemic to Tropical Asia Schisandra (25 species) - Tropical Asia- (Exception - S. glabra) Illicium (42 species) - SE Asia, SE U.S., Mexico, Greater Antilles
  • 7. WHY IS THIS PAPER INTERESTING? • First Data Ever Published On Thermogenesis & Pollination In Any Asian Illicium Species • TR-71U THERMO RECORDER
  • 8. PREVIOUS STUDIES POLLINATION BIOLOGY (5 of 90 Species) North American Schisandraceae & Asian Schisandraceae Thien et al., 1983; White and Thien, 1985; Deringer et al.,1999; Liu et al., 2007; Yuan et al., 2007, 2008
  • 9. PREVIOUS EXPERIMENTS SUPPORT: FLOWER HEATING: 1.) is a direct energy reward for ectothermic pollinators 2.) increases the volatilization of chemicals directed at pollinators 3.) helps the plant mimic mammalian feces or carrion attracting saprophytic pollinators 4.) enhances the respiratory release of CO₂, which in combination with other volatile chemicals may stimulate fly oviposition
  • 10. NORTH AMERICAN SPECIES (New World Illicium & Schisandra) -Pollinated predominantly by flies with beetles as co- pollinators Thien et al., 1983; White and Thien, 1985; Deringer et al.,1999; Liu et al., 2007
  • 11. Illicium floridanum Jack Scheper - www.Floridata.com
  • 12. Illicium parviflorum Photo By:
  • 14. ASIAN SPECIES (Old World Kadsura & Schisandra) -Exclusively Pollinated by pollen-eating, Megomatta gall midges Yuen et al., 2007, 2008
  • 15. Kadsura longipedunculata www.plant.ac.cn
  • 16. Schisandra henryii Photo By:
  • 17. Schisandra henryii PHOTO BY:
  • 18. FLOWER HEATING FOLLOWING ANTHESIS & THE EVOLUTION OF GALL MIDGE POLLINATION IN SCHISANDRACEAE
  • 19. POST ANTHESIS FLOWER HEATING Schisandraceae Old World Illicium EVOLUTION OF GALL MIDGE POLLINATION
  • 20. OBJECTIVES QUESTIONS AUTHORS ARE TRYING TO ANSWER 1.) Given that some Schisandraceae exhibit flower heating, do Asian Illicium species also possess this trait? 2.) Does any flower heating continue after a flower’s sexual function is over, and if so, what is the adaptive significance of postanthetic flower heating? 3.) Are Asian Illicium species pollinated by gall midges, or do they show “generalized” fly and/or beetle pollination similar to New World Illicium? 4.) Are flower heating and midge pollination functionally correlated?
  • 22. STUDY SPECIES 1: Illicium dunnianum DESCRIPTION: Small shrub (0.5 – 2m high) HABITAT: Riverbanks in wooded ravines (Elevations 300-750m) LOCATION: Shiheqiguan - Nan Kun Shan National Forest Park, China OBSERVED: Mid March to late April – 2008 & 2009 # OBSERVED: 53 individuals total: (38 at site A, 8 at site B, 5 at site C) STUDY SPECIES 2: Illicium tsangii DESCRIPTION: Shrub or small tree (up to 10m) HABITAT: Mixed forests or thickets (Elevations 500-800m) LOCATION: Along road - Nan Kun Shan National Forest Park, China OBSERVED: April to late May – 2008 & 2009 # OBSERVED: 56 individuals total
  • 24. FLORAL DEVELOPMENT, FUNCTION, & TEMPERATURE Open Pollinated, Male Phase, Cross Pollinated, Nursing Phase, Bagged Pre-Anthesis Female Phase • Relative position & color of tepals, stamens, and styles. • Presence or absence of secretion. • Timing of; style movements, anther dehiscence, and floral organ wilting/abscission. • Ambient and floral temperatures in open-pollinated flowers recorded for 50 h) using TR-71U thermo recorder (AWESOME!) Readings taken every 5 sec. (male phase, nursing phase, female phase) • One-way ANOVA F-tests and t-tests – using statistical package SPSS • G-test- using Microsoft Excel Poptools 3.0
  • 25. Plant Reproductive Biology -Mating Systems- 1.) Randomly selected flowers and marked as controls 2.) Flowers self pollinated from male-phase flower then bagged 3.) Flowers bagged to test for agamospermy 4.) Flowers cross pollinated then bagged
  • 26. VISITORS
&
POLLINATORS Diurnal & nocturnal observations made over 150 hours: -Recorded Kinds & Numbers of Visitors -Duration of Visits -Insect Behavior To investigate flowers functional phases • Visitor behavior was monitored inside flowers • Flowers were bagged at end of female phase • Flowers were bagged at end of male phase • Tepal tips were trimmed in male phase flowers • Tepal tips were trimmed at beginning of nursing phase
  • 27. Molecular Phylogenetics & Ancestral Trait Reconstruction • Sequenced complete internal transcribed spacer (ITS) of ribosomal DNA • Sequenced part of the chloroplast trnL region • Data set comprised 43 species – 2197 aligned nucleotides • Performed Maximum Likelihood analysis under GTR + G model of substitution • Bootstrap support values were estimated w/ 100 replicate heuristic searches
  • 28. RESULTS ♦ 1st night = Female flowers w/ fully receptive stigmas ♦ 1st day = interim phase (female to male) ♦ 2nd night = flowers entered male phase –last 2-3 nights (anther dehiscence) ♦ Day 3-7= Nursing phase begins  Inner tepals close, forming a chamber,  Adaxial filament surfaces and inner tepal bases produce a secretion,  Filaments increased in diameter in control from female to nursing phase  Filaments thickened from female to male and then wilted by nursing.
  • 29. FEMALE PHASE (Illicium dunnianum)  Pollination occurred mostly at night  Single midges entered same flower numerous times  Only one midge at a time – if occupied other midges would fly away  Midge eggs became visible & pollen grains appeared on stigmas after visits  Single midges deposited 26 (+/- 3) pollen grains  Captured midges carried numerous pollen grains on body  New species of Clinodiplosis was likely discovered
  • 30. MALE PHASE (Illicium dunnianum)  When anthers dehisced midges again visited to oviposit  20 eggs were found per flower in the male phase, but flowers with trimmed tepals caused eggs to die  The highest number of larvae/ flower were found in the bagged male-phase.  Highest number of eggs was found in male phase flowers  High temperatures beginning of female phase then dropped to ambient during male  Produced little heat during day and throughout male stages
  • 31. NURSING PHASE (Illicium dunnianum)  Onset
of
nursing
phase

chamber
temp.
increased
to
higher
than
in
female
phase
  Midges
did
not
try
to
enter
nursing
phase
flowers  Larvae
in
nursing
phase
flowers
were
seen
feeding
on
filament
and
tepal
secre=on.


  • 32. OBJECTIVES 1. Asian
Illicium
species
do
possess
postanthe:c
flower
hea:ng. 2. The
key
stage
of
thermal
warming
was
well
aEer
the
sexual
func:on
was
over
in
the
two
 studied
species.

The
only
observable
&
supported
benefit
of
post‐anthe:c
hea:ng
is
 exclusive
to
the
larvae
of
the
pollinator.

The
increase
in
temperature
provides
warm
enough
 temperatures
for
larvae
to
survive
and
the
heat
also
causes
filament
and
tepal
secre:on
 which
the
larvae
feed
on.

The
hea:ng
seems
to
be
a
reward
for
the
pollinator
in
a
plant‐ pollinator
mutualis:c
rela:onship.
 3. In
contrast
to
the
New
World
Illicium,
the
two
study
species
Illicium
dunnianum
and
I.
tsangii
 were
pollinated
only
by
Clinodiplosis
gall
midges.
 4. Despite
these
findings,
comparison
of
the
ancestral
state
reconstruc:ons
for
two
traits
 showed
that
flower
hea:ng
and
gall
midge
pollina:on
are
not
strictly
correlated.


  • 34. Thank You For Your Patience
  • 35. SOURCES • http://www.efloras.org/florataxon.aspx?flora_id=610&taxon_id=242326638 •

Editor's Notes

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