Pollen dna content and flow cytometry: improved methodology and use




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Pollen DNA content and flow cytometry: improved methodology and use


Table S1: Species tested using the filter bursting method, with maximum measured pollen diameters (long axis), filter sizes and trial numbers.


Family

 Quality

Genus

 Quality

Species

Quality

Diameter (µm)

PF

BF

Trials

Alstroemeriaceae

F

Alstroemeria

F

A. cultivar

F

138

150

50

6

Amaranthaceae

CC

Chenopodium

CC

C. album

CC

30

100

10

1

Amaryllidaceae

CC+

Allium

GS

A. schoenoprasum

GS

30

100

10

3

 




Galanthus

CC+

G. nivalis

CC+

27

150

10

3

Apiaceae

CC

Daucus

CC

D. carota

CC

27

50

10

2

Asteraceae

CC+

Achillea

CC

A. millefolium

CC

33

100

30

2*

 




Ambrosia

CC+

A. trifida

CC+

24

50

10

1*

 




Ambrosia




A. artemisifolia

CC+

22

50

10

2*







Artemisia

CC+

A. biennis

F1

n.m.

501

10

1

 




Artemisia




A. vulgaris

CC+

26

50

10

3

 




Centaurea

GS

C. nigra

GS

51

100

30

1

 




Chrysantheum

CC

C. leucanthemum

CC

33

100

10

1

 




Eupatorium

CC+

E. perfoliatum

CC+

26

50

20

4*

 




Eutrochium

CC+

E. purpureum

CC+

31

50

20

1*

 




Eutrochium




E. maculatum

CC+

29

50

20

2*

 




Rudbeckia

CC+

R. hirta

CC+

29

50

20

1*

 




Symphyotrichum

F1

S. novae-angliae

F1

28

100

20

1

 




Taraxacum

F

T. officinale

F

56

150

50

4

 




Tragopogon

GS

T. pratensis

GS

48

100

10

6

Betulaceae

CC+

Betula

CC+

B. papyrifera

CC+

41

50

10

1*

Bignoniaceae

GS

Campsis

GS

C. radicans

GS

42

100

20

2

Brassicaceae

CC+

Arabidopsis

F

A. thaliana

F

36

100

10

2

 




Brassica

CC+

B. napus

CC+

39

50

10

3*

 




Leavenworthia

GS

L. alabamica

GS

n.m.

1001

10

1

 




Thlaspi

CC+

T. arvense

CC+

30

100

10

2

 




Raphanus

CC+

R. sativus

CC+

36

50

10

2

Campanulaceae

GS

Campanula

GS

C. rotundifolia

GS

n.m.

150

10

4

Caprifoliaceae

GS

Lonicera

GS

L. tatarica

GS

68

150

10

1

Caryophyllaceae

F1

Lychnis

F1

L. noctiflora

F1

56

150

10

1

Cucurbitaceae

F1

Cucurbita

F1

C. pepo

F1

201

unf

20

1

Fabaceae

CC+

Lotus

CC+

L. corniculata

CC+

28

50

10

5*

 




Trifolium

GS

T. repens

GS

39

100

10

1

 




Vicia

CC+

V. faba

CC+

55

100

20

4*

 




Vicia




V. villosa

F1

n.m.

50

10

1

Gesneriaceae

F

Aeschynanthus

F

A. lobbianus

F

27

50

10

10

Iridaceae

F

Crocus

F

C. sativus

F

140

150

10

7

 




Iris

F1

I. reticulata

F1

161

unf

10

1

Lamiaceae

CC+

Agastache

GS

A. scrophulariifolia

GS

44

100

20

1

 




Lamium

CC+

L. maculatum

CC+

37

100

20

4

Liliaceae

CC+

Hemerocallis

F1

H. fulva

F1

144

unf

30

1

 




Lilium

CC+

L. ‘Asiatic cultivar’

F

114

unf

30

22







Lilium




L. longiflorum

CC+

135

150

30

8

Oleaceae

CC+

Fraxinus

CC+

F. americana

CC+

36

50

10

1*

Onagraceae

GS

Chamerion

GS

C. angustifolium

GS

103

150

10

22

Plantaginaceae

CC+

Plantago

CC+

P. lanceolata

CC+

38

50

10

10*

 




Plantago




P. major

CC+

28

50

10

10*

 




Veronica

GS

V. gentianoides

GS

n.m.

150

10

1

 




Veronica




V. vindobonensis

GS

50

100

10

1

Poaceae

CC+

Alopecurus

CC+

A. pratensis

CC+

43

100

10

2*

 




Elymus

CC

E. canadensis

CC

48

100

20

10

 




Elymus




E. hystrix

CC

49

100

20

12

 




Phalaris

GS

P. arundinacea

GS

43

100

30

1

 




Zea

CC

Z. mays

CC

102

150

30

2*

Polygonaceae

CC+

Rumex

CC+

R. nivalis

CC+

27

50

20

3*

Ranunculaceae

CC+

Caltha

CC+

C. palustris

CC+

32

100

10

4*

 




Ranunculus

GS

R. repens

GS

n.m.

150

10

1

Rosaceae

CC+

Amelanchier

CC+

A. arborea

CC+

37

100

10

4

 




Amelanchier




A. bartmanii

GS

48

100

10

1

 




Crataegus

GS

C. macrosperma

GS

66

150

10

1

 




Fragaria

CC+

F. vesca

CC+

23

50

10

3*

 




Malus

CC+

M. coronaria

CC+

57

100

10

7

 




Malus




M. domestica

GS

46

100

10

1*

 




Malus




M. baccata cultivar

GS

145

150

10

6*

 




Prunus

GS

P. nigra

GS

52

100

10

1

 




Rosa

GS

R. rugosa

GS

n.m.

150

10

1

Rubiaceae

CC

Galium

CC

G. mullugo

F1

n.m.

1501

10

1







Galium




G. verum

CC

n.m.

150

10

1

Salicaceae

CC+

Salix

CC+

S. babylonica

GS

29

100

10

4







Salix




S. discolor

CC+

27

100

10

2

Sapindaceae

CC+

Acer

CC+

A. saccharum

CC+

38

100

10

1

Scrophulariaceae

CC+

Buddleia

CC+

B. davidii

CC+

26

50

10

2*

Solanaceae

CC+

Lycium

GS

L. barbarum

GS

n.m.

1001

10

1

 




Solanum

CC+

S. dulcamara

CC+

19

50

10

4

 




Solanum




S. lycopersicum

CC+

29

50

10

1*

Strelitziaceae

F

Strelitzia

F

S. reginae

F

219

unf

30

2

Typhaceae

CC+

Typha

CC+

T. angustifolia

CC+

49

100

10

1

 




Typha




T. latifolia

CC+

63

100

10

1

Urticaceae

GS

Laportea

GS

L. canadensis

GS

n.m.

1001

10

1

Verbenaceae

CC+

Verbena

CC+

V. hastata

GS

41

100

20

18

 




Verbena




V. officinalis

CC+

36

100

20

5

"Quality" following a taxonomic level indicates the class assigned to the best genus, species, or individual within that taxon;



GS = genome size quality; CC = cell cycle quality; CC+ = cell cycle quality with 10,000 pollen grains; F = failed to meet minimum GS criteria; F1 = F with only one sample tested (See Materials and Methods). PF = pre-filter; BF = bursting filter; "Trial" is the number of samples tested for this survey, but when additional samples were run for other purposes, *this number is the number of samples run to first achieve the level of quality shown

1size not recorded, most likely sizes shown; n.m. = not measured microscopically; unf = unfiltered

Table S2: Protocol details for genome size tests (GS) and for comparison of three extraction methods (MC)





Filter sizes (µm)

Basic tissue amount for GS tests

Freeze time (h)

Run times (min)

Species

Pre-

Bursting

Pollen

Test plant leaf (cm2)

Standard leaf1 (cm2)

MC

MC

GS

B. napus

50

10

18 anthers

1

0.5

2.75-3

5

2.5-5

E. perfoliatum

50

20

~30 flowers

1

0.25

2.75

8

2.5-5.55

Z. mays

150

30

10 cm inflorescence

1.5

0.25

not tested

n. t.

2.5-7.5

B. davidii

50

10

20 flowers

0.25

0.375

3.25

4

3-14

S. lycopersicum

50

10

1 anther tube

0.5

0.5

2

3

2-6

V. faba

100

20

3 flowers

0.25

0.75

2

6

4-10


1Standard species are listed in Table 4


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