Seed production trait associations and inheritance …15 Will-2 Ladino-5 x BC 1 T-1 BC 3 F 1 9 Will-2 Tienshan-5 x BC 1 Cu-2 T. uniflorum 3 Az-4437 T. uniflorum 4 Az4383-11-0P T. uniflorum

10.1071/CP17048_AC © CSIRO 2017

Supplementary Material: Crop & Pasture Science, 2017, 885–892.

Seed production trait associations and inheritance in interspecific hybrids between

Trifolium repens (white clover) and Trifolium uniflorum

Muhammad NaeemA,B, I. M. VerryC, P. D. KempA, J. P. MillnerA and W. M. WilliamsA,C,D

ACollege of Sciences, Massey University, Palmerston North 4442, New Zealand.

BCurrent address: Federal Seed Certification and Registration Department, Islamabad, Pakistan.

CAgResearch Grasslands Research Centre, Private Bag 11008, Palmerston North 4442, New

Zealand.

DCorresponding author: Email: [email protected]

Supplementary Table 1: Details of the families. White clover cultivars Aran (A),

Barblanca (B), Crusader (C), Crau (Cu), Sustain (S), and Trophy (T), were crossed

with F1 (T. repens x T. uniflorum) plants to generate the 23 BC1F1 hybrids used as

starting material.

In the pedigree columns, BC1 A-1 to BC1 A-15 represent different BC1F1 plants having

Aran as the recurrent parent. Similarly, BC1 B1-B9, C1-C8, Cu1-Cu4, S1-S19 and T1-T6

represent BC1F1 plants with the designated recurrent parent. Ladino, Durana, Tienshan and

Will were added as later white clover recurrent populations to generate BC2 and BC3

generations and these are designated by name and plant number (e.g. Tienshan-5). The

abbreviation BC2F2-BI show families developed by first backcrossing BC1F1 hybrids to

white clover and then, during the next season the BC2F1 progenies were intercrossed to

produce the BC2F2 generation. Alternatively, BC1F1 hybrids were first intercrossed and

during the next season their BC1F2 progenies were backcrossed to white clover to produce

a BC2 generation derived from BC1F2 plants, here designated BC2F2-IB (although strictly

these were BC2 F(BC1F2):1 populations). Families in bold type were used in assessments for

pollen viability, ovules per ovary and selfed seed setting.

Gen Max

plants

Female parent(s) Male parent(s)

BC1F1 12 Bulk of Aran x F1 (T. repens x T. uniflorum)

BC1F1 9 Bulk of Sustain x F1 (T. repens x T. uniflorum)

BC1F1 5 BC1 S-18 [(Sustain x F1)–plant 18]

BC1F2 15 BC1 S-14 BC1 S-10

BC1F2 12 BC1 B-2 BC1 B-3

BC1F2 BC1 Cu-2 Self-pollinated

BC1F2 BC1 B-3 Self-pollinated

BC1F2 BC1 S-14 Self-pollinated

BC1F3 15 BC1 B-2 x BC1 B-3 BC1 S-10 x BC1 S-1

BC1F3 15 BC1 S-10 x BC1 S-1 BC1 B-2 x BC1 B-3

BC1F3 15 BC1 B-2 x BC1 B-3 BC1 A-11 x BC1 A-6

BC1F3 12 BC1 A-6 x BC1 A-11 BC1 S-14 x BC1 S-10

BC1F3 10 BC1 B-2 x BC1 B-3 BC1 C-8 x BC1 C-6

BC1F3 15 BC1 A-6 x BC1 A-10 BC1 C-5 x BC1 C-8

BC1F3 12 BC1 S-10 x BC1 S-12 BC1 A-6 x BC1 A-10

BC1F3 15 BC1 S-10 x BC1 S-12 BC1 S-1 x BC1 S-10

BC1F3 13 BC1 C-4 x BC1 C-8 BC1 S-12 x BC1 S-10

BC1F3 6 BC1 B-2 x BC1 B-8 BC1 S-14 x BC1 S-10

BC1F3 15 BC1 S-10 x BC1 S-1 BC1 B-2 x BC1 B-8

BC1F3 14 BC1 A-11 x BC1 A-6 BC1 S-14 x BC1 S-10

BC1F3 13 BC1 S-10 x BC1 S-12 BC1 C-5 x BC1 C-8

BC1F3 1 BC1 B-2 x BC1 B-8 BC1 C-8 x BC1 C-6

BC1F3 BC1 B-2 x BC1 B-3 Self-pollinated

BC1F3 BC1 S-14 x BC1 S-10 Self-pollinated

BC2F1 15 Tienshan-5 BC1 Cu-2

BC2F1 15 Aran-28 BC1 Cu-2

BC2F2-BI 15 Ladino-4 x BC1 S-10 Ladino-5 x BC1 T-4

BC2F2-BI 15 Durana-44 x BC1 A-11 Durana-57 x BC1 A-1

BC2F2-BI 8 Ladino-8 x BC1 A-1 Ladino-4 x BC1 A-1

BC2F2-BI 15 Tienshan-5 x BC1 Cu-2 Durana-45 x BC1 A-1

BC2F2-BI 15 Durana-44 x BC1 A-11 Ladino-3 x BC1 Cu-4

BC2F2-BI 15 Tienshan-6 x BC1 Cu-4 Ladino-4 x BC1 S-10

BC2F2-BI 15 Aran-28 x BC1 Cu-4 Ladino-4 x BC1 A-1

BC2F2-BI 15 Durana-57 x BC1 A-1 Tienshan-5 x BC1 Cu-2

BC2F2-BI 7 Ladino-4 x BC1 A-1 Ladino-8 x BC1 A-1

BC2F2-BI 15 Durana-44 x BC1 A-11 Durana-44 x BC1 A-6

BC2F2-BI 15 Ladino-4 x BC1 S-10 Tienshan-13 x BC1 Cu-1

BC2F2-BI 15 Ladino-3 x BC1 T-3 Ladino-5 x BC1 T-1

BC2F2-BI 15 Aran-28 x BC1 Cu-2 Tienshan-4 x BC1 A-1

BC2F2-BI 7 Ladino-3 x BC1 T-1 Durana-21 x BC1 A-10

BC2F2-BI 15 Durana-44 x BC1 A-11 Tienshan-5 x BC1 Cu-2

BC2F2-BI Aran-28 x BC1 Cu-2 Self-pollinated

BC2F2-BI Tienshan-5 x BC1 Cu-2 Self-pollinated

BC2F2-IB 11 Will-2 BC1 C-5 x BC1 C-8

BC2F2-IB 15 Tienshan-13 BC1 S-10 x BC1 S-12

BC2F2-IB 14 Aran-28 BC1 S-10 x BC1 S-12

BC2F2-IB 15 Tienshan-13 BC1 B-2 x BC1 B-3

BC2F2-IB 15 Will-2 BC1 S-10 x BC1 S-12

BC2F2-IB 15 Will-2 BC1 A-15 x BC1 A-6

BC2F2-IB 15 Aran-28 BC1 B-2 x BC1 B-3

BC2F2-IB 5 Aran-802 BC1 B-2 x BC1 B-3


BC2F2-IB 15 Will-2 BC1 B-2 x BC1 B-8



BC2F2-IB 15 Will-2 BC1 S-10 x BC1 S-1

BC2F2-IB 6 Will-2 BC1 C-8 x BC1 C-6

BC2F2-IB 15 Aran-802 BC1 S-10 x BC1 S-12


BC3F1 14 Will-2 Ladino-4 x BC1 S-10

BC3F1 15 Will-2 Ladino-3 x BC1 T-3

BC3F1 15 Aran-28 Ladino-3 x BC1 T-3

BC3F1 15 Will-2 Durana-44 x BC1 A-6


BC3F1 6 Tienshan-13 Durana-44 x BC1 A-11

BC3F1 15 Tienshan-13 Ladino-3 x BC1 T-3


BC3F1 15 Aran-28 Durana-44 x BC1 A-11

BC3F1 13 Aran-802 Durana-44 x BC1 A-11

BC3F1 15 Will-2 Aran-28 x BC1 Cu-2

BC3F1 13 Aran-802 Ladino-3 x BC1 T-3




BC3F1 9 Will-2 Tienshan-5 x BC1 Cu-2

T.

uniflorum

3 Az-4437

T.

uniflorum

4 Az4383-11-0P

T.

uniflorum

4 Az4383-18-OP

T. repens 11 Aran

T. repens 15 Sustain

Supplementary Table 2. Details of families used to estimate heritability. There were 10

comparisons, each with a common female parent crossed with more than one male to

produce both full- and half-sibs.

For each cross there were on average 13.9 full-sibs (n). There were 10 females (N) and, on

average, 2.3 males per female (M).

Comparison

Female

(common parent)

Male

Generation

1

BC1 S-10 x BC1 S-12 BC1 A-6 x BC1 A-10 BC1F3

BC1 S-10 x BC1 S-12 BC1 S-1 x BC1 S-10 BC1F3

2

BC1 S-10 x BC1 S-1 BC1 B-2 x BC1 B-3 BC1F3

BC1 S-10 x BC1 S-1 BC1 B-2 x BC1 B-8 BC1F3

3

BC1 B-2 x BC1 B-3 BC1 S-10 x BC1 S-1 BC1F3

BC1 B-2 x BC1 B-3 BC1 A-11 x BC1 A-6 BC1F3

4

Tienshan-5 x BC1 Cu-2 Tienshan-5 x BC1 Cu-2 BC2F2-BI

Tienshan-5 x BC1 Cu-2 Durana-45 x BC1 A-1 BC2F2-BI

5

Aran-28 x BC1 Cu-2 Aran-28 x BC1 Cu-2 BC2F2-BI

Aran-28 x BC1 Cu-2 Tienshan-4 x BC1 A-1 BC2F2-BI

6

Durana-44 x BC1 A-11 Durana-57 x BC1 A-1 BC2F2-BI

Durana-44 x BC1 A-11 Ladino-3 x BC1 Cu-4 BC2F2-BI

Durana-44 x BC1 A-11 Durana-44 x BC1 A-6 BC2F2-BI

7

Ladino-3 x BC1 T-3 Ladino-5 x BC1 T-1 BC2F2-BI

Ladino-3 x BC1 T-3 Durana-21 x BC1 A-10 BC2F2-BI

8

Aran-802 BC1 B-2 x BC1 B-3 BC2F2-IB

Aran-802 BC1 S-10 x BC1 S-12 BC2F2-IB

Aran-802 Ladino-3 x BC1 T-3 BC3F1

9

BC1 S-14 BC1 S-10 BC1F2

BC1 S-14 BC1 S-14 BC1F2

10

Will-2 BC1 A-11 x BC1 A-6 BC2F2-IB

Will-2 BC1 A-6 x BC1 A-1 BC2F2-IB

Will-2 BC1 S-10 x BC1 S-1 BC2F2-IB

Supplementary Table 3. ANOVA design and estimation of phenotypic variation (σ2p),

additive genetic variation (σ2A) and narrow sense heritability (h2).

Source DF SS Mean Square Expected MS

square

σ2p σ2

A h2

Replication r-1

Female N-1 = dff SSf SSf / dff = MSf σ2e + nσ2

m + Mnσ2f

σ2e + σ2

m + σ2f σ2

m x 4 σ2A / σ2

p Males (female) N(M-1) = dfm SSm SSm / dfm = MSm σ2e + nσ2

m

Error MNn - MN = dfe SSe SSe / dfe = MSe σ2e

Total MNn-1 = dft

r, N, M and n represent numbers of replications, females, males/female and progenies per male, respectively.

σ2e, σ2

m and σ2f represent variance components of error, male (female) and female parent, respectively.

Supplementary Table 4. Detailed results of ANOVA and estimations of the variance

components (σ2e, σ2

m and σ2f) and phenotypic variation (σ2

p), additive genetic variation

(σ2A) and narrow sense heritability (h2) calculated from transformed data of selected

traits.

Trait

Source DF SS Mean Square σ2§ σ2

p σ2A h2

Seeds/floret

Replication 2 3.34 1.67

Female 9 5.35 0.59 0.01

0.09 0.03 0.35 Males (female) 13 4.23 0.33 0.02

Error 294 19.97 0.07 0.07

Florets/head


Female 9 283.91 31.55 0.63

2.71 2.52 0.93 Males (female) 13 149.03 11.46 0.73

Error 294 397.19 1.35 1.35

Seeds/head


Female 9 169.47 18.83 0.25

2.50 1.01 0.40 Males (female) 13 140.05 10.77 0.66

Error 294 466.24 1.59 1.59

Peduncle

length

Replication 2 61666.71 30833.36

Female 9 205778.98 22864.33 98.95

3098.00 395.79 0.13 Males (female) 13 256202.70 19707.90 1298.32

Error 294 500013.68 1700.73 1700.73

Heads/plant


Female 9 1029.47 114.39 2.10

15.32 8.39 0.55 Males (female) 13 617.25 47.48 2.66

Error 292 3083.23 10.56 10.56

Florets/plant

Replication 2 1758.86 879.43

Female 9 52886.80 5876.31 81.72

1122.21 326.87 0.29 Males (female) 13 42503.30 3269.48 173.20

Error 292 253249.27 867.29 867.29

Seeds/plant


Female 9 426.71 47.41 0.69

8.82 2.74 0.31 Males (female) 13 332.17 25.55 1.35

Error 292 1978.97 6.78 6.78

Foliage dry

weight


Female 9 913.71 101.52 0.45

17.79 1.78 0.10 Males (female) 13 1134.87 87.30 5.44

Error 292 3477.86 11.91 11.91

§ σ2f , σ2

m and σ2e ,respectively (supplementary Table 3).

Supplementary Table 5. PCA results for reproductive traits based on 1008 complete

observations with three retained factors rotated with the varimax procedure.

Coefficients of the eigenvalues show relationship among traits.

Variable Factor1 Factor2 Factor3

Eigenvalue 3.41 2.1 1.46

Variance proportion 0.43 0.26 0.18

Seeds/floret 3 93* -14

Florets/head 2 57* 61*

Seeds/head 3 98* 15

Heads/plant 97* -12 -15

Florets/plant 94* 14 14

Seeds/plant 72* 67* 2

Peduncle length -3 -7 90*

Foliage dry weight 61* 2 59*

*values greater than 40.

Supplementary Figure 1. Path diagrams with coefficients and residuals (e), showing

interrelationships among (a) major vegetative and reproductive traits and (b) major

floral traits affecting seed production in white clover x T. uniflorum hybrids

a.

Seeds/plant

Heads/plant

0.64

0.69

0.02

e=0.89

e=0.23

Foliage dry weight

Peduncle length

Seeds/floret

Ovules/ovarySeed weight Seeds/head

0.79

0.43

-0.38

0.14

0.22

0.12

e=0.96 e=0.93

e=0.95

e=0.11

e=0.91

0.33

Florets/head

Leaf area

0.42

0.41

e=0.82

e=0.91

0.57

0.45

b.

Seeds/plant

Seeds/floret

Florets/head

Seeds/head

Heads/plant

0.69

0.75

0.79

0.43

e=0.11

e=0.25

Documents

Seed production trait associations and inheritance …15 Will-2 Ladino-5 x BC 1 T-1 BC 3 F 1 9 Will-2 Tienshan-5 x BC 1 Cu-2 T. uniflorum 3 Az-4437 T. uniflorum 4 Az4383-11-0P T. uniflorum