seed production trait associations and inheritance …15 will-2 ladino-5 x bc 1 t-1 bc 3 f 1 9...
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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 A-11 x BC1 A-6
BC2F2-IB 15 Will-2 BC1 B-2 x BC1 B-8
BC2F2-IB 15 Will-2 BC1 A-6 x BC1 A-11
BC2F2-IB 15 Will-2 BC1 A-6 x BC1 A-1
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
BC2F2-IB 13 Will-2 BC1 A-6 x BC1 A-10
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 15 Will-2 Durana-44 x BC1 A-11
BC3F1 6 Tienshan-13 Durana-44 x BC1 A-11
BC3F1 15 Tienshan-13 Ladino-3 x BC1 T-3
BC3F1 15 Will-2 Ladino-5 x BC1 T-4
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 15 Will-2 Durana-57 x BC1 A-1
BC3F1 15 Will-2 Durana-21 x BC1 A-10
BC3F1 15 Will-2 Ladino-5 x BC1 T-1
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
Replication 2 14.54 7.27
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
Replication 2 83.53 41.77
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
Replication 2 30.47 15.24
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
Replication 2 136.94 68.47
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
Replication 2 107.24 53.62
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