Supplementary information Strong phylogenetic signals and phylogenetic niche conservatism in ecophysiological traits across divergent lineages of Magnoliaceae

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Supplementary information
Strong phylogenetic signals and phylogenetic niche conservatism in ecophysiological traits across divergent lineages of Magnoliaceae

Hui Liu, Qiuyuan Xu, Pengcheng He, Louis S. Santiago, Keming Yang and Qing Ye

Table S1. Species list with subgenus, section, life forms and accession numbers of DNA sequences from GenBank. The taxonomic system of Magnoliaceae follows Figlar & Nooteboom (2004).

Species	Subgenus	Section	Growth form	Leaf form	matK	trnH
Liriodendron chinense (Hemsl.) Sargent.	-	-	Tree	Deciduous	JN050093	AB021046
Magnolia championii (Bentham) N. H. Xia & C. Y. Wu	Magnolia	Gwillimia	Shrub	Evergreen	JN050033	JN050197
Magnolia coco (Loureiro) N. H. Xia & C. Y. Wu	Magnolia	Gwillimia	Shrub	Evergreen	JN050035	AB021034
Magnolia henryi (Dunn) N. H. Xia & C. Y. Wu	Magnolia	Gwillimia	Tree	Evergreen	JN050050	AY009027
Magnolia paenetalauma Dandy	Magnolia	Gwillimia	Tree	Evergreen	this study	this study
Magnolia decidua Q. Y. Zheng	Magnolia	Manglietia	Tree	Deciduous	AB055542	AB055565
Magnolia fordiana (Oliver) Hu var. forrestii (W. W. Smith ex Dandy) V. S. Kumar	Magnolia	Manglietia	Tree	Evergreen	this study	this study
Magnolia grandis (Hu & W. C. Cheng) V. S. Kumar	Magnolia	Manglietia	Tree	Evergreen	JN050048	AY009022
Magnolia fordiana var. hainanensis (Dandy) Nooteboom, comb. et stat. nov.	Magnolia	Manglietia	Tree	Evergreen	AF123478	-
Magnolia lucida (B. L. Chen & S. C. Yang) V. S. Kumar	Magnolia	Manglietia	Tree	Evergreen	this study	this study
Magnolia megaphylla (Hu & W. C. Cheng) V. S. Kumar	Magnolia	Manglietia	Tree	Evergreen	this study	this study
Magnolia kwangtungensis Merrill	Magnolia	Manglietia	Tree	Evergreen	JN050052	JN050170
Magnolia fordiana (Oliver) Hu	Magnolia	Manglietia	Tree	Evergreen	this study	this study
Magnolia baillonii Pierre	Yulania	Michelia	Tree	Evergreen	-	AY009017
Magnolia cavaleriei (Finet & Gagnepain) Figlar	Yulania	Michelia	Tree	Evergreen	JN050073	AY009009
Magnolia chapensis (Dandy) Sima	Yulania	Michelia	Tree	Evergreen	JN050074	JN050193
Magnolia figo var. crassipes (Y. W. Law) Figlar & Nooteboom	Yulania	Michelia	Shrub	Evergreen	JN050096	JN050212
Magnolia figo (Loureiro) Candolle	Yulania	Michelia	Shrub	Evergreen	JN050021	AB021045
Magnolia foveolata (Merrill ex Dandy) Figlar	Yulania	Michelia	Tree	Evergreen	JN050070	AB623298
Magnolia maudiae (Dunn) Figlar	Yulania	Michelia	Tree	Evergreen	JN050083	HQ415444
Magnolia odora (Chun) Figlar & Nooteboom	Yulania	Michelia	Tree	Evergreen	JN050056	AY009013
Magnolia shiluensis (Chun & Y. F. Wu) Figlar	Yulania	Michelia	Tree	Evergreen	JN050084	JN050202
Magnolia biondii Pampanini	Yulania	Yulania	Tree	Deciduous	JN050031	-
Magnolia cylindrica E. H. Wilson	Yulania	Yulania	Tree	Deciduous	JN050067	AY009020
Magnolia denudata Desrousseaux in Lamarck	Yulania	Yulania	Tree	Deciduous	JN050037	AB021037
Magnolia glabrata Law et R.Z.Zhou ined.	Yulania	Yulania	Tree	Deciduous	this study	this study
Magnolia jigongshanensis T. B. Chao, D.L. Fu & W.B. Sun	Yulania	Yulania	Shrub	Deciduous	this study	this study
Magnolia liliiflora Desrousseaux in Lamarck	Yulania	Yulania	Shrub	Deciduous	JN050098	JN050214

Table S2. Primers used for the amplifications of matK and trnH genes in this study.

Primer	Sequence
matK_for	5’-CGTACAGTACTTTTGTGTTTACGAG-3’
matK_rev	5’-ACCCAGTCCATCTGGAAATCTTGGTTC-3’
trnH_for	5’-CGCGCATGGTGGATTCACAATCC-3’
trnH_rev	5’-GTTATGCATGAACGTAATGCTC-3’

Table S3. Values and multiple comparison results of plant traits and environmental variables of the four main sections in Magnoliaceae. Data are mean±s.e.m. for species within each section (species numbers are given in brackets). There are 3~5 replicates for each species as described in Methods, and then mean values for each species are calculated, which are used for the multiple comparisons. Superscript letters are results of Tukey HSD. Lowest values with significant results in multiple comparative tests are in bold. See abbreviations in Table 1.

Section	Michelia (9)	Yulania (6)	Manglietia (8)	Gwillimia (4)
Height (m)	9.9 ± 1.59 ^ab	5.2 ± 0.91 ^a	11.7 ± 1.02 ^b	4.6 ± 0.77 ^a
DBH (cm)	22.4 ± 3.62 ^b	12.2 ± 1.74 ^ab	23.1 ± 2.70 ^b	6.2 ± 2.11 ^a
WD (g cm^–3)	0.5 ± 0.02 ^b	0.5 ± 0.01 ^b	0.4 ± 0.01 ^a	0.6 ± 0.01 ^c
A_L/A_S (m² cm^–2)	0.9 ± 0.11 ^b	0.4 ± 0.08 ^a	0.8 ± 0.05 ^b	0.9 ± 0.12 ^b
K_S (kg m^–1 s^–1MPa^–1)	1.7 ± 0.29 ^ab	2.3 ± 0.18 ^bc	2.8 ± 0.23 ^c	0.9 ± 0.20 ^a
K_L×10^–4(kg m^–1 s^–1MPa^–1)	2.0 ± 0.30 ^ab	8.0 ± 1.42 ^c	3.9 ± 0.49 ^b	1.0 ± 0.22 ^a
SLA (cm² g^–1)	126.8 ± 10.42 ^a	192.9 ± 20.41 ^b	135.9 ± 11.99 ^a	94.1 ± 8.65 ^a
DMC (%)	34.6 ± 1.76 ^b	27.5 ± 1.40 ^a	28.1 ± 0.92 ^a	43.0 ± 2.19 ^c
SPI (%)	22.8 ± 1.37 ^a	28.6 ± 1.83 ^b	21.5 ± 1.06 ^a	16.4 ± 3.02 ^a
Ψ_tlp (MPa)	-1.3 ± 0.06 ^b	-1.4 ± 0.09 ^ab	-1.2 ± 0.13 ^b	-1.9 ± 0.20 ^a
A_area (µmol m^–2s^–1)	8.7 ± 0.57 ^b	8.0 ± 0.73 ^ab	9.3 ± 0.53 ^b	5.6 ± 1.13 ^a
A_mass (nmol g^-1s^-1)	109.1 ± 10.13 ^b	150.8 ± 14.65 ^b	127.0 ± 14.64 ^b	54.1 ± 13.14 ^a
g_s (mol m^–2s^–1)	0.14 ± 0.01 ^b	0.17 ± 0.03 ^b	0.17 ± 0.01 ^b	0.07 ± 0.01 ^a
E (mmol m^–2s^–1)	2.9 ± 0.31 ^a	2.9 ± 0.44 ^a	3.2 ± 0.19 ^a	2.0 ± 0.11 ^a
WUE_i (µmol mol^–1)	67.8 ± 8.01 ^a	51.4 ± 6.38 ^a	57.2 ± 5.54 ^a	78.2 ± 20.94 ^a
Leaf N (%)	1.7 ± 0.09 ^ab	2.1 ± 0.06 ^c	2.0 ± 0.05 ^bc	1.5 ± 0.14 ^a
Leaf P (%)	0.10 ± 0.01 ^a	0.15 ± 0.01 ^b	0.10 ± 0.01 ^a	0.10 ± 0.02 ^a
Leaf N/P	18.4 ± 0.89 ^ab	14.9 ± 1.12 ^a	19.9 ± 1.13 ^b	15.0 ± 1.38 ^a
PNUE (µmol mol^–1s^–1)	87.9 ± 7.65 ^b	99.5 ± 8.54 ^b	91.0 ± 10.13 ^b	48.8 ± 8.42 ^a
PPUE (mmol mol^–1s^–1)	3.6 ± 0.38 ^b	3.3 ± 0.45 ^ab	4.0 ± 0.47 ^b	1.6 ± 0.37 ^a
MAT_mean(°C)	17.2 ± 1.03 ^ab	14.0 ± 0.95 ^a	17.8 ± 0.88 ^b	19.5 ± 1.27 ^b
MAT_min(°C)	12.3 ± 1.33 ^a	8.8 ± 1.46 ^a	14.1 ± 1.37 ^a	15.2 ± 2.26 ^a
MAT_max (°C)	21.6 ± 0.92 ^ab	18.7 ± 0.72 ^a	21.0 ± 0.91 ^ab	23.7 ± 0.97 ^b
MAT_range (°C)	9.2 ± 1.16 ^a	9.9 ± 2.0 ^a	7.0 ± 1.42 ^a	8.5 ± 1.93 ^a
MAP_mean (mm)	1457 ± 53 ^b	1193 ± 62 ^a	1422 ± 68 ^ab	1494 ± 90 ^b
MAP_min (mm)	1208 ± 69 ^b	881 ± 81 ^a	1231 ± 91 ^b	1269 ± 119 ^b
MAP_max (mm)	1755 ± 47 ^a	1555 ± 96 ^a	1653 ± 56 ^a	1778 ± 119 ^a
MAP_range (mm)	547 ± 82 ^a	674 ± 121 ^a	422 ± 88 ^a	509 ± 133 ^a

Table S4. Phylogenetic principal component analysis (PPCA) for the first two principal components based on 20 plant traits of 27 Magnoliaceae species. PC loadings and the percentage of variance explained by the first two PCs are reported. For each PC, the first six variables with highest loadings are in bold. See trait abbreviations in Table 1.

	PC1	PC2
A_mass	-0.90	0.39
PNUE	-0.88	0.11
WD	0.78	0.23
PPUE	-0.78	-0.03
A_area	-0.66	0.25
LDMC	0.63	-0.33
WUE_i	-0.02	0.87
K_S	-0.34	-0.71
Leaf N	-0.37	0.68
g_s	-0.53	-0.62
SPI	-0.40	0.57
Leaf P	-0.30	0.52
Total variance %	29%	20%
Cumulative variance %	29%	49%

Figure S1. Histograms of (a) Blomberg’s K and (b) Pagel’s λ values from 300 phylogenetic trees for indices in Table 1. K values are between 0 to 0.8, λ values are between 0 to 1.

(a)

(b)

Figure S2. Range of mean annual temperature (MAT, black bars) and precipitation (MAP, grey bars) for each species of the four Magnoliaceae sections in their naturally distributed areas in China. Horizontal lines are the means (solid lines) and standard deviations (dashed lines) for MAT (black lines) and MAP (grey lines) in the study site, respectively.

Supplementary Methods for calculating climatic data in Figure S2.

Geographical background included: (1) the administrative map of 348 municipalities of China, from National Geomatics Center of China (http://ngcc.sbsm.gov.cn/); (2) the altitude grid map of China at one square kilometre resolution, from DIVA-GIS (http://www.diva-gis.org/); (3) the grid maps of 1961-1999 MAT and MAP of China at one square kilometre resolution, from the Thematic Database for Human-earth System (http://www.data.ac.cn/).

The distribution of altitude range and municipalities for each species in China was recorded in “Magnolias of China” , the corresponding climatic data were calculated in three steps: (1) Extraction of municipalities for each species from administrative maps, followed by exaction of exact areas within municipalities based on altitudinal range. (2) Use of exact areas to mask and retrieve MAT/MAP maps, resulting in specific corresponding MAT/MAP data composed of 1 × 1 kilometre grid squares. (3) Summarizing of values of all grids for mean, maximum, minimum and range values for altitude, MAT and MAP. All processes were carried out in ArcGIS (version 9.3, Esri) for the 27 species.