Corresponding author: Xing-Jin He (
Academic editor: A. Sennikov
Xiao Y-P, Guo X-L, Price M, Gou W, Zhou S-D, He X-J (2021) New insights into the phylogeny of
Palynological study of
Previous studies have shown that fruit characteristics play a key role in the classification of subfamily
Similarly, previous molecular studies have been limited and results ambiguous.
Therefore, the objective of this study was to estimate the phylogenetic placement of
Samples were obtained from type localities and adjacent areas of
Voucher details and GenBank accession numbers of taxa used in this study.
Taxa | Voucher | Locality |
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T2010093003( |
Muli, Sichuan, China |
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XZ2011081741( |
Xizang, China |
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T2010100602( |
Xiangcheng, Sichuan, China |
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ZJ810826(KUN) | Shangri-La, Yunnan, China |
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– | |
ZJ0566(KUN) | Daocheng-Litang, Sichuan, China |
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ZJ0744(KUN) | Habahe, Xinjiang, China |
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J101(KUN) | Jiangsu Institute of Botany, China |
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J105(KUN) | Xinlong, Sichuan, China |
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J034(KUN) | Lijiang, Yunnan, China |
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ZJ0536(KUN) | Derong, Sichuan, China |
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ZJ810846(KUN) | Shangri-La, Yunnan, China |
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Pimenov & Kljuykov 200 (MW) | Khasan distr., Primorsk Terr., Russia | – | – | ||
666939( |
– |
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– | – | |
ZJ0697(KUN) | KIB nursery, Yunnan, China |
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ZJ0748(KUN) | Luquan, Yunnan, China |
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T2012052603 ( |
Baoxing, Sichuan, China |
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– | – | |
ZJ0521(KUN) | Shangri-La, Yunnan, China |
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Pimenov & Kljuykov 438 (MW) | Lorestan, Iran | – | – | ||
– | – |
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ZJ810841(KUN) | Shangri-La, Yunnan, China |
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Pimenov & Kljuykov 28 (MW) | Langtang National Park, Nepal | – | – | ||
ZJ0673(KUN) | Fumin, Yunnan, China |
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ZJ0519(KUN) | Shangri-La, Yunnan, China |
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– | – | |
Farille 81-421 (G) | N Annapurna, Nepal |
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– | – | |
J111(KUN) | Taiwan, China |
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– |
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YL757(KUN) | Lijiang, Yunnan, China |
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Watson & Gilbert 1580 (E, EBH) | Madoi, Qinghai, China, |
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– |
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J066(KUN) | Lijiang, Yunnan, China |
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ZJ810822(KUN) | Sichuan, China |
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ZJ0686(KUN) | Ninglang, Yunnan, China |
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FED 378 (E) | Shangri-La, Yunnan, China | – | – | ||
ZJ0678(KUN) | Ninglang, Yunnan, China |
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ZJ0503(KUN) | Lijiang, Yunnan, China |
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ZJ0524(KUN) | Shangri-La, Yunnan, China |
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ZJ0527(KUN) | Shangri-La, Yunnan, China |
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ZJ0573(KUN) | Yajiang-Kangding, Sichuan, China |
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ZJ0545(KUN) | Sichuan, China |
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ZJ0669(KUN) | Shangri-La, Yunnan, China |
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ZJ091033(KUN) | Shangri-La, Yunnan, China |
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ZJ04(KUN) | Suijiang, Yunnan, China |
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ZJ0581(KUN) | Luding-Mianning, Sichuan, China |
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ZJ06(KUN) | Suijiang, Yunnan, China |
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– | – |
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– | – | |
Pimenov & Kljuykov 169 (MW) | Primorsk Terr., Russia | – | – | ||
Skvortzov & Proskurjakova (MHA) | West Bengal, India |
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– | – | |
XYP19071901( |
Lijiang, Yunnan, China | ||||
YL561(KUN) | Lijiang, Yunnan, China |
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– | – |
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– | – | |
XYP19080301( |
Dali, Yunnan, China | ||||
ZJ0672(KUN) | Shangri-La, Yunnan, China |
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– | – | – | – | ||
ZJ0548(KUN) | Sichuan, China |
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XYP19080302( |
Dali, Yunnan, China | ||||
XYP19091803( |
Dali, Yunnan, China | ||||
XYP19080401( |
Dali, Yunnan, China | ||||
XYP19080402( |
Dali, Yunnan, China | ||||
Pimenov & |
Yumthang, Sikkim, India |
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– | – | |
0465919(KUN) | Xizang, China |
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ZJ0520(KUN) | Shangri-La, Yunnan, China |
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– | – | |
– | – | – | – | ||
T2012091505 ( |
Songxian, Henan, China |
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– | – | |
T2012093001 ( |
Anshan, Liaoning, China |
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– | – | |
Pimenov & Kljuykov 139 (MW) | Saghalien, Russia | – | – | ||
ZJ0623(KUN) | Hongyuan, Sichuan, China |
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Pimenov |
Hongyuan-Barkam, Sichuan, China | – | – | ||
ZJ0554 (KUN) | Daocheng, Sichuan, China |
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– | – | |
ZJ0501(KUN) | Lijiang, Yunnan, China |
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ZJ810821(KUN) | Sichuan, China |
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J075(KUN) | Lijiang, Yunnan, China |
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J091(KUN) | Lijaing, Yunnan, China |
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ZJ0564(KUN) | Daocheng-Litang, Sichuan, China |
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– unavailable sequences. * Newly generated sequences; otherwise, sequences were obtained from GenBank.
The related specimens in A, BM, CDBI, E, GB, GH, HNWP, IBSC, JAY, K, KATH, KUN, NAS, NWFC, NY, P, PE,
Species | Type specimens | Additional specimens examined |
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Without specific locality, |
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Total genomic DNA was extracted from silica gel-dried leaves and herbarium materials according to the protocols of plant genomic DNA kit (Tiangen Biotech, Beijing, China). Nuclear ribosomal DNA (
We used 53
SeqMan (
The plant morphological characteristics of
Comparison of plant morphological characteristics of
Characteristics |
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Root | taproot elongate, thickened at apex, branched | rootstock short, thick; roots torulose | taproot elongate, stout, often branched | taproot conic | taproot elongate, thick, often branched |
Stems | characteristically purplish | green | green | green | green |
Basal petioles | 2–10 cm | 5–7 cm | 8–15 cm, sparsely pubescent | 5–8 cm | 5–18 cm |
Sheath | oblong-ovate | oblong-ovate | broadly ovate | broadly lanceolate | ovate |
Basal leaves | blade ovate-lanceolate in outline, 1–2-pinnate; pinnae 4–5 pairs | triangular in outline, ternate-1–2-pinnate; pinnae 3–5 pairs | triangular in outline, 2-pinnate; pinnae 3–7 pairs | triangular in outline, ternate to 1- or 2-pinnate | triangular in outline, ternate-2–3-pinnate; pinnae 4–6 pairs |
Ultimate segments of blade | linear-lanceolate | linear-lanceolate | oblong-ovate, margins serrate, abaxially sparsely pubescent along veins | oblong-lanceolate | elongate-linear |
Cauline leaves | similar to the basal leaves | elongate-linear, reduced upwards becoming 1-pinnate or 3-lobed | similar to the basal leaves; upper leaves 1-pinnate | similar to the basal leaves | elongate-linear, 1–2-pinnate, reduced upwards |
Bracts | absent or occasionally 1–2, linear | absent, occasionally 1 | 1–4, linear-lanceolate | absent, occasionally 1, linear-lanceolate | absent or occasionally 1 |
Bracteoles | absent, rarely 1, linear | absent, occasionally 1 | 5–8, linear-lanceolate | 3–5, linear-lanceolate | absent |
Rays | 5–8(–12), unequal | 4–7(–10), subequal | 2–8, 1–3 cm, unequal | (3–)5–6(–8), unequal | 10–12, unequal |
Petals | ovate or broadly obovate, apex usually entire, occasionally 2–3-lobed, white | oblong-ovate or broadly obovate, apex obtuse to subacute, greenish-white | ovate or broadly obovate, apex subacute, white | apex palmately 3–4-lobed, lobes lanceolate or oblanceolate, white or violet | oblong-ovate or broadly obovate, apex subacute, white |
Specimens of
The fruit morphological and anatomical characteristics of
Mature fruit morphological and anatomical characteristics of
Species | Fruit shape | Transection | Endosperm concrescence | Development degree of ribs | Vittae number | ||
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L × W (mm) | Shape | Furrow | Commissure | ||||
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4.08 × 2.35 | ovoid | semicircle | cordate concave | unobvious | 2–3 | 4 |
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2.09 × 1.16 | ovoid | sub-pentagon | flat | obvious | 1–3 | 2–4 |
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2.41 × 0.96 | oblong-ovoid | sub-pentagon | flat | obvious | 1–3 | 2–4 |
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2.81 × 1.97 | ovoid | semicircle | sub-cordate concave | unobvious | 2–3 | 4 |
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1.43 × 1.36 | broad-ovoid | semicircle | slightly concave | unobvious | 1–3 | 2 |
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2.28 × 0.90 | oblong-ovoid | sub-pentagon | flat | obvious | 1–3 | 2–4 |
Morphological and anatomical characteristics of mature fruit
The pollen morphology of the five
Pollen morphology of
Species | Type | Shape | Size(μm) | Polar axis/Equatorial axis(P/E) | Size index ( |
Exine ornamentation | ||
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Equatorial view | Polar view | Equatorial area | Polar area | |||||
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super-rectangular | super-rectangular | trilobate circular | (27.20~32.90)30.36× (11.72~13.95)12.78 | 2.38 (2.16~2.59) | 19.69 (18.51~20.00) | cerebro reticulate | cerebroid with a few perforations |
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super-rectangular | super-rectangular | trilobate circular | (26.07~32.15)28.84× (11.42~15.33)13.24 | 2.19 (1.80~2.56) | 19.52 (17.25~20.92) | cerebro reticulate | cerebroid with a few perforations |
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super-rectangular | super-rectangular | trilobate circular | (22.46~27.08)24.95× (11.12~13.57)12.56 | 1.99 (1.66~2.28) | 17.69 (15.80~18.68) | cerebro reticulate | striate reticulate with a few perforations |
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sub-rhomboidal | sub-rhomboidal | obtuse triangle | (18.27~20.81)19.61× (10.40~13.31)11.66 | 1.69 (1.56~1.92) | 15.11 (14.15~16.62) | pitted reticulate | cerebroid |
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super-rectangular | super-rectangular | trilobate circular | (25.83~29.30)27.44× (10.92~14.62)13.58 | 2.03 (1.82~2.37) | 19.29 (16.79~20.20) | cerebro reticulate | cerebroid with a few perforations |
Pollen morphology in scanning electron microscope (
The characteristics of the three DNA regions are summarized in Table
Statistical summary for
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East Asia clade | |||||
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No. of accessions | 59 | 44 | 45 | 34 | 52 | |
Aligned length | 472 | 476 | 1897 | 444 | 2128 | |
No. variable characters | 349 (73.94%) | 288 (60.50%) | 318 (16.76%) | 252 (56.76%) | 278 (13.06%) | |
No. parsimony informative characters | 276 (58.47%) | 205 (43.07%) | 143 (7.54%) | 184 (41.44%) | 163 (7.66%) | |
Model |
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GTR+G | GTR+G | GTR+G | GTR+G | GTR+G |
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– | GTR+G | GTR+I+G | GTR+G | GTR+G |
After phylogenetic analyses with comprehensive sequence data, we confirmed that the collected
Phylogenetic relationships inferred from maximum likelihood (
The
Bayesian 50% strict consensus trees of 44
Bayesian 50% strict consensus tree of 34
We have studied the plant morphology, fruit morphological and anatomical characteristics, and palynology of five species of
Previous studies have shown that
This study’s phylogeny results indicated that there was a close and complex relationship between
Our
The results showed that the two populations of
Our phylogenetic results showed that
This work was supported by the National Natural Science Foundation of China (Grant Nos. 31872647, 32070221), National Specimen Information Infrastructure, Educational Specimen Sub-Platform (Grant No. 2005DKA21403-JK), the fourth national survey of traditional Chinese medicine resources (Grant No. 2019PC002). The authors thank Q. P. Jiang for her help in material collection; curators and staff of the following herbaria: A, BM, CDBI, E, GB, GH, HNWP, IBSC, JAY, K, KATH, KUN, NAS, NWFC, NY, P, PE,