Research Article |
Corresponding author: Atsuko Takano ( takano@hitohaku.jp ) Academic editor: Eberhard Fischer
© 2017 Atsuko Takano.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Takano A (2017) Taxonomic study on Japanese Salvia (Lamiaceae): Phylogenetic position of S. akiensis, and polyphyletic nature of S. lutescens var. intermedia. PhytoKeys 80: 87-104. https://doi.org/10.3897/phytokeys.80.11611
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Both Salvia akiensis and S. lutescens (Lamiaceae) are endemic to Japan. Salvia akiensis was recently described in 2014 in the Chugoku (= SW Honshu) region, and each four varieties of S. lutescens distributed allopatrically. Among varieties in S. lutescens, var. intermedia show a disjunctive distribution in the Kanto (=E Honshu) and Kinki (= W Honshu) regions. Recent field studies of S. lutescens var. intermedia revealed several morphological differences between the Kanto and Kinki populations. Here, I evaluated these differences among Salvia lutescens var. intermedia and its allies with morphological analysis and molecular phylogenetic analyses of nuclear ribosomal DNA (internal and external transcribed spacer regions) and plastid DNA (ycf1-rps15 spacer, rbcL, and trnL-F) sequences. Both morphological analysis and molecular phylogenetic analyses showed that S. lutescens var. intermedia from the Kinki region and var. lutescens were closely related to each other. However, var. intermedia from the Kanto region exhibited an association with S. lutescens var. crenata and var. stolonifera, which also grew in eastern Japan, rather than var. intermedia in the Kinki region. These results indicated that S. lutescens var. intermedia is not a taxon with a disjunctive distribution, but a combination of two or more allopatric taxa. Present study also suggested that S. akiensis was most closely related to S. omerocalyx.
cpDNA, Lamiaceae , nrDNA, Phylogenetics, Salvia akiensis , Salvia lutescens
The genus Salvia L. (tribe Mentheae) is the largest genus in Lamiaceae; it comprises nearly 1,000 species. Salvia has radiated extensively into three regions of the world: Central and South America (500 spp.), West Asia (200 spp.), and East Asia (100 spp.) (Alziar, 1988–1993). In Japan, twelve species, eight varieties, and one putative hybrid have been described since
There are four varieties known in S. lutescens (Koidz.) Koidz.: var. crenata, var. intermedia, var. lutescens, and var. stolonifera (
Recently, a new species of Japanese Salvia, S. akiensis A.Takano, T.Sera et Kurosaki has been described from Shimane and Hiroshima Prefectures (
As a step toward taxonomic revision of variety of S. lutescens and to confirm monophyly and phylogenetic position of S. akiensis, morphological and molecular phylogenetic analyses were conducted.
The protocols for DNA extraction, polymerase chain reaction (PCR), purification, and DNA sequencing were described previously by
Raw sequence data were assembled and edited manually, with BioEdit software (ver. 7.2.5
DNA sequences were aligned with the CLUSTALW 1.83 software package, with default settings and multiple alignments (
Compared to
Taxa, Genbank accession number, and voucher specimens/references used in this study. Newly sequecned data are shown bold.
Name | Pop. Code | rbcL | trnL-F | ycf1-rps15 | ETS | ITS | Voucher / References |
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S. akiensis A.Takano, T.Sera et Kurosaki | HIR (Hirohsima Pref.) | LC124176 | LC124188 | LC060529 | LC060825 | LC060729 | A.Takano and N.Kurosaki with T.Sera 130606-1( |
S1(Shimane Pref.) | LC124177 | LC124189 | LC060530 | LC060826 | LC060728 | M.Sakoda et al. 1 ( |
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S. arisanensis Hayata | AB295063 | AB295074 | LC060531 | LC060827 | AB295085 |
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S. glabrescens (Franch. et Sav.) Makino | |||||||
var. glabrescens | FS (Wakasa, Fukui) | AB541134 | AB541148 | LC060532 | LC060828 | AB541120 |
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var. repens (Koidz.) Kurosaki | KY (Kyoto) | AB295064 | AB295075 | LC060533 | LC060829 | AB295086 |
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S. isensis Nakai ex Hara | MIE | AB266221 | AB266231 | LC060534 | LC060830 | AB266241 |
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AICHI | LC124178 | LC124190 | LC060535 | LC060831 | LC060730 | A-200933 (living material at Hiroshima Bot.Gard. Originally from Owariasahi city, Aichi Pref.) | |
S. japonica Thunb. | |||||||
f. albiflora Hiyama | AB266220 | AB266230 | LC060536 | LC060832 | AB260240 |
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f. japonica | Osaka | AB266219 | AB266229 | LC060537 | LC060833 | AB266239 |
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f. japonica | Gotenba | LC124179 | LC124191 | LC060538 | LC060834 | LC060731 | A.Takano 140806-5 ( |
f. longipes (Nakai) Sugimoto | AB266218 | AB266228 | LC060539 | LC060835 | AB266238 |
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S. koyamae Makino | AB541128 | AB541142 | LC060540 | LC060836 | AB541114 |
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S. lutescens Koidz. | |||||||
var. crenata (Makino) Murata | AICHI | AB266223 | AB266233 | LC060541 | LC060837 | AB266243 |
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Yushin | AB353202 | AB353198 | LC060542 | LC060838 | AB353206 |
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Akita | LC124180 | LC124193 | LC124205 | LC124201 | LC124203 | Y. Horhii, S. Nishida et al. 2015026 ( |
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Fukui | LC124181 | LC124194 | LC124204 | LC124200 | LC124202 | A.Takano 150702-1a ( |
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var. intermedia (Makino) Murata | Nara | LC124182 | LC124195 | LC060544 | LC060840 | AB295097 |
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Shiga | LC124183 | LC124196 | LC060546 | LC060842 | LC060735 | A.Takano 140821-1 ( |
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Mt.Mikuni | LC124184 | LC124197 | LC060547 | LC060843 | LC060733 | A.Takano 140806-4 ( |
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Tanzawa | LC124185 | LC124198 | LC060548 | LC060844 | LC060734 | A.Takano 140622-2 ( |
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var. lutescens Koidz. | MIE | AB266222 | AB266232 | LC060549 | LC060845 | AB266242 |
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Aoyama | LC124186 | LC128192 | LC060550 | LC060846 | LC060737 | a201241 (living material at Hiroshima Bot.Gard. Originally from Aoyama Kogen, Mie Pref.) | |
var. stolonifera G.Nakai | AB541139 | AB541153 | LC060551 | LC060847 | AB541125 |
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S. nipponica Miq. | |||||||
var. nipponica | TOKU (Tokushima) | AB541132 | AB541146 | LC060552 | LC060848 | AB541118 |
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KUMA (Kumamoto) | AB541127 | AB541141 | LC060553 | LC060849 | AB541113 |
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var. kisoensis K.Imai | NAK | AB541136 | AB541150 | LC060554 | LC060850 | AB541122 |
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S. omerocalyx Hayata | |||||||
var. omerocalyx | HI (Hidaka, Hyogo) | AB353204 | AB353196 | LC060555 | LC060851 | AB353200 |
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AB353205 | AB353197 | LC060556 | LC060852 | AB353201 |
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var. prostrata Satake | AB541138 | AB541152 | LC060557 | LC060853 | AB541124 |
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S. pygmaea Matsum. | |||||||
var. pygmaea | AB295072 | AB295083 | LC060558 | LC060854 | AB295094 |
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var. simplicior Hatus. ex T.Yamaz. | AB541140 | AB541154 | LC060559 | LC060855 | AB541126 |
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S. ranzaniana Makino | AB287373 | AB287374 | LC060560 | LC060856 | AB287375 |
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S. × sakuensis Naruh. et Hihara | AB541129 | AB541143 | LC060561 | LC060857 | AB541116 |
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Outgroup | |||||||
S. plebeia R.Br. | KIZU | AB295073 | AB295084 | LC060563 | LC060858 | AB295095 |
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KAMI | LC124187 | LC124199 | LC060562 | LC060859 | LC060738 | A.Takano and N.Kurosaki 090607-2 ( |
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S. Polystachya M.Martens et Galeotti | AY570435 | JF301399 | JF289067 | JF301334 | JF301356 | Drew and Systma (2011) |
The incongruence length difference (ILD) test (
Among the 89 specimens of S. lutescens var. intermedia examined, 52 specimens from the Kinki region were pilose at the base of the anther connective (Fig.
Photographs of S. lutescens var. intermedia flowers. a Flower of A. Takano 140806-4-2 (
Totally, 18 specimens of S. lutescens var. lutescens were deposited at
A likelihood analysis using the concatenate cpDNA datasets (rbcL+trnL-F+ycf1-rps15 spacer) for 36 individuals in 23 taxa resulted in a ML tree with –lnL = 5295.264. The ML and Bayesian trees had similar topology; the Bayesian maximum clade credibility tree is shown with ML bootstrap (ML-BS) and Bayesian posterior probability (BI-PP) in Figure
A concatenate nrDNA datasets (ETS+ITS) yielded a ML tree with –lnL = 3789.114. The ML and Bayesian trees had similar topology; the Bayesian maximum clade credibility tree is shown with ML-BS and BI-PP in Figure
This study suggests that S. lutescens var. intermedia is polyphyletic. Four individuals of var. intermedia, two from the Kanto and two from the Kinki region fell into different subclades in both molecular phylogenetic trees using cpDNA and nrDNA datasets, although the two from the Kinki region were always in the same subclade (Figs
The Bayesian maximum clade credibility tree derived from plastid DNA (concatenate dataset of rbcL, trnL-F, ycf1-rps15). ML-bootstrap/Bayesian PP numbers are shown near the corresponding branch. Thick lines denote a clade that was strongly supported, with ML- bootstrap and/or Bayesian-PP greater than 95 %. ML: maximum likelihood; PP: posterior probability.
The Bayesian maximum clade credibility tree derived from nuclear ribosomal DNA (concatenate dataset of ETS and ITS). ML-bootstrap/Bayesian-PP numbers are shown above or below the corresponding branch. Thick lines denote a clade that was strongly supported with ML-bootstrap and/or Bayesian-PP values greater than 95 %. ML: maximum likelihood; PP: posterior probability.
On the contrary, present morphological and molecular phylogenetic analyses indicated that S. lutescens var. lutescens and var. intermedia from the Kinki region are closely related to each other. In molecular phylogenetic analysis, they formed a cluster in both cpDNA- and nrDNA trees, though ML-BP/BI-PP support was not strong in cpDNA tree. The morphological study revealed var. lutescens is pilose at the base of the anther connective: therefore, S. lutescens var. intermedia in the Kinki region share the same morphological status with var. lutescens. The distribution of var. lutescens is very near to that of var. intermedia in the Kinki region (Mie, Shiga, Nara Prefs.), although var. lutescens and populations of the Kinki regions of var. intermedia have never been found to grow together.
Salvia lutescens var. intermedia in the Kanto region may be more closely related to var. crenata and var. stolonifera.
The phylogenetic analyses also suggest that S. akiensis comprises a monophyletic group, as indicated by nrDNA tree, and that most of the species allied to S. akiensis was the S. omerocalyx group. Salvia akiensis and S. omerocalyx group comprised a subclade in nrDNA (ML-BS/BI-PP: 89/1.00). These two taxa did not form a subclade in cpDNA, but it may be of introgression/chloroplast capture /hybridization as mentioned above. In contrast, S. akiensis and S. omerocalyx share following characters: bearing the largest flowers among species in the subg. Allagospadonopsis, flower from May to June, and exhibit gynodioecy (
I am grateful to Teruo Katsuyama (
Specimens examined Salvia lutescens var. intermedia. And var. lutescens.
Salvia lutescens var. intermedia
Specimens glabrous at base of anther connective (27 sheets)
KANTO Region. Yamagata Pref.: Mt. Kushigata, S.Kigawa s.n., July 10, 2001 (
KINKI Region. Nara Pref.; Yoshino, Yamato, Y.Kato s.n., Aug. 8. 1936 (
Specimens showed one to several hairs at base of anther connective (10 sheets)
KANTO Region. Pref.Sizuoka: Mt. Higane, Prov. Izu, S. Shimazu s.n., July 18, 1920(
Pref.Kanagawa: Sirogane rindo, Yugawara-cho, Y.Hasegawa 14263 (
Specimens which showed long pilose at base of anther connective (52 sheets)
KINKI Region. Kyoto Pref.: Rakuhoku, Ohara, Otonashi W.F., S.Hajacava s.n., Aug. 1896 (
Salvia lutescens var. lutescens
Specimens which showed long pilose at base of anther connective (13 sheets)
Mie Pref.: Itaya, Kata, Kameyama, S.Tsugaru & T.Sawada 34155 (
Shiga Pref.: Nasugahara, Ohara-Mura, Kouga gun, G.Koizumi s.n., 2 July, 1939 (