Biodiversity Science ›› 2016, Vol. 24 ›› Issue (3): 296-303.doi: 10.17520/biods.2015259

• Orginal Article • Previous Article     Next Article

Genetic relationships of buckwheat species based on the sequence analysis of ITS and ndhF-rpl32

Yani Hu1,2, Zongwen Zhang2,3,*(), Bin Wu2, Jia Gao2, Yanqin Li1,*()   

  1. 1 Institute of Biotechnology, Shanxi University, Taiyuan 030006
    2 Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081
    3 China Office of Biodiversity International, Beijing 100081
  • Received:2015-09-21 Accepted:2016-03-02 Online:2016-04-05
  • Zhang Zongwen,Li Yanqin E-mail:zhangzongwen@caas.cn;yanqin@sxu.edu.cn

Buckwheat was originated in China and evolved into abundant species and genetic diversity. In order to effectively use germplasm resources of buckwheat and its wild relatives, the genetic relationships of buckwheat species were analyzed on 71 Fagopyrum samples including 10 species (subspecies, varieties and complex), which were collected mainly from Sichuan, Guizhou and Gansu provinces. Through the analysis of ITS and ndhF-rpl32 sequences and the construction of phylogenetic trees, the inter- and intra-specific relationships of these species were discussed. Results showed that the sequence length of ITS was 725 bp in the matrix, while the parsimony informative sites was 150, accounting for 20.7% of the total length. In the matrix of ndhF-rpl32 sequence, the length was 940 bp, while the parsimony informative sites was 158, accounting for 16.8% of total length. On one hand, the phylogenetic trees based on both ITS and ndhF-rpl32 sequencing data clearly separated the big-achene-group buckwheat (F. tataricum, F. cymosum complex, F. esculentum and F. esculentum ssp. ancestralis) from the small-achene-group buckwheat (F. gracilipes var. odontopterum, F. leptopodum var. grossii, F. leptopodum, F. densovillosum, F. gracilipes, and F. urophyllum). On the other hand, the phylogenetic tree based on ndhF-rpl32 sequences can be used to distinguish F. esculentum ssp. ancestralis and F. esculentum, which indicated that the ndhF-rpl32 sequence was more effective in analyzing the phylogenetic relationships of buckwheat species. These results provide a scientific basis for studies of taxonomy and establishing barcodes of Fagopyrum species.

Key words: buckwheat, genetic relationships, ITS, ndhF-rpl32

Table 1

The Latin names and sources of the tested Fagopyrum materials"

编号
Collecting number
原产地
Origin
代号
Code
编号
Collecting number
原产地
Origin
代号
Code
齿翅野荞 Fagopyrum gracilipes var. odontopterum 细柄野荞 F. gracilipes
YXCC201211030 四川越西 Yuexi, Sichuan a1 XDXB201211028 四川喜德 Xide, Sichuan g1
ZJCC2012102601 四川昭觉 Zhaojue, Sichuan a2 ZJXB2012102602 四川昭觉 Zhaojue, Sichuan g2
ZJCC2012102603 四川昭觉 Zhaojue, Sichuan a3 ZJXB2012102605 四川昭觉 Zhaojue, Sichuan g3
ZJCC2012102604 四川昭觉 Zhaojue, Sichuan a4 ZJXB2012102703 四川昭觉 Zhaojue, Sichuan g4
ZJCC2012102704 四川昭觉 Zhaojue, Sichuan a5 MGXB2012102901 四川美姑 Meigu, Sichuan g5
BTCC2012102706 四川布拖 Butuo, Sichuan a6 HLXB2012110303 四川会理 Huili, Sichuan g6
ZJCC2012102801 四川昭觉 Zhaojue, Sichuan a7 HLXB2012110306 四川会理 Huili, Sichuan g7
MGCC2012102902 四川美姑 Meigu, Sichuan a8 HDXB2012110404 四川会东 Huidong, Sichuan g8
LBCC2012103002 四川雷波 Leibo, Sichuan a9 HDXB2012110501 四川会东 Huidong, Sichuan g9
HLCC2012110304 四川会理 Huili, Sichuan a10 HDCC2012110504 四川会东 Huidong, Sichuan g10
HLCC2012110305 四川会理 Huili, Sichuan a11 HDXB2012110504 四川会东 Huidong, Sichuan g11
HDCC2012110502 四川会东 Huidong, Sichuan a12 HLXB2012110301 四川会理 Huili, Sichuan g12
HDCC2012110503 四川会东 Huidong, Sichuan a13 MNXB201210006 四川冕宁 Mianning, Sichuan g13
YXCC201211040 四川越西 Yuexi, Sichuan a14 XDXB201210014 四川喜德 Xide, Sichuan g14
YXCC201211044 四川越西 Yuexi, Sichuan a15 XDXB201211046 四川喜德 Xide, Sichuan g15
YYCC201210012 四川盐源 Yanyuan, Sichuan a16 YXXB201211031 四川越西 Yuexi, Sichuan g16
HDCC2012110402 四川越西 Yuexi, Sichuan a17 金荞复合体 F. cymosum complex
栽培苦荞 F. tataricum XDJQ201210004 四川喜德 Xide, Sichuan h1
XDKQ201210003 四川喜德 Xide, Sichuan b1 MNJQ201210007 四川冕宁 Mianning, Sichuan h2
XDKQ201210005 四川喜德 Xide, Sichuan b2 XDKQ201210017 西川喜德 Xide, Sichuan h3
XDKQ201210013 四川喜德 Xide, Sichuan b3 XDJQ201210022 西川喜德 Xide, Sichuan h4
XDKQ201210019 四川喜德 Xide, Sichuan b4 YXJQ201211029 四川越西 Yuexi, Sichuan h5
XDKQ201210020 四川喜德 Xide, Sichuan b5 YXJQ201211039 四川越西 Yuexi, Sichuan h6
YXKQ201211032 四川越西 Yuexi, Sichuan b6 XDJQ201211045 四川喜德 Xide, Sichuan h7
YXKQ201211033 四川越西 Yuexi, Sichuan b7 DCJQ2012110302 四川德昌 Dechang, Sichuan h8
疏穗小野荞 F. leptopodum var.grossii PGJQ2012110601 四川普格 Puge, Sichuan h9
GLSS201211037 四川甘洛 Ganluo, Sichuan c1 DCJQ2012110202 四川德昌 Dechang, Sichuan h10
GLSS201211038 四川甘洛 Ganluo, Sichuan c2 HDJQ2012110403 四川会东 Huidong, Sichuan h11
GLSS201211050 四川甘洛 Ganluo, Sichuan c3 XDKQ201210018 四川喜德 Xide, Sichuan h12
GLSS201211052 四川甘洛 Ganluo, Sichuan c4 YXJQ201211043 四川越西 Yuexi, Sichuan h13
YXSS201211051 四川越西 Yuexi, Sichuan c5 硬枝万年荞 F. urophyllum
野生甜荞 F. esculentum ssp. ancestralis YYYZ201210011 四川盐源 Yanyuan, Sichuan i1
XCYTQ201210008 四川西昌 Xichang, Sichuan d1 LBYZ2012103004 四川雷波 Leibo, Sichuan i2
LBTJ2012103001 四川雷波 Leibo, Sichuan d2 MGYZ2012102903 四川美姑 Meigu, Sichuan i3
小野荞 F. leptopodum 栽培甜荞 F. esculentum
XDXY201210021 四川喜德 Xide, Sichuan e1 00000664 甘肃武威 Wuwei, Gansu j1
MNXY201210024 四川冕宁 Mianning, Sichuan e2 00000906 贵州威宁 Weining, Guizhou j2
密毛野荞 F. densovillosum 米苦荞 F. tataricum
MGMM2012102804 四川美姑 Meigu, Sichuan f1 YXXMQ201211034 四川越西 Yuexi, Sichuan k1
LBMM2012103003 四川雷波 Leibo, Sichuan f2 YXXMQ201211042 四川越西 Yuexi, Sichuan k2

Table 2

The G+C contents of ITS and ndhF-rpl32 sequences in Fagopyrum species"

种名 Species ITS (%) ndhF-rpl32 (%)
齿翅野荞 F. gracilipes var. odontopterum 65.57 24.17
栽培苦荞 F. tataricum 67.65 22.79
疏穗小野荞 F. leptopodum var. grossii 65.96 22.01
野生甜荞 F. esculentum ssp. ancestralis 66.46 24.47
小野荞 F. leptopodum 66.82 23.43
密毛野荞 F. densovillosum 65.34 24.77
细柄野荞 F. gracilipes 65.81 23.17
金荞复合体 F. cymosum complex 65.80 22.75
硬枝万年荞 F. urophyllum 63.93 24.68
栽培甜荞 F. esculentum 65.47 24.13
米苦荞 F. tataricum 67.95 23.40

Table 3

Distance within Fagopyrum species based on ITS and ndhF-rpl32 sequences"

种名 Species ITS ndhF-rpl32
齿翅野荞 F. gracilipes var. odontopterum 0.001 0.182
栽培苦荞 F. tataricum 0.003 0.000
疏穗小野荞 F. leptopodum var. grossii 0.002 0.002
野生甜荞 F. esculentum ssp. ancestralis 0.107 0.001
小野荞 F. leptopodum 0.062 0.021
密毛野荞 F. densovillosum 0.003 0.018
细柄野荞 F. gracilipes 0.001 0.099
金荞复合体 F. cymosum complex 0.028 0.026
硬枝万年荞 F. urophyllum 0.165 0.014
栽培甜荞 F. esculentum 0.110 0.000
米苦荞 F. tataricum 0.000 0.001

Table 4

Distance between Fagopyrum species based on ITS (below diagonal) and ndhF-rpl32 (above diagonal) sequences"

CC KQ SS TJ XY MM XB JQ YZ TQ MKQ
齿翅野荞(CC) F. gracilipes var. odontopterum 0.191 0.024 0.198 0.001 0.001 0.001 0.191 0.027 0.197 0.191
栽培苦荞(KQ)
F. tataricum
0.157 0.185 0.037 0.191 0.190 0.191 0.007 0.188 0.039 0.001
疏穗小野荞(SS)
F. leptopodum var. grossii
0.034 0.169 0.191 0.025 0.024 0.025 0.186 0.021 0.194 0.186
野生甜荞(TJ)
F. esculentum ssp. ancestralis
0.193 0.145 0.209 0.199 0.198 0.199 0.039 0.194 0.003 0.038
小野荞(XY)
F. leptopodum
0.031 0.176 0.038 0.204 0.001 0.000 0.192 0.028 0.198 0.192
密毛野荞(MM)
F. densovillosum
0.003 0.160 0.036 0.196 0.033 0.001 0.191 0.027 0.197 0.191
细柄野荞(XB)
F. gracilipes
0.001 0.157 0.034 0.193 0.031 0.003 0.191 0.027 0.198 0.192
金荞复合体(JQ)
F. cymosum complex
0.144 0.059 0.156 0.144 0.161 0.147 0.144 0.188 0.041 0.008
硬枝万年荞(YZ)
F. urophyllum
0.111 0.238 0.117 0.267 0.118 0.114 0.112 0.217 0.197 0.189
栽培甜荞(TQ)
F. esculentum
0.211 0.170 0.233 0.133 0.226 0.214 0.211 0.168 0.279 0.040
米苦荞 (MKQ)
F. tataricum
0.156 0.002 0.168 0.146 0.175 0.159 0.156 0.060 0.237 0.071

Fig. 1

Neighbor-joining trees of the genus Fagopyrum resulted from (A) ITS sequences; (B) ndhF-rpl32 sequences."

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