Preliminary study on identification of Dendrobium officinale from Zhejiang based on ITS2 and SNP technology

doi:10.3969/j.issn.1004-1524.2021.01.09

Acta Agriculturae Zhejiangensis ›› 2021, Vol. 33 ›› Issue (1): 69-76.DOI: 10.3969/j.issn.1004-1524.2021.01.09

• Horticultural Science • Previous Articles Next Articles

Preliminary study on identification of Dendrobium officinale from Zhejiang based on ITS2 and SNP technology

CHEN Wenqiang¹(), WANG Xiaofu²^,^*(), CHEN Xiaoyun², PENG Cheng², XU Junfeng², CAI Jian¹^,^*()

1. College of Biological and Food Engineering, Fuyang Normal University, Fuyang 236037, China
2. State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products (in Prepared), Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China

Received:2020-05-07 Online:2021-01-25 Published:2021-01-25
Contact: WANG Xiaofu,CAI Jian

Abstract

Abstract:

Dendrobium officinale is favored by consumers for its nutritional value and health benefits. However, it is difficult to distinguish and identify different varieties and different qualities of D. officinale. It is urgent to establish a fast, accurate and efficient identification method. In this study, 12 D. officinale from Zhejiang Province and 10 other Dendrobium were used, the DNA barcode ITS2 was selected, the amplification system for ITS2 was optimized, then the amplified products were sequenced. After sequence comparison and analysis, the specific SNP sites were obtained from DNA barcode ITS2 of D. officinale; specific primers designed for the SNP sites were verified by PCR, and a rapid identification method for SNP sites based on high-resolution melting curve (HRM) technology was established. The results showed that DNA barcode ITS2 could distinguish D. officinale from other Dendrobium. The rapid detection based on SNP realized the rapid identification of D. officinale, which provided a new concept and technical path for identification and quality control of D. officinale.

Key words: ITS2, SNP technology, Dendrobium officinale, DNA barcode, quick identification

CLC Number:

S567.23⁺9

CHEN Wenqiang, WANG Xiaofu, CHEN Xiaoyun, PENG Cheng, XU Junfeng, CAI Jian. Preliminary study on identification of Dendrobium officinale from Zhejiang based on ITS2 and SNP technology[J]. Acta Agriculturae Zhejiangensis, 2021, 33(1): 69-76.

Figures/Tables 7

References 16

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编号No.	品种Specie name	样品部位Plant part	产地Locality
1	铁皮石斛D. officinale	叶、茎Leaves, stems	金华(浙江)Jinhua, Zhejiang
2	铁皮石斛D. officinale	叶、茎Leaves, stems	金华(浙江)Jinhua, Zhejiang
3	铁皮石斛D. officinale	叶、茎Leaves, stems	金华(浙江)Jinhua, Zhejiang
4	铁皮石斛D. officinale	叶、茎Leaves, stems	金华(浙江)Jinhua, Zhejiang
5	铁皮石斛D. officinale	叶、茎Leaves, stems	金华(浙江)Jinhua, Zhejiang
6	铁皮石斛D. officinale	叶、茎Leaves, stems	金华(浙江)Jinhua, Zhejiang
7	铁皮石斛D. officinale	茎Stems	杭州(浙江)Hangzhou, Zhejiang
8	铁皮石斛D. officinale	茎Stems	临安(浙江)Lin'an, Zhejiang
9	铁皮石斛D. officinale	茎Stems	杭州(浙江)Hangzhou, Zhejiang
10	铁皮石斛D. officinale	茎Stems	武义(浙江)Wuyi, Zhejiang
11	铁皮石斛D. officinale	茎Stems	磐安(浙江)Pan'an, Zhejiang
12	铁皮石斛D. officinale	茎Stems	磐安(浙江)Pan'an, Zhejiang
13	金钗石斛D. nobile	叶、茎Leaves, stems	红河(云南)Honghe, Yunnan
14	重唇石斛D. hercoglossum	叶、茎Leaves, stems	红河(云南)Honghe, Yunnan
15	钩状石斛D. aduncum	叶、茎Leaves, stems	来宾(广西)Laibin , Guangxi
16	铜皮石斛D. moniliforme	叶、茎Leaves, stems	来宾(广西)Laibin , Guangxi
17	大包鞘石斛D. wardianum	叶、茎Leaves, stems	宏德(云南)Hongde, Yunnan
18	束花石斛D. chrysanthum	叶、茎Leaves, stems	宏德(云南)Hongde, Yunnan
19	兜唇石斛D. aphyllum	叶、茎Leaves stems	宏德(云南)Hongde, Yunnan
20	喇叭唇石斛D. lituiflorum	叶、茎Leaves, stems	宏德(云南)Hongde, Yunnan
21	肿节石斛D. pendulum	叶、茎Leaves, stems	宏德(云南)Hongde, Yunnan
22	鼓槌石斛D. chrysotoxum	叶、茎Leaves, stems	宏德(云南)Hongde, Yunnan

编号No.	品种Specie name	样品部位Plant part	产地Locality
1	铁皮石斛D. officinale	叶、茎Leaves, stems	金华(浙江)Jinhua, Zhejiang
2	铁皮石斛D. officinale	叶、茎Leaves, stems	金华(浙江)Jinhua, Zhejiang
3	铁皮石斛D. officinale	叶、茎Leaves, stems	金华(浙江)Jinhua, Zhejiang
4	铁皮石斛D. officinale	叶、茎Leaves, stems	金华(浙江)Jinhua, Zhejiang
5	铁皮石斛D. officinale	叶、茎Leaves, stems	金华(浙江)Jinhua, Zhejiang
6	铁皮石斛D. officinale	叶、茎Leaves, stems	金华(浙江)Jinhua, Zhejiang
7	铁皮石斛D. officinale	茎Stems	杭州(浙江)Hangzhou, Zhejiang
8	铁皮石斛D. officinale	茎Stems	临安(浙江)Lin'an, Zhejiang
9	铁皮石斛D. officinale	茎Stems	杭州(浙江)Hangzhou, Zhejiang
10	铁皮石斛D. officinale	茎Stems	武义(浙江)Wuyi, Zhejiang
11	铁皮石斛D. officinale	茎Stems	磐安(浙江)Pan'an, Zhejiang
12	铁皮石斛D. officinale	茎Stems	磐安(浙江)Pan'an, Zhejiang
13	金钗石斛D. nobile	叶、茎Leaves, stems	红河(云南)Honghe, Yunnan
14	重唇石斛D. hercoglossum	叶、茎Leaves, stems	红河(云南)Honghe, Yunnan
15	钩状石斛D. aduncum	叶、茎Leaves, stems	来宾(广西)Laibin , Guangxi
16	铜皮石斛D. moniliforme	叶、茎Leaves, stems	来宾(广西)Laibin , Guangxi
17	大包鞘石斛D. wardianum	叶、茎Leaves, stems	宏德(云南)Hongde, Yunnan
18	束花石斛D. chrysanthum	叶、茎Leaves, stems	宏德(云南)Hongde, Yunnan
19	兜唇石斛D. aphyllum	叶、茎Leaves stems	宏德(云南)Hongde, Yunnan
20	喇叭唇石斛D. lituiflorum	叶、茎Leaves, stems	宏德(云南)Hongde, Yunnan
21	肿节石斛D. pendulum	叶、茎Leaves, stems	宏德(云南)Hongde, Yunnan
22	鼓槌石斛D. chrysotoxum	叶、茎Leaves, stems	宏德(云南)Hongde, Yunnan

引物名称 Primer name	序列 Sequence(5'-3')	退火温度 Annealing temperature/℃	扩增长度 Length/bp
ITS2	F:CATTGTCGAGACCGAAACA	45	490
	R:ACCAACGAGAAGATGACCC
ARMS PCR	F:CGCAAGTTGCGCCAAAC	56	128
	R:GCACATCCGAGCCTTAGTAACAC
HRM 180	F:TGAAGAGCGCAGCGAAAT	58	180
	R:ACCAAGGGGCACGAGGAG
HRM 53	F:GAGTCTTTGAACGCAAGTTG	58	53
	R:GCGGACGTGCCCTTAGCC

引物名称 Primer name	序列 Sequence(5'-3')	退火温度 Annealing temperature/℃	扩增长度 Length/bp
ITS2	F:CATTGTCGAGACCGAAACA	45	490
	R:ACCAACGAGAAGATGACCC
ARMS PCR	F:CGCAAGTTGCGCCAAAC	56	128
	R:GCACATCCGAGCCTTAGTAACAC
HRM 180	F:TGAAGAGCGCAGCGAAAT	58	180
	R:ACCAAGGGGCACGAGGAG
HRM 53	F:GAGTCTTTGAACGCAAGTTG	58	53
	R:GCGGACGTGCCCTTAGCC

Preliminary study on identification of Dendrobium officinale from Zhejiang based on ITS2 and SNP technology

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