Study on honey identification based on fluorescence quantitative PCR method

doi:10.16178/j.issn.0528-9017.20240068

Abstract

Abstract:

This study explored the applicability of real-time fluorescence quantitative PCR in the identification of honey. The pollen DNA and bee DNA in honey samples were extracted by CTAB method. The DNA in honey was detected by real-time fluorescence quantitative PCR using five plants and bee DNA primer probes to determine the pollen composition, production time and production location of different honey samples. The feasibility of the method was verified by 17 kinds of honey from unknown sources, and the results showed that the method was feasible. This study plays a role in promoting and guiding the use of molecular genetics for honey identification.

Key words: honey, honey identification, pollen DNA, real-time fluorescence quantitative PCR, bees

CLC Number:

S896.1

WANG Xiaoyu, WU Pan, WANG Chengli, YUE Wanfu. Study on honey identification based on fluorescence quantitative PCR method[J]. Journal of Zhejiang Agricultural Sciences, 2025, 66(3): 730-736.

Figures/Tables 7

References 26

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样品编号	样本名称	生产厂家/产地
1	百花蜜(2022年)	浙江省遂昌风采中蜂养殖场
2	无患子	浙江省遂昌青云岭中蜂养殖场
3	野桂	浙江省遂昌风采中蜂养殖场
4	百花蜜	浙江省林氏家庭农场
5	百花蜜(2023年)	浙江省遂昌风采中蜂养殖场
6	土蜂蜜	浙江省潘石玄根家庭农场
7	土蜂蜜	浙江省家旺家庭农场
8	土蜂蜜	浙江省青云岭中蜂养殖场
9	张蜂霸	浙江省遂昌甜蜜园蜂业专业合作社
10	皇浆花粉蜜	浙江省老山里蜂蜜(意蜂)
11	野桂蜜	浙江省开化县
12	春桂蜜	福建省闽侯县
13	野桂蜜	浙江省金华市
14	五倍子蜜	浙江省松阳县
15	野桂蜜	浙江省松阳县
16	春桂蜜	浙江省松阳县
17	野桂蜜	浙江省青田县
18	五倍子蜜	浙江省景宁县
19	柯树	浙江省金华市
20	野桂	浙江省庆元县
21	野桂	浙江省云和县
22	五倍子	浙江省缙云县
23	山花原蜜	浙江省兰溪蜂农自产自销店
24	冬青原蜜	浙江省兰溪蜂农自产自销店
25	百花原蜜	浙江省兰溪蜂农自产自销店

样品编号	样本名称	生产厂家/产地
1	百花蜜(2022年)	浙江省遂昌风采中蜂养殖场
2	无患子	浙江省遂昌青云岭中蜂养殖场
3	野桂	浙江省遂昌风采中蜂养殖场
4	百花蜜	浙江省林氏家庭农场
5	百花蜜(2023年)	浙江省遂昌风采中蜂养殖场
6	土蜂蜜	浙江省潘石玄根家庭农场
7	土蜂蜜	浙江省家旺家庭农场
8	土蜂蜜	浙江省青云岭中蜂养殖场
9	张蜂霸	浙江省遂昌甜蜜园蜂业专业合作社
10	皇浆花粉蜜	浙江省老山里蜂蜜(意蜂)
11	野桂蜜	浙江省开化县
12	春桂蜜	福建省闽侯县
13	野桂蜜	浙江省金华市
14	五倍子蜜	浙江省松阳县
15	野桂蜜	浙江省松阳县
16	春桂蜜	浙江省松阳县
17	野桂蜜	浙江省青田县
18	五倍子蜜	浙江省景宁县
19	柯树	浙江省金华市
20	野桂	浙江省庆元县
21	野桂	浙江省云和县
22	五倍子	浙江省缙云县
23	山花原蜜	浙江省兰溪蜂农自产自销店
24	冬青原蜜	浙江省兰溪蜂农自产自销店
25	百花原蜜	浙江省兰溪蜂农自产自销店

引物名称	引物序列(5'→3')
枇杷蜜-F	GCCCGGCGTCCCCTTATC
枇杷蜜-R	ATCGCATTTCGCTACGTTCTTCAT
五倍子蜂蜜-F	AAACCTGCCTAGCAGAACGA
五倍子蜂蜜-R	TAAGATTTCCTTGGCGCAAT
柃属植物-F	AATCTCGAGGCTTGTGCAGT
柃属植物-R	TGTTAGGCCCTGCTATTTGG
中蜂-F	ATTCTTGCACAGCATCTTTGCACAT
中蜂-R	CTTTTCGATACTGCTTTAGCCAAT
意蜂-F	GTTAGTCAAAATTCAACAGTTCTTT
意蜂-R	TGGAAACACTTACTTAGATACTGCA

引物名称	引物序列(5'→3')
枇杷蜜-F	GCCCGGCGTCCCCTTATC
枇杷蜜-R	ATCGCATTTCGCTACGTTCTTCAT
五倍子蜂蜜-F	AAACCTGCCTAGCAGAACGA
五倍子蜂蜜-R	TAAGATTTCCTTGGCGCAAT
柃属植物-F	AATCTCGAGGCTTGTGCAGT
柃属植物-R	TGTTAGGCCCTGCTATTTGG
中蜂-F	ATTCTTGCACAGCATCTTTGCACAT
中蜂-R	CTTTTCGATACTGCTTTAGCCAAT
意蜂-F	GTTAGTCAAAATTCAACAGTTCTTT
意蜂-R	TGGAAACACTTACTTAGATACTGCA

样品编号	提取浓度/ (ng·μL^-1)	C_t值
样品编号	提取浓度/ (ng·μL^-1)	中蜂	意蜂	柃属	五倍子	枇杷
1	1 912.36	34.01±0.99	—	35.24±1.21	32.36±0.99	33.00±0.49
2	973.73	30.22±0.35	—	34.33±0.90	29.75±0.67	31.13±0.66
3	54.11	30.85±0.32	—	34.78±0.28	34.65±0.36	32.31±0.68
4	91.37	31.27±0.62	—	34.19±0.87	30.70±0.41	35.79±0.68
5	297.91	28.98±0.68	—	35.41±0.39	31.14±0.57	35.27±0.18
6	133.41	29.89±0.50	—	35.47±1.54	25.40±0.47	36.27±0.65
7	88.59	30.16±0.93	—	30.24±0.54	26.16±0.18	35.06±1.44
8	175.25	30.21±0.55	—	36.22±1.14	24.56±0.45	31.85±0.38
9	2 132.09	29.85±0.77	—	34.25±1.42	30.85±1.11	33.30±1.00
10	2 510.32	35.42±0.74	32.63±1.34	—	32.42±0.35	—
11	152.37	28.99±0.60	—	33.91±1.54	28.37±1.13	32.63±1.46
12	138.52	28.34±1.42	—	33.08±1.31	28.98±0.59	32.63±0.51
13	267.13	25.66±1.54	—	33.22±1.49	29.53±0.8	31.55±0.66
14	97.53	32.91±1.38	—	—	32.12±0.74	33.04±1.07
15	308.71	29.98±1.18	—	33.38±1.05	26.64±1.48	31.51±0.53
16	72.00	34.73±0.80	—	34.60±0.16	31.57±1.11	32.87±1.12
17	2 175.53	33.54±1.33	—	32.13±1.52	29.95±1.35	33.01±0.68
18	413.01	28.46±0.94	—	—	22.23±0.89	33.84±1.70
19	348.69	30.23±0.11	—	33.98±0.67	—	27.27±0.18
20	208.46	30.19±0.46	—	34.76±0.20	32.44±0.52	28.99±0.75
21	2 102.93	34.06±0.39	—	33.27±1.10	36.14±1.29	33.60±0.62
22	188.07	31.30±0.69	—	34.12±0.08	—	31.34±0.84
23	332.70	34.16±0.24	35.61±0.70	33.74±0.54	—	—
24	152.82	32.00±0.28	31.50±0.15	34.63±1.09	—	33.32±0.27
25	60.86	32.94±0.75	30.84±0.16	33.79±0.60	35.58±0.29	33.88±0.53
NC	0	—	—	—	—	—