该研究利用TaqMan特异性探针实时荧光PCR检测手段，建立了高效、精准鉴别贵州特色（中蜂）蓝莓蜜的方法。对贵州白竹林、麻卡和乌卡坪三个地域的蓝莓种植区蓝莓蜜进行采集（包括意蜂蓝莓蜜），同时购买市售蜂蜜及加拿大蓝莓蜜，该方法通过采集贵州麻江县白竹林地区蓝莓种植园及蜂场周围蓝莓同花期26种植物样本，基于植物基因组中trnL基因序列的多序列比对，设计蓝莓trnL基因特异性引物，并进行TaqMan探针验证。结果表明，本研究设计的TaqMan探针特异性强，建立的TaqMan探针实时荧光PCR检测方法能检测到蓝莓花粉DNA最低浓度为0.3 ng/μL；利用建立的方法对11种市售蜂蜜和贵州蓝莓蜜样本进行检测，发现贵州蓝莓蜜的Ct值为24~26，蓝莓花粉数在800~1 700颗，其余蜂蜜Ct值在30以上，表明该方法能够有效实现贵州蓝莓蜜和其他蜂蜜的区分，具有良好的应用前景。

In this study, a highly efficient and accurate method was developed for identifying Guizhou characteristic blueberry honey (from honey bee Apis cerana) based on specific TaqMan probe of real-time fluorescent PCR. Blueberry honey samples (including Apis mellifera) were collected from the blueberry growing areas of Bai zhulin, Maka and Wukaping in Guizhou, meanwhilw, commercial local honey and Canadian blueberry honey were also purchased. Twenty-six blueberry samples at the same flowering stage were collected from the blueberry gardens and the areas around the bee farms in the Baizhulin region of Majiang Town, Guizhou Province. The trnL gene in the sampling plant genome were sequenced. A trnL gene-specific primer of blueberry was designed based on the multiple sequence alignments of trnL gene sequences in the plant genome. TaqMan probe was used for validation. The results showed that the TaqMan probe had high specificity, and the minimum concentration of DNA as low as 0.3 ng/μL can be detected was in blueberry sample using the established TaqMan probe real-time fluorescent PCR detection method. Such an established method was used to test the samples of 11 kinds of commercial honey and Guizhou blueberry honey. It was found that the Ct value of Guizhou blueberry honey was 24~26, the number of blueberry pollen was 800~1 700, and the Ct values of other honey samples were over 30, indicating that this method can effectively distinguish Guizhou blueberry honey from other honeys, thereby having a good application prospect.

财政部和农业农村部：国家现代农业产业技术体系资助项目（CARS-44-SYZ4）；贵州喀斯特山区优势特色产业提质增效技术集成与示范（2021YFD1100300）；黔科合支撑[（2021）160]