Wu Jinlong, Chen Lin, Lin Dingbo, Ma Zhaocheng, Deng Xiuxin
Key Laboratory of Horticultural Plant Biology (Ministry of Education), Huazhong Agricultural University , Wuhan 430070, China.
Department of Nutritional Sciences, Oklahoma State University , 419 Human Sciences, Stillwater, Oklahoma 74078, United States.
J Agric Food Chem. 2016 Nov 30;64(47):9089-9098. doi: 10.1021/acs.jafc.6b03410. Epub 2016 Nov 14.
The effects of tissue type, harvest maturity, and genetic factors on the expression of genes that related to citrus fruit allergies remain poorly understood. In the present study, a multiplex real-time PCR assay was developed to monitor the expression of citrus allergen genes individually with the advantages of much fewer sample requirements and simultaneously multiple target genes detection. Gene specific primer pairs and Taqman probes of three citrus allergen genes Cit s 1.01, Cit s 2.01, and Cit s 3.01 and the house-keeping gene β-actin were designed based on gene sequence differences. The PCR results showed that differential expression patterns were found during the ripening process. The expression levels of Cit s 3.01 were much higher than those of Cit s 1.01 and Cit s 2.01 in both peel and pulp tissues among 10 citrus cultivars. Data suggested that Kao Phuang Pummelo could be safely consumed with a potential low risk in allergenicity. Considering that assessing allergenicity is one of the tests in food safety, this assay might also facilitate the breeding and production of "allergy-friendly" citrus fruits.
组织类型、收获成熟度和遗传因素对与柑橘类水果过敏相关基因表达的影响仍知之甚少。在本研究中，开发了一种多重实时PCR检测方法，以单独监测柑橘过敏原基因的表达，该方法具有样品需求量少得多和同时检测多个靶基因的优点。基于基因序列差异，设计了三种柑橘过敏原基因Cit s 1.01、Cit s 2.01和Cit s 3.01以及管家基因β-肌动蛋白的基因特异性引物对和Taqman探针。PCR结果表明，在成熟过程中发现了差异表达模式。在10个柑橘品种的果皮和果肉组织中，Cit s 3.01的表达水平远高于Cit s 1.01和Cit s 2.01。数据表明，高盘柚食用时潜在过敏风险较低，可以安全食用。考虑到评估致敏性是食品安全检测之一，该检测方法也可能有助于“低过敏”柑橘类水果的育种和生产。
