Abstract：
The kiwi plant is dioecious, and its sex is generally identified from flower morphology at blossoming, which takes several years. It is quite necessary but challenging to on-spot identify the plant sex in juvenile stage. Here the target DNA was obtained by screening the Friendly boy (FrBy) gene which is sex-related for different kiwi plant species. Its complementary sequence was divided into two parts as primer DNA and further attached to different gold nanoparticles (GNPs). The connection between target DNA and primer DNA will promote the formation of plasmonic dimers. Dark field microscopy (DFM) can distinguish particles in different aggregation states. Various conditions were optimized based on the standard of increasing the proportion of dimers while reducing that of large aggregates. Furthermore, two Raman reporters (RR) are separately labeled on the nanoprobes, and the plasmonic dimers lead to a tremendous Raman enhancement of two reporters located at the dimer nanogap. Double-blind tests proved the feasibility of this method on the actual samples of kiwi plant leaves. Our SERS method is sensitive, specific, and reliable for rapid sex identification analysis at the kiwi seeding stage, with great promise for decision-making in field management.
摘要：
猕猴桃植物是雌雄异株的，它的性别通常从开花时的花朵形态中确定，这需要几年的时间。在幼年期进行现场识别是非常必要但具有挑战性的。在此,通过筛选与不同猕猴桃植物物种性别相关的友好男孩(FrBy)基因获得靶DNA。其互补序列分为两部分作为引物DNA，并进一步连接到不同的金纳米颗粒（GNP）。靶DNA和引物DNA之间的连接将促进等离子体二聚体的形成。暗视野显微镜（DFM）可以区分不同聚集状态的颗粒。基于增加二聚体比例同时减少大聚集体比例的标准,优化了各种条件。此外，两个拉曼报道分子（RR）分别标记在纳米探针上，和等离子体二聚体导致位于二聚体纳米间隙的两个报道分子的巨大拉曼增强。双盲试验证明了该方法在猕猴桃植物叶片实际样品上的可行性。我们的SERS方法很灵敏,具体,在猕猴桃播种阶段进行快速性别鉴定分析，在现场管理决策方面大有可为。
