SUMMARYThe environmental concern about diffuse pollution from nitrogen (N) fertilizers has led to increased research on the diagnosis of crop N status. The SPAD chlorophyll (Chl) meter is the most commonly used tool for rice (Oryza sativa L.) N status diagnosis, but measurements are conducted at a specific point and readings are affected by different leaf positions. Many measurements per plant must be taken in order to increase the accuracy of N status diagnosis, which limits its application. The present paper attempts to determine rice N status at the canopy level using Multiplex®, a new hand-held optical fluorescence sensor. The fluorescence emission of rice leaves under light excitation was utilized by Multiplex® to non-destructively assess rice leaf Chl and phenolic compound content. A field experiment was conducted in 2011 using a completely randomized split-plot design, with main-plot treatments being six N fertilizer application rates and subplot treatments being different plant densities. Leaf Chl and phenolic compounds were evaluated using the ratio of far-red fluorescence (FRF) to red fluorescence (RF) emission under red light excitation (simple fluorescence ratio, SFR_R) (R2 = 0·35, P < 0·01) and the ratio of decadic logarithm of red to ultra-violet (UV) fluorescence emission (R2 = 0·30, P < 0·01), respectively. Both SPAD reading and fluorescence-based indices including flavonoids (FLAV), nitrogen balance index (NBI_R) and SFR_R could be used to predict rice leaf N contents. The canopy FLAV, SFR_R and NBI_R were all highly correlated to average SPAD readings (R2 > 0·70 in most cases, P < 0·01). Therefore, Multiplex® can be used as an alternative to SPAD to determine rice N status in paddy fields.

Accurate and reliable detection of exosomal miRNA can serve as a promising method for early diagnosis of disease and evaluation of therapeutic effects. However, current exosomal miRNA detection methods commonly involve exosome enrichment, containing RNA extraction, and qRT-PCR based quantification, which are expensive and time-consuming. Herein, we develop a DNA zipper-mediated membrane fusion approach for rapid exosomal miRNA detection and cancer diagnosis. First, a lipid vesicle probe containing miR21-targeting molecular beacons (MBs) is constructed and further loaded with zipper DNA constructs (ZDCs) on its surface. Meanwhile, complementary zipper DNA constructs (cZDCs) are introduced on the exosome of interest. Upon mixing them together, zipping between ZDC and cZDC induces the membrane fusion of exosomes and vesicle probes, triggering the recognition of exosomal miR21 by contained MBs and fluorescence emission that can be conveniently detected within 30 min. Importantly, with the assistance of flow cytometry, miR21-overexpressed tumor exosomes derived from either cell culture medium or clinical patient serums can be distinguished from exosomes secreted from normal cells. This approach provides a convenient way to accurately detect the exosomal miRNA, which may hold great potential in liquid biopsy for early cancer diagnosis and monitoring the therapeutic effects during the treatments.

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