To diminish the time required for some diagnostic assays including reverse transcription PCR (RT-PCR), reverse transcription loop-mediated isothermal amplification (RT-LAMP; due to mainly RNA extraction step) and also DAS-ELISA into a minimum level, an innovative immunocapture RT-LAMP (IC-RT-LAMP) and immunocapture reverse transcription (IC/RT-PCR) protocol on the basis of Potato Leafroll virus (PLRV) genome were used and optimized. In this regard, all six IC-RT-LAMP primers (i.e. F3, B3, FIP, BIP, LF and LB) together with IC/RT-PCR primers were designed on the basis of the highly conserved sequence (ORF3) of coat protein gene (GenBank accession number: U73777) of PLRV genome. Even though DAS-ELISA, IC/RT-PCR and IC-RT-LAMP assays could successfully detect positive infected plant samples, considering the time, safety, sensitivity, cost and simplicity, the last one was overall superior. Meanwhile, among five different visual dyes to accurately detect IC-RT-LAMP products, both hydroxynaphthol blue and GeneFinder™ could produce long stable colour change and brightness in a close tube-based approach to prevent cross-contamination risk, concluded eventually as the best ones. Altogether, as IC-RT-LAMP is sensitive, cost-effective, fairly user friendly and also can generate more accurate results than previous diagnostic procedures, we accordingly propose this colorimetric assay as a highly reliable alternative viral recognition system regarding PLRV recognition and probably other viral-based diseases.

Total flavonoid content (TFC) and cyanidin-3-glucoside (Cyd-3-glu) of seed and seed coat extract of 16 genotypes from five species of Carthamus were studied, and their major polyphenolic compounds and antioxidant activity of the seed coat extracts were determined using HPLC analysis and DPPH assay, respectively. Additionally, fatty acids composition of the seed oil was analyzed by GC. In general, TFC and Cyd-3-glu content of seed coat extracts were higher than those of seed extracts. A novel breeding line with black seed coat (named A82) depicted lower TFC (3.79 mg QUE/g DW) but higher Cyd-3-glu (24.64 mg/g DW) compared to the white and other seed-pigmented genotypes. DPPH radical scavenging activity showed a strong association with Cyd-3-glu content (r = 0.84), but no correlation with TFC (r = -0.32). HPLC analysis of seed coat extracts revealed that four compounds were dominant constituents including rutin (7.23 - 117.95 mg/100 g DW), apigenin (4.37 - 64.88 mg/100 g DW), quercetin (3.09 - 14.10 mg/100 g DW), and ferulic acid (4.49 - 30.41 mg/100 g DW). Interestingly, the genotype A82 with an appropriate polyunsaturated/saturated fatty acids index (5.46%) and a moderate linoleic fatty acid content (64.70%) had higher nutritional and pharmaceutical value than all the other genotypes.

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In the present study, the interactive effect of irrigation and nitrogen (N) on biomass and physiological characteristics of amaranth and the influence of zeolite in this interaction were investigated. A 2-year field experiment was conducted as split-plot factorial in a randomized complete block design in three replicates. The main factor consisted of different levels of deficit irrigation (DI) such as irrigation after 40 (DI 1), 60 (DI 2), and 75% (DI 3) depletion of soil available water. The subplots derived from a combination of different factors including nitrogen rates (zero (N 0), 80 (N 1), 160 (N 2), and 240 (N 3) kg urea ha −1) and zeolite levels (zero (Z 0) and 10 (Z 1) ton ha −1). Reduction of soil moisture resulted in a decrease in the amount of chlorophylls, carotenoids, protein, maximum quantum yield of photosystem II (F v /F m ratio), and biomass of amaranth. In contrast, it resulted in the increase of antioxidant enzyme activity, soluble carbohydrates, and proline. However, nitrogen application led to an increase in the contents of chlorophyll, carotenoids, protein, antioxidant enzyme activity, F v /F m ratio, and biomass of amaranth and a decrease in the soluble carbohydrates and proline. Zeolite treatment by increasing the content of chlorophylls (16%), carotenoids (19%), protein (25%), F v /F m ratio (11%) as well as enchantment of soluble carbohydrate content and antioxidant enzyme activity and by decreasing the proline content (40%) moderated the adverse consequences of water-deficit stress and improved biomass of amaranth. In general, the use of zeolite along with the lowest rate of nitrogen fertilizer (80 kg urea ha −1) reduced the harmful effects of the oxidative stress caused by water scarcity and improved the growth conditions for the plant and thus produced maximum biomass of amaranth plant under moderate and severe deficit irrigations.