Functional Assessment of an Overexpressed Arabidopsis Purple Acid Phosphatase Gene (Atpap26) in Tobacco Plants

Abstract

BackgroundOverexpression of known genes encoding key phosphate (Pi)-metabolizing enzymes, such as acid phosphatases (APases), is presumed to help plants with Pi availability and absorption as they are mostly exposed to suboptimal environmental conditions for this vital element.ObjectivesIn this study, the overexpression effect of AtPAP26, one of the main contributors in retrieving Pi from intracellular and extracellular compounds, was evaluated from various viewes in tobacco plant.Materials and MethodsAs a heterologous expression system, the encoding cDNA sequence of AtPAP26 was transferred into tobacco plants.ResultsA high growth rate of the transgenic lines was observed which could be due to an increased APase activity, leading to the high total phosphorus as well as the free Pi content of the transgenic plants. Interestingly, a significant increased activity of the other APases was also noticed, indicating a networking among them. These were accompanied by less branched and short primary roots and a decreased lateral root numbers grown in Pi-starvation condition compared to the wild type seedlings. Besides, a delayed germination and dwarf phenotype indicates the possible reduction in gibberellic acid biosynthesis in the transgenic lines.ConclusionsSuch transgenic plants are of interest not only for increased yield but also for the reduced need for chemical fertilizers and removal of excessive Pi accumulation in soils as a consequence of fertilizers’ or poultry wastes’ over-usage.

Effects of ethyl acetate extract of peony (Paeonia suffruticosa) seed coat on the proliferation and apoptosis of oral squamous carcinoma cells through miR-424-3p/STAT3/Survivin pathway

“…MicroRNAs play an important role in this regard (31). There are some factors that stimulate the expression of genes in plants, or by producing transgenic plants and using the engineering method to create metabolites (32)(33)(34)(35). In this research, we investigate the effects of ethyl acetate extract of peony (Paeonia suffruticosa) seed coat on the proliferation and apoptosis of oral squamous carcinoma cells through miR-424-3p/ STAT3/Survivin pathway.…”

The Critical Role of AtPAP17 and AtPAP26 Genes in Arabidopsis Phosphate Compensation Network

“…PCR amplification was performed under following conditions: 95°C for 3 min, followed by 40 cycles at 95°C for 20 s, 58°C for 20 s, and 72°C for 20 s, and a final step at 72°C for 5 min. Semi-quantitative RT-PCR was performed as described previously ( Zamani et al., 2012 ; Sabet et al., 2018 ). Also, actin and α- tubulin genes were used as internal controls to normalize gene expression in quantitative real time-PCR and semi-quantitative RT-PCR, respectively.…”

“…Afterwards, they were placed in growth chamber with a 16-h-light/8-h-dark photoperiod at 25°C and fertilized twice weekly by sub-irrigation with Hoagland's nutrient solution. Also, transformed Arabidopsis plants carrying CaMV-35S: AtPAP17 and CaMV-35S:AtPAP26 constructs were used as overexpressing lines of AtPAP17 and AtPAP26 genes, respectively (Sabet et al, 2018). More than 10 single-copy transgenic lines were generated for each construct.…”

“…Pi starvation inducible (PSI) extracellular APases are involved in Pi scavenging and hydrolysis from external phosphoesters or organic P, whereas intracellular APases remobilize and recycle Pi from its compound in cytoplasm and organelles ( Veljanovski et al., 2006 ; Tran et al., 2010a ; Wang et al., 2014 ; Zhang et al., 2014 ; Wang and Liu, 2018 ). Several PSI APases have been biochemically and molecularly characterized in some plant species such as Arabidopsis thaliana ( Veljanovski et al., 2006 ; Lohrasebi et al., 2007 ; Kuang et al., 2009 ; Tran et al., 2010b ; Wang et al., 2011 ; Zamani et al., 2014 ; Liu et al., 2016 ; Bhadouria et al., 2017 ; Kong et al., 2018 ; Sabet et al., 2018 ). Among APases, purple acid phosphatases (PAPs) are the most important class of plant PSI APases which appear purple or pink color in water solution due to a bimetallic active center ( Olczak et al., 2003 ; Tran et al., 2010a ).…”

Identification and Functional Analysis of Two Purple Acid Phosphatases AtPAP17 and AtPAP26 Involved in Salt Tolerance in Arabidopsis thaliana Plant

“…For cDNA synthesis, 1 μg of DNase-treated RNA was reverse transcribed by M-MLV reverse transcriptase (Fermentas, Lithuania) as stated by the manufacturing instructions. A series of semi-quantitative RT-PCR was conducted to evaluate the level of relative gene transcript variants ( Zamani et al, 2012 ; Sabet et al, 2018 ), and their band intensity in agarose gel was quantified by TotalLab software (Phoretix International, New Castle, United Kingdom). The RT-PCR conditions were as follows: 94°C for 5 min; 35 cycles of 94°C for 1 min, specific annealing temperature for each primer pair for 1 min and 72°C for 1 min; and a final extension step at 72°C for 10 min.…”