Abstract
A primary cDNA library of Aspergillus niger H1 was constructed using SMART (switching mechanism at the 5' end of RNA transcript) technique. A total of 169 clones had halos on the insoluble phosphate medium, and clone H-47 had clear halos. The full-length cDNA of clone H-47 was 625 bp, with a complete open reading frame (ORF) of 390 bp, encoding a protein of 129 amino acids. Multiple alignment revealed a high degree of homology between the ORF of the clone and other fungi cellulosome-integrating protein (CipC-like). The expression vector of ORF was constructed and transformed into Escherichia coli DH-5a. The transformant (ORF-1) with the CipC-like gene secreted more organic acid when grown in tricalcium phosphate (TCP) medium, with TCP as the sole source of phosphate. E. coli DH5a containing the cipc-like gene secreted methanoic acid, acetic acid, malic acid, and citric acid reached 81.2, 93.3, 50.6, and 147.7 ug mL-1, respectively, within 28 h. These results showed that the expression of the A. niger H1 CipC-like gene in E. coli could enhance organic acid secretion and improve phosphate solubilizing ability.
Highlights
Phosphorus is one of the major essential macronutrients for plants, and it is applied to soil in the form of phosphate (P)
E. coli DH5α containing the cipc-like gene secreted methanoic acid, acetic acid, malic acid, and citric acid reached 81.2, 93.3, 50.6, and 147.7 μg mL-1, respectively, within 28 h. These results showed that the expression of the A. niger H1 CipC-like gene in E. coli could enhance organic acid secretion and improve phosphate solubilizing ability
We demonstrated that overexpression of CipC-like in E. coli DH5α increased the secretion of organic acids and enhanced P solubilization ability
Summary
Phosphorus is one of the major essential macronutrients for plants, and it is applied to soil in the form of phosphate (P). Most P are immobile and the available content of P is below the limit of plant growth in soils. Microorganisms play an important role in rendering insoluble P into soluble forms that are utilized by plants and prompt plant growth (Mehta et al, 2013; Xiao et al, 2009). Numerous studies have focused on the organic acid secretion of microorganisms to solubilize P in soil. The main organic acids known to solubilize P in soil are gluconic, oxalic, citric, 2-ketoglutaric, lactic, acetic, formic, and succinic acids (Kumar et al, 2013; Bianco et al, 2010; Werra et al, 2009; Khan et al, 2007; Fomina et al, 2004; Altomare et al, 1999; Banik et al, 1982)
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