Abstract
BackgroundIndole-3-acetic acid (IAA) extraction and purification are of great importance in auxin research, which is a hot topic in the plant growth and development field. Solid-phase extraction (SPE) is frequently used for IAA extraction and purification. However, no IAA-specific SPE columns are commercially available at the moment. Therefore, the development of IAA-specific recognition materials and IAA extraction and purification methods will help researchers meet the need for more precise analytical methods for research on phytohormones.ResultsSince the AUXIN RESISTANT/INDOLE-3-ACETIC ACID INDUCIBLE (Aux/IAA) proteins show higher specific binding capability with auxin, recombinant IAA1, IAA7 and IAA28 proteins were used as sorbents to develop an IAA extraction and purification method. A GST tag was used to solidify the recombinant protein in a column. Aux/IAA proteins solidified in a column have successfully trapped trace IAA in aqueous solutions. The IAA7 protein showed higher IAA binding capability than the other proteins tested. In addition, expression of the IAA7 protein in Drosophila Schneider 2 (S2) cells produced better levels of binding than IAA7 expressed in E. coli.ConclusionThis work validated the potential of Aux/IAA proteins to extract and purify IAA from crude plant extracts once we refined the techniques for these processes.
Highlights
Indole-3-acetic acid (IAA) extraction and purification are of great importance in auxin research, which is a hot topic in the plant growth and development field
To ensure that an adequate level of recombinant protein was produced, the vector containing IAA7. The cells produced 0.019 (IAA1), IAA7 and IAA28 segments was transformed into three expression strains of E. coli, and the expression was compared in BL21, Tuner and Rosetta (Fig. 1b–d)
IAA1, IAA7 and IAA28 were all highly expressed in different strains when they were induced by 0.4 mM IPTG at 25 °C
Summary
Indole-3-acetic acid (IAA) extraction and purification are of great importance in auxin research, which is a hot topic in the plant growth and development field. Solid-phase extraction (SPE) is frequently used for IAA extraction and purification. The development of IAA-specific recognition materials and IAA extraction and purification methods will help researchers meet the need for more precise analytical methods for research on phytohormones. As one of the most important categories of phytohormones, auxin contributes to virtually all aspects of plant growth and development [1, 2]. IAA is extracted from plant tissues and highly purified using standardized protocols [3, 4]. IAA extraction from plant tissues involves liquid nitrogen freezing/lyophilization, grinding/homogenization, treatment with organic solvents and the removal of solid impurities. Organic solvents with polarity close to IAA are the most effective
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