ABSTRACT
Auxin is the first discovered plant hormone and is essential for many aspects of plant growth and development. Indole-3-acetic acid (IAA) is the main auxin and plays pivotal roles in intercellular communication through polar auxin transport. Phenylacetic acid (PAA) is another natural auxin that does not show polar movement. Although a wide range of species have been shown to produce PAA, its biosynthesis, inactivation and physiological significance in plants are largely unknown. In this study, we demonstrate that overexpression of the CYP79A2 gene, which is involved in benzylglucosinolate synthesis, remarkably increased the levels of PAA and enhanced lateral root formation in Arabidopsis. This coincided with a significant reduction in the levels of IAA. The results from auxin metabolite quantification suggest that the PAA-dependent induction of GRETCHEN HAGEN 3 (GH3) genes, which encode auxin-amido synthetases, promote the inactivation of IAA. Similarly, an increase in IAA synthesis, via the indole-3-acetaldoxime pathway, significantly reduced the levels of PAA. The same adjustment of IAA and PAA levels was also observed by applying each auxin to wild-type plants. These results show that GH3 auxin-amido synthetases can alter the ratio of IAA and PAA in plant growth and development.
Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/metabolism , Cytochrome P-450 Enzyme System/metabolism , Indoleacetic Acids/metabolism , Phenylacetates/metabolism , Plant Growth Regulators/metabolism , Arabidopsis/genetics , Arabidopsis/growth & development , Cytochrome P-450 Enzyme System/genetics , Gene Expression Regulation, Plant , Indoles , Ligases/metabolism , Oximes , Thiocyanates/metabolism , Thioglucosides/biosynthesisABSTRACT
Consumption of cruciferous vegetables is associated with reduced risk of developing cancer, a phenomenon attributed to glucosinolates, which are characteristic of these vegetables. We report production of the bioactive benzylglucosinolate in the noncruciferous plant Nicotiana benthamiana through metabolic engineering. The study includes identification of gamma-glutamyl peptidase 1 (GGP1), which substantially increased glucosinolate production by metabolizing an accumulating glutathione conjugate, an activity not previously described for glucosinolate biosynthesis or for proteins containing glutamine amidotransferase domains.
Subject(s)
Glucosinolates/biosynthesis , Nicotiana/enzymology , Plant Proteins/genetics , Plants, Genetically Modified/enzymology , Thioglucosides/biosynthesis , Open Reading Frames , Plant Leaves/enzymology , Plant Leaves/genetics , Plant Leaves/metabolism , Plants, Genetically Modified/genetics , Plants, Genetically Modified/metabolism , Thiocyanates , Nicotiana/genetics , Nicotiana/metabolism , Transformation, GeneticABSTRACT
Mutants and wild type plants of Arabidopsis thaliana were analysed for differences in glucosinolate accumulation patterns, indole-3-acetic acid (IAA) biosynthesis and phenotype. A previously identified series of mutants, termed TU, with altered glucosinolate patterns was used in this study. Only the line TU8 was affected in shoot phenotype (shorter stems, altered branching pattern). Synthesis of IAA and metabolism were not much affected in the TU8 mutant during seedling development, although the content of free IAA peaked earlier in TU8 during plant development than in the wild type. Indole glucosinolates and IAA may, however, be involved in the development of clubroot disease caused by the obligate biotrophic fungus Plasmodiophora brassicae since the TU3 line had a lower infection rate than the wild type, and lines TU3 and TU8 showed decreased symptom development. The decline in clubroot formation was accompanied by a reduced number of fungal structures within the root cortex and slower development of the fungus. Indole glucosinolates were lower in infected roots of TU3 and TU8 than in control roots of these lines, whereas in wild-type plants the differences were not as prominent. Free IAA and indole-3-acetonitrile (IAN) were increased in infected roots of the wild type and mutants with normal clubroot symptoms, whereas they were reduced in infected roots of mutants TU3 and TU8. These results indicate a role for indole glucosinolates and IAN/IAA in relation to symptom development in clubroot disease.