Indole-3-acetamide(IAM)is the first confirmed auxin biosynthetic intermediate in some plant pathogenic bacteria.Exogenously applied IAM or production of IAM by overexpressing the bacterial iaaM gene in Arabidopsis cau...Indole-3-acetamide(IAM)is the first confirmed auxin biosynthetic intermediate in some plant pathogenic bacteria.Exogenously applied IAM or production of IAM by overexpressing the bacterial iaaM gene in Arabidopsis causes auxin overproduction phenotypes.However,it is still inconclusive whether plants use IAM as a key precursor for auxin biosynthesis.Herein,we reported the isolation IAM HYDROLASE 1(IAMH1)gene in Arabidopsis from a forward genetic screen for IAM-insensitive mutants that display normal auxin sensitivities.IAMH1 has a close homolog named IAMH2 that is located right next to IAMH1on chromosomeⅣin Arabidopsis.We generated iamh1 iamh2 double mutants using our CRISPR/Cas9gene editing technology.We showed that disruption of the IAMH genes rendered Arabidopsis plants resistant to IAM treatments and also suppressed the iaaM overexpression phenotypes,suggesting that IAMH1 and IAMH2 are the main enzymes responsible for converting IAM into indole-3-acetic acid(IAA)in Arabidopsis.The iamh double mutants did not display obvious developmental defects,indicating that IAM does not play a major role in auxin biosynthesis under normal growth conditions.Our findings provide a solid foundation for clarifying the roles of IAM in auxin biosynthesis and plant development.展开更多
Both tryptophan （Trp） and auxin are essential for plant growth and Trp is a precursor for auxin biosynthesis. Concentrations of Trp and auxin need to be tightly con- trolled to ensure optimal growth and development....Both tryptophan （Trp） and auxin are essential for plant growth and Trp is a precursor for auxin biosynthesis. Concentrations of Trp and auxin need to be tightly con- trolled to ensure optimal growth and development. It has been very difficult to study the homeostasis of these two essential and inter-dependent compounds. Auxin is mainly synthesized from Trp via a two-step pathway using indole- 3-pyruvate （IPA） as the intermediate. Here we used a bacterial Trp oxidase RebO, which does not exist in Ara- bidopsis and which converts Trp to the imine form of IPA, to modulate IPA levels in Arabidopsis. Our results demonstrate that Arabidopsis plants use two strategies to ensure that no excess IPA is made from Trp. IPA is made from Trp by the TAA family of aminotransferases, which we show catalyzes the reverse reaction when IPA level is high. Moreover, excess IPA is converted back to Trp by the YAS1 aminotransferase. We show that the VASl-cat- alyzed reaction is very important for Trp homeostasis. This work not only elucidates the intricate biochemical mech- anisms that control the homeostasis of Trp, IPA, and auxin, but also provides novel tools for further biochemical studies on Tip metabolism and auxin biosynthesis in plants.展开更多
文摘Indole-3-acetamide(IAM)is the first confirmed auxin biosynthetic intermediate in some plant pathogenic bacteria.Exogenously applied IAM or production of IAM by overexpressing the bacterial iaaM gene in Arabidopsis causes auxin overproduction phenotypes.However,it is still inconclusive whether plants use IAM as a key precursor for auxin biosynthesis.Herein,we reported the isolation IAM HYDROLASE 1(IAMH1)gene in Arabidopsis from a forward genetic screen for IAM-insensitive mutants that display normal auxin sensitivities.IAMH1 has a close homolog named IAMH2 that is located right next to IAMH1on chromosomeⅣin Arabidopsis.We generated iamh1 iamh2 double mutants using our CRISPR/Cas9gene editing technology.We showed that disruption of the IAMH genes rendered Arabidopsis plants resistant to IAM treatments and also suppressed the iaaM overexpression phenotypes,suggesting that IAMH1 and IAMH2 are the main enzymes responsible for converting IAM into indole-3-acetic acid(IAA)in Arabidopsis.The iamh double mutants did not display obvious developmental defects,indicating that IAM does not play a major role in auxin biosynthesis under normal growth conditions.Our findings provide a solid foundation for clarifying the roles of IAM in auxin biosynthesis and plant development.
基金supported by the NIH(R01GM114660 to YZ)supported by NIH(GM52413to JC)HHMI
文摘Both tryptophan （Trp） and auxin are essential for plant growth and Trp is a precursor for auxin biosynthesis. Concentrations of Trp and auxin need to be tightly con- trolled to ensure optimal growth and development. It has been very difficult to study the homeostasis of these two essential and inter-dependent compounds. Auxin is mainly synthesized from Trp via a two-step pathway using indole- 3-pyruvate （IPA） as the intermediate. Here we used a bacterial Trp oxidase RebO, which does not exist in Ara- bidopsis and which converts Trp to the imine form of IPA, to modulate IPA levels in Arabidopsis. Our results demonstrate that Arabidopsis plants use two strategies to ensure that no excess IPA is made from Trp. IPA is made from Trp by the TAA family of aminotransferases, which we show catalyzes the reverse reaction when IPA level is high. Moreover, excess IPA is converted back to Trp by the YAS1 aminotransferase. We show that the VASl-cat- alyzed reaction is very important for Trp homeostasis. This work not only elucidates the intricate biochemical mech- anisms that control the homeostasis of Trp, IPA, and auxin, but also provides novel tools for further biochemical studies on Tip metabolism and auxin biosynthesis in plants.
