摘要
The HPLC-MS/MS method was developed to profile the dynamics of abscisic acid （ABA） and ABA-glucose ester （ABA-GE） after cloning glycosyltransferase enzyme family gene AtUGT71C5 into Arabidopsis thaliana. By constructing over-expression lines （OE） and down-expression lines （DN）, we acquired mutant strains to analyze the function of AtUGT71C5. The multiple-reaction monitoring （MRM） was used for quantitative determination in negative mode. The transition was m/z 263.1→153.0 for ABA （[M-H]^＋）, m/z 425.1→263.0 for ABA-GE （[M-H]^＋）, and m/z 321.0-152.0 for chloramphenicol. The linear range was 0.8684-217.1 ng/mL for ABA and 0.3920-196.0 ng/mL for ABA-GE. The accuracy was 88.0-109.0% for ABA and 86.6-113.0% for ABA-GE; the inter-day and intra-day precisions were less than 5.4%for ABA and 8.9%for ABA-GE, respectively. This method is simple and sensitive enough for determination of ABA and ABA-GE in A. thaliana leaves. All the evidence confirmed the speculation that AtUGT71C5 can mediate abscisic acid homeostasis.
The HPLC-MS/MS method was developed to profile the dynamics of abscisic acid （ABA） and ABA-glucose ester （ABA-GE） after cloning glycosyltransferase enzyme family gene AtUGT71C5 into Arabidopsis thaliana. By constructing over-expression lines （OE） and down-expression lines （DN）, we acquired mutant strains to analyze the function of AtUGT71C5. The multiple-reaction monitoring （MRM） was used for quantitative determination in negative mode. The transition was m/z 263.1→153.0 for ABA （[M-H]^＋）, m/z 425.1→263.0 for ABA-GE （[M-H]^＋）, and m/z 321.0-152.0 for chloramphenicol. The linear range was 0.8684-217.1 ng/mL for ABA and 0.3920-196.0 ng/mL for ABA-GE. The accuracy was 88.0-109.0% for ABA and 86.6-113.0% for ABA-GE; the inter-day and intra-day precisions were less than 5.4%for ABA and 8.9%for ABA-GE, respectively. This method is simple and sensitive enough for determination of ABA and ABA-GE in A. thaliana leaves. All the evidence confirmed the speculation that AtUGT71C5 can mediate abscisic acid homeostasis.
