Expression Of MAP18 In Arabidopsis NO/SA/Ubiquitin Mutants

Microtubule-associated proteins(MAPs)play important roles in the regulation of microtubule function in plant cells.Map18 is an important factor in regulating the microtubule(MT)dynamics.Arabidopsis thaliana MAP18,a unstable protein,has been reported in plant growth and development.The functions of Microtubule-associated proteins(MAPs)is mainly realized through the interaction with microtubules.As an important part of cytoskeleton in eukaryotic cells,microtubules(MT)play an important role in plant growth and development,cell morphogenesis and cytoplasmic matrix.The mechanism of NO,SA and ubiquitin involved in the regulation of map18 on plant growth and development is not clear.In this study,we considered wild-type Arabidopsis(Columbia ecotype,Col-0),sad2-1,sad2-2,eds1,npr1,NOA,hub2-2,hub1-4,hub1-5,ubc1-1,ubc2-1,ubc3-1.Arabidopsis mutants and nia1nia2 as experimental materials to do Genetic analysis.And we studied the changes of morphological,cytological and molecular between mutants and wild type.This paper attempts to explain the mechanism of map18 in Arabidopsis NO,SA and ubiquitin mutants.The main results of this experiment are as follows:1.The germination rate and leaf hair of different mutants were analyzed.Arabidopsis wild type(Columbia ecotype,Col-0)and sad2-1,sad2-2,eds1,npr1,No A,nia1nia2,hub2-2,hub1-4,hub1-5,ubc1-1,ubc2-1,ubc3-1 Arabidopsis mutants were cultured in a dish containing MS solid medium.Temperature 20 ± 2?,Light cycle 16 hours light / 8hours dark,and Illumination intensity 5000 LX.The result about statistics of Arabidopsis germination rate after 5 days show that Arabidopsis wild-type germination percentage was100%,while that of sad2-1 and sad2-2 were more than 95%,and that of NOA mutant was98%,which was not significantly different from that of wild-type;the germination rate of nia1nia2 Arabidopsis mutant was about 80%,which was significantly different.Ten days later,observe the plant phenotype,the results show that The second pair lesf and metaphyll surface of Arabidopsis wild-type and sad2-2,eds1,npr1,NOA,nia1nia2 mutants had leaf hairs,while sad2-1 had no leaf hairs.2.Analzing the seed germination dynamics of Arabidopsis wild type and related mutants by High flux dynamic imaging analyzer(Dyna Plant)?Arabidopsis seeds were seeded in MS solid culture dish,vernalization treatment,and then put them in High flux dynamic imaging analyzer(Dyna Plant)(parameters: temperature 20 ± 2?,light cycle reference nature 16 h light / 8h dark,light intensity 5000LX).To observe the growth and development of Arabidopsis.The results showed that the growth trend of wild type Arabidopsis was better than that of mutants,and the growth pattern of mutants was different from that of wild type.3.Analyzing the expression of MAP18 in Arabidopsis mutants.Cultivated Arabidopsis wild type and Arabidopsis sad2-1,sad2-2,eds1,npr1,NOA,nia1nia2,hub2-2,hub1-4,hub1-5,ubc1-1,ubc2-1,ubc3-1 mutants,after 17 days,collecting plants' samples,the total RNA of each Arabidopsis strain was extracted,obtain c DNA by reverse transcription,carried real-time quantitative PCR,then the expression profiles of Arabidopsis wild type and mutant MAP18 were obtained.The results showed that the expression level of MAP18 in sad2-1,eds1 and nia1nia2 was higher than that in wild type,while that in sad2-2,npr1 and NOA was lower than that in wild type.