Preliminary Research On The Expression Pattern And Function Of AtPRGL In Arabidopsi Thaliana

AtPRGL (At5g14920) is an unknown gene which has two domains GASA andPRP. In this study, we conducted primarily research on the expression pattern andfunction of AtPRGL (At5g14920) in Arabidopsis wild type (Col) and Salk₀54982,Salk₀69495,Salk₁12197 (mutated by T-DNA insertion). The details are as follows:1. We have confirmed that three T-DNA insertions (Salk- 054982, Salk-069495,Salk-112197) were located at different sites, and obtained the homozygous lines bymeans of Kan resistance selection and PCR with 3 pairs of related primers.Overexpression vector has also been constructed and transformed into wild type inorder to obtain the homozygous AtPRGL overexprssion lines and analyze thephenotype with the mutants and wild type.2. There were no obvious morphological differences between mutants and wildtype under nomarl green house conditions. It was interesting that the germination ratioof mutants were lower than wild type when germinated on MS with 5μM PAC, butdid not have difference when germinated on MS. The seeds which couldn'tgerminated on MS with 5μM PAC also could germinate on MS,MS+PAC (5μM),MS+GA (10μM). When replaced the seeds that did not germinated on MS with 5μMPAC on MS or MS+GA, the rescuing germination ratio of Col is higher than themutants.3. Roots, inflorescence stems, rosette leaves, cauline leaves and flowers ofseedlings under LD condition were used to conduct RT-PCR. The results indicatedAtPRGL expressed in all the organs tested above, and the AtPRGL mRNA abundancein rosette leaves was the highest and with the lowest expression in roots. AtPRGLpromoter/β-glucuronidase fusion has been constructed and transformed into the wildtype. GUS histochemical analysis showed AtPRGL expressed in root apex, hypocotyls,the vein of leaves and petals, inflorescence stems, stamen filaments. The intensity ofGUS staining in cotyledon and hypocotyl was enhanced after treated with GA and wasweakened after treated with PAC, which indicated that AtPRGL expression may be induced by GA₃. The intensity of GUS staining in stem was enhanced after hurted instem, and there is no obvious change of the intensity of GUS staining in all leavessuch as roette leaves, cauline leaves when hurted.4,To study the subcellular localization of AtPRGL, AtPRGL:: GFP (greenfluorescent protein) fusion has been constructed for transformation into wild typeArabidopsis. Kan resistance plants have been obtained.