Functional Analysis Of GsALS3 Of Glycine Soja

Acid soils significantly limit soybean yield because of Aluminum toxicity.Wild soybean has broader genetic background compared with cultivated soybean,and provide more candidate gene for soybean improvement.In this study,ALS3-like genes was cloned from the wild soybean cultivar Bw69 and named as GsALS3.We overexpressed GsALS3 in the Arabidopsis wild type and cultivated soybean cultivar “Huachun6” to analyze their regulatory mechanism of plant adaption to acid soils.The results were shown as follows:ALS3 plays an important role in the adaptation to the low pH and aluminum toxicity.We got ALS3 homologous genes by BLAST in the NCBI and named as GsALS3.The full length of cDNA was 1163 bp,including 876 bp CDS and encoding 291 amino acids.Bioinformatic analysis showed that Gs ALS3 has high homology to ALS3.Diverse cis-acting promoter elements involved in light,heat,temperature,salt tolerance were discovered in the1500 bp upstream region of GsALS3,such as ACE,HSE,P-box,G-box and other elements.Protein structure prediction showed that it has transmembrane but did not have region signal-peptide.Predict of subcellular localization showed that GsALS3 protein is located in the plasma membrane.Predict of SMART showed that GsALS3 interact with STOP1?The tissue localization results of qRT-PCR showed that GsALS3 was highly expressed in stem and compound leaf.Under control condition,GsALS3 was found to express higher in 2-6cm root tip.However,in the Al stress condition,there was high expression in 0-2cm root tip.The transcription abundance of GsALS3 in the transgenic soybean was affected by aluminumion concentration,and when the Al concentration was 50?mol/L,transgenic soybean has highest expression.Temporal expression patterns of GsALS3 showed that GsALS3 expression has highest expression in 12 h.The CDS ALS3-like fragments were cloned by PCR and constructed into the vector pTP101.1.The recombinant overexpression vector was transformed into soybean cotyledonby Agrobacterium EHA101-mediated method,and then we got three T0 transgenic.In this study,transgenic soybean and “Huachun6” were grown in the Aluminum treatments.The important root traits,including total root length,root surface area,total root volume,relative root elongation were investigated.Then we found that traits of transgenic soybean both increased,and Gs ALS3 in transgenic soybean enhance aluminum tolerance.In summary,we have preliminarily elucidated the functional of GsALS3 which may be involved in the molecular mechanism of Glycine Soja aluminum tolerance.This finding provides a theoretical basis to select and improve soybean genotypes adapting to acid soils.