Isolation and Homology Analysis of Alanine Aminotransferase Gene of Barley, Foxtail Millet, Cucumber, and Tomato

Abstract

<p>Overexpression of <em>alanine aminotransferase<strong> </strong></em>(<em>AlaAT</em>) gene can improve nitrogen use efficiency (NUE) in plants. The previous isolated <em>AlaAT</em> genes cannot be freely applied to generate NUE plants due to IPR restriction. Therefore, isolation of the gene from targeted monocot and dicot plants is necessary. The objectives of this study were to isolate <em>AlaAT</em> genes from barley, foxtail millet, cucumber, and tomato and analyze their homology to other isolated <em>AlaAT</em> genes in sequence databases (gene bank). Total RNA was isolated from roots of barley, foxtail millet, cucumber, and tomato, and then converted into cDNA using reverse transcription method. The cDNA was then cloned into <em>pGEM®-T Easy</em> plasmid and the verified clones were sequenced. The amino acid sequences were analyzed for their homologies using Clustal Omega software and phylogenetic tree was constructed. The results showed that the amino acids of <em>AlaAT</em> gene from barley was different from <em>AlaAT</em> genes of tomato and cucumber with homology level less than 80%. Phylogenetic tree predicted that <em>AlaAT</em> genes clustered into three groups with <em>AlaAT</em> genes of foxtail millet and barley clustered in one group together with other monocots in group I. <em>AlaAT</em> genes derived from dicots clustered into two groups<em> </em>in which<em> AlaAT</em> gene of tomato clustered in group II, while that of cucumber was in group III. The identity differences of <em>AlaAT</em> gene of tomato and that of cucumber as well as the estimates of the same enzymatic functions can open up enormous opportunities in genetic engineering research for the development of NUE rice.</p>

Assessment of zygosity and nitrogen used efficacy in Mekongga transgenic rice

“…AlaAT genes from tomato (LeAlaAT) and cucumber (CsAlaAT2) have been successfully isolated by Sisharmini et al (2020). The transformation of the LeAlaAT and CsAlaAT2 genes under the control of tissue-specific promoter OsAnt1 improved the NUE of transgenic rice.…”