31. Phenotypic analysis of Arabidopsis thaliana arginine-deficient mutants

Main Article Content

Shahbaz Anwar, Muhammad Amjad Ali, Erich Inselsbacher, Krzysztof Wieczorek

Abstract

Arabidopsis thaliana has been successfully used as a model organism for agriculture research. This model plant gives the opportunity to find out novel ways for the manipulation of gene function to improve commercial crop production. To elucidate gene function, reverse genetics got ample attention. Reduced gene expression in model plant and its implications as phenotypic response aids large scale investigation of gene functions. To find out the effect of the reduced arginine content on plant growth and development, we did a complete phenotypic analysis of arginine-deficient mutants. We generated artificial microRNA (amiRNA)-based mutants for two genes in arginine biosynthesis, argininosuccinate synthase (AS) and argininosuccinate lyase (AL). Quantitative real time PCR (qRT-PCR) of mutant plants showed reduced gene expression. In case of AS mutant lines, as3-3 and as2-6 showed almost 60% decrease in gene expression. Whereas two AL mutants al1-5 and al4-3 showed more than 50% decrease in gene expression. Amino acid analyses of mutant plant lines by Ultra High-Performance Liquid Chromatography (UPLCTM) showed reduced arginine production as compared to wild type plants. Arginine content of 20-day-old as2-6 and al4-3 lines was reduced to 46% and 54%, respectively. However, in 30-day-old plants arginine levels tend to increase and in as2-6 and al4-3 lines the arginine content was 50% and 57% respectively, finally, the phenotypic analysis revealed altered rosette growth and delayed flowering response in mutant plants as compared to the wild type plants. Our results provide insights on the role of arginine metabolism in plant rosette growth and initiation of flowering.


Keywords: Arabidopsis thaliana; Arginine-deficient mutants; Argininosuccinate synthase; Argininosuccinate lyase; Phenotypic analysis; Quantitative RT-PCR


http://dx.doi.org/10.19045/bspab.2022.110032

Most read articles by the same author(s)

Obs.: This plugin requires at least one statistics/report plugin to be enabled. If your statistics plugins provide more than one metric then please also select a main metric on the admin's site settings page and/or on the journal manager's settings pages.