PCR-based suppression subtractive hybridization analyses of enantioselective gene expression in root tips of wheat treated with optically active urea compounds

• Published in: Pesticide biochemistry and physiology Vol. 98; no. 3; pp. 359 - 369
• Main Authors: Kojima, Hisahiro, Numata, Takako, Tadaki, Ryota, Omokawa, Hiroyoshi
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier Inc 01.11.2010; Elsevier
• Subjects: Acetohydroxyacid synthase; Agronomy. Soil science and plant productions; Biological and medical sciences; Chiral response; Fundamental and applied biological sciences. Psychology; Mode of action; Optically active urea compounds; PCR-based suppression subtractive hybridization; Plant growth regulator; Quantitative real-time RT-PCR; Triticum aestivum; Triticum aestivum L; Chiral response; Optically active urea compounds; Triticum aestivum L; Plant growth regulator; PCR-based suppression subtractive hybridization; Quantitative real-time RT-PCR; Mode of action; Molecular hybridization; Monocotyledones; Enantioselectivity; Suppression; hybridization; Organic nitrogen compounds; Cereal crop; Suppression subtractive hybridization; Gramineae; Chirality; Angiospermae; Plant growth substance; Ureas; Mechanism of action; C3-Type; Apex; PCR-based suppression subtractive; Organic compounds; Vegetals; Root; Pesticides; Gene expression; Polymerase chain reaction; Response; Urea; Analysis; Vegetative apparatus; Optical activity; Spermatophyta; Triticum aestivum
• ISSN: 0048-3575; 1095-9939
• DOI: 10.1016/j.pestbp.2010.08.001

[Display omitted] ► R-/ S-MBTU show dichotomic and enantioselective effects on Gramineae plants. ► S-MBTUs mode of action was explored using SSH and qRT-PCR technique. ► S-MBTU down-regulated gene expression of GS, AHAS, a-TUB2, and ACT, enantioselectively. The optically active urea compounds R- and S-1-α-methylbenzyl-3- p-tolylurea ( R- and S-MBTU) have qualitatively different effects on plant physiology, especially in Gramineae plants. To elucidate the mechanisms by which S-MBTU inhibits root growth of wheat ( Triticum aestivum L. var. Chinese Spring), we used PCR-based suppression subtractive hybridization (SSH) and quantitative real-time RT-PCR (qPCR) analyses. Genes related to amino acid biosynthesis, cell cycle and skeleton, protein synthesis, cell wall biosynthesis, glycolysis, signaling, DNA modification, and detoxification were enantioselectively regulated. These expression profiles suggested that the putative mode of action of S-MBTU is disruption of primary amino acid biosynthesis, especially the glutamine synthetase (GS)- and acetohydroxyacid synthase (AHAS)-enzyme systems, and disruption of cell membrane skeleton homeostasis related to a putative α-tubulin 2 (α-TUB2), a putative β-actin (ACT) and β-tubulin 1 (β-TUB1). Other genes specifically affected by S-MBTU may result in disruption of metabolic processes, leading to inhibition of root growth.