Molecular characterization and functional analysis of Glycine max sterol methyl transferase 2 genes involved in plant membrane sterol biosynthesis.
Sterol C24 methyltransferase (SMT2) genes governing the pattern of phytosterols synthesized in higher plants have been studied in Glycine seedlings and wild-type and engineered Arabidopsis thaliana plants. The SMT2 genes of soybean (SMT2-1 and SMT2-2) previously cloned and characterized (Neelakandan et al. 2009) were shown to complement the SMT deficient cvp1 mutant Arabidopsis plants, consistent with their role in regulation of 24-alkyl sterol-controlled plant physiology. Further analysis of these genes showed that environmental cues, including dehydration, cold, and abscisic acid induced differential changes in transcript levels of the SMT2 during soybean seedling growth. Sterol analyses of transgenic Arabidopsis seeds originating in variant constructs of AtHMGR1, GmSMT1, and GmSMT2 engineered in seeds showed relevant modifications in the ratio of 24-methyl to 24-ethyl sterol in the direction of sitosterol formation. To provide insight into the structural features of the sterol gene that affects transcript regulation, the upstream promoter sequences of soybean SMT2 genes were cloned and characterized. Sequence analysis revealed several important cis-elements and transcription factor binding sites. The analysis of promoter-GUS fusions in transgenic Arabidopsis plants revealed shared and distinct expression features in different developmental stages and tissues. The data are interpreted to imply that SMT2 is an important contributor to normal plant growth and development.
Neelakandan
Anjanasree K
AK
Division of Plant Sciences and National Center for Soybean Biotechnology, University of Missouri, Columbia, MO 65211, USA.
Nguyen
Hanh T M
HT
Kumar
Rajesh
R
Tran
Lam-Son Phan
LS
Guttikonda
Satish K
SK
Quach
Truyen Ngoc
TN
Aldrich
Donovan L
DL
Nes
W David
WD
Nguyen
Henry T
HT
D000040:Abscisic Acid / Q000494:pharmacology; D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000235:genetics; D003001:Cloning, Molecular; D003080:Cold Temperature; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D008780:Methyltransferases / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D008969:Molecular Sequence Data; D010840:Phytosterols / Q000096:biosynthesis* / Q000737:chemistry; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D030821:Plants, Genetically Modified / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012639:Seeds / Q000378:metabolism; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D020539:Sequence Analysis, Protein; D012965:Sodium Chloride / Q000494:pharmacology; D013025:Glycine max / Q000187:drug effects / Q000201:enzymology* / Q000502:physiology; D013312:Stress, Physiological; D014867:Water / Q000378:metabolism
D010840:Phytosterols; D010940:Plant Proteins; D014867:Water; D012965:Sodium Chloride; D000040:Abscisic Acid; D008780:Methyltransferases