Plants from Violaceae produce cyclotides, peptides characterized by a circular peptide backbone and a
cystine knot. This signature motif gives stability that can harness a wide spectrum of biological activities,
with implications in plant defense and with applications in medicine and biotechnology. In the current
work, cyclotide expressing in vitro cultures were established from Viola uliginosa. These cultures are
useful models for studying biosynthesis of cyclotides and can also be used in their production. The cyclotide
expression pattern is shown to be dependent on exogenous plant growth regulators, both on peptide
and gene expression levels. The highest yields of cyclotides were obtained on media containing only a
cytokinin and were correlated with storage material accumulation. Exposure to auxins decreased cyclotide
production and caused shifting of the biosynthesis pattern to root specific cyclotides. The response to
stimuli in terms of cyclotide expression pattern appears to be developmental, and related to polar auxin
transportation and the auxin/cytokinin ratio regulating tissue differentiation. By the use of whole
transcriptome shotgun sequencing (WTSS) and peptidomics, 20 cyclotide sequences from V. uliginosa
(including 12 new) and 12 complete precursor proteins could be identified. The most abundant cyclotides
were cycloviolacin O3 (CyO3), CyO8 and CyO13. A suspension culture was obtained that grew exponentially
with a doubling time of approximately 3 days. After ten days of growth, the culture provided a yield
of more than 4 mg CyO13 per gram dry mass.
พืชจาก Plants from Violaceae produce cyclotides, peptides characterized by a circular peptide backbone and a
เปปไทด์ที่โดดเด่นด้วยกระดูกสันหลังเปปไทด์และวงกลมปมซีสตีน cystine knot. This signature motif gives stability that can harness a wide spectrum of biological activities,
บรรทัดฐานลายเซ็นนี้จะช่วยให้ความมั่นคงที่สามารถใช้ประโยชน์คลื่นความถี่กว้างของกิจกรรมทางชีวภาพที่มีผลกระทบในการป้องกันพืชและการประยุกต์ใช้ในการแพทย์และเทคโนโลยีชีวภาพ with implications in plant defense and with applications in medicine and biotechnology. In the current
work, cyclotide expressing in vitro cultures were established from Viola uliginosa. These cultures are
useful models for studying biosynthesis of cyclotides and can also be used in their production. The cyclotide
expression pattern is shown to be dependent on exogenous plant growth regulators, both on peptide
and gene expression levels. The highest yields of cyclotides were obtained on media containing only a
cytokinin and were correlated with storage material accumulation. Exposure to auxins decreased cyclotide
production and caused shifting of the biosynthesis pattern to root specific cyclotides. The response to
stimuli in terms of cyclotide expression pattern appears to be developmental, and related to polar auxin
transportation and the auxin/cytokinin ratio regulating tissue differentiation. By the use of whole
transcriptome shotgun sequencing (WTSS) and peptidomics, 20 cyclotide sequences from V. uliginosa
(including 12 new) and 12 complete precursor proteins could be identified. The most abundant cyclotides
were cycloviolacin O3 (CyO3), CyO8 and CyO13. A suspension culture was obtained that grew exponentially
with a doubling time of approximately 3 days. After ten days of growth, the culture provided a yield
of more than 4 mg CyO13 per gram dry mass.
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