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Molecular characterization of carotenoid biosynthetic genes and carotenoid accumulation in Scutellaria baicalensis Georgi.

Tuan PA, Kim YB, Kim JK, Arasu MV, Al-Dhabi NA, Park SU - EXCLI J (2014)

Bottom Line: SbCHXB and SbZEP transcripts were expressed at relatively high levels in the roots, stems, and flowers and were expressed at low levels in the leaves, where carotenoids were mostly distributed.The predominant carotenoids in S.b aicalensiswere lutein and β-carotene, with abundant amounts found in the leaves (517.19 and 228.37 μg g(-1) dry weight, respectively).Our study on the biosynthesis of carotenoids in S. baicalensis will provide basic data for elucidating the contribution of carotenoids to the considerable medicinal properties of S. baicalensis.

View Article: PubMed Central - PubMed

Affiliation: Department of Crop Science, Chungnam National University, 99 Daehak-Ro, Yuseong-gu, Daejeon 305-764, Korea.

ABSTRACT
Scutellaria baicalensis has a wide range of biological activities and has been considered as an important traditional drug in Asia and North America for centuries. A partial-length cDNA clone encoding phytoene synthase (SbPSY) and full-length cDNA clonesencoding phytoene desaturase (SbPDS), ξ-carotene desaturase (SbZDS), β-ring carotene hydroxylase (SbCHXB), and zeaxanthin epoxidase (SbZEP)were identifiedin S. baicalensis. Sequence analyses revealed that these proteins share high identity and conserved domains with their orthologous genes. SbPSY, SbPDS, SbZDS, SbCHXB, and SbZEP were constitutively expressed in the roots, stems, leaves, and flowers of S.b aicalensis. SbPSY, SbPDS, and SbZDS were highly expressed in the stems, leaves, and flowers and showed low expression in the roots, where only trace amounts of carotenoids were detected. SbCHXB and SbZEP transcripts were expressed at relatively high levels in the roots, stems, and flowers and were expressed at low levels in the leaves, where carotenoids were mostly distributed. The predominant carotenoids in S.b aicalensiswere lutein and β-carotene, with abundant amounts found in the leaves (517.19 and 228.37 μg g(-1) dry weight, respectively). Our study on the biosynthesis of carotenoids in S. baicalensis will provide basic data for elucidating the contribution of carotenoids to the considerable medicinal properties of S. baicalensis.

No MeSH data available.


Primers used for real-time PCR
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T1: Primers used for real-time PCR

Mentions: QRT-PCR primers amplified five target carotenoid biosynthetic genes and Actin housekeeping gene (HQ847728) of S. baicalensis were designed using the Primer3 website (http://frodo.wi.mit.edu/primer3/) (Table 1(Tab. 1)) and then tested for the specificity by PCR. For quantification of standard, the PCR products amplified from cDNA were purified, and the concentration of the products was measured to calculate the number of cDNA copies. The expression of five carotenoid biosynthetic genes was calculated by the method of relative quantification to Actin as the reference. Real-time PCR reactions were carried out in a 20-µl reaction mix containing 5 µl of template cDNA, 10 µl of 2× SYBR Green Realtime PCR Master Mix (Toyobo, Japan), 0.5 µl of each primer (10 µM), and DEPC water. Thermal cycling conditions were as follows: 95 °C for 5 min; 40 cycles of 95 °C for 15 s, 56 °C for 15 s, 72 °C for 20 s. PCR products were analyzed using Bio-Rad CFX Manager 2.0 software. Three replications for each sample were used for real-time analysis.


Molecular characterization of carotenoid biosynthetic genes and carotenoid accumulation in Scutellaria baicalensis Georgi.

Tuan PA, Kim YB, Kim JK, Arasu MV, Al-Dhabi NA, Park SU - EXCLI J (2014)

Primers used for real-time PCR
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4556017&req=5

T1: Primers used for real-time PCR
Mentions: QRT-PCR primers amplified five target carotenoid biosynthetic genes and Actin housekeeping gene (HQ847728) of S. baicalensis were designed using the Primer3 website (http://frodo.wi.mit.edu/primer3/) (Table 1(Tab. 1)) and then tested for the specificity by PCR. For quantification of standard, the PCR products amplified from cDNA were purified, and the concentration of the products was measured to calculate the number of cDNA copies. The expression of five carotenoid biosynthetic genes was calculated by the method of relative quantification to Actin as the reference. Real-time PCR reactions were carried out in a 20-µl reaction mix containing 5 µl of template cDNA, 10 µl of 2× SYBR Green Realtime PCR Master Mix (Toyobo, Japan), 0.5 µl of each primer (10 µM), and DEPC water. Thermal cycling conditions were as follows: 95 °C for 5 min; 40 cycles of 95 °C for 15 s, 56 °C for 15 s, 72 °C for 20 s. PCR products were analyzed using Bio-Rad CFX Manager 2.0 software. Three replications for each sample were used for real-time analysis.

Bottom Line: SbCHXB and SbZEP transcripts were expressed at relatively high levels in the roots, stems, and flowers and were expressed at low levels in the leaves, where carotenoids were mostly distributed.The predominant carotenoids in S.b aicalensiswere lutein and β-carotene, with abundant amounts found in the leaves (517.19 and 228.37 μg g(-1) dry weight, respectively).Our study on the biosynthesis of carotenoids in S. baicalensis will provide basic data for elucidating the contribution of carotenoids to the considerable medicinal properties of S. baicalensis.

View Article: PubMed Central - PubMed

Affiliation: Department of Crop Science, Chungnam National University, 99 Daehak-Ro, Yuseong-gu, Daejeon 305-764, Korea.

ABSTRACT
Scutellaria baicalensis has a wide range of biological activities and has been considered as an important traditional drug in Asia and North America for centuries. A partial-length cDNA clone encoding phytoene synthase (SbPSY) and full-length cDNA clonesencoding phytoene desaturase (SbPDS), ξ-carotene desaturase (SbZDS), β-ring carotene hydroxylase (SbCHXB), and zeaxanthin epoxidase (SbZEP)were identifiedin S. baicalensis. Sequence analyses revealed that these proteins share high identity and conserved domains with their orthologous genes. SbPSY, SbPDS, SbZDS, SbCHXB, and SbZEP were constitutively expressed in the roots, stems, leaves, and flowers of S.b aicalensis. SbPSY, SbPDS, and SbZDS were highly expressed in the stems, leaves, and flowers and showed low expression in the roots, where only trace amounts of carotenoids were detected. SbCHXB and SbZEP transcripts were expressed at relatively high levels in the roots, stems, and flowers and were expressed at low levels in the leaves, where carotenoids were mostly distributed. The predominant carotenoids in S.b aicalensiswere lutein and β-carotene, with abundant amounts found in the leaves (517.19 and 228.37 μg g(-1) dry weight, respectively). Our study on the biosynthesis of carotenoids in S. baicalensis will provide basic data for elucidating the contribution of carotenoids to the considerable medicinal properties of S. baicalensis.

No MeSH data available.