Cloning and Characterization of Chalcone Synthase Gene from Pueraria candollei var. mirifica

Wiriyaampaiwong, Piyachat; Thanonkeo, Pornthap; Thanonkeo, Sudarat

doi:10.1016/j.jbiotec.2010.09.724

Cited by 6 publications

(7 citation statements)

References 61 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The leucine zipper motif, L 310 , L 317 , L 319 and L 331 (Claudot et al, 1999) play an important role as the functional active site and was also found in the PcCHS gene product (Wiriyaampaiwong et al, 2012). The results are shown in Figure 3 that all residues remained unaltered in the two sequences of P. lobata and P. mirifica.…”

Section: Amplification Of Chs Genes By Using Rt-pcrmentioning

confidence: 89%

“…Especially, Wani concluded that the two isoforms GaCHS1 (an ORF of 1176 bp) and GaCHS2 (an ORF of 1170 bp) (G. asiatica L) have appeared the differential expression pattern at different stages (Wani et al, 2017). Besides that, the P. mirifica CHS gene, with 1170 bp in length encoding 389 amino acid residues is also demonstrated its characteristics belonging to a multigene family (Wiriyaampaiwong et al, 2012). In a recent study, Suntichaikamolkul and his colleagues (2019) based on transcriptome and phylogenetic analysis of P. mirifica show that two CHS genes (CHS1, CHS2 encoding chalcone synthase) were involved in isoflavone biosynthesis in a total of 14 putative genes (Suntichaikamolkul et al, 2019).…”

Section: Amplification Of Chs Genes By Using Rt-pcrmentioning

confidence: 98%

See 1 more Smart Citation

Study on characterization of chalcone synthase gene from \(\textit{Pueraria lobata}\) and \(\textit{Pueraria mirifica}\) in Vietnam

Hue

Phương

Canh

2021

AJB

View full text Add to dashboard Cite

Two species of genus Pueraria ((Pueraria lobata (synonym: Pueraria montana var. lobata) and Pueraria mirifica (synonym: Pueraria candollei var. mirifica)) are traditional plants used in medicine since ancient times. These plants have been used and became commercially crucial indigenous medicinal plants. Currently, both roots and flowers of P. mirifica are used as a dietary supplement and functional food for women because of their rich source of phytoestrogen and nutrition. However, little information of genes on both species of Pueraria genus (P. lobata and P. mirifica) are known in Vietnam. The purpose of this research is to support more understanding about Chalcone synthase (CHS) genes by determining and sequence analyzing an encoding region of CHS genes that were isolated from P. lobata and P. mirifica. The full-length open reading frame (ORF) sequence CHS was identified with 1170 bp which encodes 389 amino acids by Sanger sequencing. The isolated CHS gene of P. lobata has no difference in sequence with CHS reported on GenBank (D10223.1), whereas a difference of 26 nucleotide positions in CHS sequence of P. mirifica compared with the published gene sequence (JQ409456.1) as consequential having 97.78% genetic similarity. The CHS genes sequence of P. lobata and P. mirifica are homologous with 98.4% because of having 19 nucleotide differences. Chalcone-stilbene synthase N-C terminal, PLN03173, CHS-like, BH0617, fabH are some important domains predicting the CHS genes. Especially, the family signature ‘GVLFGFGPGLTI’ motif of CHS gene as a part of the active-site scaffold contributes to decide the product of cyclization reactions performing the stereochemistry of cyclization which was also observed in P. lobata and P. mirifica, but it was not included for all members in Fabaceae family. With in sillico analysis, the P. lobata and P. mirifica CHS sequences have highly conserved regions to maintain their structure and function, so that it needs further studies to clarify these points.

show abstract

Section: Amplification Of Chs Genes By Using Rt-pcrmentioning

confidence: 89%

Section: Amplification Of Chs Genes By Using Rt-pcrmentioning

confidence: 98%

Study on characterization of chalcone synthase gene from \(\textit{Pueraria lobata}\) and \(\textit{Pueraria mirifica}\) in Vietnam

Hue

Phương

Canh

2021

AJB

View full text Add to dashboard Cite

show abstract

“…The highest root chs gene expression was also observed in another local medicinal plant, Polygonum minus (Roslan et al 2013). Conversely, Pueraria candollei displayed the highest chs expression in the leaves, followed by the roots and stems (Wiriyaampaiwong et al 2012). High chs expression in roots may be attributed to the roots' central role in plant interactions with microorganisms, which include symbiotic, virulent or avirulent pathogens (Dhawale et al 1989).…”

Section: Relative Expression Of the Pks Genes In Different Tissuesmentioning

confidence: 99%

Molecular cloning and characterization of a cDNA encoding a polyketide synthase from Melastoma decemfidum

et al. 2014

View full text Add to dashboard Cite

A famous plant polyketide synthase (PKS) is a chalcone synthase (CHS), which catalyzes the formation of naringenin chalcone, a limiting factor for flavonoid biosynthesis. Flavonoids such as naringenin and kaempferol-3-O-(2 ,6 -di-O-p-trans-coumaroyl glucoside) have been identified from the flower extracts of Melastoma decemfidum RoxB. Therefore, a study was conducted to clone and characterize the pks gene from M. decemfidum. A newly isolated full-length cDNA containing the pks sequence (designated MdCHS; GenBank accession No. KF234569) was found to harbour a 1,355-bp open reading frame encoding 392 amino acids. The deduced MdCHS protein, which had a predicted molecular weight of approximately 42.6 kDa, exhibited 89% sequence identity to CHS from Anthurium andraeanum (GenBank accession No. AY232492) and 91% to those from Camellia sinensis (D26594) and Camellia chekiangoleosa (ADW11243). Southern blot analysis indicated that at least five pks genes are present in M. decemfidum. Additionally, transcription analysis revealed that pks gene expression was higher in roots than in other tissues (shoots, leaves, stems, flower buds and flowers). In conclusion, the characterization and cloning of the MdCHS gene further elucidates its role in flavonoid biosynthesis.

show abstract

“…1) [12]. CHS and IFS have been identified in P. mirifica several years ago [13, 14], however, their enzymatic functions have not been elucidated. Udomsuk et al [15] suggested that miroestrol biosynthesis may share a common pathway with isoflavonoid biosynthesis due to their similar backbone structure.…”

Section: Introductionmentioning

confidence: 99%

Transcriptome analysis of Pueraria candollei var. mirifica for gene discovery in the biosyntheses of isoflavones and miroestrol

et al. 2019

View full text Add to dashboard Cite

BackgroundPueraria candollei var. mirifica, a Thai medicinal plant used traditionally as a rejuvenating herb, is known as a rich source of phytoestrogens, including isoflavonoids and the highly estrogenic miroestrol and deoxymiroestrol. Although these active constituents in P. candollei var. mirifica have been known for some time, actual knowledge regarding their biosynthetic genes remains unknown.ResultsMiroestrol biosynthesis was reconsidered and the most plausible mechanism starting from the isoflavonoid daidzein was proposed. A de novo transcriptome analysis was conducted using combined P. candollei var. mirifica tissues of young leaves, mature leaves, tuberous cortices, and cortex-excised tubers. A total of 166,923 contigs was assembled for functional annotation using protein databases and as a library for identification of genes that are potentially involved in the biosynthesis of isoflavonoids and miroestrol. Twenty-one differentially expressed genes from four separate libraries were identified as candidates involved in these biosynthetic pathways, and their respective expressions were validated by quantitative real-time reverse transcription polymerase chain reaction. Notably, isoflavonoid and miroestrol profiling generated by LC-MS/MS was positively correlated with expression levels of isoflavonoid biosynthetic genes across the four types of tissues. Moreover, we identified R2R3 MYB transcription factors that may be involved in the regulation of isoflavonoid biosynthesis in P. candollei var. mirifica. To confirm the function of a key-isoflavone biosynthetic gene, P. candollei var. mirifica isoflavone synthase identified in our library was transiently co-expressed with an Arabidopsis MYB12 transcription factor (AtMYB12) in Nicotiana benthamiana leaves. Remarkably, the combined expression of these proteins led to the production of the isoflavone genistein.ConclusionsOur results provide compelling evidence regarding the integration of transcriptome and metabolome as a powerful tool for identifying biosynthetic genes and transcription factors possibly involved in the isoflavonoid and miroestrol biosyntheses in P. candollei var. mirifica.

show abstract

Cloning and Characterization of Chalcone Synthase Gene from Pueraria candollei var. mirifica

Cited by 6 publications

References 61 publications

Study on characterization of chalcone synthase gene from \(\textit{Pueraria lobata}\) and \(\textit{Pueraria mirifica}\) in Vietnam

Study on characterization of chalcone synthase gene from \(\textit{Pueraria lobata}\) and \(\textit{Pueraria mirifica}\) in Vietnam

Molecular cloning and characterization of a cDNA encoding a polyketide synthase from Melastoma decemfidum

Transcriptome analysis of Pueraria candollei var. mirifica for gene discovery in the biosyntheses of isoflavones and miroestrol

Contact Info

Product

Resources

About