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적색 또는 백색 뽕나무 열매에서 High-Throughput Sequencing을 이용한 Anthocyanin과 Betulinic Acid 생합성의 분자적 특성 이해
Shicheng Zhao,Chang Ha Park,Thanislas Bastin, Baskar,Jin Jeon,Sang Un Park 한국약용작물학회 2016 한국약용작물학술대회 발표집 Vol.2016 No.10
Background : Mulberry (Morus alba L.), renowned for their medicine benefits and the leave as the sole food for silkworm (Bombyx mori). To understanding the molecular mechanism of color formation and nutritive value in different mulberry fruit varieties, we use high-throughput transcriptome sequencing technique to investigated the anthocyanin and betulinic biosynthesis pathway related functional genes. In addition, the total antosyanin and betuinic acid contend were also measured. Methods and Results : The resulting cDNA library was then sequenced using an Illumina HiSeq™ 2000 system. The clean reads were assembled using Trinity software, Then perform gene family clustering to get final unigenes. The pH differential method was used to determine the total anthocyanin content (TAC) of methanol extract from the red and white mulberry, and High-performance liquid chromatography (HPLC) analysis was used to quantify the triterpenes content. In this study, total 50,149 unigenes with an average length of 1,125 nt and N50 equaling 1,861 nt were generated. Using these transcriptome sequecing, cDNAs encoding anthocyanin biosynthetic genes and triterpene biosynthetic genes were isolated. In addition, total anthocyanins and betulinic acid content were analyzed. A great amount of total anthocyanins (59.16 mg/g) were found in fully ripe fruit of Cheongil. Accumulation of betulin and betulinic acid were also detected in all stages of Cheongil and Turkey fruits with small amount. Conclusion : The results of transcriptome sequencing provide useful information at molecular lever in mulberry research, such as interesting gene discovering, marker assisted molecular breeding, and interesting metabolic pathway investigate. The gene expression results could help us understanding of the molecular mechanisms of different fruit color determining factor.
Zhao, Shicheng,Park, Chang Ha,Yang, Jingli,Yeo, Hyeon Ji,Kim, Tae Jin,Kim, Jae Kwang,Park, Sang Un Elsevier 2019 Food chemistry Vol.279 No.-
<P><B>Abstract</B></P> <P>To better understand the molecular mechanism of color formation in different varieties of the mulberry fruit, we investigated the functional genes related to anthocyanin and betulinic acid biosynthesis using high-throughput transcriptome sequencing and detected the primary and secondary metabolites in the white (<I>Morus alba</I> L. cv. ‘Turkey’) and red (<I>Morus alba</I> L. cv. ‘Cheongil’) mulberry cultivars. We obtained 171,702,058 high-quality reads with an average read length of 125 bp. These reads were assembled into 51,272 and 51,159 unigenes in Turkey and Cheongil, respectively. We also identified the genes related to anthocyanin and triterpene biosynthesis and investigated their expression and metabolite profiles. Overall, our transcriptome sequencing provides valuable information that could be used in gene discovery, marker-assisted selection, and investigation of metabolic pathways in mulberry. Additionally, gene expression and metabolite profiles provide new insights into the underlying mechanism of anthocyanin and betulinic acid biosynthesis and relationship between primary and secondary metabolites.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Transcriptomic and metabolic analysis of white and red fruits of <I>M. alba</I>. </LI> <LI> A total of 46 primary and secondary metabolites were detected in fruits of <I>M. alba</I>. </LI> <LI> Anthocyanin and betulinic acid biosynthetic genes were identified in transcriptome. </LI> <LI> Some secondary metabolites were positively correlated with its biosynthetic genes. </LI> </UL> </P>
Park, Chang Ha,Zhao, Shicheng,Yeo, Hyeon Ji,Park, Ye Eun,Baskar, Thanislas Bastin,Arasu, Mariadhas Valan,Al-Dhabi, Naif Abdullah,Park, Sang Un NATURAL PRODUCT COMMUNICATIONS 2017 Natural product communications Vol.12 No.4
<P>Three Agrobacterium rhizogenes strains were tested for their ability to transform the plant Morus alba L. and to induce production of the secondary metabolites betulin and betulinic acid. All the tested strains of A. rhizogenes (R1601, LBA9402 and R1000) were able to induce hairy root formation in leaf tissue explants. Strain LBA9402 had the highest rate of infection (92.7% +/- 8.8%), whereas strain R1601 had the lowest rate (87.4% +/- 9.3%). The highest number of hairy roots per explant (5.6 +/- 0.5) and the greatest root length (2.4 +/- 0.2 mm) were obtained with strain LBA9402. We also evaluated dry weight (a measure of growth) and betulin and betulinic acid production in hairy roots and found that the highest growth (167.8 +/- 14.5 mg/flask) occurred after infection with strain LBA9402. Furthermore the highest production of betulin (5.4 +/- 0.4 mg/g dry weight) and betulinic acid (2.3 +/- 0.2 mg/g dry weight) was noted using strain LBA9402. Among three elicitors, yeast extract showed the highest induction of betulin production (8.7 +/- 0.4 mg/g) and silver nitrate induced the highest yield of betulinic acid (4.1 +/- 0.2 mg/g). Our study showed that A. rhizogenes strain LBA9402 was the most effective of the three tested strains for production of transformed root cultures and betulin and betulinic acid.</P>