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Cloning and characterization of an aromatic amino acid aminotransferase (ArAT) gene involved in tropane alkaloid biosynthesis from Hyoscyamus niger
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Xiao LI1, 2, Wei QIANG1, 2, Fei QIU1, 2, Min CHEN3, Xiao-zhong LAN4, Zhi-hua LIAO1, 2, Xiao-qiang LIU1, 2, *
Acta Pharmaceutica Sinica | 2017, 52(1) : 172 - 179
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Acta Pharmaceutica Sinica | 2017, 52(1): 172-179
• ORIGINAL ARTICLES •
Cloning and characterization of an aromatic amino acid aminotransferase (ArAT) gene involved in tropane alkaloid biosynthesis from Hyoscyamus niger
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Xiao LI1, 2, Wei QIANG1, 2, Fei QIU1, 2, Min CHEN3, Xiao-zhong LAN4, Zhi-hua LIAO1, 2, Xiao-qiang LIU1, 2, *
Affiliations
  • 1. Key Laboratory of Eco-environments in Three Gorges Reservoir Region, Ministry of Education of PRC, School of Life Sciences, Southwest University, Chongqing 400715, China
  • 2. SWU-TAAHC Medicinal Plant Joint R & D Centre, Chongqing 400715, China
  • 3. School of Pharmaceutical Sciences, Southwest University, Chongqing 400715, China
  • 4. Agricultural and Husbandry College, Tibet University, Tibet 850000, China
Published: 2017-01-12 doi: 10.16438/j.0513-4870.2016-0601
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Tropane alkaloids are anticholinergic drugs widely used clinically. Biosynthesis of tropane alkaloids in planta involves a step of transamination of phenylalanine. Based on the sequenced transcriptomes of lateral roots and leaves of Hyoscyamus niger, we found three annotated aromatic amino acid aminotransferases, which were respectively named HnArAT1, HnArAT2 and HnArAT3. Sequence analysis showed that HnArAT3 had highest similarity with the reported Atropa belladonna AbArAT4, which was involved in tropane alkaloid (TA) to provide the precursor of the phenyllactic acid moiety. Tissue expression pattern analysis indicated that HnArAT3 was specifically expressed in lateral roots, where is the organ synthesizing tropane alkaloids. Then, method of virus induced gene silencing (VIGS) was used to characterize the function of HnArAT3 in H. niger. Gene expression analysis given by real-time quantitative PCR showed that all the transgenic lines had lower expression levels of HnArAT3 than the non-transgenic control, and HPLC analysis of alkaloids demonstrated significant decrease in the contents of hyoscyamine, anisodamine and scopolamine in planta. These results suggested that HnArAT3 was involved in the phenyllactic acid branch of TA biosynthetic pathway. Molecular cloning and functional identification of HnArAT3 laid the foundation for further understanding of TA biosynthesis and metabolic regulation, and also provided a new candidate gene for engineering biosynthetic pathway of tropane alkaloids.

Hyoscyamus niger  /  aromatic amino acid aminotransferase  /  virus induced gene silencing  /  tropane alkaloids
Xiao LI, Wei QIANG, Fei QIU, Min CHEN, Xiao-zhong LAN, Zhi-hua LIAO, Xiao-qiang LIU. Cloning and characterization of an aromatic amino acid aminotransferase (ArAT) gene involved in tropane alkaloid biosynthesis from Hyoscyamus niger[J]. Acta Pharmaceutica Sinica, 2017 , 52 (1) : 172 -179 . DOI: 10.16438/j.0513-4870.2016-0601
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Article Info
doi: 10.16438/j.0513-4870.2016-0601
  • Receive Date：2016-06-21
  • Online Date：2026-01-14
  • Published：2017-01-12
Article Data
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History
  • Received：2016-06-21
  • Revised：2016-08-04
Funding
Affiliations
    1. Key Laboratory of Eco-environments in Three Gorges Reservoir Region, Ministry of Education of PRC, School of Life Sciences, Southwest University, Chongqing 400715, China
    2. SWU-TAAHC Medicinal Plant Joint R & D Centre, Chongqing 400715, China
    3. School of Pharmaceutical Sciences, Southwest University, Chongqing 400715, China
    4. Agricultural and Husbandry College, Tibet University, Tibet 850000, China
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表12种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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