Dissection of Tomato Lycopene Biosynthesis through Virus-Induced Gene Silencing

被引:195
作者
Fantini, Elio [1 ,2 ]
Falcone, Giulia [1 ]
Frusciante, Sarah [1 ,3 ]
Giliberto, Leonardo [2 ]
Giuliano, Giovanni [1 ]
机构
[1] Italian Natl Agcy New Technol Energy & Sustainabl, Casaccia Res Ctr, I-00123 Rome, Italy
[2] Genelab, Palazzolo Acreide 96010, SR, Italy
[3] Univ Roma La Sapienza, I-00185 Rome, Italy
关键词
ZETA-CAROTENE DESATURASE; PHYTOENE SYNTHASE GENE; BETA-CAROTENE; CIS ISOMERS; IN-VITRO; FRUIT; PATHWAY; DIVERSIFICATION; IDENTIFICATION; ACCUMULATION;
D O I
10.1104/pp.113.224733
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
Lycopene biosynthesis in tomato (Solanum lycopersicum) fruits has been proposed to proceed through a poly-cis pathway catalyzed by phytoene synthase (PSY), two desaturases (phytoene desaturase [PDS] and zeta-carotene desaturase [ZDS]), and two cis-trans isomerases (zeta-carotene isomerase [ZISO] and prolycopene isomerase [CrtISO]). The mechanism of action of these enzymes has been studied in Escherichia coli, but a systematic study of their in vivo function is lacking. We studied the function of nine candidate genes (PSY1, PSY2, PSY3, PDS, ZDS, ZISO, CrtISO, CrtISO-Like1, and CrtISO-Like2) using virus-induced gene silencing (VIGS) coupled to high-resolution liquid chromatography coupled with diode array detector and mass spectrometry, which allowed the identification and quantitation of 45 different carotenoid isomers, including linear xanthophylls. The data confirm the confinement of the VIGS signal to the silenced fruits and the similarity of the phenotypes of PSY1- and CrtISO-silenced fruits with those of the yellow flesh and tangerine mutants. Light was able to restore lycopene biosynthesis in ZISO-silenced fruits. Isomeric composition of fruits silenced at different metabolic steps suggested the existence of three functional units, comprising PSY1, PDS/ZISO, and ZDS/CrtISO, and responsible for the synthesis of 15-cis-phytoene, 9,9'-di-cis-zeta-carotene, and all-trans-lycopene, respectively. Silencing of a desaturase (PDS or ZDS) resulted in the induction of the isomerase in the same functional unit (ZISO or CrtISO, respectively). All-trans-zeta-carotene was detectable in nonsilenced fruits, greatly increased in ZDS-silenced ones, and disappeared in CrtISO-Like1-/CrtISO-Like2-silenced ones, suggesting the existence of a metabolic side branch, comprising this compound and initiated by the latter enzymes.
引用
收藏
页码:986 / 998
页数:13
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