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Plant J ; 108(1): 81-92, 2021 10.
Article in English | MEDLINE | ID: mdl-34273198

ABSTRACT

Steroidal glycoalkaloids (SGAs) are toxic specialized metabolites found in members of the Solanaceae, such as Solanum tuberosum (potato) and Solanum lycopersicum (tomato). The major potato SGAs are α-solanine and α-chaconine, which are biosynthesized from cholesterol. Previously, we have characterized two cytochrome P450 monooxygenases and a 2-oxoglutarate-dependent dioxygenase that function in hydroxylation at the C-22, C-26 and C-16α positions, but the aminotransferase responsible for the introduction of a nitrogen moiety into the steroidal skeleton remains uncharacterized. Here, we show that PGA4 encoding a putative γ-aminobutyrate aminotransferase is involved in SGA biosynthesis in potatoes. The PGA4 transcript was expressed at high levels in tuber sprouts, in which SGAs are abundant. Silencing the PGA4 gene decreased potato SGA levels and instead caused the accumulation of furostanol saponins. Analysis of the tomato PGA4 ortholog, GAME12, essentially provided the same results. Recombinant PGA4 protein exhibited catalysis of transamination at the C-26 position of 22-hydroxy-26-oxocholesterol using γ-aminobutyric acid as an amino donor. Solanum stipuloideum (PI 498120), a tuber-bearing wild potato species lacking SGA, was found to have a defective PGA4 gene expressing the truncated transcripts, and transformation of PI 498120 with functional PGA4 resulted in the complementation of SGA production. These findings indicate that PGA4 is a key enzyme for transamination in SGA biosynthesis. The disruption of PGA4 function by genome editing will be a viable approach for accumulating valuable steroidal saponins in SGA-free potatoes.


Subject(s)
4-Aminobutyrate Transaminase/metabolism , Solanine/analogs & derivatives , Solanum tuberosum/genetics , 4-Aminobutyrate Transaminase/genetics , Gene Editing , Hydroxylation , Ketocholesterols/biosynthesis , Ketocholesterols/chemistry , Plant Proteins/genetics , Plant Proteins/metabolism , Plant Tubers/enzymology , Plant Tubers/genetics , Plant Tubers/physiology , Saponins/biosynthesis , Saponins/chemistry , Solanine/chemistry , Solanine/metabolism , Solanum tuberosum/enzymology , Solanum tuberosum/physiology
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