5.4.99.8 H477N mutated enzyme produces lanosterol instead of cycloartenol 653405 5.4.99.8 H477N produces 88% lanosterol and 12% parkeol 661481, 661939 5.4.99.8 H477N/I481V produces 99% lanosterol and 1% parkeol 661939 5.4.99.8 H477Q mutated enzyme produces parkeol instead of cycloartenol 653405 5.4.99.8 H477Q produces 22% lanosterol, 73% parkeol, and 5% 9-beta-delta-7-lanosterol 661481, 661939 5.4.99.8 H477Q/I481V produces 94% lanosterol and 6% parkeol 661939 5.4.99.8 I481A production of lanosterol, cycloartenol, parkeol, acilleol A and camelliol C 653402 5.4.99.8 I481F inactive 653402 5.4.99.8 I481G production of lanosterol, cycloartenol, parkeol, acilleol A and camelliol C 653402 5.4.99.8 I481L production of cycloartenol and parkeol 653402 5.4.99.8 I481V produces 54% cycloartenol, 25% lanosterol, and 21% parkeol 661481, 661939 5.4.99.8 additional information construction of three RNAi gene-silencing constructs in backbone of RNAi vector pGSA and a full-length overexpression construct. The expression of WsCAS gene is considerably downregulated in stable transgenic silenced Withania somnifera lines compared to non-transformed control. Transgenic plants overexpressing CAS gene display enhanced level of CAS transcript and withanolide content compared to non-transformed controls 749078 5.4.99.8 additional information mutation N478H/V482I in lanosterol synthase is a change of amino acids crucial for lanosterol specificity to the cycloartenol synthase type. Mutant produces 4% lanosterol, 83% parkeol, and 13% cycloartenol from substrate (S)-2,3-epoxysqualene, while wild-type lanosterol synthase produces only lanosterol 682344 5.4.99.8 additional information RNA interference (RNAi) of the cycloartenol synthase (CAS) gene and simultanous recombinant expression of farnesyl diphosphate synthase (FPS) gene in Panax notoginseng cells leads to both higher expression levels of FPS and lower expression levels of CAS compared to the wild-type cells. Transgenic cell lines provide a higher accumulation of total triterpene saponins, and a lower amount of phytosterols compared to wild-type cells. Overexpression of FPS can break the rate-limiting reaction catalyzed by FPS in the triterpene saponins biosynthetic pathway, and inhibition of CAS expression can decrease the synthesis metabolic flux of the phytosterol branch. Thus, more precursors flow in the direction of triterpene synthesis, and ultimately promote the accumulation of Panax notoginseng saponins. Silencing and overexpression of key enzyme genes simultaneously is more effective than just manipulating one gene in the regulation of saponin biosynthesis 748718 5.4.99.8 additional information upon galactose-induced recombinant expression of NtCAS1, yeast cells grown in the presence of exogenous ergosterol display a sterol profile that includes cycloartenol and four other sterols in addition to ergosterol. The major sterol is 24-methylene pollinastanol that accounts for 53% of the total and represents the most probable pathway end-product generated by the yeast steroidogenic machinery, apparently being versatile enough to accept cycloartenol as a substrate. The sterol biosynthetic features of erg7::NtCAS1 are identical to the 9beta,19-cyclopropylsterol biosynthetic segment of protists, algae, or plants. In the presence of ergosterol, erg7 or erg7::NtCAS1 cells display a closely similar growth profile along a range of dilutions, whereas in the absence of ergosterol, only erg7::NtCAS1 is able to grow significantly when compared to yeast mutant erg7 749137 5.4.99.8 additional information virus-induced gene silencing (VIGS) of CAS1 in Nicotianan benthamiana based on gene specific sequences from a Nicotiana tabacum CAS1. A morphogenetic inhibition is developed by PVX::CAS1 Nicotiana benthamiana leaves 749137 5.4.99.8 Y410T produces 65% lanosterol, 2% parkeol, and 33% 9-beta-delta-7-lanosterol 661481, 661939 5.4.99.8 Y410T/H477N/I481V produces 78% lanosterol and 22% 9-beta-delta-7-lanosterol 661481, 661939 5.4.99.8 Y410T/H477Q/I481V produces 78% lanosterol and 22% 9-beta-delta-7-lanosterol 661481, 661939 5.4.99.8 Y410T/I481V mutated enzyme produces lanosterol instead of cycloartenol 653405 5.4.99.8 Y410T/I481V produces 78% lanosterol, less than 1% parkeol, and 22% 9-beta-delta-7-lanosterol 661481, 661939 5.4.99.8 Y532H mutated enzyme produces a mixture of parkeol, achilleol and lanosterol instead of cycloartenol 653405