Isolation and Identification of the Phenols of Paul's Scarlet Rose Stems and Stem-Derived Suspension Cultures

Abstract

The phenols of Paul's Scarlet rose stems and stem-derived cell cultures have been analyzed using Cl,rreversed-phase high performance liquid chromatography.Rose stems were found to contain pllic acid, (+)catechin, (-)epicatechin, the dimers (-)epicatechin-(+)catechin and (+)catechin-(+)catechin, a polymeric procyanidin, ferulic acid, and several gallotannins. In contrast, a cell suspension of Paul's Scarlet rose which has been maintained in culture for over 25 years contained only low levels of gallic acid and (-)epicatechin(+)catechin. The phenol content of a second rose cell line which was started from the same initial isolate in 1957, but which was maintained in a laboratory other than our own was quantitatively and qualitatively similar to the cell line kept in our laboratory for the last 20 years. A third cell line which we started 6 months ago contained a wide variety of phenols, most of which were in common with those of rose stems.Selective subculturing of smaller cell clumps of our oldest cell line failed to enhance either the quantities or the diversity of phenols which accumulated in these cultured cells. Possible reasons for the failure of selective subculturing to enhance phenol levels in this long-established cell line are discussed.Several investigators have studied the synthesis of phenols in cultures of Paul's Scarlet rose (1,6,7,15,16,18); however, only tentative identifications were made of a few of the phenols, and no effort was made to compare the phenol content of cultured cells with that of intact rose plants. In Stem tissue used for phenol analysis was removed from a 14-year-old Paul's Scarlet rose bush. Tissue designated as 'young stem' tissue in this report represents new spring growth removed from the tips of older canes, whereas 'old stem' tissue represents 0.5-to 1.5-cm diameter sections of stem, removed from lower portions of older canes which had experienced considerable secondary growth.Isolation of Phenols. Phenols from either rose stem or suspension cultures were extracted into 70% (v/v) acetone (grinding ratio 1 g:2 ml) using a mortar and pestle. The grindings were centrifuged at 20,000g for 10 min and the resulting pellets were discarded. Acetone was removed from the supernatants under vacuum at 30°C. The concentrated extract was collected and the evaporation flask was rinsed with 2.5 ml of 25% methanol. The extract and wash were then pooled. Stem samples were partitioned three times against petroleum ether to remove Chl, with the petroleum ether fractions being subsequently discarded. (Preliminary testing indicated that the omission of the petroleum ether partitioning step for the achlorophyllous cell cultures had no effect on the phenol analysis of this tissue source; consequently, this step was omitted from the phenol isolation procedure for cultured cells). The concentrated extracts plus washes were then passed through a Teflon filter (Millipore Corp., pore size = 45 ism) and a Water Associates C,8 Sep-Pack cartridge.The cartridge was subsequently washed with 2 to...

LC‐DAD‐ESI‐MS and HPLC‐DAD phytochemical investigation and in vitro antioxidant assessment of Rosa sp. stem pruning products from different northern areas in Tunisia

“…Very little is known about the phenolic profile and content of stems from Rosa spp. ; Muhitch and Fletcher in 1984 had indicated the presence of several flavan‐3‐ols in the stems of Paul's Scarlet Rose, whilst Riffault et al ., 30 years later (2014), identified around 60 compounds in different organs from Rosa hybrida cv. “Jardin de Granville”, with catechin and several quercetin derivatives as the main compounds present in the stems.…”

Rosa spp. (Roses): In Vitro Culture, Micropropagation, and the Production of Secondary Products

“…Therefore it would seem that extending the life Fletcher (1984, 1985) Mubitch and Fletcher (1984and Fletcher ( , 1985 Mubitch and Flecther (1984) Amorim et al (1977) Mubitch and Fletcher (1984) Wegg and Townsley (1983) span of mature nondividing cell cultures offers a means of increasing the yield of secondary products.…”

Natural products from plant tissue culture

“…Cultures of Derris elliptica rapidly lost their ability to accumulate rotenoids, but a low ( 1 60 pg per gram of dry weight) level of production resumed when root differentiation was i n d ~c e d . " ~ Both rotenone (90) and deguelin (91) could be identified. By comparison, main roots in the plant accumulate > 8 mg of rotenoids per gram of dry weight.…”