The WPM medium was also the best for the growth of transformed roots of other plant species like as Trigonella foenum-graecum [ 15 ] or Dracocephalum moldavica [ 16 ]. The transformed roots maintained in SH and B5 media with full and half-strength content of superhero porn and micronutrients were thick with an average root diameter hairy 0.
Generally, exposure to light increased the growth of hairy roots of R. Hairy these media fresh weights of the roots grown under photoperiod were lower than those achieved under darkness.
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However, the differences were not statistically significant at. The physical culture conditions affected also the morphology of R. Greening was absent in the dark-grown roots which were beige.
The previous report results showed that exposure of hairy root hairy to light can induce greening due to enhanced chlorophyll biosynthesis [ 1718 ]. The genetic transformation of the R. PCR analysis was carried out using primers specific to vir G to confirm that hairy roots were not contaminated with A. The detected compounds were identified by comparing their retention times, UV-Vis spectra, and fragmentation patterns in MS spectra with those of caged lesbians reference compounds and the hairy data [ 19 — 22 ].
The UPLC-PDA study showed that the caffeoylquinic acids and their derivatives constitute the major phenolic class occurring in both the hairy roots and the roots of soil-grown plants. Their preliminary identification was facilitated by analysis of structure-diagnostic hierarchical keys proposed by Clifford et al. Finally, the unequivocal identification of compounds 1 — 3 was confirmed by comparison of their retention time and MS data with the commercially available standard of CHA as well as with the qualitative standards of NCHA and CCHA prepared in our laboratory according to Clifford et al.
A comparison of the elution order and the fragmentation patterns of the product ions described above to those reported in the literature for dicaffeoylquinic acids [ 2124 ] suggests that the detected isomers 4 and 11 — 14 were 1,3-; 3,4-; 3,5-; 1,5-; and 4,5- O -dicaffeoylquinic acids, respectively. Finally, compound 4 was compared with the commercial standard of cynarin 1,3-di -O- caffeoylquinic acid. Based on these results, the detected compounds 16 and 17 might be only tentatively identified as two isomers of tricaffeoylquinic acid substituted with an unidentified logan pierce xxx. Due to the lack of suitable reference standards and literature data, the complete identification of these compounds needs isolation and full spectral characterization.
A comparison of these spectral data with those obtained from the authentic standard allowed to confirm the identification of hydroxyecdysone compound 8 Table 3. All compounds 5 — 7 and 9 - 10 were identified as flavonoid hexosides due to neutral losses of mass units in their MS 2 spectra. Likewise, compound 10 was tentatively identified as patuletin hexoside by comparing its UV-Vis spectrum and fragmentation pattern of the aglycone hairy in MS 3 spectrum with the literature [ 22 ].
Compounds 6 and 7 had identical MS profiles: Thus, according to the observed MS 3 fragmentation Table 3the aglycones of compounds 6 and 7 were assigned as quercetin. The comparison of the UV-Vis spectrum of 9 and the fragmentation pattern of its aglycone in MS 3 spectrum with the literature [ 222829 ] indicated the presence of luteolin.
The hairy interest of caffeoylquinic acids and their derivatives is based on their diverse biological activities which include anti-inflammatory, analgesic, antipyretic, and anticarcinogenic effects [ 3031 ]. Caffeoylquinic acids are free radical and metal scavengers and have been shown to modulate the gene expression of antioxidant enzymes [ 32 ].
Hairy root culture for mass-production of high-value secondary metabolites.
Also, they have neuroprotective, neurotrophic [ 33 ], and hepatoprotective activity [ 34 ]. The results indicate that the total concentration of caffeoylquinic acids and their derivatives calculated as the sum hairy compounds 1 — 4 and 11 — 17 Table 4 was about 2-times hairy in hairy roots cultured in photoperiod HR-L The mean individual caffeic acid derivative content followed a similar pattern.
It suggests a regulation response to light of the phenylpropanoid biosynthetic pathway. Asa akira wicked positive effect of light on the biosynthesis of caffeic acid derivatives has been observed in transformed roots of some other plant species, such as Echinacea purpurea [ 35 ] and Cichorium intybus [ 36 ]. Its content ranged from 1. The amounts were much higher than chlorogenic acid level in transformed roots of Echinacea purpurea [ 3537 ], Fagopyrum tataricum hairy 38 ], or Polygonum multiflorum [ 39 ].
In transformed roots of R. The amount of the compound was 3. Additionally, in extracts of HR-L and HR-D four other dicaffeoylquinic acids were found but at considerably lower amounts compared with 3,5- O -dicaffeoylquinic acid Table 4.
In both types of R. The tricaffeoylquinic acids and their derivatives are less common in plants than mono- and dicaffeoyl ones [ 34 ].
This type of isabella soprano sex was earlier identified in other species of family Asteraceae, such as Arnica montana [ 21 ] and Erigeron breviscapus [ 20 ]. To date there have been no reports on tricaffeoylquinic acid production in R. Considerable differences in qualitative and quantitative profiles of phytochemicals between transformed roots and normal roots of soil-grown plants of R.
The production of caffeoylquinic acids and their derivatives was times higher in SR roots than in transformed roots. The most euro milf component of SR roots was chlorogenic acid The SR roots accumulated 17—23 times less of compound 16 tricaffeoylqunic acid derivative than transformed roots, which was dominant component in the latter.
Moreover, 1,4,5-tri- O -caffeoylquinic acid compound 15 Table 4 was identified only in the hairy root cultures. The differences between transformed roots and nontransformed roots of R.
Only hairy roots hairy able to produce the flavonoid glycosides quercetagetin, quercetin, luteolin, and patuletin hexosides when they were cultured in the light conditions. The total flavonoid content in this sample was 2. A comparative study of the ecdysteroids of transformed and normal roots of R. The results of the present study showed that the transformation by A. Differences in the chemical profiles of transformed and normal roots have been also reported in other plant species [ 4041 ] which indicate that the insertion of Ri T-DNA interferes with the biosynthesis of the secondary metabolites.
However, the differences observed between transformed and normal roots of R. The present study demonstrates that hairy roots of R. The establishment of hairy root culture with highly increased levels of tricaffeoylquinic acids and their derivatives observed in the present study indicates that the hairy roots can be used as potential sources of these secondary metabolites instead of the normal roots of soil-grown plants.
This is especially important because tricaffeoylquinic acids have been shown to possess antimutagenic, antihyperglycemic, strong hairy, and radical scavenging effects.
However, it has been found that the tricaffeoylquinic acids have more biological activity rectal rooter 3 mono- and dicaffeoylquinic acid derivatives [ 42 — 44 ]. The antioxidant activity of hairy hairy is currently under investigation. The authors declare that there is no conflict of interests regarding the publication of this paper. BioMed Research International.
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Abstract The aim of the study was to obtain transformed roots of Rhaponticum carthamoides and evaluate their phytochemical profile. Introduction Rhaponticum carthamoides Willd. Material and Methods 2. Induction and Establishment of Hairy Root Culture The leaf explants were wounded with a sterile needle immersed in the bacterial culture.
Table 1: Induction of hairy roots of R. Figure 1: The hairy roots of R. Table 2: Specific pairs of primers used for PCR analysis of transformed roots of R. Figure 2: Comparative growth of R. Means followed by the same letter at the columns individually to fresh and dry weight are not significantly different at the hairy of the Mann-Whitney test. Figure 3: Table 3: Table 4: Figure 4: Sample and peak codes are given in Table 4.
References L. Kokoska and D.
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