Isolation of Total RNA from a Freshwater Green Alga, Zygnema cruciatum, Containing High Levels of Pigments

Abstract

Conventional methods for the isolation and purification of mRNA from Zygnema were unsuccessful because of its high amount of pigments and RNA interactive molecules. In particular, pigments were difficult to remove using conventional protocols because they interacted with RNA during pulverization of the materials. This resulted in total degeneration of RNA in two to three hours. To alleviate this problem, we developed an isolation method that utilized DEAE-cellulose resin. The pigments bound to DEAE anion exchange resin and separated from the RNA. Purified total RNA showed an yield of 50 µg per 100 mg of tissue with this method. The amplified 2 nd strand cDNA was distributed 300 bp and over.

Unusual phenolic compounds contribute to ecophysiological performance in the purple‐colored green alga Zygogonium ericetorum (Zygnematophyceae, Streptophyta) from a high‐alpine habitat

“…While phenolics are less common in non‐streptophycean green algae, in Zygnematophyceae they have been repeatedly reported (Han et al. , Remias et al. ,b, Pichrtová et al.…”

“…Other abiotic stresses such as desiccation and temperature fluctuations could also be responsible for increasing the content of phenolics, due to their high-antioxidative potential. While phenolics are less common in non-streptophycean green algae, in Zygnematophyceae they have been repeatedly reported (Han et al 2007, Remias et al 2012a,b, Pichrtov a et al 2013.…”

Characterization of an UV- and VIS-absorbing, purpurogallin-derived secondary pigment new to algae and highly abundant in Mesotaenium berggrenii (Zygnematophyceae, Chlorophyta), an extremophyte living on glaciers

“…Pinkish vacuoles are also known from Mesotaenium macrococcum (Ettl & Gärtner, 1995). The tannin-like precipitation capability of phenols from Zygnema cruciatum was depicted by Han et al (2007), who reported that the pigments effectively interact with RNA during grinding, thus denaturising not only proteins but also nucleotides, despite using established extraction protocols. The tannin-like precipitation capability of phenols from Zygnema cruciatum was depicted by Han et al (2007), who reported that the pigments effectively interact with RNA during grinding, thus denaturising not only proteins but also nucleotides, despite using established extraction protocols.…”

Changes in Phenolic Compounds and Cellular Ultrastructure of Arctic and Antarctic Strains of Zygnema (Zygnematophyceae, Streptophyta) after Exposure to Experimentally Enhanced UV to PAR Ratio

“…In contrast, these compounds have been found in members of the Zygnematophyceae [12, 60], which might be explained by their close relationship to land plants [6, 77]. In addition, a high amount of uncharacterised pigments was detected when attempting to isolate RNA from Z. cruciatum [22]. It could be speculated that these pigments were also phenolics, but there are many other compounds in the aqueous phase.…”

“…The UV-screening ability of phenolic substances found in some members of the class Zygnematophyceae has already been discussed [18, 60]. Moreover, high amounts of water-soluble pigments, probably also phenolics, have been reported to occur in Zygnema cruciatum [22], but their role in UV screening has not been tested. In this study, we examined if the production of phenolics in Zygnema is enhanced by experimental UV exposure.…”