Overexpression of 3β-hydroxysteroid dehydrogenases in Digitalis lanata shoot cultures

2021 ◽  
Author(s):  
T Leykauf ◽  
J Klein ◽  
M Lödel ◽  
W Kreis ◽  
J Munkert
1993 ◽  
Vol 48 (9-10) ◽  
pp. 713-721 ◽  
Author(s):  
Ursel Stuhlemmer ◽  
Werner Haussmann ◽  
Frank Milek ◽  
Wolfgang Kreis ◽  
Ernst Reinhard

Abstract Putative intermediates of cardenolide biosynthesis, namely progesterone, pregnenolone, 5β-pregnane-3,20-dione or 5β-pregnan-3β-ol-20-one, were administered to light- or darkgrown shoot cultures of Digitalis lanata. The unsaturated com pounds were reduced to their respective 5 a-pregnanes, 5β-pregnane-3,20-dione was reduced to 5β-pregnan-3α-ol-20-one and 5β-pregnan-3β-ol-20-one was isomerized to the respective 3α-pregnane. Suspension cultures of Digitalis lanata, on the other hand, accumulated both the 3α- and the 3β-isom er of 5β-pregnan-3-ol-20-one when incubated in the presence of 5β-pregnane- 3.20-dione. When 5β-pregnan-3α-ol-20-one was administered the cultured cells accumulated large amounts of the 3β-isomer together with small amounts of 5β-pregnane-3,20-dione, which may be regarded as an intermediate during the isomerization reaction. Cell-free, buffered extracts from light-grown shoots were shown to reduce 5β-pregnane- 3.20-dione almost exclusively to 5β-pregnan-3α-ol-20-one when 0.05 m MgCl2 were present in the incubation mixture. Under these conditions the formation of 5β-pregnan-3β-ol-20-one was inhibited. The enzyme activity could be recovered from m em brane-free supernatants. Optimum enzyme activity occurred at pH 7.0 and 42 °C. The energy of activation was 56.2 kJ/mol and the enzyme reaction was found to be NADPH -dependent. SH reagents were essential for enzyme activity. The enzyme seems to be specific for 5β-pregnan-3-ones since neither 5 a-pregnane-3-ones nor Δ4/Δ5-pregnenes were reduced. The NADPH : 5β-pregnane 3α-hydroxysteroid-5β-oxidoreductase described here may play a role in the regulation of cardenolide biosynthesis by removing precursors, such as 5β-pregnane-3,20-dione, from the pathway


1999 ◽  
Vol 37 (1) ◽  
pp. 13-23 ◽  
Author(s):  
Marina Eisenbeiß ◽  
Wolfgang Kreis ◽  
Ernst Reinhard

HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 506d-506
Author(s):  
Robert R. Tripepi ◽  
Holly J. Schwager ◽  
Mary W. George ◽  
Joseph P. McCaffrey

Two insecticides, acephate or azadirachtin, were added to tissue culture media to determine their effectiveness in controlling onion thrips (Thrips tabaci Lindeman.) and to determine if these insecticides could damage the plant shoot cultures. To test for insecticide phytotoxicity, microshoots from European birch (Betula pendula), American elm (Ulmus americana), `Pink Arola' chrysanthemum (Dendranthema grandiflora), `America' rhododendron (Rhododendron catawbiense), `Golden Emblem' rose (Rosa hybrida), and `Gala' apple (Malus domestica) were placed in 130-ml baby food jars containing 25 ml of medium supplemented with 6.5, 13, or 26 mg/l Orthene® (contained acephate) or 0.55, 1.1, or 2.2 ml/l Azatin® (contained azadirachtin). Control jars lacked insecticide. To test for thrips control, 13 mg/l Orthene® or 0.55 ml/l Azatin® was added to Murashige and Skoog medium, and 10 thrips were placed on `Gala' apple microshoots in each jar. Jars were sealed with plastic wrap. In both studies, microshoot dry weight and heights were determined. In the second study, the total number of thrips per jar was also determined 3 weeks after inoculation. Microshoots on Orthene®-treated media lacked phytotoxicity symptoms, regardless of the concentration used. In contrast, Azatin® hindered plant growth, decreasing shoot height or dry weight by up to 85% depending on the species. Both insecticides prevented thrips populations from increasing, since less than 10 thrips were found in jars with insecticide-treated medium. Control jars, however, contained an average of almost 70 thrips per jar. This study demonstrated that both Orthene® and Azatin® were effective for eradicating thrips from plant tissue cultures, but Orthene® should probably be used because Azatin® was phytotoxic to all species tested.


2021 ◽  
Vol 105 (3) ◽  
pp. 1215-1226
Author(s):  
Wojciech Makowski ◽  
Aleksandra Królicka ◽  
Anna Nowicka ◽  
Jana Zwyrtková ◽  
Barbara Tokarz ◽  
...  

Abstract The Venus flytrap (Dionaea muscipula J. Ellis) is a carnivorous plant able to synthesize large amounts of phenolic compounds, such as phenylpropanoids, flavonoids, phenolic acids, and 1,4-naphtoquinones. In this study, the first genetic transformation of D. muscipula tissues is presented. Two wild-type Rhizobium rhizogenes strains (LBA 9402 and ATCC 15834) were suitable vector organisms in the transformation process. Transformation led to the formation of teratoma (transformed shoot) cultures with the bacterial rolB gene incorporated into the plant genome in a single copy. Using high-pressure liquid chromatography, we demonstrated that transgenic plants were characterized by an increased quantity of phenolic compounds, including 1,4-naphtoquinone derivative, plumbagin (up to 106.63 mg × g−1 DW), and phenolic acids (including salicylic, caffeic, and ellagic acid), in comparison to non-transformed plants. Moreover, Rhizobium-mediated transformation highly increased the bactericidal properties of teratoma-derived extracts. The antibacterial properties of transformed plants were increased up to 33% against Staphylococcus aureus, Enterococcus faecalis, and Escherichia coli and up to 7% against Pseudomonas aeruginosa. For the first time, we prove the possibility of D. muscipula transformation. Moreover, we propose that transformation may be a valuable tool for enhancing secondary metabolite production in D. muscipula tissue and to increase bactericidal properties against human antibiotic-resistant bacteria. Key points • Rhizobium-mediated transformation created Dionaea muscipula teratomas. • Transformed plants had highly increased synthesis of phenolic compounds. • The MBC value was connected with plumbagin and phenolic acid concentrations.


Plants ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 670
Author(s):  
Katalin Magyar-Tábori ◽  
Nóra Mendler-Drienyovszki ◽  
Alexandra Hanász ◽  
László Zsombik ◽  
Judit Dobránszki

In general, in vitro virus elimination is based on the culture of isolated meristem, and in addition thermotherapy, chemotherapy, electrotherapy, and cryotherapy can also be applied. During these processes, plantlets suffer several stresses, which can result in low rate of survival, inhibited growth, incomplete development, or abnormal morphology. Even though the in vitro cultures survive the treatment, further development can be inhibited; thus, regeneration capacity of treated in vitro shoots or explants play also an important role in successful virus elimination. Sensitivity of genotypes to treatments is very different, and the rate of destruction largely depends on the physiological condition of plants as well. Exposure time of treatments affects the rate of damage in almost every therapy. Other factors such as temperature, illumination (thermotherapy), type and concentration of applied chemicals (chemo- and cryotherapy), and electric current intensity (electrotherapy) also may have a great impact on the rate of damage. However, there are several ways to decrease the harmful effect of treatments. This review summarizes the harmful effects of virus elimination treatments applied on tissue cultures reported in the literature. The aim of this review is to expound the solutions that can be used to mitigate phytotoxic and other adverse effects in practice.


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