Abstract
Culinary rhubarb is a popular vegetable crop, valued for its long, thickened stalks, very rich in different natural bioactive ingredients. Tissue cultures are a useful tool for vegetative propagation of virus-free rhubarb plants and rapid multiplication of valuable selected genotypes. The aim of this study was to develop an effective method for in vitro propagation of selected genotypes of Polish rhubarb ‘Malinowy’ characterized by high yield and straight, thick and intensive red stalks. Identification and quantification of anthocyanins and soluble sugars by the HPLC method in shoot cultures and ex vitro established plantlets were also performed. Shoot cultures were established from axillary buds isolated from dormant, eight-year-old rhizomes. Effective shoot multiplication of rhubarb ‘Malinowy’ was obtained in the presence of 6.6 µM benzylaminopurine or 12.4 µM meta-topolin. Both cytokinins stimulated shoot formation in a manner that depended on sucrose concentration. Increasing the sucrose concentration from 59 to 175 mM decreased the production of shoots and outgrowth of leaves by 3-fold but enhanced shoot length, single shoot mass and callus formation at the base of shoots. This coincided with increased accumulation of soluble sugars (fructose, glucose) and anthocyanins-cyanidin-3-O-rutinoside (max. 208.2 mg·100 g−1 DM) and cyanidin-3-O-glucoside (max. 47.7 mg·100 g−1 DM). The highest rooting frequency (94.9%) and further successful ex vitro establishment (100%) were observed for shoots that were earlier rooted in vitro in the presence of 4.9 µM indole-3-butyric acid. Our results indicated that anthocyanin contents in leaf petioles were influenced by developmental stage. Under in vitro conditions, it is possible to elicit those pigments by sucrose at high concentration and meta-topolin.
Highlights
Rhubarb (Rheum) is a herbaceous perennial of the Polygonaceae family
The donor plants were tested by enzyme-linked immunosorbent assay (DAS-ELISA) with commercially available antibodies against Arabis mosaic virus (ArMV), Turnip mosaic virus (TuMV), Cucumber mosaic virus (CMV), Cherry leaf roll virus (CLRV), Tobacco ring spot virus (TRSV), Tomato ring spot virus (ToRSV), Tomato spotted wilt virus (TSWV), Tomato black ring virus (TBRV), Strawberry latent ringspot virus (SLRV) (LOEWE Biochemica, Sauerlach, Germany) and Tobacco mosaic virus (TMV) (Agdia, Elkhart, IN) by the method of
We developed a practical protocol for mass propagation of the selected genotype of rhubarb with a high content of anthocyanins
Summary
Rhubarb (Rheum) is a herbaceous perennial of the Polygonaceae family. Rheum includes about 60 species, and most of them are native to the northern and central regions of Asia [1]. Rheum has been cultivated in China for medicinal purposes. The dried rhizomes and roots of medicinal species (R. palmatum L., R. officinale Baill, R. tanguticum Maxim.) have been used to treat constipation, inflammation and ulcers [2,3]. Culinary rhubarb is popular as a vegetable crop, valued for its long, thickened stalks [4]. They are used in the production of desserts, cakes, jam, juices, fruit teas and wine. The use of rhubarb petioles for food was discovered at the beginning of the 18th century in Great Britain, but widespread consumption of rhubarb stalks began in the early
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