Effects of 4-CPPU on in vitro multiplication and sustainable generation of Hibiscus rosa-sinensis L. ‘White Butterfly’

Abstract

There are more than nine thousand cultivars of Hibiscus rosa-sinensis L., with a series of flowers with shapes, colors and new cultivars continues as generated through both traditional and modern breeding techniques. In this study, advanced biotech methods of in vitro culture have been used to identify a technique for the efficient mass multiplication of H. rosa-sinensis ‘White Butterfly’, using phenyl urea, N-(2-Chloro-4-pyridyl)-N′-phenylurea (4-CPPU). For the first time, the effects of 4-CPPU for stimulating axillary shoot proliferation and multiple shoot regenerations from nodal explants were evaluated, and the optimal nutrient media deduced. From the diverse concentrations as 0.1, 0.5, 2.5, 5.0 & 10.0 µM of 4-CPPU, the highest frequency of shoots was recorded at 2.5 µM supplied in Murashige and Skoog (MS, pH-5.8) medium. After eight-weeks of culture, on an average of 6.7 shoot were obtained on this media with shoot heights of 4.2 cm from each explant. With the involvement of 0.5 µM-IBA (indole-3-butyric acid) in MS medium the regenerated shoots were rooted and followed by successful acclimation to ex vitro conditions. The ploidy consistency among the micro-plants was analyzed using flow cytometry and compared with ex vitro grown plants. No differences in the ploidy levels were observed among the 4-CPPU induced plants, when compared with the donor plants.

Preliminary study on in vitro shoot culture of Hibiscus coddii subsp. barnardii, an indigenous South African flowering plant

“…This response might be linked to the excessive callus formation on explants from these BAP treatments that could lead to limited nutrient supply and subsequent death of explants. Faisal et al (2020) also reported that high cytokinin concentrations stimulated callus growth and decreased the regeneration potential of H. rosa-sinensis explants. More frequent transferring onto fresh MS medium could alleviate the problem (Bhalla et al, 2009;Davies et al, 2017).…”

“…Cytokinins are known to enhance cell division, cell elongation and induction of shoots from the nodal meristem of explants cultured in vitro (Sami et al, 2016;Wybouw and De Rybel, 2019). 6-Benzyl-aminopurine (BAP) is reported to be the most effective cytokinin for shoot induction, since it can cause proliferation of more axillary buds in a shorter period of time (Bhalla et al, 2009;Bekircan et al, 2018;Faisal et al, 2020) than others. Micropropagation studies of some Hibiscus species, such as Hibiscus sabdariffa, H. moscheutos and H. rosa-sinensis showed that the shoot induction response depended on the BAP concentration, plant species and explant type (Kumar et al, 2016;Faisal et al, 2020;Lobodina et al, 2020).…”

“…6-Benzyl-aminopurine (BAP) is reported to be the most effective cytokinin for shoot induction, since it can cause proliferation of more axillary buds in a shorter period of time (Bhalla et al, 2009;Bekircan et al, 2018;Faisal et al, 2020) than others. Micropropagation studies of some Hibiscus species, such as Hibiscus sabdariffa, H. moscheutos and H. rosa-sinensis showed that the shoot induction response depended on the BAP concentration, plant species and explant type (Kumar et al, 2016;Faisal et al, 2020;Lobodina et al, 2020). For instance, Ayadi et al (2011) reported a better shoot induction response when Hibiscus cannabinus explants were cultured on media without cytokinins.…”

Micropropagation of Hardy Hibiscus (Hibiscus Moscheutos L.)

“…The genus Hibiscus has more than 250 species, including annual and perennial herbaceous forms and ending with small woody shrubs [1]. Representatives of various species of the genus Hibiscus, due to their attractiveness, are used for landscaping for various purposes, including in coastal areas with a high salt content, and some species are used as raw materials for the preparation of medicines [2][3][4].…”

“…The publications on clonal micropropagation of hibiscus in vitro are extremely rare. There are some results on the reproduction of Hibiscus syriacus L. [12], Hibiscus sabdariffa L. [13], Hibiscus srosa-sinensis L. [1,14]. However, there is no information on in vitro cultivation of Hardy hibiscus.…”