Manipulating Some Culturing Conditions Enhances Production of Solanine in Microshoots, Callus and Cell suspension Cultures of Solanum nigrum L.: A Wild Medicinal Plant

Abstract

Solanum nigrum L. is a medicinal plant of solanaceae family with distinguished therapeutic properties. Traditionally, S. nigrum. had been used as an anti-tumorgenic, antioxidant, hepatoprotective, diuretic, and antipyretic agent. The most important alkaloid member in this plant is solanine. Therefore, this study was conducted to utilize tissue culture techniques for the enhancement of solanine production in the in vitro grown cultures of this promising neglected plant. For callus growth and development experimental part, the highest callus growth parameters (callus diameter (21.4 mm) and callus fresh weight (2202.4 mg) were obtained in callus grown on Murashige & Skoog MS media supplemented with 2,4-Dichlorophenoxyacetic acid (2.0 mg·L-1) plus 1.5 mg·L-1 Thidiazuron. Similar trend was also obtained in cell suspension culture experiment, as maximum growth was recorded at similar hormone combination. Moreover, High-performance liquid chromatography analysis revealed that, solanine was affected by growth regulator type and concentration. The highest solanine levels were obtained when the explants were treated with 6-benzylaminopurine at level of 2.0 mg·L-1, as solanine content reached up to (2.61, 1.53 mg.g-1) for callus and cell suspension, respectively, while, microshoot contained the highest solanine (4.52 mg.g-1 DW) at 6-benzylaminopurine level of 1.6 mg.L-1. Additionally, carbon source had positively affected solanine level, where 0.2 M sucrose resulted in production of the highest amounts (3.13, 2.03 and 1.20 mg.g-1 DW) of solanine in microshoots, callus and cell suspension, respectively. Also, exposing microshoots and callus to light intensity of (100 µmol.m -2 s-1) yielded the highest solanine content (4.03 and 1.26 mg.g-1 DW, respectively),while the lowest solanine levels (1.50 and 0.48 mg.g-1 DW) were observed in plant material exposed to the lowest light intensity treatment (25 µmol.m -2 s-1) .

Evaluation of genetic diversity in Jordanian Solanum nigrum plants and genetic stability of invitro grown plant using (AFLP) technique

“…Also, mature seeds were collected from a mother plant grown at Jerash -Jordan (N: 32.27372, S: 35.89464) and in vitro propagated plants were established from mother plants that are grown in Jerash-Jordan location according to Al-Kiyyam et al (2019). The microshoots were developed in MS media (Murashige and Skoog, 1962) plus 1.2 mg.L -1 Thidiazuron (TDZ), while the callus and cell suspension were established on MS media plus2.0 mg.L -1 of 2,4-Dichlorophenoxyacetic acid (2,4-D) and1.5 mg.L -1 TDZ.…”

“…In vitro propagation system was only applied to the Jerash population. The in vitro propagation protocol was mentioned in detail previously in (Al-Kiyyam et al, 2019). The establishment of the in vitro material was from seeds collected from the Jerash population.…”

“…Solanum nigrum is one of the Solanaceae family members and is known as the black nightshade (Mandal et al, 2023). S. nigrum is considered as a weed and can be found in neglected areas and waste lands (Al-Kiyyam et al, 2019). According to Jordan's red list (Taifour, 2016), S. nigrum grows wild in different parts of Jordan in the north, such as Jerash and Ajloun, until reaching the south parts such as Aqaba and Petra.…”

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“…The establishment of the in vitro material was from seeds collected from the Jerash population. After seed germination; micro shoots were established from in vitro germinated seeds and further callus and cell suspension cultures were produced from microshoots according to (Al-Kiyyam et al, 2019); and each culture was maintained in the determined media for four weeks.…”

“…Solanum nigrum is one of the Solanaceae family members and is known as the black nightshade (Mandal et al, 2023). S. nigrum is considered as a weed and can be found in neglected areas and waste lands (Al-Kiyyam et al, 2019). According to Jordan's red list (Taifour, 2016), S. nigrum grows wild in different parts of Jordan in the north, such as Jerash and Ajloun, until reaching the south parts such as Aqaba and Petra.…”