SOME FACTORS AFFECTING MICROPROPAGATION OF Haplophyllum tuberculatum L

Abstract

Effect of three medium types, Murashige and Skoog (MS), Linsmaier and Skoog (LS) and Lloyd and McCown (WPM), two explant types (shoot-tip and single node cutting), three cytokinin types (Benzyladenine, Kinetin and 2-isopenteny1 adenine), two additives; viz., s (thiamine and pyridoxine hydrochloride) and amino acids (asparagine and arginine) and two gelling agents (guar seed powder and locust bean seed powder) combined with agar on micropropagation of Haplophyllum tuberculatum were investigated. For rooting, the effect of three auxins (Indole acetic acid, Indol butyric acid and Naphthalene acetic acid) were studied in combination with three medium strengths (full, half and quarter) combined with the best mixture (0.50 mgl-1 Kin + 6 gl-1 agar with 2 gl-1 guar seed powder) of gelling agents. Obtained results showed that Murashige and Skoog (MS) was the best medium with single node explant as starting material during establishment stage. Multiple shoots were obtained on MS medium supplemented with 1.0 mgl-1 Kinetin (Kin) combined with 0.1 mgl-1 α-naphthalene acetic acid (NAA). Thiamine and arginine were the best vitamins and amino acid at 0.1 mgl-1 and 0.50 mgl-1 , respectively. Agar at 6 gl-1 + guar seed powder at 2 gl-1 was the best gelling agent treatment. Half strength MS medium with 0.50 mgl-1 Kin + 0.5 mgl-1 NAA and 6 gml-1 agar with 2 gml-1 guar seed powder was the best combination for rooting were successfully (85% survival) acclimatized in mixture of peatmoss, vermiculite, washed sterilized sand and perlite (1:1:1:1, V/V/V/V).

Efficient shoot regeneration of medicinal plant Haplophyllum tuberculatum by direct and indirect organogenesis and genetic fidelity assessment using Inter Simple Sequence Repeats markers

“…Several disinfectants, such as Ethanol, Ca(ClO)2, NaOCl, H2O2, AgNO3, HgCl2, Silver nitrate, etc., are commonly used for surface sterilization of explants at the different immerse times ( Hesami et al, 2019 ). In this study, the surface sterilization method was developed based on the method described by Abdallah et al (2019) but was slightly modified by excluding the treatment of the explant with mercuric chloride (HgCl2) due to its toxicity ( Abdallah et al, 2019 ). Despite not using HgCl2, we were able to achieve a nearly 100% efficacy of disinfection on the explants.…”

“…In other Haplophyllum species such as Haplophyllum virgatum var. virgatum, optimal callus generation was achieved with a combination of 0.1 mg/l Kinetin and 5 mg/l IAA in a B5 medium ( Abdallah et al, 2019 ). In Haplophyllum gilesii callus generation was achieved in MS media containing 0.25 mg/l kinetin, 2.5 mg/l BAP and 2.5 mg/l NAA ( Almazrooei, 2018 ).…”

“…Surface sterilization was done according to the method described by Abdallah et al (2019) with some modifications. In short, unopened immature seed pods were immediately collected into a beaker filled with tap water to maintain the freshness of the explants.…”

“…Surface sterilization and evaluation of the effect of 2,4-D (2,4-Dichlorophenoxyacetic acid) and BAP (6-Benzylaminopurine) on Haplophyllum tuberculatum immature seeds Surface sterilization was done according to the method described by Abdallah et al (2019) with some modifications. In short, unopened immature seed pods were immediately collected into a beaker filled with tap water to maintain the freshness of the explants.…”

“…In-vitro regeneration of H. tuberculatum has not received adequate attention in the literature. Only few studies evaluated the micropropagation potential of H. tuberculatum using axillary buds, shoot tips, and single-node cuttings ( Almazrooei, 2018 ; Abdallah et al, 2019 ).…”