Range of factors in the reduction of hyperhydricity associated with in vitro shoots of Salvia santolinifolia Bioss.

T Jan,S Pervez,R Azam,A Rahim,M A Nawaz,H Ullah,M S Ahmad,A Noor,A Khan,S Bibi,S Gul,H Amin

doi:10.1590/1519-6984.246904

Abstract

Hyperhydricity is a serious physiological disorder and affects In vitro propagation of many plants and as well of Salvia santolinifolia. The donor material to initiate the in vitro culture was the callus taken from the in vitro shoots produced on Murashig and Skoogs (MS) medium at 4.0 mg/l BA. This callus formed numerous hyperhydric shoots on culturing upon the medium of the same composition. The aim was to systematically evaluate the effect of cytokinins (Benzyladnine (BA) and N6-(-2-isopentenyl) adenine (2iP), culture vessels magnitude, medium solidification, source of nitrogen and calcium chloride for the alleviation of hyperhydricity. In the tissue cultures of S. santolinifolia BA and 2iP induced severe hyperhydricity, when other factors i.e. culture vessels magnitude and a suitable concentration of agar, ammonium nitrate (NH4NO3), potassium nitrate (KNO3) & calcium chloride (CaCl2.2H2O) were not optimized. After 30 days' culture, we observed 83.82% hyperhydric shoots at increased level (1.5 mg/l 2iP) and 81.59% at decreased levels (1.0 mg/l 2iP). On the other hand, hyperhydricity percentage at decreased (0.4%) and at increased (0.8%) levels of agar were 72.37% and 39.08%, respectively. MS medium modification with NH4NO3 (412 mg/l), KNO3 (475 mg/l) and CaCl2.2H2O (880 mg/l) was found the best medium to reduced hyperhydricity (23.6%).

Highlights

Hyperhydricity is a physiological disorder and hampered the micropropagation of many plants. Phillips and Matthews (1964) reported it for the first time in carnation shoot tip culture
The greatest collective reason of the hyperhydricity existence is the culture in liquid medium (Silva et al, 2013) and the use of high concentrations of cytokinins causes the accumulation of ethylene in the culture vessels (Isah, 2015)
To study the scope of factors on hyperhydricity of S. santolinifolia, we evaluated the effects of cytokinin types and concentration, culture vessels magnitude, medium solidification and source of nitrogen with calcium chloride on hyperhydricity and regeneration of adventitious shoots from callus

Summary

Introduction

Hyperhydricity is a physiological disorder and hampered the micropropagation of many plants. Phillips and Matthews (1964) reported it for the first time in carnation shoot tip culture. Hyperhydric individual obstructs the industrial production of plants regeneration and caused economic loss. Numerous factors can encourage the hyperhydricity an in vitro shoots, large quantities of ammonium ions (Brand 1993), high level of exogenous cytokinins (Ivanova et al, 2006; Oliveira et al, 2010), vessel aeration (Lai et al, 2005), cultivar (Carvalho et al, 2013) and agar concentrations in the medium (Abdoli et al 2007). The greatest collective reason of the hyperhydricity existence is the culture in liquid medium (Silva et al, 2013) and the use of high concentrations of cytokinins causes the accumulation of ethylene in the culture vessels (Isah, 2015)

Methods

Results

Discussion

Conclusion