Abstract: Crop improvement through genetic engineering depends on effective and reproducible plant regeneration systems. Immature embryos are the most widely used explant source for in vitro regeneration in barley (Hordeum vulgare L.). However, immature embryos require the continuous growth of donor plants and the suitable stage for their culture is also certainly limited. On the other hand, mature embryos can be procured and stored easily; they can be studied throughout the year. In this study, an effective callus induction and plant regeneration were aimed to develop from mature embryos of different barley genotypes. The effect of medium (MS1 and MS2), auxin type (2,4-D, dicamba, picloram and 2,4,5-T) and concentrations (2, 4, 6 mg/l) on callus formation and effect of cytokinin type (TDZ, BAP) and concentrations (0.2, 0.5, 1.0 mg/l) on green plant regeneration were evaluated in mature embryo culture of barley. Callus and shoot formation was successful for all genotypes. By depending on genotype, MS1 is the best medium, 4 mg/l dicamba is the best growth regulator in the callus induction and MS1 is the best medium, 1 mg/l BAP is the best growth regulator in the shoot formation were determined.
Abstract: This research aims to investigate callus induction,
somatic embryogenesis and indirect plant regeneration of Crassula
ovata (Mill.) Druce – the famous ornamental plant. Experiment no.1:
Callus induction was obtained from leaf and stem explants on
Murashige and Skoog (MS) medium supplemented with various plant
growth regulators (PGRs). Effects of different PGRs, plant
regeneration and subsequent plantlet conversion were also assessed.
Indirect plant regeneration was achieved from the callus of stem
explants by the addition of 1.5 mg/L Kinetin (KN) alone. Best shoot
induction was achieved (6.5 shoots/per explant) after 60 days. For
successful rooting, regenerated plantlets were sub-cultured on the
same MS media supplemented with 1.5 mg/L KN alone. The rooted
plantlets were acclimatized and the survival rate was 90%.
Experiment no.2: Results revealed that 0.5 mg/L 2,4-D alone and in
combination with 1.0 mg/L 6-Benzyladenine (BA) gave 89.8% callus
from the stem explants as compared to leaf explants. Callus
proliferation and somatic embryo formation were also evaluated by
‘Double Staining Method’ and different stages of somatic
embryogenesis were revealed by scanning electron microscope. Full
Strength MS medium produced the highest number (49.6%) of
cotyledonary stage somatic embryos (SEs). Mature cotyledonary
stage SEs developed into plantlets after 12 weeks of culture. Wellrooted
plantlets were successfully acclimatized at the survival rate of
85%. Indirectly regenerated plants did not show any detectable
variation in morphological and growth characteristics when
compared with the donor plant.
Abstract: The extract of milk thistle contains a mix of flavonolignans termed silymarine.. In order to analysis influence of growth regulators, genotype, explant and subculture on the accumulation of flavonolignans, a study was carried out by using two genotype (Budakalszi and Noor abad moghan cultivars), cotyledon and hypocotyle explants, solid media of MS supplemented by different combinations of two growth regulators; Kinetin (0.1, 1 mg/l) and 2,4-D (1, 2 mg/l). Seeds of the plant were germinated in MS media whitout growth regulators in growth chamber at 26°C and darkness condition. In order to callus induction, the culture media was supplemented whit different concentrations of 2,4-D and kinetin. Calli obtained from explants were sub-cultured four times into the fresh media of the first experiment. flavonoides was extracted from calli in four subcultures. The flavonoid components were determined by high- performance liquid choromatography (HPLC) and separated into Taxifolin, Silydianin+Silychristin, Silybin A+B and Isosilybin A+B. Results showed that with increasing callus age, increased accumulation of silybin A+B, but reduced Isosilybin A+B content. Highest accumulation of Taxifolin was observed at first calli. Calli produced from cotyledon explant of Budakalszi cultivar were superior for Silybin A+B, where calli from hypocotyl explant produced higher amount of Taxifolin and Silydianin+Silychristin. The best cultivar for Silymarin production in this study was Budakalszi cultivar. High amount of SBN A+B and TXF were obtained from hypocotil explant.
Abstract: Stevia rebaudiana Bertoni (natural sweetener) belongs
to Asteraceae family and can be used as substitute of artificial
sweeteners for diabetic patients. Conventionally, it is cultivated by
seeds or stem cutting, but seed viability rate is poor. A protocol for
callus induction and multiplication was developed to produce large
no. of calli in short period. Surface sterilized nodal, leaf and root
explants were cultured on Murashige and Skoog (MS) medium with
different concentrations of plant hormone like, IBA, kinetin, NAA,
2,4-D, and NAA in combination with 2,4-D. 100% callusing was
observed from leaf explants cultured on combination of NAA and
2,4-D after three weeks while with 2,4-D, only 10% callusing was
observed. Calli obtained from leaf and root explants were shiny green
while with nodal explants it was hard and brown. The present
findings deal with induction of callusing in Stevia to achieve the
rapid callus multiplication for study of steviol glycosides in callus
culture.