Callogenic Response Research Articles

Morphogenic and biochemical variations under different spectral lights in callus cultures of Artemisia absinthium L.

Through its impact on morphogenesis, light is the key environmental factor that alters plant architectural development; however, the understanding that how light controls plant growth and developmental processes is still poor and needs further research. In this study, we monitored the effect of various monochromatic lights and plant growth regulators (PGRs) combinations on morphogenic and biochemical variation in wild grown-leaf derived callus cultures of Artemisia absinthium L. Combination of α-naphthalene acetic acid (NAA 1.0mg/l) and Thidiazuron (TDZ 2.0mg/l) resulted in optimum callogenic frequency (90%) when kept under fluorescent light for 4weeks (16/8h). In contrast to the control (white spectrum), red spectrum enhanced peroxidase activity, protease activity, total protein content and chlorophyll a/b ratio. Green spectrum was found to be more supportive for total phenolics, total flavonoids and antioxidant activity. Yellow light enhanced MDA content while white and green light improved total chlorophyll content and carotenoid content. A positive correlation among callogenic response, antioxidant activities and set of antioxidative enzyme activities was also observed in the current report. This study will help in understanding the influence of light on production of commercially important secondary metabolites and their optimization in the in vitro cultures of A. absinthium L.

The Study of Ascorbate Peroxidase, Catalase and Peroxidase During In Vitro Regeneration of Argyrolobium roseum

Here, we demonstrate the micropropagation protocol of Argyrolobium roseum (Camb.), an endangered herb exhibiting anti-diabetic and immune-suppressant properties, and antioxidant enzymes pattern is evaluated. Maximum callogenic response (60%) was observed from leaf explant at 1.0mg L(-1) 1-nephthalene acetic acid (NAA) and 0.5mg L(-1) 6-benzyl aminopurine (BA) in Murashige and Skoog (MS) medium using hypocotyl and root explants (48% each). Addition of AgNO3 and PVP in the culture medium led to an increase in callogenic response up to 86% from leaf explant and 72% from hypocotyl and root explants. The best shooting response was observed in the presence of NAA, while maximum shoot length and number of shoots were achieved based on BA-supplemented MS medium. The regenerated shoots were rooted and successfully acclimatized under greenhouse conditions. Catalase and peroxidase enzymes showed ascending pattern during in vitro plant development from seed while ascorbate peroxidase showed descending pattern. Totally reverse response of these enzymes was observed during callus induction from three different explants. During shoot induction, catalase and peroxidase increased at high rate while there was a mild reduction in ascorbate peroxidase activity. Catalase and peroxidase continuously increased; on the other hand, ascorbate peroxidase activity decreased during root development and acclimatization states. The protocol described here can be employed for the mass propagation and genetic transformation of this rare herb. This study also highlights the importance and role of ascorbate peroxidase, catalase, and peroxidase in the establishment of A. roseum in vitro culture through callogenesis and organogenesis.

Piper nigrum: Micropropagation, Antioxidative enzyme activities, and Chromatographic Fingerprint Analysis for Quality Control

A reliable in vitro regeneration system for the economical and medicinally important Piper nigrum L. has been established. Callus and shoot regeneration was encouraged from leaf portions on Murashige and Skoog (MS) medium augmented with varied concentrations of plant growth regulators. A higher callus production (90 %) was observed in explants incubated on MS medium incorporated with 1.0 mg L(-1) 6-benzyladenine (BA) along with 0.5 mg L(-1) gibberellic acid after 4 weeks of culture. Moreover, a callogenic response of 85 % was also recorded for 1.0 mg L(-1) BA in combination with 0.25 mg L(-1) α-naphthalene acetic acid (NAA) and 0.25 mg L(-1) 2,4-dichlorophenoxyacetic acid or 0.5 mg L(-1) indole butyric acid (IBA) along with 0.25 mg L(-1) NAA and indole acetic acid. Subsequent sub-culturing of callus after 4 weeks of culture onto MS medium supplemented with 1.5 mg L(-1) thiodiazoran or 1.5 mg L(-1) IBA induced 100 % shoot response. Rooted plantlets were achieved on medium containing varied concentrations of auxins. The antioxidative enzyme activities [superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX)] revealed that significantly higher SOD was observed in regenerated plantlets than in other tissues. However, POD, CAT, and APX were higher in callus than in other tissues. A high-performance liquid chromatography (HPLC) fingerprint analysis protocol was established for quality control in different in vitro-regenerated tissues of P. nigrum L. During analysis, most of the common peaks represent the active principle "piperine." The chemical contents, especially piperine, showed variation from callus culture to whole plantlet regeneration. Based on the deviation in chromatographic peaks, the in vitro-regenerated plantlets exhibit a nearly similar piperine profile to acclimated plantlets. The in vitro regeneration system and HPLC fingerprint analysis established here brought a novel approach to the quality control of in vitro plantlets, producing metabolites of interest with substantial applications for the conservation of germplasm.

Callogenesis and Shoot Organogenesis from Flowers of Stevia rebaudiana (Bert.)

The callogenic and shoot organogenic potential of the medicinaly important plant, S. rebaudiana were investigated. Callus induction and shoot regeneration were induced from flower explants of greenhouse plants incubated on Murashige and Skoog (MS)-medium supplemented with different plant growth regulators (PGRs). The best callogenic response was observed on explants incubated on MS-medium supplemented with 2.0 mg/l 6-benzyladenine (BA) along with 2.0 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) after 30-days of culture. Subsequent transfer of callogenic explants onto MS-medium supplemented with 2.0 mg/l BA gave 85.2% shoot organogenesis after 30-days following culture. Maximum number (21.6) of shoots/explant was recorded for 2.0 mg/l BA. Subsequent sub culturing of shoots after 3 weeks of culture, on medium with similar compositions of PGRs gave 5.1 cm longest shoot. The palynology of S. rebaudiana has also been investigated.

Efficient regeneration and antioxidant potential in regenerated tissues of Piper nigrum L.

The organogenic potential and antioxidant potential (1, 1-diphenyl-2-picrylhydrazyl-scavenging activity) of the medicinal plant Piper nigrum L. (black pepper) were investigated. Callus induction and shoot regeneration were induced from leaf explants of potted plants cultured on MS medium supplemented with different plant growth regulators. The best callogenic response was observed on explants cultured for 30 days on MS medium supplemented with either 0.5 or 1.5 mg l−1 6-benzyladenine (BA) + 1.0 mg l−1 α-naphthaleneacetic acid. Subsequent transfer of the callogenic explants onto MS medium supplemented with 1.5 mg l−1 BA + 1.0 mg l−1 gibberellic acid (GA3) achieved 85% shoot organogenesis after 30 days of culture. The maximum number (7.2) of shoots/explant was recorded for explants cultured in MS medium supplemented with 1.0 mg l−1 BA. Following the transfer of shoots to an elongation medium, the longest shoots (5.4 cm) were observed on MS medium supplemented with 1.0 mg l−1 BA + 1.0 mg l−1 GA3. The elongated shoots were rooted on MS medium supplemented with different concentrations of indole butyric acid. An assay of the antioxidant potential of the in vitro-grown tissues revealed that the antioxidant activity of the regenerated shoots was significantly higher than that of callus and the regenerated plantlets.

Callogenic Studies of Achyranthes aspera Leaf Explant at Different Hormonal Combinations

With the objective to promote in vitro callus induction, leaf segments of Achyranthes aspera were inoculated on basal MS medium supplemented with 3.0% sucrose and 0.8% agar with different concentrations of 2,4-D alone and in combination with NAA, BAP, IAA, IBA and Zeatin. The explants were maintained in growth room at 25 +/- 1 degrees C and 16 h light cycle. The best callus induction was obtained with 2,4-D (1.0 and 2.0 mg L(-l)) in combination with NAA (0.5 mg L(-1)). Callus induction and good texture from leaf explant was also observed at 2,4-D with BAP. On these combinations morphologically, light green, soft, compact and non-embryogenic callus (Type III callus) was observed. While morphology of callus and callogenic response was poor at 2,4-D alone or in combination with other hormones at different concentrations.

In vitro Plant Regeneration from Direct and Indirect Organogenesis of Memordica charantia

The study was conducted to analyze the effect of plant growth regulators on callogenesis and direct and indirect organogenesis of Memordica charantia. Callus cultures were induced from leaf, stem and cotyledonary explants of Momordica charantia, at different auxin and cytokinin concentrations either in single or in combination in MS medium. The best callogenic response was observed from all three explants (leaf, stem and cotyledon) on MS medium supplemented with 1.0 and 1.5 mg L(-1) BAP with 1.5 mg L(-1) NAA and 1.0 mg L(-1) 2,4-D, respectively. The callus produced was hard, green and compact. These totipotent cells were failed to give rise shooting response when transferred to same or different growth regulator containing medium as second subculture. Indirect organogenesis response was very low or absent due to hardening of callus and habitutation. Best shooting was observed at 1.0 mg L(-1) BAP + 0.1 TDZ and 1.5 mg L(-1) BAP + 0.2 mg L(-1) NAA from shoot tip and cotyledonary node explants. While root formation was achieved when generated shoots were transferred to MS medium both full and half strength supplemented with different auxin concentrations.