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Effect of Silver Nitrate and Amino Acids on High Frequency Plants Regeneration in Barley (hordeum Vulgare L.)

Mozidul Haque
Plant Genetic Engineering Laboratory, Institute of Biological Sciences, University of Rajshahi, Rajshahi‐6205, Bangladesh

Abu Baker Siddique
Plant Genetic Engineering Laboratory, Institute of Biological Sciences, University of Rajshahi, Rajshahi‐6205, Bangladesh

S. M. Shahinul Islam*
Plant Genetic Engineering Laboratory, Institute of Biological Sciences, University of Rajshahi, Rajshahi‐6205, Bangladesh

Abstract/ Executive Summary:

Three Bangladeshi barley genotypes viz. BARI barley‐1, 3 and 6 were selected for this study to evaluate the efficiency of callus induction and plant regeneration. The effect of five doses of AgNO₃ singly, and combined with amino acids (L‐ proline, L‐glutamine) on callus induction and plant regeneration efficiency was evaluated using BARI barley‐3 and 6. The maximum values of callus induction were recorded at 49.20 and 32.66% for BARI barley‐6 and 3, respectively when 2.0 mg/l AgNO₃ and 200 mg/l L‐glutamine were added to the callus induction medium. Moreover, plant regeneration remarkably increased on MS + 1.0 mg/l BAP + 1.5 mg/l AgNO₃ + 150 mg/l L‐glutamine as 37.20% in BARI barley‐6 and 16.13% in BARI barley‐3. For rooting AgNO₃ singly affect positively, whereas negative influence was observed in combinations with any amino acids. However, by using AgNO₃ and amino acids, around < 4, < 27 and < 5 fold increase in callus induction were obtained. Regeneration and rooting were also found to increase considerably.

Keywords: Barley, Immature embryos, Somatic embryogenesis

Location: Experimental field of the Institute of Biological Sciences, University of Rajshahi, Bangladesh.

Start Date:

End Date:

Program Area: Crop-Soil-Water Management

Commodity: Barley

Objectives:

this study has been conducted to provide a suitable and efficient protocol for callus induction and regeneration using AgNO3 and amino acids through immature embryos of barley in Bangladesh.

Methodology:

Mature seeds of three barley genotypes viz., BARI barley‐1, 3, and 6 were collected from Bangladesh Agricultural Research Institute (BARI), Gazipur, Bangladesh and grown in the experimental field of the Institute of Biological Sciences, University of Rajshahi, Bangladesh. The seeds of milky phase, containing immature embryos were collected and used as explants. Spikes, containing seeds of milky phase, approximately 14‐16 days after anthesis were sterilized for 45 sec using 70% ethanol in a laminar air flow cabinet. To induce callus the embryos were aseptically dissected from the seeds, and inoculated on callus induction medium CIM containing MS supplemented with different concentrations of 2, 4‐D (1.0, 1.5, 2.0, 2.5 and 3.0 mg/l). The inoculated Petri dishes were sealed and cultures were incubated in the dark chamber at 25 ± 2°C for callus induction. When the age of the calli was around three weeks, they were transferred to the regeneration medium (RM = MS + 1.0 mg/l BAP) and cultured under low light conditions at 25 ± 2°C together with 14/10 hrs (light/dark) photoperiods. The regenerated shoots length around 2 ‐ 3 cm were transferred to root formation medium (RFM) that was GM (modified from MS, Islam 2000) + 1.0 mg/l IAA for better roots. The well developed rooted plantlets were transferred to pots, and acclimatized plants were transferred to field’s growing up to maturity for seed collection. To observe the effect of AgNO₃ and amino acids (AA) on callus induction (CI), plant regeneration (PR) and root formation (RF), five different concentrations of AgNO₃ singly and combinations with L‐proline and/or L‐ glutamine were added to related media. The combinations were added to CIM for callusing, RM for regeneration and RFM for rooting. The media without AgNO₃ and amino acids considered as control. Sucrose (30%), gelling agent agar (7%) were added to medium and pH adjusted 5.8. The average or mean values were computed from five replicates with standard error (SE) and each experiment was repeated thrice. ANOVA and post hoc DMRT were done using SPSS 16.0 software.

Source of Information: Plant Tissue Cult. & Biotech. 25(1): 37‐50, 2015 (June)

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

In present study, 2‐5‐folds increased rooting was recorded when AgNO₃ was individually added in various concentrations to rooting medium. On the contrary, the efficiency of AgNO3 to influence the root induction was reduced when any amino acids were combined with AgNO₃. Silva et al. (2011) mentioned that although the effect of AgNO₃ is not well understood, it is supposed that the silver ion binds a possible ethylene receptor at the plasma membrane, thus inhibiting the binding of ethylene to this receptor and consequently triggering the specific action of the hormone.

Knowledge Management: Journal