Muhammad Ahmad1*, Abu Bakar Siddique1, Abbas Shoukat2, Hafiza Hadiqa Anum3, Faisal Mushtaq4, Ahmad Bilal5, Muhammad Hamza Ishaq Rao1 and Muhammad Mubashar6Corresponding author: Muhammad Ahmad (Email: ahmad31710@gmail.com)
Submitted | Accepted | Published |
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Jun 08,2018 | Dec 19,2018 | Jan 10,2019 |
2019 / Vol: 2 / Issue: 1
Abstract
The Amaranthus genus has extraordinary morphological and biological features, which enable this group of plants to adapt a variety of environmental conditions. Moreover, shorter lifecycle made them more competitive than many other crops. The establishment of an efficient in vitro regeneration protocol for Amaranth spp. Initially, surface sterilization of seeds was carried using ethanol and mercuric chloride. The protocols were found effective for controlling microbial contamination however, Amaranth seeds were found to be highly sensitive to even very dilute regents. Time of treatment was reduced to only five minutes, yet the %age germination remained zero. Hence, it was decided that seeds without surface sterilization should be germinated and plants without contamination should be rescued soon after germination. Seeds of different Amaranth lines germinated in this way yielded 88% germination in A-20, 69% in A-96, 62% in A-42 and 58% in A-21 line. The seeds were germinated in petri plates on MSO (MS medium without plant growth regulators) and shifted to jars having MS medium supplemented with 30, 45, 60, 75 and 90 g/L of sucrose for further proliferation. Vigorous growth of plants was observed on MSO medium having 30% sucrose. Later, epicotyl region of Amaranth plants with vigorous growth were used as explant for in vitro callus induction and subsequent regeneration. For callus induction MS media supplemented with 2, 4-D and kinetin were used and all Amaranth lines exhibited variable response with 100% callus induction in A-21, 82% in A-42, 58% in A96 and in A-20 there was no callus induction at all. The calli were shifted to regeneration media having 2.4 µM naphthalene acetic acid (NAA) and 4.4 µM benzyl amino purine (BAP). In vitro regenerated plants were shifted to plastic pots having peat moss for acclimatization and further seed setting.
Key words: Amaranthus, in vitro regeneration, callus.
Key words: Amaranthus, in vitro regeneration, callus.
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