Botanica Pacifica. A journal of plant science and conservation Preprint
Article first published online: 08 JUL 2017 | DOI: 10.17581/bp.2017.06105
Optimization of in vitro and ex vitro regeneration and micromorphological studies of Micrococca mercurialis (L.) Benth.
Mani Manokari1 & Mahipal S. Shekhawat2
1 Department of Botany, K.M. Center for Postgraduate Studies, Pondicherry, India
2 Biotechnology Laboratory, Department of Plant Science, M.G.G.A.C., Mahe, Pondicherry, India
A regeneration protocol has been successfully developed using nodal segments and leaf explants of Micrococca mercurialis (L.) Benth. The surface sterilized leaf explants were induced to proliferative green colored callus on Murashige and Skoog (MS) medium supplemented with 0.5 mg l-1 of 2,4-dichlorophenoxyacetic acid (2,4-D). Shoots were regenerated from the callus on MS medium supplemented with 0.5 mg l-1 6-benzylaminopurine (BAP). The nodal shoot segments responded on MS medium augmented with 1.0 mg l-1 BAP. The shoots were multiplied on MS medium fortified with 0.5 mg l-1 each of BAP, kinetin (Kin) and 0.1 mg l-1 α-naphthalene acetic acid (NAA). The maximum 17.3 shoots with 8.58 cm average length were resulted on this media combination. The shoots were rooted ex vitro when pulse treated with 100 mg l-1 indole-3 butyric acid (IBA). The plantlets were successfully established in the field with 96% success rate. Micromorphological studies were conducted for the successive stages of in vitro, ex vitro and in vivo grown plants to analyze the foliar developmental changes taken place across the environments. It has been concluded that the abnormal morphophysiology of in vitro plantlets were repaired while acclimatization under the greenhouse conditions before transferring to the field, which speed up the micropropagation process and increase the survival rate of the plantlets.
Манокари М., Шехават М.С. Оптимизация процесса регенерации in vitro и ex vitro и микроморфологические исследования Micrococca mercurialis (L.) Benth. Протокол регенерации был успешно разработан с использованием узловых сегментов и эксплантов листьев Micrococca mercurialis (L.) Benth. Из эксплантов листьев был получен пролиферативный зеленый каллус на среде Murashige и Skoog (MS) с добавлением 0,5 мг/л 2,4-дихлорфеноксиуксусной кислоты (2,4-D). Побеги регенерировали из каллюса на среде MS с добавлением 0,5 мг/л 6-бензиламинопурина (BAP). Сегменты узловых побегов отреагировали на среду MS, дополненную 1 мг/л BAP. Побели были размножены на среде MS, обогащенной 0,5 мг/л кинетином (Kin) и 0,1 мг/л α-нафталин уксусной кислотой (NAA). Максимальное число побегов – 17,3, со средней длиной 8.58 см было получено в этой комбинации сред. Побеги были укоренены ex vitro при периодической обработке 100 мг/л индол-3 масляной кислотой (IBA). Ростки были получены, коэффициент прорастения – 96 %. Микроморфологические исследования проводились для последовательных стадий развития растений in vitro, ex vitro и in vivo для анализа изменений развития листьев в разных средах. Сделан вывод о том, что аномальная морфофизиология проростков in vitro была компенсирована при акклиматизации в тепличных условиях до переноса в природную среду, что ускоряет процесс микроразмножения и увеличивает выживаемость проростков.
Keywords: Micrococca mercurialis, regeneration, ex vitro rooting, micromorphological studies, organogenesis, регенерация, укоренение ex vitro, микроморфологические исследования, органогенез
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