Article (Scientific journals)
Improved Agrobacterium-mediated transformation and direct plant regeneration in four cultivars of finger millet (Eleusine coracana (L.) Gaertn.)
Lakkakula , Satish; Stanislaus, Antony Ceasar; Manikandan , Ramesh
2017In Plant Cell, Tissue and Organ Culture, 131 (3), p. 547-565
Peer reviewed
 

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Keywords :
Agrobacterium; Direct plant regeneration; Finger millet ; Shoot apical meristems; Southern blotting; Transgenic plants
Abstract :
[en] We have developed an improved Agrobacterium-mediated transformation and rapid regeneration system for four cultivars (‘CO(Ra)-14’, ‘PR-202’, ‘Try-1’ and ‘Paiyur-2’) of finger millet using optimized transformation and direct plant regeneration conditions. The shoot apical meristems (SAMs) were used as explants in this study. Agrobacterium strain EHA105 carrying binary vector pCAMBIA1301 was used to optimize the transformation conditions. Concentration of hygromycin, the optical density of the culture, infection time, age of the explants, co-cultivation period, the concentrations of acetosyringone and antibiotics were optimized to improve the transformation frequency. The highest frequency of mean transient gus expression (85.1%) was achieved in cultivar ‘CO(Ra)-14’. The entire transformation procedure, from initiating SAMs to planting putative transgenic plantlets in the greenhouse, was completed within 45 days with the highest stable transformation frequency of 11.8% for ‘CO(Ra)-14’. PCR, gus staining and Southern blot analyses were performed in T0 and T1 generations to confirm the gene integration. Six events from T0 had a single copy of the transgene and showed a normal Mendelian pattern of segregation. To our knowledge, this is the first report on the high frequency transformation of finger millet by Agrobacterium and subsequent recovery of transgenic plants via direct plant regeneration without a callus phase, in short duration (45 days). The proposed protocol could be supportive in breaking through the bottleneck in transformation and regeneration of finger millet cultivars.
Disciplines :
Biotechnology
Author, co-author :
Lakkakula , Satish
Stanislaus, Antony Ceasar ;  Université de Liège - ULiège > Département des sciences de la vie > Génomique fonctionnelle et imagerie moléculaire végétale
Manikandan , Ramesh
Language :
English
Title :
Improved Agrobacterium-mediated transformation and direct plant regeneration in four cultivars of finger millet (Eleusine coracana (L.) Gaertn.)
Publication date :
September 2017
Journal title :
Plant Cell, Tissue and Organ Culture
ISSN :
0167-6857
eISSN :
1573-5044
Publisher :
Kluwer Academic Publishers, Netherlands
Volume :
131
Issue :
3
Pages :
547-565
Peer reviewed :
Peer reviewed
Funders :
The author L. Satish sincerely thanks the University Grants Commission, New Delhi, India for financial support in the form of UGC BSR JRF and SRF (UGC order no. F.4-1/2006 (BSR)/7-326/2011/BSR) for the Ph.D. program.
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