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An efficient Agrobacterium-mediated genetic transformation method for foxtail millet (Setaria italica L.)

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Title An efficient Agrobacterium-mediated genetic transformation method for foxtail millet (Setaria italica L.)
 
Creator Sood, Priyanka
Singh, Roshan Kumar
Prasad, Manoj
 
Subject Setaria italica
Mature seed-derived calli
Transformation
Regeneration
Agrobacterium tumefaciens
 
Description Accepted date: 30 December 2019
Foxtail millet (Setaria italica L.) is a model crop to study C4 photosynthesis, abiotic stress tolerance, and bioenergy traits. Advances in molecular genetics and genomics had identified several potential genes in this crop that would serve as candidates for imparting climate-resilient traits in related millets, cereals, and biofuel crops. However, the lack of an efficient genetic transformation system has been impeding the functional characterization of these genes in foxtail millet per se. Given this, an easy and efficient regeneration and transformation protocol was optimized using mature seeds as a choicest explant. The suitability of secondary embryogenic calli over primary calli is underlined due to their high competence. The use of perfect combinations of plant growth regulators together with the ionic strength of organic and inorganics salts was found to influence regeneration and genetic transformation. We studied and optimized various crucial factors that affect the genetic transformation of foxtail millet calli using Agrobacterium tumefaciens-mediated approach. Secondary embryogenic calli and LBA44404 strain were found to be the best targets for transformation. The use of high sucrose and glucose, together with freshly prepared tobacco leaves extract, Silwet L-77 and acetosyringone, improved the efficiency of the genetic transformation of foxtail millet. Moreover, the use of an in vitro regeneration system with 84% callusing efficiency and 70–74% regeneration frequency led to a high recovery of transformants. Altogether, the present study reports a highly efficient (~ 27%) transformation system in foxtail millet that will expedite forward and reverse genetic studies in this important crop.
Authors research in the area of foxtail millet genomics is supported by Science and Engineering Research Board (SERB), Department of Science and Technology (DST), Govt. of India [Grant no. EMR/2015/000464]. PS acknowledges the Young Scientist Award from DST-SERB, Govt. of India [File No. YSS/2014/000870/LS]. RKS acknowledges the research fellowship received from the Council of Scientific and Industrial Research, Govt. of India. Dr. Muthamilarasan, DST INSPIRE Faculty, ICAR-NIPB, New Delhi is acknowledged for his critical inputs during this study. The authors are also thankful to DBT-eLibrary Consortium (DeLCON) for providing access to e-resources.
 
Date 2020-01-17T08:59:20Z
2020-01-17T08:59:20Z
2020
 
Type Article
 
Identifier Plant Cell Reports, (In Press)
1432-203X
https://doi.org/10.1007/s00299-019-02507-w
https://link.springer.com/article/10.1007%2Fs00299-019-02507-w
http://223.31.159.10:8080/jspui/handle/123456789/1031
 
Language en_US
 
Format application/pdf
 
Publisher Springer Nature Publishing AG