Abstract
Agrobacterium-mediated transformation is an important research tool for the genetic improvement of cassava. The induction of friable embryogenic callus (FEC) is considered as a key step in cassava transformation. In the present study, the media composition was optimized for enhancing the FEC induction, and the effect of the optimized medium on gene expression was evaluated. In relative comparison to MS medium, results demonstrated that using a medium with reducing nutrition (a 10-fold less concentration of nitrogen, potassium, and phosphate), the increased amount of vitamin B1 (10 mg/L) and the use of picrolam led to reprogram non-FEC to FEC. Gene expression analyses revealed that FEC on modified media increased the expression of genes related to the regulation of polysaccharide biosynthesis and breakdown of cell wall components in comparison to FEC on normal CIM media, whereas the gene expression associated with energy flux was not dramatically altered. It is hypothesized that we reprogram non-FEC to FEC under low nitrogen, potassium and phosphate and high vitamin B1. These findings were more effective in inducing FEC formation than the previous protocol. It might contribute to development of an efficient transformation strategy in cassava.
Highlights
Cassava (Manihot esculenta Crantz) is grown throughout the tropics for its production of starchy roots
Gresshoff & Doy (GD) medium has been used in various studies as the standard medium for friable embryogenic callus (FEC) induction (Table 1), the induction of somatic embryos (SE), FEC and non-FEC depends on the components of other macronutrients [7]
We developed an improved medium for the efficient production of FEC in cassava, which is an essential need for the genetic transformation of cassava
Summary
Cassava (Manihot esculenta Crantz) is grown throughout the tropics for its production of starchy roots. It is widely used in Africa and Asia for human consumption, animal feed, and as an industrial material [1,2,3]. Cassava is an important tropical crop, conventional breeding programs, based on sexual hybridization, have not been well developed. Cassava breeding is problematic due to the high degree of genetic heterozygosity, genetic overloading, separation of progeny, few flowers, low pollen fertility, self-incompatibility, and low fruit set [4, 5]. Optimization of a genetic transformation system for the molecular breeding.
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