Volume 12, Issue 43 (spring 2008)
jwss 2008, 12(43): 117-126 |
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Najafi B, Seifi A, Babaeian Jelodar N, Shakib A. A Study on Effective Factors in Gene Transformation to Dioecious Plant. jwss 2008; 12 (43) :117-126
URL: http://jstnar.iut.ac.ir/article-1-826-en.html
URL: http://jstnar.iut.ac.ir/article-1-826-en.html
Abstract: (32856 Views)
Sorrel (Rumex acetosa L.) is a model dioecious plant in genetic and molecular studies for sex determination. In this research, gus reporter gene transformation to leaf disks of this plant via Agrobacterium tumefaciens was evaluated based on transient expression of this gene. Three strains of Agrobacterium( LBA4404, C58 and EHA101) and two kinds of bacterial suspensions (suspension I: Agrobacterium cells were grown in LB medium with PH 7 and suspension II: the cells were resuspended in MS medium containing 100 mM acetosyringone with PH 5.2) were analyzed in a factorial experiment with 3 replications in Complete Randomized Design (CRD). Also, the effects of three different times of pre-culture of explants (0, 2 and 4 days) were examined. Results showed no significant difference among bacterial strains, whereas suspension I and explants without pre-culture had higher efficiency in gene transformation to this plant. Based on the best treatment, stable transformation with different days of cocultivation was carried out and revealed that the 4-day cocultivation is more efficient than the 2-day cocultivation in gene transformation to Sorrel. The histochemical GUS assay and PCR analysis were done on putative transgenic plants. Results showed that some of these plants contain at least one copy of the transferred gene or genes in comparison with control plants .
Keywords: Rumex acetosa, Dioecious, Transformation, gusgene, Transient expression, Stable expression
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