Identification of differentially expressed genes involved in spine formation on seeds ofDaucus carota L. (carrot), using annealing control primer (ACP) system

Abstract

Carrot seeds normally have surface spines. The availability of a spineless mutant would be agronomically beneficial, eliminating the current efforts to remove those spines pre-sowing. Furthermore, the identification of spine-specific genes would provide insights into spine development in wild-type carrot seed. This effort could be facilitated through the use of an annealing control primer (ACP) system. Here, we employed a new and accurate reverse transcriptionpolymerase chain reaction (RT-PCR) that involves ACPs for identifying genes of interest. With these techniques, 11 expressed sequence tags (ESTs) were obtained for cloning and sequencing the genes that are differentially expressed in wild-type spiny seeds, but not in the spineless mutant. In all, 7 cDNAs exhibited significant sequence similarity with known genes from other species. These included cell wall-associated hydrolase, tail fiber assembly protein, transcriptional regulatory protein, berberine bridge enzyme, S-adenosyl methionine synthase, transketolase, and phenylalanyl t-RNA synthetase beta chain. Four other cDNA sequences had no significant identities with known genes. As revealed by RT-PCR, these genes regulate spine formation during the developmental stage. Our results suggest that PCR-based differential display RT-PCR techniques are a very useful tool for identifying spine-specific genes from carrot seeds.

Different Gene Expressions of Resistant and Susceptible Hop Cultivars in Response to Infection with a Highly Aggressive Strain of Verticillium albo-atrum

“…ACP systems have been widely used in differential gene expression studies since the development of improved systems and commercially available kits (Kim et al 2004 ). Although more studies have been reported for animals, the ACP differential display has been successfully used for identification of genes involved in spine formation on seeds of carrots (Park et al 2006 ), Cd-responsive genes in Solanum nigrum (Xu et al 2009 ), drought induced genes in barley (Lee et al 2011 ) and induced genes in cotton after inoculation with Aspergillus flavus (Lee et al 2012 ). We found ACP-based differential display more advantageous than cDNA-AFLP because of its methodological simplicity, due to the use of agarose gels instead of long AFLP polyacrylamide gels and due to obtaining much longer TDFs with good matches to known sequences.…”

Bacterial feeding induces changes in immune-related gene expression and has trans-generational impacts in the cabbage looper (Trichoplusia ni)

“…Like all differential display techniques, the GeneFishing method can be expected to reveal a subset of expression differences; and may be biased towards more abundant transcripts. Although relatively new, it has already been applied to studies of animal neurophysiology [ 10 ], cancer [ 11 ] and plant allellopathy [ 12 ] and development [ 13 ]. So far a quantitative evaluation of the technique compared with others such as cDNA-AFLP, RAPD-differential display, or microarray analysis has not been carried out.…”

Identification of differentially expressed genes in a pistillody common wheat mutant using an annealing control primer system

“…Therefore, this method has a high reproducibility rate and eliminates false-positive results. The ACP system has been widely used for studies of cell differentiation during animal and plant development (Park et al, 2006;Yang et al, 2011). In the present study, ACP was used to identify potential genes related to the pistillody trait.…”