| Title |
Genotype-independent Agrobacterium rhizogenes-mediated root transformation of chickpea: a rapid and efficient method for reverse genetics studies
|
|---|---|
| Published in |
Plant Methods, July 2018
|
| DOI | 10.1186/s13007-018-0315-6 |
| Pubmed ID | |
| Authors |
Pooja Rani Aggarwal, Papri Nag, Pooja Choudhary, Niranjan Chakraborty, Subhra Chakraborty |
| Abstract |
Chickpea (Cicer arietinum L.), an important legume crop is one of the major source of dietary protein. Developing an efficient and reproducible transformation method is imperative to expedite functional genomics studies in this crop. Here, we present an optimized and detailed procedure for Agrobacterium rhizogenes-mediated root transformation of chickpea. Transformation positive roots were obtained on selection medium after two weeks of A. rhizogenes inoculation. Expression of green fluorescent protein further confirmed the success of transformation. We demonstrate that our method adequately transforms chickpea roots at early developmental stage with high efficiency. In addition, root transformation was found to be genotype-independent and the efficacy of our protocol was highest in two (Annigiri and JG-62) of the seven tested chickpea genotypes. Next, we present the functional analysis of chickpea hairy roots by expressing Arabidopsis TRANSPARENT TESTA 2 (AtTT2) gene involved in proanthocyanidins biosynthesis. Overexpression of AtTT2 enhanced the level of proanthocyanidins in hairy roots that led to the decreased colonization of fungal pathogen, Fusarium oxysporum. Furthermore, the induction of transgenic roots does not affect functional studies involving infection of roots by fungal pathogen. Transgenic roots expressing genes of interest will be useful in downstream functional characterization using reverse genetics studies. It requires 1 day to perform the root transformation protocol described in this study and the roots expressing transgene can be maintained for 3-4 weeks, providing sufficient time for further functional studies. Overall, the current methodology will greatly facilitate the functional genomics analyses of candidate genes in root-rhizosphere interaction in this recalcitrant but economically important legume crop. |
X Demographics
The data shown below were collected from the profiles of 5 X users who shared this research output. Click here to find out more about how the information was compiled.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| India | 2 | 40% |
| United Kingdom | 1 | 20% |
| Unknown | 2 | 40% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 2 | 40% |
| Scientists | 2 | 40% |
| Science communicators (journalists, bloggers, editors) | 1 | 20% |
Mendeley readers
The data shown below were compiled from readership statistics for 41 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 41 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 9 | 22% |
| Researcher | 5 | 12% |
| Student > Master | 5 | 12% |
| Student > Bachelor | 4 | 10% |
| Student > Doctoral Student | 3 | 7% |
| Other | 0 | 0% |
| Unknown | 15 | 37% |
| Readers by discipline | Count | As % |
|---|---|---|
| Agricultural and Biological Sciences | 15 | 37% |
| Biochemistry, Genetics and Molecular Biology | 6 | 15% |
| Environmental Science | 1 | 2% |
| Immunology and Microbiology | 1 | 2% |
| Medicine and Dentistry | 1 | 2% |
| Other | 1 | 2% |
| Unknown | 16 | 39% |
Attention Score in Context
This research output has an Altmetric Attention Score of 3. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 11 July 2018.
All research outputs
#13,542,652
of 23,577,654 outputs
Outputs from Plant Methods
#617
of 1,120 outputs
Outputs of similar age
#163,889
of 328,762 outputs
Outputs of similar age from Plant Methods
#18
of 34 outputs
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