Highly efficient Agrobacterium-mediated transformation and plant regeneration system for genome engineering in tomato

Sandhya, Dulam; Jogam, Phanikanth; Venkatapuram, Ajay Kumar; Savitikadi, Pandarinath; Peddaboina, Venkataiah; Allini, Venkateswar Rao; Abbagani, Sadanandam

doi:10.1016/j.sjbs.2022.103292

Cited by 16 publications

(5 citation statements)

References 63 publications

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“…As a matter of fact, our initial attempts to regenerate alfalfa plant using the existing procedures (a number of published studies) did not succeed or only obtained several numbers of unhealthy regenerated plants (even no regenerated plants). We could understand that the poor repeatability is probably due to the differences of genotype/cultivar, explant type, or hormone ration compared to the previous studies (Rajesh et al 2016;Kumar et al 2017;Sandhya et al 2022). Again, unfortunately our request to obtain the original alfalfa material and the detailed transformation procedures from the researchers who have already published the study was declined.…”

Section: Discussionmentioning

confidence: 75%

“…Since Deak et al (1986) rstly reported the successful alfalfa transformation mediated by Agrobacterium tumefaciens, many researchers conducted studies on optimization of the alfalfa transformation conditions (i.e., suitable genotype and explant, hormone component and ratio) and improvement of transformation technologies (Koul et Sandhya et al 2022). Nevertheless, several of the procedures are characterized by a low transformation frequency or the procedures described vaguely.…”

Section: Introductionmentioning

confidence: 99%

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Optimization and establishment of Agrobacterium-mediated transformation in alfalfa (Medicago sativa L.) using eGFP as a visual reporter

Gao,

Zhang,

et al. 2023

Plant Cell Tiss Organ Cult

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Efforts are still required for establishing an e cient and standard transformation procedure in alfalfa, an important worldwide forage crop, due to the unstable repeatability of the published genetic transformation. The objectives of this study were to optimize and establish an effective genetic transformation system in alfalfa (Medicago sativa L.) using eGFP as a visual reporter, and test the feasibility of established procedure in other Medicago species. Comparisons of percent callus induction using three different explants of the four alfalfa cultivars showed that hypocotyl explant had greater callus induction (70-98.6% across four cultivars) than that of cotyledon (17.1%-73.2%) and cotyledonary node (39.7%-70.2%). Explants of 'Qingshui' showed the greater mean value of callus induction (80.6%) compared to other cultivars (mean: 46.9-55.5% across three cultivars). Using hypocotyl of 'Qingshui', the optimal callus induction and bud differentiation mediums were determined. The established procedure showed the transformation rate of eGFP to 'Qingshui' was about 72% with the time taken 85-120 days.Fluorescence and PCR detections evidenced the integration of the eGFP in regenerated plant genome. The procedure also yielded the transformation rate > 64% in M. falcata and M. polymorpha. The eGFPtagged pollens produced by regenerated alfalfa could be an enabling tool for tracking pollen movement (by wind or insect) to study the pollen distribution patterns and pollinator behavior, thus assess potential risks of transgene ow in genetically modi ed alfalfa. Additionally, the established procedure would serve a useful approach for introducing agronomically important traits into alfalfa, and thus contribute to breeding program of Medicago species. Key MessageWe established an e cient agrobacterium-mediated transformation using eGFP as a visual reporter in alfalfa and successfully applied this procedure to other Medicago species.

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Section: Discussionmentioning

confidence: 75%

Section: Introductionmentioning

confidence: 99%

Optimization and establishment of Agrobacterium-mediated transformation in alfalfa (Medicago sativa L.) using eGFP as a visual reporter

Gao,

Zhang,

et al. 2023

Plant Cell Tiss Organ Cult

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“…Regenerating calli resulted in the formation of multiple shoots after 3 weeks on medium containing 3 mg/L BAP and 0.1 mg/L IAA. Furthermore, Sandhya et al (2012) reported that 2.0 mg/L ZT in combination with 0.1 mg/L IAA produced the highest number of shoots from the cotyledon explants of two different tomato cultivars, Arka Vikas and Pusa Early Dwarf [ 23 ]. Shorter explant-to-plantlet regeneration time is always desirable, as it reduces labor and resource costs and allows high-throughput approaches for early recovery and evaluation of transgenic lines.…”

Section: Discussionmentioning

confidence: 99%

Optimization of Callus Induction and Shoot Regeneration from Tomato Cotyledon Explants

Yaroshko

Pasternak

Larriba

et al. 2023

Plants

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Cultivated tomato (Solanum lycopersicum L.) is one of the most important horticultural crops in the world. The optimization of culture media for callus formation and tissue regeneration of different tomato genotypes presents numerous biotechnological applications. In this work, we have analyzed the effect of different concentrations of zeatin and indole-3-acetic acid on the regeneration of cotyledon explants in tomato cultivars M82 and Micro-Tom. We evaluated regeneration parameters such as the percentage of callus formation and the area of callus formed, as well as the initiation percentage and the number of adventitious shoots. The best hormone combination produced shoot-like structures after 2–3 weeks. We observed the formation of leaf primordia from these structures after about 3–4 weeks. Upon transferring the regenerating micro-stems to a defined growth medium, it was possible to obtain whole plantlets between 4 and 6 weeks. This hormone combination was applied to other genotypes of S. lycopersicum, including commercial varieties and ancestral tomato varieties. Our method is suitable for obtaining many plantlets of different tomato genotypes from cotyledon explants in a very short time, with direct applications for plant transformation, use of gene editing techniques, and vegetative propagation of elite cultivars.

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“…However, there is still a gap in the use of Agrobacterium -mediated transformation for the delivery of CRISPR-Cas9 systems in plants that must be overcome, or some alternative methods need to be applied. Efforts have been made for the optimization of Agrobacterium -mediated transformation for GE of multiple plants, such as cotton, wheat, tomato, Maize, soybean, and rice [ 17 , 30 , 31 , 32 , 33 , 34 , 35 , 36 ]. The Agrobacterium -mediated delivery of CRISPR-Cas9 has been shifted to other field crops, such as Hemp [ 37 ].…”

Section: Ways To Deliver Crispr/cas Reagents To Plantsmentioning

confidence: 99%

Plant Virus-Derived Vectors for Plant Genome Engineering

Mahmood

Naqvi

Rahman

et al. 2023

Viruses

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Advances in genome engineering (GE) tools based on sequence-specific programmable nucleases have revolutionized precise genome editing in plants. However, only the traditional approaches are used to deliver these GE reagents, which mostly rely on Agrobacterium-mediated transformation or particle bombardment. These techniques have been successfully used for the past decades for the genetic engineering of plants with some limitations relating to lengthy time-taking protocols and transgenes integration-related regulatory concerns. Nevertheless, in the era of climate change, we require certain faster protocols for developing climate-smart resilient crops through GE to deal with global food security. Therefore, some alternative approaches are needed to robustly deliver the GE reagents. In this case, the plant viral vectors could be an excellent option for the delivery of GE reagents because they are efficient, effective, and precise. Additionally, these are autonomously replicating and considered as natural specialists for transient delivery. In the present review, we have discussed the potential use of these plant viral vectors for the efficient delivery of GE reagents. We have further described the different plant viral vectors, such as DNA and RNA viruses, which have been used as efficient gene targeting systems in model plants, and in other important crops including potato, tomato, wheat, and rice. The achievements gained so far in the use of viral vectors as a carrier for GE reagent delivery are depicted along with the benefits and limitations of each viral vector. Moreover, recent advances have been explored in employing viral vectors for GE and adapting this technology for future research.

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Highly efficient Agrobacterium-mediated transformation and plant regeneration system for genome engineering in tomato

Cited by 16 publications

References 63 publications

Optimization and establishment of Agrobacterium-mediated transformation in alfalfa (Medicago sativa L.) using eGFP as a visual reporter

Optimization and establishment of Agrobacterium-mediated transformation in alfalfa (Medicago sativa L.) using eGFP as a visual reporter

Optimization of Callus Induction and Shoot Regeneration from Tomato Cotyledon Explants

Plant Virus-Derived Vectors for Plant Genome Engineering

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