A simple and efficient Agrobacterium-mediated in planta transformation protocol for horse gram (Macrotyloma uniflorum Lam. Verdc.)

Amal, Thomas Cheeran; Karthika, Palanisamy; Gurusamy, Dhandapani; Selvakumar, S.; Vasanth, Krishnan

doi:10.1186/s43141-020-00023-z

Cited by 11 publications

(7 citation statements)

References 46 publications

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“…To increase the number of viable embryos and ovaries via in vitro techniques, future work should entail various hormones and concentrations of the different embryo and ovary ages. Among other varieties of horse gram paiyur-2 variety exhibited a maximum germination percentage of 93% 26 . So, this study was carried out in the paiyur-2 variety.…”

Section: Discussionmentioning

confidence: 98%

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Embryo and Ovary Culture of Important Legume Horse Gram, Macrotyloma uniflorum (Lam.) Verdc

Karthika¹,

Venkatacha²,

Gurusamy³

et al. 2022

Asian J. of Crop Science

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Background and Objective: Macrotyloma uniflorum is nutritious and a good source of bioactive substances with high biological processes. The high frequency and efficient in vitro regeneration protocol have developed first time by utilizing embryo and ovary explants in Horsegram, Macrotyloma uniflorum. Materials and Methods: This research was carried out from horse gram paiyur-2 variety seeds derived embryo and ovary explants used by MS medium supplemented with cytokinins and auxins. Results: The phytohormone 0.4 mg LG 1 zeatin induced a maximum of 12 and 9 shoots from an embryo and ovary explants, respectively. The maximum frequency of rooting with about 9 roots was procured from the shoots cultured from embryonic explants using 0.2 mg LG 1 NAA and 7 roots were propagated from shoots derived from ovary explants. Overall studies observed that the embryonic explants show a high percent frequency of shoot and root induction when compared to ovary culture and further plantlets rose using embryonic explants with shoot growth period than the ovary explants. Conclusion: In vitro legume regeneration utilizing embryo and ovary is efficient, dependable and has fewer problems. Embryo explants were used to induce high-frequency plant regeneration, indicating that the ovaries are useful in genetic transformation.

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Section: Discussionmentioning

confidence: 98%

“…Earlier works have been able to induce the horse gram plant regeneration and organogenesis of plantlets using shoot tip 25 , leaf 22 , immature cotyledon 23 , cotyledonary nodal 26 explants and recently 27 done the embryo culture. But no attempt was made to culture the embryo and ovary explants.…”

Section: Discussionmentioning

confidence: 99%

Embryo and Ovary Culture of Important Legume Horse Gram, Macrotyloma uniflorum (Lam.) Verdc

Karthika¹,

Venkatacha²,

Gurusamy³

et al. 2022

Asian J. of Crop Science

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“…Moreover, even when vacuum agroinfiltration is performed on the entire plant, observing the expression in the petioles and stem is often not possible [ 29 , 30 ]. However, studies focused on the seedling expression have potential relevance for future in planta transformation procedures [ 31 , 32 ]. An example of this potential was demonstrated in a study by Hu et al [ 33 ] with radishes, who obtained mutants through an in planta transformation procedure in seedlings.…”

Section: Discussionmentioning

confidence: 99%

Comparative Evaluation of Transient Protein Expression Efficiency in Tissues across Soybean Varieties Using the Tsukuba System

Fuhrmann-Aoyagi,

Igarashi,

Miura

2024

Plants

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Transient protein expression is a versatile tool with diverse applications and can be used in soybeans to study gene function, obtain mutants, and produce proteins for commercial use. However, soybeans are considered recalcitrant for agroinfiltration. Subsequent studies on soybeans have demonstrated a green fluorescent protein (GFP) expression in seedpods, but not in leaves, using syringe agroinfiltration. To evaluate agroinfiltration-based transient protein expression levels in plant cells, we used the transient expression vector pTKB3 harboring the GFP gene. Using Agrobacterium tumefaciens, vacuum agroinfiltration of the leaves and needle agroinfiltration of the seedlings of different soybean varieties were performed. GFP was transiently expressed in all of the samples. However, the Enrei and Williams 82 varieties presented better results than the other varieties in the leaf tissue, with results confirmed by immunoblot analysis, demonstrating that both varieties are good candidates for molecular biological studies. GFP expression in the seedlings was less extensive than that in the leaves, which may be due to the tissue characteristics, with Enrei showing the best results. Based on this observation, we conclude that the Tsukuba system is an effective tool that can be used for different tissues and soybean varieties.

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“…As such, multiple highly efficient in planta research articles performing callus regeneration under in vivo conditions have been published over the years [16][17][18][19][20]. Similarly, several effective in planta protocols using minimal in vitro steps have also been published [21][22][23][24][25]. Conceptually speaking, these published methods all fall within the scope of in planta transformation and were self-described as in planta by their authors; however, their methods do not strictly follow the definitions mentioned above.…”

Section: Definition Of In Planta Stable Transformationmentioning

confidence: 99%

A comprehensive review of in planta stable transformation strategies

Bélanger,

Copley,

Hoyos-Villegas

et al. 2024

Plant Methods

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Plant transformation remains a major bottleneck to the improvement of plant science, both on fundamental and practical levels. The recalcitrant nature of most commercial and minor crops to genetic transformation slows scientific progress for a large range of crops that are essential for food security on a global scale. Over the years, novel stable transformation strategies loosely grouped under the term “in planta” have been proposed and validated in a large number of model (e.g. Arabidopsis and rice), major (e.g. wheat and soybean) and minor (e.g. chickpea and lablab bean) species. The in planta approach is revolutionary as it is considered genotype-independent, technically simple (i.e. devoid of or with minimal tissue culture steps), affordable, and easy to implement in a broad range of experimental settings. In this article, we reviewed and categorized over 300 research articles, patents, theses, and videos demonstrating the applicability of different in planta transformation strategies in 105 different genera across 139 plant species. To support this review process, we propose a classification system for the in planta techniques based on five categories and a new nomenclature for more than 30 different in planta techniques. In complement to this, we clarified some grey areas regarding the in planta conceptual framework and provided insights regarding the past, current, and future scientific impacts of these techniques. To support the diffusion of this concept across the community, this review article will serve as an introductory point for an online compendium about in planta transformation strategies that will be available to all scientists. By expanding our knowledge about in planta transformation, we can find innovative approaches to unlock the full potential of plants, support the growth of scientific knowledge, and stimulate an equitable development of plant research in all countries and institutions.

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A simple and efficient Agrobacterium-mediated in planta transformation protocol for horse gram (Macrotyloma uniflorum Lam. Verdc.)

Cited by 11 publications

References 46 publications

Embryo and Ovary Culture of Important Legume Horse Gram, Macrotyloma uniflorum (Lam.) Verdc

Embryo and Ovary Culture of Important Legume Horse Gram, Macrotyloma uniflorum (Lam.) Verdc

Comparative Evaluation of Transient Protein Expression Efficiency in Tissues across Soybean Varieties Using the Tsukuba System

A comprehensive review of in planta stable transformation strategies

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