Choice of Explant for Plant Genetic Transformation

Chakraborty, N B; Chakraborty, Priyanka; Sen, Moutushi; Bandopadhyay, Rajib

doi:10.1007/978-1-0716-0356-7_5

Cited by 7 publications

(3 citation statements)

References 73 publications

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“…Explants are small fragments of living tissue that play an important role in the efficiency of transformation and can be removed from many different parts of a plant: shoots, leaves, stems, roots, etc. [ 81 ]. A variety of explants are used in rice cultivation, with the most frequent being embryogenic calli (26) and protoplasts (6).…”

Section: Discussionmentioning

confidence: 99%

Gene Editing for Plant Resistance to Abiotic Factors: A Systematic Review

Nascimento

Rocha

Soares

et al. 2023

Plants

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Agricultural crops are exposed to various abiotic stresses, such as salinity, water deficits, temperature extremes, floods, radiation, and metal toxicity. To overcome these challenges, breeding programs seek to improve methods and techniques. Gene editing by Clustered Regularly Interspaced Short Palindromic Repeats—CRISPR/Cas—is a versatile tool for editing in all layers of the central dogma with focus on the development of cultivars of plants resistant or tolerant to multiple biotic or abiotic stresses. This systematic review (SR) brings new contributions to the study of the use of CRISPR/Cas in gene editing for tolerance to abiotic stress in plants. Articles deposited in different electronic databases, using a search string and predefined inclusion and exclusion criteria, were evaluated. This SR demonstrates that the CRISPR/Cas system has been applied to several plant species to promote tolerance to the main abiotic stresses. Among the most studied crops are rice and Arabidopsis thaliana, an important staple food for the population, and a model plant in genetics/biotechnology, respectively, and more recently tomato, whose number of studies has increased since 2021. Most studies were conducted in Asia, specifically in China. The Cas9 enzyme is used in most articles, and only Cas12a is used as an additional gene editing tool in plants. Ribonucleoproteins (RNPs) have emerged as a DNA-free strategy for genome editing without exogenous DNA. This SR also identifies several genes edited by CRISPR/Cas, and it also shows that plant responses to stress factors are mediated by many complex-signaling pathways. In addition, the quality of the articles included in this SR was validated by a risk of bias analysis. The information gathered in this SR helps to understand the current state of CRISPR/Cas in the editing of genes and noncoding sequences, which plays a key role in the regulation of various biological processes and the tolerance to multiple abiotic stresses, with potential for use in plant genetic improvement programs.

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Section: Discussionmentioning

confidence: 99%

Gene Editing for Plant Resistance to Abiotic Factors: A Systematic Review

Nascimento

Rocha

Soares

et al. 2023

Plants

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“…The results of your research suggest that the root explants used by Bolouri et al were less effective compared to the hypocotyl explants employed in your study. Indeed, the choice of explant is a critical factor influencing the success of in vitro culture [47,48].…”

Section: Discussionmentioning

confidence: 99%

Modeling Callus Induction and Regeneration in Hypocotyl Explant of Fodder Pea (Pisum sativum var. arvense L.) Using Machine Learning Algorithm Method

Türkoğlu,

Bolouri,

Haliloğlu

et al. 2023

Agronomy

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A comprehensive understanding of genetic diversity and the categorization of germplasm is important to effectively identify appropriate parental candidates for the goal of breeding. It is necessary to have a technique of tissue culture that is both effective and reproducible to perform genetic engineering on fodder pea genotypes (Pisum sativum var. arvense L.). In this investigation, the genetic diversity of forty-two fodder pea genotypes was assessed based on their ability of callus induction (CI), the percentage of embryogenic callus by explant number (ECNEP), the percentage of responding embryogenic calluses by explant number (RECNEP), the number of somatic embryogenesis (NSE), the number of responding somatic embryogenesis (RSE), the regeneration efficiency (RE), and the number of regenerated plantlets (NRP). The findings of the ANOVA showed that there were significant differences (p < 0.001) between the genotypes for all in vitro parameters. The method of principal component analysis (PCA) was used to study the correlations that exist between the factors associated with tissue culture. While RE and NRP variables were most strongly associated with Doğruyol, Ovaçevirme-4, Doşeli-1, Yolgeçmez, and Incili-3 genotypes, RECNEP, NSE, RDE, and RECNEP variables were strongly associated with Avcılar, Ovaçevirme-3, and Ardahan Merkez-2 genotypes. The in vitro process is a complex multivariate process and more robust analyses are needed for linear and nonlinear parameters. Within the scope of this study, artificial neural network (ANN), random forest (RF), and multivariate adaptive regression spline (MARS) algorithms were used for RE estimation, and these algorithms were also compared. The results that we acquired from our research led us to the conclusion that the employed ANN-multilayer perceptron (ANN-MLP) model (R2 = 0.941) performs better than the RF model (R2 = 0.754) and the MARS model (R2 = 0.214). Despite this, it has been shown that the RF model is capable of accurately predicting RE in the early stages of the in vitro process. The current work is an inquiry regarding the use of RF, MARS, and ANN models in plant tissue culture, and it indicates the possibilities of application in a variety of economically important fodder peas.

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“…Transgenic plants are commonly used in molecular biology to understand the function of specific genes ( Chakraborty et al, 2020). The establishment of an efficient, stable and regenerating receptor system is the basis for genetic transformation.…”

Section: Introductionmentioning

confidence: 99%

Optimization of callus induction and proliferation of Paeonia lactiflora Pall. and Agrobacterium-mediated genetic transformation

et al. 2022

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Paeonia lactiflora Pall. is an important ornamental plant with high economic and medicinal value, which has considerable development prospects worldwide. The lack of efficient tissue culture techniques and genetic transformation systems has become a master obstacle for P. lactiflora research. The purpose of the present study focuses on obtaining an efficient and stable genetic transformation method using callus as the receptor and exploring an efficient protocol for callus induction and proliferation associated with P. lactiflora. Callus induction and proliferation were performed using MS medium with various concentrations of 2,4-Dichlorophenoxyacetic acid (2,4-D), 1-Naphthaleneacetic acid (NAA), 6-Benzylaminopurine (6-BA) and thidiazuron (TDZ). The sensitivity of callus to kanamycin and cefotaxime was determined. Several parameters such as Agrobacterium cell density, infection time and co-culture duration were studied to optimize transformation efficiency. Agrobacterium strains EHA105 and pBI121 binary vector harboring the β-glucuronidase (GUS) gene were used for transformation. Expression of the GUS reporter gene was detected by GUS assay, polymerase chain reaction (PCR) and Quantitative Real-time PCR (RT-qPCR). The MS medium containing 1.0 mg·L-1 NAA, 0.5 mg·L-1 2,4-D and 0.5 mg·L-1 TDZ was optimal for callus induction and MS medium containing 0.5 mg·L-1 NAA, 1.0 mg·L-1 2,4-D and 0.5 mg·L-1 TDZ was the best for callus proliferation. The concentrations of kanamycin and cefotaxime used for screening positive callus were 125 mg·L-1 and 200 mg·L-1, respectively. Among various combinations analyzed, the best transformation result was obtained via the 25 min of infection of Agrobacterium at 0.6 OD600 and 3 d of co-culture. Overall, this study provided technical support and theoretical guidance for improving the callus induction and proliferation efficiency and the study of gene function in P. lactiflora.

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Choice of Explant for Plant Genetic Transformation

Cited by 7 publications

References 73 publications

Gene Editing for Plant Resistance to Abiotic Factors: A Systematic Review

Gene Editing for Plant Resistance to Abiotic Factors: A Systematic Review

Modeling Callus Induction and Regeneration in Hypocotyl Explant of Fodder Pea (Pisum sativum var. arvense L.) Using Machine Learning Algorithm Method

Optimization of callus induction and proliferation of Paeonia lactiflora Pall. and Agrobacterium-mediated genetic transformation

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