Agrobacterium-mediated genetic transformation of commercially elite rice restorer line using nptII gene as a plant selection marker

Chakraborty, M.; Reddy, P. Sreedhara; Narasu, M. Laxmi; Krishna, Gaurav; Rana, Debashis

doi:10.1007/s12298-015-0334-y

Cited by 17 publications

(9 citation statements)

References 39 publications

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“…Hygromycin resistance is an effective selection marker in regenerating rice plants. An alternative selection marker such as G418/kanamycin resistance is required, however, when introducing another transgene or the CRISPR/Cas9 cassette into the existing hygromycin-resistant transgenic rice plants (Chakraborty et al 2016 ; Park et al 2016 ). In the 23 vectors suitable for rice transgenic experiments, 13 contain the HptII gene and 10 contain the NptII gene (Additional file 1 : Table S1, Figure S2).…”

Section: Discussionmentioning

confidence: 99%

A Versatile Vector Toolkit for Functional Analysis of Rice Genes

Zhang

Sun

et al. 2018

Rice

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BackgroundRice (Oryza sativa) is the main food for half of the world’s population, and is considered the model for molecular biology studies of monocotyledon species. Although the rice genome was completely sequenced about 15 years ago, the function of most rice genes is still unknown.ResultsIn this study, we developed a vector toolkit that contains 42 vectors for transient expression studies in rice protoplasts and stable expression analysis in transgenic rice. These vectors have been successfully used to study protein subcellular localization, protein-protein interaction, gene overexpression, and the CRISPR/Cas9-mediated gene editing. A novel feature of these vectors is that they contain a universal multiple cloning site, which enables more than 99% of the rice coding sequences to be conveniently transferred between vectors.ConclusionsThe versatile vectors represent a highly efficient and high-throughput toolkit for functional analysis of rice genes.Electronic supplementary materialThe online version of this article (10.1186/s12284-018-0220-7) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 99%

A Versatile Vector Toolkit for Functional Analysis of Rice Genes

Zhang

Sun

et al. 2018

Rice

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“…Histochemical data in this study showed that more than 90% of the transformed calli were evidently positive with GUS stain, and about 36.7% of the calli exhibited a deep blue staining intensity (scoring = XXX). Previous studies reported that transformation using different vector backbones such as the pPZP200 and pCAMBIA2201 produced less than 65% of GUS-positive transgenic rice calli [31,32]. This indicates that the newly constructed pMDC140-wwin2 vector in this study provides a higher transformation success rate and is able to express GUS consistently in the transformed calli.…”

Section: Discussionmentioning

confidence: 68%

Agrobacterium-Mediated Transformation of pMDC140 Plasmid Containing the Wheatwin2 Gene into the Tadong Rice Genome

et al. 2021

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Blast disease resulting from Magnaporthe oryzae fungal infection reduces annual rice yield by up to 30% globally. The wheatwin2 (wwin2) is a pathogenesis-related (PR) gene that encodes for a PR-4 protein with chitinase properties that is capable of degrading chitin, a major constituent of certain fungal cell walls. However, the potential for wwin2 to contribute to M. oryzae resistance in rice is unclear. This study reports the construction of a pMDC140 vector carrying the wwin2 gene and its Agrobacterium-mediated transformation into the Tadong rice genome. In brief, the wwin2 gene was synthesized and integrated into a pMDC140 vector using Gateway cloning technology and was transformed into the Tadong rice genome. Our results show a promising high transformation rate, with more than 90% of the transformed rice calli expressing β-glucuronidase (GUS), the reporter gene marker. The expression of the wwin2 gene in transformed rice calli was further confirmed using quantitative real-time polymerase chain reaction. In conclusion, a pMDC140-wwin2 vector was constructed, which had a high transformation rate and could consistently induce expression of the GUS and wwin2 genes in Tadong rice. Data of this study is beneficial for subsequent in vitro and M. oryzae-infected field experiments to confirm the defense mechanism of the wwin2 gene towards blast disease in rice.

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“…A amplificação do fragmento do gene nptII, nas amostras de raízes correspondentes, sugeriram a integração do T-DNA no genoma da pimenta longa (Figura 3). O gene nptII é o gene marcador de seleção mais utilizado em transformação genética de plantas (BHATTACHARYYA et al, 2014CHAKRABORTY et al, 2016. Ele codifica a enzima neomicina fosfotransferase II (nptII), também chamada aminoglicosídeo 3'-fosfotransferase II, que atua transferindo o grupamento  fosfato do ATP para um grupo 3-hidroxil da porção amino-hexose dos antibióticos aminoglicosados, como a canamicina, a neomicina, a geneticina e a paromicina, que assim são detoxificados por fosforilação (NYABOGA et al, 2014).…”

Section: Resultsunclassified

Produção De Raízes Transformadas De Pimenta-Longa Por Infecção Com Agrobacterium Rhizogenes

Júnior¹,

Ribeiro

Berbara

et al. 2017

RSA

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Resumo -Este trabalho teve por objetivo a obtenção de raízes transgênicas (hairy roots) de Piper hispidinervium (pimenta-longa), espécie reconhecida pela produção de óleo essencial rico em Safrol, metabólito secundário utilizado com fins terapêuticos e industrial. As raízes foram produzidas a partir da infecção de explantes in vitro de pimentalonga com Agrobacterium rhizogenes (Estirpe R1601). Os resultados mostraram que a estirpe R1601 de A. rhizogenes foi eficiente para inoculação baseado na imersão de explantes em meio de cultura liquido, com a bactéria em crescimento, o que representou maior praticidade, menor índice de contaminação, maior taxa de infecção e menores danos aos explantes. A resposta morfológica foi a mesma entre os clones, não permitindo a distinção dos mesmos. Explantes de folha de pimenta-longa, inoculados com a estirpe R1601, formaram raízes que apresentaram crescimento nos subcultivos subsequentes. Palavras-chave -Piper hispidinervium, cultura de raízes, metabolitos secundários, cultivo in vitro.Abstract -This work aimed to obtain transformed roots of Piper hispidinervium (long pepper) specie recognized for the production of Safrol-rich essential oil, a substance already used for therapeutic and industrial purposes. The roots were obtained from the infection of the plants with strain R1601 of the bacterium Agrobacterium rhizogenes. The results show that A. rhizogenes strain R1601 was very efficient for inoculation based on the immersion of explants in liquid culture medium, with the growing bacterium, because it presents greater practicality, lower contamination index, higher infection rate and what causes less damage to the explants. A morphological response was observed that did not allow the clones to be distinguished from each other. Long pepper leaf explants, inoculated with strain R1601, formed roots that showed growth in subsequent subcultures.Keywords -Piper hispidinervium, root culture, second metabolites, in vitro culture..

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Agrobacterium-mediated genetic transformation of commercially elite rice restorer line using nptII gene as a plant selection marker

Cited by 17 publications

References 39 publications

A Versatile Vector Toolkit for Functional Analysis of Rice Genes

A Versatile Vector Toolkit for Functional Analysis of Rice Genes

Agrobacterium-Mediated Transformation of pMDC140 Plasmid Containing the Wheatwin2 Gene into the Tadong Rice Genome

Produção De Raízes Transformadas De Pimenta-Longa Por Infecção Com Agrobacterium Rhizogenes

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