High-throughput transformation pipeline for a Brazilian japonica rice with bar gene selection

Dedičová, Beata; Bermúdez, C.; Prías, Mónica; Zuniga, Esthefani G.; Brondani, C.

doi:10.1007/s00709-014-0741-x

Cited by 9 publications

(7 citation statements)

References 98 publications

(88 reference statements)

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“…A commonly used for selection of transformed shoots is bar and applied bialaphos (glufosinate ammonium) assay is used Our results showed that 3.0 mg L -1 was a lethal concentration of bialaphos and this is consistent with those obtained by Dedicova et al (2015) who emphasis that the selection agent should completely suppress growth of untransformed plant cells. Herouet et al (2005) indicated that the bialaphos resistance gene bar encode for the protein phosphinothricin acethyl transferase (PAT) that provides resistance to Phosphinothricin (PPT) by catalyzing the addition of an acetyl group to the free amino group and this applies to other herbicide derivatives such as bialaphos, which are competitive inhibitors of glutamine synthesis.…”

Section: Discussionsupporting

confidence: 80%

Producing Transgenic Thompson Seedless Grape (Vitis vinifera) Plants using Agrobacterium tumefaciens

Metwali¹,

Soliman²,

Almaghrabi³

et al. 2016

IJAB

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Under osmotic stress plant avoid it by different modification in their metabolisms such as increasing the synthesis of mRNA of delta 1-pyrroline-5-carboxylate synthase (P5CS), where P5CS is a target gene for increasing proline production and expect to improve the resistance to abiotic stress. To test this hypothesis, grape (Vitis vinifera L.) cv. Thompson Seedless was conducted to develop a protocol for high frequency regeneration and Agrobacterium mediated P5CS gene transfer. Callus induction was achieved by culture leaf explants on Nitsch and Nitsch (NN) basal medium including 2.0 mg L -1 2,4-di-chloro-phenoxy-aceticacid + 0.5 mg L -1 Thiadiazuron solidified with 2.5 g L -1 phytagel. While for in vitro proliferation of plant, the calli were cultured on NN medium supplemented with 3.0 mg L -1 Zeatin riboside + 0.5 mg L -1 Thiadiazuron. Agrobacterium-mediated transformation using the strain LB4404 harbouring the binary vector pBI121 with the P5CS gene under CaMV 35S promoter and the bar gene as a plant selectable marker was used for transforming grape explants. A P5CS specific band (2100 bp) was amplified from DNA extracted only from the transgenic grape plants and 24% for P5CS gene was positive for the bar gene. Over expression of the abiotic stress P5CS gene was enhanced synthesis of proline to over 6 times higher in transgenic plants compared to controls. The successful transformation of genetically diverse grape cv. Thompson Seedless indicated that the transformation system may have general application to an even wider range of grapes cultivars.

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Section: Discussionsupporting

confidence: 80%

Producing Transgenic Thompson Seedless Grape (Vitis vinifera) Plants using Agrobacterium tumefaciens

Metwali¹,

Soliman²,

Almaghrabi³

et al. 2016

IJAB

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“…oats [ 55 ] and rye [ 56 ] still required improvements. Modifications and successful development of the existing transformation protocols including also completely new techniques and approaches [ 57 – 61 ]. Moreover, there exists plant species such as grasses where this technique has not been tested effectively or widely implemented.…”

Section: Introductionmentioning

confidence: 99%

Protocol development for somatic embryogenesis, SSR markers and genetic modification of Stipagrostis pennata (Trin.) De Winter

et al. 2021

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Background Stipagrostis pennata (Trin.) De Winter is an important species for fixing sand in shifting and semi-fixed sandy lands, for grazing, and potentially as a source of lignocellulose fibres for pulp and paper industry. The seeds have low viability, which limits uses for revegetation. Somatic embryogenesis offers an alternative method for obtaining large numbers of plants from limited seed sources. Results A protocol for plant regeneration from somatic embryos of S. pennata was developed. Somatic embryogenesis was induced on Murashige & Skoog (MS) medium supplemented with 3 mg·L–1 2,4-D subsequently shoots were induced on MS medium and supplemented with 5 mg·L–1 zeatin riboside. The highest shoots induction was obtained when embryogenic callus derived from mature embryos (96%) in combination with MS filter-sterilized medium was used from Khuzestan location. The genetic stability of regenerated plants was analysed using ten simple sequence repeats (SSR) markers from S. pennata which showed no somaclonal variation in regenerated plants from somatic embryos of S. pennata. The regenerated plants of S. pennata showed genetic stability without any somaclonal variation for the four pairs of primers that gave the expected amplicon sizes. This data seems very reliable as three of the PCR products belonged to the coding region of the genome. Furthermore, stable expression of GUS was obtained after Agrobacterium-mediated transformation using a super binary vector carried by a bacterial strain LBA4404. Conclusion To our knowledge, the current work is the first attempt to develop an in vitro protocol for somatic embryogenesis including the SSR marker analyses of regenerated plants, and Agrobacterium-mediated transformation of S. pennata that can be used for its large-scale production for commercial purposes.

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“…The 3,916 base pair (bp) sequence of the homologous gene of the Rubisco large subunit (rbcL) precursor, located in nuclear genome LOC_Os12g10580, was used in the transformation of the BRSMG Curinga rice cultivar, as described by Dedicova et al (2015). In this study, the rice transformation generated 23 independent events (E1 to E23), from which the evaluated plants were obtained, and generation advance was carried out from T1 onwards.…”

Section: Methodsmentioning

confidence: 99%

Drought tolerance induced by the overexpression of the nuclear rbcL gene in rice

Oliveira,

Vianello,

Lanna

et al. 2023

Pesq. agropec. bras.

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The objective of this work was to determine whether the overexpression of the nuclear Rubisco large subunit (rbcL) improves the drought tolerance of the genetically modified (GM) BRSMG Curinga upland rice (Oryza sativa) cultivar. GM and non-genetically modified (NGM) plants of the same cultivar were compared under the two following water treatments: well watered (WW) and water deficit (WD). The performance of the agronomic traits of GM plants, including grain yield, was superior to that of NGM plants in both treatments. By quantitative polymerase chain reaction, GM plants show a significantly higher expression of the rbcL gene in both WW and WD, as well as a larger amount of abscisic acid. With the RNAseq analysis, almost three times more upregulated genes are identified in GM plants in stage 2 after water restriction, indicating a greater protection against water deficit. The higher expression of genes related to the protection of the cellular metabolism and a series of physiological alterations may be involved in the increase in the drought tolerance of GM rice plants overexpressing the rbcL gene.

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High-throughput transformation pipeline for a Brazilian japonica rice with bar gene selection

Cited by 9 publications

References 98 publications

Producing Transgenic Thompson Seedless Grape (Vitis vinifera) Plants using Agrobacterium tumefaciens

Producing Transgenic Thompson Seedless Grape (Vitis vinifera) Plants using Agrobacterium tumefaciens

Protocol development for somatic embryogenesis, SSR markers and genetic modification of Stipagrostis pennata (Trin.) De Winter

Drought tolerance induced by the overexpression of the nuclear rbcL gene in rice

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