Optimization of particle bombardment parameters for the genetic transformation of Brazilian maize inbred lines

Petrillo, Caroline Pereira; Carneiro, N. P.; Purcino, A. A. C.; Carvalho, C. H. S.; Alves, José Donizeti; Carneiro, A. A.

doi:10.1590/s0100-204x2008000300012

Cited by 35 publications

(26 citation statements)

References 20 publications

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“…However, the Hi-II hybrid presents poor agronomic performance (Ishida et al, 2007;Que et al, 2014). The use of a genotype showing good in vitro cultivation response and good agronomic characteristics, such as the BR 451 (Grando et al, 2013;Petrillo et al, 2008), accelerates commercial transgenic plant production, since the gene inserted in the transgenic event can be transferred to elite lines without the transfer of additional unwanted traits. Therefore, improvement in the genetic transformation of BR 451 opens the possibility of implementing its use in genetic engineering.…”

Section: Resultsmentioning

confidence: 99%

“…However, the poor agronomic traits of the Hi-II hybrid (Ishida, Hiei, & Komari, 2007;Que et al, 2014) motivate the search for other maize genotypes that respond well to transformation. Since the Brazilian BR 451 maize variety presents high protein quality, good agronomical performance and adaptation to different regions of Brazil (Guimarães, Parentoni, & Pacheco, 2004), as well as a suitable in vitro response (Carvalho, Bohorova, & Bordallo, 1997;Petrillo et al, 2008), it offers a Brazilian alternative to the American Hi-II hybrid.…”

Section: Introductionmentioning

confidence: 99%

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Increased transient genetic transformation in immature embryos of Brazilian BR 451 maize co-cultivated with Agrobacterium tumefaciens

et al. 2018

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ABSTRACT. Genetic engineering has amplified the possibilities of crop breeding and supported sustainability ideals. Agrobacterium tumefaciens is the preferred transformation system, since it produces transgenic plants with more stable transgene expression and inheritance. The agrobacteria and plant tissue must be co-cultivated in conditions that allow gene transfer. This study aimed to evaluate how cocultivation time and temperature affect the transformation of immature maize embryos of the Hi-II hybrid (model genotype) and the Brazilian BR 451 variety with A. tumefaciens EHA101:pTF102. The pTF102 plasmid carries a uidA reporter gene that enables transient transformation to be quickly verified by GUS histochemical assays. Increasing the co-cultivation period from three to five days at 20°C resulted in a higher number of GUS positive embryos and blue spots per embryo in the BR 451 Brazilian variety, indicating better bacterial T-DNA transfer into the target explant cells. This condition raised the BR 451 response level to match the response level of the Hi-II control genotype, indicating that this Brazilian variety is suitable for genetic transformation.Keywords: agrobacteria; GUS histochemical assay; genetic engineering; Zea mays.Aumento da transformação genética transiente em embriões imaturos do genótipo brasileiro de milho BR 451 co-cultivados com Agrobacterium tumefaciens RESUMO. A engenharia genética ampliou as possibilidades do melhoramento genético vegetal, apoiando ideais de sustentabilidade. A Agrobacterium tumefaciens é o sistema de transformação preferido, uma vez que permite a produção de plantas transgênicas com maior estabilidade na expressão gênica e na herdabilidade. A Agrobactéria e o tecido vegetal devem ser co-cultivados em condições que permitam a transferência de genes. Este estudo teve como objetivo avaliar como o tempo e a temperatura durante o co-cultivo afetam a transformação de embriões imaturos de milho do híbrido Hi-II (genótipo modelo) e da variedade brasileira BR 451 com A. tumefaciens EHA101:pTF102. O plasmídeo pTF102 carrega o gene repórter uidA permitindo que a transformação transiente seja prontamente verificada pelo ensaio histoquímico de GUS. O aumento do período de co-cultivo de três para cinco dias a 20°C resultou num maior número de embriões GUS positivos e maior número de pontos azuis por embrião na variedade brasileira BR 451, indicando uma melhor transferência do T-DNA da bactéria para as células do explante alvo. Esta condição elevou o nível de resposta da BR 451 chegando a coincidir com o nível de resposta do controle Hi-II, indicando que esta variedade brasileira é adequada para transformação genética.Palavras-chave: agrobactéria; ensaio histoquímico de GUS; engenharia genética; Zea mays.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Increased transient genetic transformation in immature embryos of Brazilian BR 451 maize co-cultivated with Agrobacterium tumefaciens

et al. 2018

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“…Usually, conditions which allow a high level of expression of this reporter gene are associated with a high rate of production of transgenic events (Hiei at al. 1994, Petrillo et al 2008, Vega et al 2008). …”

Section: Resultsmentioning

confidence: 99%

“…Embryogenic calli are normally formed when immature zygotic maize embryos are cultured in these basal media supplemented with auxins, such as 2,4-D or Dicamba (Green and Phillips 1975, Armstrong and Green 1985, Frame et al 2006, Petrillo et al 2008). …”

Section: Rav Souza Et Almentioning

confidence: 99%

Agrobacterium-mediated genetic transformation of a tropical elite maize line

Souza

Alves

Carneiro

et al. 2017

Crop Breed. Appl. Biotechnol.

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“…Teknik rekayasa genetika melalui penembakan partikel pada tanaman jagung sudah dilaporkan oleh beberapa peneliti (Klein et al, 1989;Petrillo et al, 2008;Shou et al, 2004;Wang et al, 2006) dan merupakan teknik yang efisien untuk menghasilkan tanaman transgenik jagung. Abdel-Rhman (2015) melakukan transformasi embrio muda jagung dengan gen anthranilate synthase (ASA2) yang berasal dari tembakau melalui penembakan partikel.…”

Section: Pendahuluanunclassified

Induksi dan Regenerasi Kalus Jagung yang Ditransformasi dengan Gen CsNitr1-L melalui Penembakan Partikel

Ambarwati¹,

Listanto²,

Slamet³

et al. 2016

J. AgroBiogen

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The success in development of transgenic plants is influenced by the regeneration system. The objective of the study was to assess the response of maize genotypes to regeneration system of organogenesis and embryogenesis, after transformed with CsNitr1-L gene through particle bombardment. Induction and callus regeneration of maize immature embryos of inbred lines Ult:cm.1#, ARC 178-123-112-XB3, and AZ2 were conducted through organogenesis, whereas those inbred lines AZ1, AZ2, P4G19(S)C2.59.3.3.1.3 and P4S3.29.4.4.1 were conducted through embryogenesis somatic. Transformation of CsNitr1-L gene was done with the distance of bombardment of 7 cm and 9 cm and calli were then selected using 10 mg/l hygromycin. All explants (100%) of inbred lines Ult:cm.1# formed organogenic callus, while callus formation of ARC 178-123-112-XB3 was 94.3% and AZ2 was 60.5%. Ult:cm.1# was the most responsive line to the regeneration of organogenesis and produced 24 green shoots, compared with ARC 178-123-112-XB3 which produced one green shoot and AZ2 that did not produce green shoots. The highest percentage of embryogenic calli formed through somatic embryogenesis was obtained on inbred lines AZ1 (85.4%) and the lowest was on P4S3.29.4.4.1 (18.9%). Inbred lines AZ1 had the highest percentage of regeneration (50.7%) and produced 62 plants, followed by P4G19(S)C2.59.3.3.1.3 that produced 17 plants (2.8%) and P4S3.29.4.4.1 which produced two plants. Preliminary identification on 31 putative transgenic plants through PCR analysis produced 22 plants (70.96%) that contained nptII gene. Keywords:Zea mays, organogenesis, somatic embryogenesis, transformant identification. ABSTRAKKeberhasilan perakitan tanaman transgenik dipengaruhi oleh proses regenerasi. Tujuan penelitian adalah mengetahui respons genotipe jagung terhadap sistem regenerasi organogenesis dan embriogenesis somatik setelah ditransformasi dengan gen CsNitr1-L melalui penembakan partikel. Induksi dan regenerasi kalus dari eksplan embrio muda jagung galur inbred Ult:cm.1#, ARC 178-123-112-XB3, dan AZ2 dilakukan secara organogenesis, sedangkan galur inbred AZ1, AZ2, P4G19(S)C2.59.3.3.1.3, dan P4S3.29.4.4.1 secara embriogenesis somatik. Transformasi gen CsNitr1-L dilakukan dengan jarak penembakan 7 cm dan 9 cm, kemudian kalus diseleksi dengan higromisin 10 mg/l. Semua (100%) eksplan galur Ult:cm.1# dapat membentuk kalus organogenik, sedangkan galur ARC 178-123-112-XB3 dan AZ2 masing-masing dapat membentuk kalus sebesar 94,3% dan 60,5%. Galur Ult:cm.1# paling responsif terhadap regenerasi secara organogenesis dan menghasilkan 24 tunas hijau dibanding dengan ARC 178-123-112-XB3 yang menghasilkan satu tunas hijau dan AZ2 yang tidak menghasilkan tunas hijau. Persentase terbentuknya kalus embriogenik yang tertinggi diperoleh pada galur AZ1 (85,4%) dan paling rendah pada galur P4S3.29.4.4.1 (18,9%). Galur AZ1 mempunyai daya regenerasi yang tertinggi (50,7%) dan menghasilkan 62 tanaman, diikuti galur P4G19(S)C2.59.3.3.1.3 yang menghasilkan 17 tanaman (2,8%) dan galur P4S3.29.4.4.1 yan...

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Optimization of particle bombardment parameters for the genetic transformation of Brazilian maize inbred lines

Cited by 35 publications

References 20 publications

<b>Increased transient genetic transformation <i>in immature</i> embryos of Brazilian BR 451 maize co-cultivated with <i>Agrobacterium tumefaciens

<b>Increased transient genetic transformation <i>in immature</i> embryos of Brazilian BR 451 maize co-cultivated with <i>Agrobacterium tumefaciens

Agrobacterium-mediated genetic transformation of a tropical elite maize line

Induksi dan Regenerasi Kalus Jagung yang Ditransformasi dengan Gen CsNitr1-L melalui Penembakan Partikel

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