Optimizing callus induction and proliferation for <i>Agrobacterium-</i>mediated transformation of <i>Brachypodium distachyon</i>

Behpouri, A.; Perochon, Alexandre; Doohan, Fiona M.; Ng, Carl

doi:10.1556/0806.46.2018.04

Cited by 4 publications

(3 citation statements)

References 26 publications

(60 reference statements)

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“…Addition of KH2PO4 which was found to increase the percentage of water in the callus [22] was also observed to contribute to reduction of phenolics [13]. Earlier studies [23,24] described an embryogenic callus further to be a white compact or creamy to yellowish embryogenic callus. As has been the case with induction, sorghum calli regeneration has not been reproducible.…”

Section: Introductionmentioning

confidence: 90%

Improved Culture Media for Embryogenic Callus Generation in Sorghum [Sorghum bicolor (L.) Moench]

Chege¹,

Palágyi²,

Lantos³

et al. 2020

Phyton

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Many attempts on optimization of sorghum [Sorghum bicolor (L.) Moench] tissue culture induction media have been made, but the culture system remains with some bottlenecks compared to that of other crops. This study aimed at assessing the suitability of various induction media to produce embryogenic callus (yellow and friable) with high induction rates and reduced phenolic exudation. The six culture medium modifications: 3 based on Murashige and Skoog (MS) medium and one each based on Chu N6, Gamborg B5 and 190-2 media respectively were applied in the culture of mature embryos from 10 sorghum genotypes. Although there was a genotype influence on the attainment of a yellow callus, friability of the callus was determined to be dependent on the culture medium and not the genotype. Half strength MS medium with 0.2 mg/l 2,4-D with 2.8 g/l Gelrite® as the gelling agent modified with 1.0 g/l KH2PO4, 1.0 g/l L-proline, 1.0 g/l L-asparagine and 0.16 mg/l CuSO4•5H2O (type E) was found to be the most effective resulting in about 60% yellow coloured callus induction with 25% friability. Addition of CuSO4•5H2O, KH2PO4, L-proline and L-asparagine significantly reduced the phenolic production. Half strength MS medium was observed to contribute to quality callus production when compared to full strength MS media modified with the compounds. The half strength MS medium was also observed to suppress phenolic production. Medium 190-2 produced the highest regeneration frequency (40%) among the 3-regeneration media tested. The results provide information on a suitable sorghum callus induction medium necessary for embryogenesis.

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Section: Introductionmentioning

confidence: 90%

Improved Culture Media for Embryogenic Callus Generation in Sorghum [Sorghum bicolor (L.) Moench]

Chege¹,

Palágyi²,

Lantos³

et al. 2020

Phyton

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“…Embryogenic calli are typically induced on an Murashige and Skoog medium (MS) or Linsmaier and Skoog Medium (LS) medium that has been supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) in different concentrations [33,38,46,48]. The optimal concentration of (2,4-D) for both callus induction and proliferation in B. distachyon is 2.5 mg l −1 [46,49]. Similar conditions can be used for other members of the genus, including B. sylvaticum, B. stacei and B. hybridum [42,48].…”

Section: Callus Inductionmentioning

confidence: 99%

In Vitro Tissue Culture in Brachypodium: Applications and Challenges

Betekhtin

Hus

Rojek-Jelonek

et al. 2020

IJMS

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Brachypodium distachyon has become an excellent model for plant breeding and bioenergy grasses that permits many fundamental questions in grass biology to be addressed. One of the constraints to performing research in many grasses has been the difficulty with which they can be genetically transformed and the generally low frequency of such transformations. In this review, we discuss the contribution that transformation techniques have made in Brachypodium biology as well as how Brachypodium could be used to determine the factors that might contribute to transformation efficiency. In particular, we highlight the latest research on the mechanisms that govern the gradual loss of embryogenic potential in a tissue culture and propose using B. distachyon as a model for other recalcitrant monocots.

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“…sativa, Hordeum vulgare and Brachypodium distachyon(Kumar et al 2005;Shrawat et al 2007;Behpouri et al 2018). The number of shoot apices expressing GUS at OD 600 = 1.0 was observed to be 15 ± 2.9 and 18 ± 3.7 for pCAMBIA 1301 and pCAMBIA 1305.1, respectively.…”

mentioning

confidence: 99%

Agrobacterium–mediated genetic engineering system for the C4 apomictic grass model species Cenchrus ciliaris L.

Dwivedi,

Abhishek

2024

Preprint

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Cenchrus ciliaris L. (buffel grass) is a popular tropical range grass known for its perenniality and high palatability. Owing to its apomictic mode of reproduction, breeding this polymorphic grass is restricted to in vitro approaches of genetic manipulation. Apomixis facilitates genetic improvement by fixing heterosis as well as the changes made to the genome through transgenesis. Towards its genetic improvement, a robust protocol of plant transformation is desirable. Hence, first attempt to develop a rapid and efficient Agrobacterium– mediated transformation system for Cenchrus ciliaris genotype IG-3108 has been made. In this study, direct multiple shoot induction protocol using shoot apex explants was found to be suitable for transformation. Several factors such as inoculum of bacteria, co–culture time, co–cultivation duration, concentration of acetosyringone and effect of vacuum infiltration were optimized for achieving high transformation frequency using shoot apex explants. The shoot apex explants were co-cultured with EHA 105 harbouring the binary vectors pCAMBIA 1301 and 1305.1 containing the hptII gene as a selectable marker and GUSA as a reporter gene. The highest transformation frequency with pCAMBIA 1301 vector was 1.42% while with pCAMBIA 1305.1 vector, it was 1.37% when the explants were co-cultured for 30 min with cells of Agrobacterium at OD600 = 1.0 under vacuum (0.5 X 105 Pa) followed by co-cultivation for 3 days on MS with 3 mg/L TDZ and 400 µM acetosyringone. The status of transgene and its integration in the genome of regenerated putative transformed plants of Cenchrus were confirmed by PCR and Southern blot analysis.

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Optimizing callus induction and proliferation for Agrobacterium-mediated transformation of Brachypodium distachyon

Cited by 4 publications

References 26 publications

Improved Culture Media for Embryogenic Callus Generation in Sorghum [Sorghum bicolor (L.) Moench]

Improved Culture Media for Embryogenic Callus Generation in Sorghum [Sorghum bicolor (L.) Moench]

In Vitro Tissue Culture in Brachypodium: Applications and Challenges

Agrobacterium–mediated genetic engineering system for the C4 apomictic grass model species Cenchrus ciliaris L.

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