Genetic Transformation Studies on Avocado Cultivar “Hass” (Persea americana)

Ahmed, Muhammad Faisal; Kantharajah, Arumugam S.; Holford, Paul

doi:10.4236/ajps.2012.39148

Cited by 7 publications

(4 citation statements)

References 19 publications

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“…Jourbert et al (2004 found that the transformation efficiency for tobacco leaves was decreased after addition of 250 lM acetosyringone into inoculation and co-cultivation media. A similar finding was reported by Ahmed et al (2012) where the transformation frequency in avocado cells was decreased drastically at 400 lM acetosyringone. Several studies also indicated that acetosyringone is not an essential factor for Agrobacterium-mediated transformation, particularly in barley (Shrawat et al 2007) and rice (Aananthi et al 2010).…”

Section: Resultssupporting

confidence: 85%

Enhancement of Agrobacterium-mediated transformation efficiency of model plant using quorum sensing molecule, N-3-oxo-octanoyl-l-homoserine-lactone

Chen

Chan

Tan

2015

Plant Cell Tiss Organ Cult

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Limitations of genetic transformation throughAgrobacterium tumefaciens infection have been reported in many plant species, mainly due to low virulence of Agrobacterium to plant host and inefficient integration of T-DNA into plant genome. In this study, we investigated the effect of quorum sensing molecules, N-acylhomoserine-lactones (AHLs), on Agrobacterium-mediated transformation of leaf explants of Nicotiana tabacum cv. SR1. Our results revealed that AHLs significantly influenced the transformation frequency in N. tabacum. The highest percentage of transformation (92.2 %) was recorded when leaf explants infected with Agrobacterium were co-cultivated for 3 days on Murashige and Skoog medium supplemented with 100 lM N-3-oxo-octanoyl-L-homoserine-lactone. The presence of reporter genes in regenerated plants was determined by GUS histochemical staining, gfp visualization and PCR analysis. Our results indicated the promotive effect of AHLs in facilitating the transfer of desirable genes into N. tabacum. This improved protocol could be applicable to various species for functional genomics studies and biotechnological applications including crop improvement programs.

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Section: Resultssupporting

confidence: 85%

Enhancement of Agrobacterium-mediated transformation efficiency of model plant using quorum sensing molecule, N-3-oxo-octanoyl-l-homoserine-lactone

Chen

Chan

Tan

2015

Plant Cell Tiss Organ Cult

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“…Among the three Agrobacterium strains assessed, we observed that the LBA4404 is suitable for transient expression in avocado leaves. In two previous avocado transformation studies comparing different Agrobacterium strains, the LBA4404 strain did not give the best results, though this may be because the tissue they agroinfiltrated were globular somatic embryonic masses (Palomo-Ríos et al 2012), and embryonic shoot axes (Ahmed et al 2012).…”

Section: Discussionmentioning

confidence: 86%

“…Currently the molecular analyses are hindered by the lack of efficient and rapid transformation protocols. In the last two decades there has only been a single report using biolistics for transient transformation of embryogenic callus (Chaparro-Pulido et al 2014), one report using Agrobacterium rhizogenes to produce composite avocado plants (Prabhu et al 2017), and few reports detailing the use of Agrobacterium to generate stable transformants (Cruz-Hernández et al 1998;Raharjo et al 2003Raharjo et al , 2008Palomo-Ríos et al 2011Ahmed et al 2012). These latter reports employed embryogenic masses derived from zygotic embryos, due to their high potential and competence for regeneration.…”

Section: Introductionmentioning

confidence: 99%

In-planta transient transformation of avocado (Persea americana) by vacuum agroinfiltration of aerial plant parts

Salazar-González

Castro-Medina

Bernardino-Rivera

et al. 2023

Plant Cell Tiss Organ Cult

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Avocado, Persea americana Mill, is one of the most traded tropical fruits in the international market. To date, stable and transient transformation has only been achieved for of zygotic embryos and not of adult plant tissue, which limits functional genomics research. We provide the first transient Agrobacterium-mediated transformation methodology in avocado leaves that overcomes the recalcitrance to transformation of this species. We investigated the effect of Agrobacterium strain, leaf stage, wounding pre-treatment, the phytohormone jasmonic acid, and vacuum infiltration on transient transformation of avocado leaves. Using the Agrobacterium strain LBA4404 and the RUBY reporter a transformation frequency of up to 27% was obtained for avocado detached leaves. The transformation efficiency depended on the age of the leaf, with an intermediate stage of leaf development showing the highest efficiency of transient reporter gene expression. Microwounding pre-treatment facilitates agroinfiltration and coupled with leaf age are the primary factors influencing competence for transient transformation. Jasmonic acid did not significantly affect transient transformation in the absence of microwounding. However, microwounding and 250 µM of jasmonic acid acted synergistically to significantly enhance transient expression. Using this methodology with localized vacuum agroinfiltration, transient transformation of attached avocado leaves was achieved. This method unlocks the use of Agrobacterium-mediated transient transformation as a tool for explore gene function and metabolic pathways in both, detached and attached avocado leaves.

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“…Certain combinations of host genotypes and bacterial strains have been reported to be more efficient in rooting and transformation in other plant systems as well [ 49 ]. In addition, interactions between the genetic background of the Agrobacterium , binary vectors, acetosyringone concentrations and pH have been shown to have an impact on avocado transformation efficiencies [ 12 ]. Highest transformation efficiencies were achieved in A0.74 upon H+ARqua1(+pBINUbiGUSint) infection.…”

Section: Discussionmentioning

confidence: 99%

Generation of composite Persea americana (Mill.) (avocado) plants: A proof-of-concept-study

et al. 2017

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Avocado (Persea americana (Mill.)), an important commercial fruit, is severely affected by Phytophthora Root Rot in areas where the pathogen is prevalent. However, advances in molecular research are hindered by the lack of a high-throughput transient transformation system in this non-model plant. In this study, a proof-of-concept is demonstrated by the successful application of Agrobacterium rhizogenes-mediated plant transformation to produce composite avocado plants. Two ex vitro strategies were assessed on two avocado genotypes (Itzamna and A0.74): In the first approach, 8-week-old etiolated seedlings were scarred with a sterile hacksaw blade at the base of the shoot, and in the second, inch-long incisions were made at the base of the shoot (20-week-old non-etiolated plants) with a sterile blade to remove the cortical tissue. The scarred/wounded shoot surfaces were treated with A. rhizogenes strains (K599 or ARqua1) transformed with or without binary plant transformation vectors pRedRootII (DsRed1 marker), pBYR2e1-GFP (GFP- green fluorescence protein marker) or pBINUbiGUSint (GUS- beta-glucuronidase marker) with and without rooting hormone (Dip 'N' Grow) application. The treated shoot regions were air-layered with sterile moist cocopeat to induce root formation. Results showed that hormone application significantly increased root induction, while Agrobacterium-only treatments resulted in very few roots. Combination treatments of hormone+Agrobacterium (-/+ plasmids) showed no significant difference. Only the ARqua1(+plasmid):A0.74 combination resulted in root transformants, with hormone+ARqua1(+pBINUbiGUSint) being the most effective treatment with ~17 and 25% composite plants resulting from strategy-1 and strategy-2, respectively. GUS- and GFP-expressing roots accounted for less than 4 and ~11%, respectively, of the total roots/treatment/avocado genotype. The average number of transgenic roots on the composite plants was less than one per plant in all treatments. PCR and Southern analysis further confirmed the transgenic nature of the roots expressing the screenable marker genes. Transgenic roots showed hyper-branching compared to the wild-type roots but this had no impact on Phytophthora cinnamomi infection. There was no difference in pathogen load 7-days-post inoculation between transformed and control roots. Strategy-2 involving A0.74:ARqua1 combination was the best ex vitro approach in producing composite avocado plants. The approach followed in this proof-of-concept study needs further optimisation involving multiple avocado genotypes and A. rhizogenes strains to achieve enhanced root transformation efficiencies, which would then serve as an effective high-throughput tool in the functional screening of host and pathogen genes to improve our understanding of the avocado-P. cinnamomi interaction.

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Genetic Transformation Studies on Avocado Cultivar “Hass” (Persea americana)

Cited by 7 publications

References 19 publications

Enhancement of Agrobacterium-mediated transformation efficiency of model plant using quorum sensing molecule, N-3-oxo-octanoyl-l-homoserine-lactone

Enhancement of Agrobacterium-mediated transformation efficiency of model plant using quorum sensing molecule, N-3-oxo-octanoyl-l-homoserine-lactone

In-planta transient transformation of avocado (Persea americana) by vacuum agroinfiltration of aerial plant parts

Generation of composite Persea americana (Mill.) (avocado) plants: A proof-of-concept-study

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