Agroinfiltration of intact leaves as a method for the transient and stable transformation of saponin producing Maesa lanceolata

Faizal, Ahmad; Geelen, Danny

doi:10.1007/s00299-012-1266-4

Cited by 16 publications

(12 citation statements)

References 40 publications

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“…1c). The A. tumefaciens strain has been shown to affect the efficiency of transient expression in grapevine (Zottini et al 2008), rose (Yasmin and Debener 2010), Maesa lanceolata (Faizal and Geelen 2012), and spinach (Cao et al 2017). In coffee, Mishra et al (2008) demonstrated that the Agrobacterium strain influenced the transient expression of uidA and gfp in embryogenic tissues.…”

Section: Effect Of the A Tumefaciens Strain On Transient Expression mentioning

confidence: 99%

“…In coffee, Mishra et al (2008) demonstrated that the Agrobacterium strain influenced the transient expression of uidA and gfp in embryogenic tissues. The virulence of Agrobacterium strains can be dependent on the target plant species (Faizal and Geelen 2012). This may be due to the ability of the bacteria to attach to plant cells or to differences in the T-DNA transfer mechanism (Wroblewski et al 2005).…”

Section: Effect Of the A Tumefaciens Strain On Transient Expression mentioning

confidence: 99%

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A simple and efficient agroinfiltration method in coffee leaves (Coffea arabica L.): assessment of factors affecting transgene expression

et al. 2018

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The establishment of a simple, rapid and efficient transient expression system is a necessary tool for the functional validation of candidate genes in coffee biotechnology. The effects of Agrobacterium strain, age of the donor plant, infiltration method, and infiltration medium on transgene expression in detached coffee leaves were evaluated. Regarding the effect of Agrobacterium strain, the expression of uidA was higher in GV3101-treated coffee disks than in LBA4404 and ATHV-treated samples. On the other hand, transient expression of uidA was significantly higher in leaf disks from young plants (6-weeks-old) (13.1 ± 1.4%) than in mature tissue (12-weeks-old) (1.6 ± 1.2%). Transient uidA expression was higher in detached coffee leaf disks from young plants infiltrated with one injection of 15 µL of Agrobacterium strain GV3101::1303 suspended in MS salts supplemented with 30 g/L sucrose, 1.9 g/L MES and 200 uM AS with subsequent sanding of the abaxial epidermis. Using the optimized protocol, expression of the uidA gene was observed 6, 24 and 48 h and 5 weeks after bacterial injection. DNA was extracted from coffee disks with positive GUS expression and specific mgfp5 and uidA fragments were amplified 5 weeks post-agroinfiltration. On the other hand, using the optimized protocol, a specific cry10Aa (500 bp) fragment was amplified in the agro-infiltrated coffee leaf disks 5 weeks post-agroinfiltration with the plasmid pB427-35S-cry10Aa. Moreover, the expression of the gene cry10Aa in two infiltrated coffee leaf disks was verified by RT-PCR and an expected 500 bp fragment was amplified.

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Section: Effect Of the A Tumefaciens Strain On Transient Expression mentioning

confidence: 99%

Section: Effect Of the A Tumefaciens Strain On Transient Expression mentioning

confidence: 99%

A simple and efficient agroinfiltration method in coffee leaves (Coffea arabica L.): assessment of factors affecting transgene expression

et al. 2018

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“…Another study has been conducted on the transformation of Maesa lanceolata through Agrobacterium-infection in planta transformation for both transient and stable gene expression. It was concluded that optimum bacterial density (OD 600 nm = 1.0) significantly increased TE (Faizal and Geelen 2012). Trieu et al (2000) modelled a novel system of in planta transformation via seedlings infiltration in M. truncatula and argued that OD 600 nm of 1.6 was optimum for securing efficient TE, ranged from 2.9 to 27.6 %.…”

Section: Discussionmentioning

confidence: 98%

Piercing and incubation method of in planta transformation producing stable transgenic plants by overexpressing DREB1A gene in tomato (Solanum lycopersicum Mill.)

Shah

Ali²,

Jan

et al. 2014

Plant Cell Tiss Organ Cult

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Cold is a major constraint for tomato growth and productivity; as it is a cold sensitive crop. DREB1A plays a key role in generating cold tolerance in tomato by regulating the response of multiple genes under chilling stress. In this study, cold tolerant gene (DREB1A) driven by Lip9 promoter, was transformed in three tomato genotypes through Agrobacterium tumefaciens, employing tissue culture independent transformation strategy. Overnight imbibed seeds of tomato were surface sterilized, 3-day old shoot apical meristem of the developing seedling were pierced and incubated for twenty minutes with A. tumefaciens strain EHA-105 having OD 600 nm of 1.0. The treated seeds were co-cultivated with Agrobacterium for 2 days. The regenerated shoots were subjected to 35 mg/l hygromycin as a selection for a period of 2-3 weeks. The presence of DREB1A and hpt genes in the hygromycin resistant (T 0 ) transgenic plants was evaluated by PCR analysis. The transgene activity was detected in T 1 plants by Reverse Transcriptase Polymerase Chain Reaction and Southern blotting that showed the stable integration of the transgene to the next generation. Physiological analysis of T 2 transgenic plants depicted that increased expression of DREB1A induced only during chilling stress. After various chilling stresses, stomatal conductance, transpiration rate and relative water contents of transgenic lines were significantly higher than those of NT plants. While leaf osmotic potential of transgenic plants was lower as compared to NT plants. The established procedure is novel and can produce stable transgenic tomato plants in efficient manner by saving potential resources in terms of cost and time.

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“…This result suggested that improvement of this process was effective. Indeed, improvement of T-DNA transfer increased stable transformation 46 47 . Inevitable, we focused on improvement of T-DNA transfer through new bred A. tumefaciens strain.…”

Section: Discussionmentioning

confidence: 99%

An Agrobacterium tumefaciens Strain with Gamma-Aminobutyric Acid Transaminase Activity Shows an Enhanced Genetic Transformation Ability in Plants

Nonaka

Someya

Zhou

et al. 2017

Sci Rep

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Agrobacterium tumefaciens has the unique ability to mediate inter-kingdom DNA transfer, and for this reason, it has been utilized for plant genetic engineering. To increase the transformation frequency in plant genetic engineering, we focused on gamma-aminobutyric acid (GABA), which is a negative factor in the Agrobacterium-plant interaction. Recent studies have shown contradictory results regarding the effects of GABA on vir gene expression, leading to the speculation that GABA inhibits T-DNA transfer. In this study, we examined the effect of GABA on T-DNA transfer using a tomato line with a low GABA content. Compared with the control, the T-DNA transfer frequency was increased in the low-GABA tomato line, indicating that GABA inhibits T-DNA transfer. Therefore, we bred a new A. tumefaciens strain with GABA transaminase activity and the ability to degrade GABA. The A. tumefaciens strain exhibited increased T-DNA transfer in two tomato cultivars and Erianthus arundinacues and an increased frequency of stable transformation in tomato.

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Agroinfiltration of intact leaves as a method for the transient and stable transformation of saponin producing Maesa lanceolata

Abstract: We have developed a transformation protocol for saponin producing Maesa lanceolata. Using the protocol reported here, now we are able to generate the tools for the modification of saponin production.

Cited by 16 publications

References 40 publications

A simple and efficient agroinfiltration method in coffee leaves (Coffea arabica L.): assessment of factors affecting transgene expression

A simple and efficient agroinfiltration method in coffee leaves (Coffea arabica L.): assessment of factors affecting transgene expression

Piercing and incubation method of in planta transformation producing stable transgenic plants by overexpressing DREB1A gene in tomato (Solanum lycopersicum Mill.)

An Agrobacterium tumefaciens Strain with Gamma-Aminobutyric Acid Transaminase Activity Shows an Enhanced Genetic Transformation Ability in Plants

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