Technologies of Agrobacterium plant transformation In planta

Chumakov, M. I.; Moiseeva, E. M.

doi:10.1134/s0003683812080017

Cited by 16 publications

(10 citation statements)

References 47 publications

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“…Despite optimal conditions provided for regeneration and transformation, incidents like somaclonal variation, morphological deformities, chromosomal aberration, anomalies in plant reproductive characteristics etc. have been reported for tissue culture dependent transformation studies (10). Alternatively, in planta transformation offers several benefits as it can avoid somaclonal variation and there is no report of morphological change, chromosomal alteration and loss of plant reproductivity (10).…”

Section: Introductionmentioning

confidence: 99%

“…have been reported for tissue culture dependent transformation studies (10). Alternatively, in planta transformation offers several benefits as it can avoid somaclonal variation and there is no report of morphological change, chromosomal alteration and loss of plant reproductivity (10). Previously, in planta transformation methods have been reported for tomato using floral dip method, whole fruit injection method, smearing Agrobacterium suspension on floral buds but efficiency of seeds as explant in in planta transformation is yet to be explored (11,12).…”

Section: Introductionmentioning

confidence: 99%

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Establishment of in planta transformation protocol of tomato (Solanum lycopersicum L.) through antiporter gene for improved salinity tolerance

2022

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Tomato stands as the world’s third most consumed vegetable, but its production has been suffering due to climate vulnerability, notably for saline sensitivity. Despite its economic importance, developing salinity tolerant tomato has not been prioritized lately. Current study was aimed to establish a simple and efficient Agrobacterium-mediated in planta transformation protocol to transform Na+/H+ antiporter gene into 5 Bangladeshi tomato varieties, namely BARI tomato 2, BARI tomato 3, BINA tomato 2, BINA tomato 3 and Bahar, to improve their salt tolerance, through optimization of crucial transformation factors like optical density, infection time, co-cultivation period etc. Two vectors were constructed by cloning Na+/H+ antiporter gene from Arabidopsis (pK7WG2_AtNHX1_1.6) and Rice (pK7WG2_OsNHX1_1.6) individually to gateway vector pENTR/D-TOPO and electroporated to Agrobacterium while another vector pBI121 was used as control. Non-pricked seeds were found optimum for achieving more than 90% efficiency for GUS expression and germination percentages under conditions of OD600 1.1-1.4 with 30 min of infection time followed by 24 hrs co-cultivation period during transformation using the 3 vectors. Transformed plantlets were screened through resistance to Kanamycin 50 mg/l in germination medium while Cefotaxime 100 mg/l was applied to prevent Agrobacterium overgrowth during co-cultivation. Tolerance of 100 mM NaCl for 14 days has been observed in putative transformants in Leaf Disc Bioassay. No significant morphological changes were observed during the acclimatization of putatively transformed plantlets. This established protocol is novel and can efficiently produce genotype-independent transgenic tomato plants obviating intervening tissue culture. Hence, this study provides scope for climate-resilient crop improvement to ensure nutritional security.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Establishment of in planta transformation protocol of tomato (Solanum lycopersicum L.) through antiporter gene for improved salinity tolerance

2022

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“…It has been proven that embryogenic friable calliwere essential for the development of cell suspension cultures that have played an important role in successful somatic hybridization (Davey et al, 2005;Chumakov and Moiseeva, 2012). In the present study, embryogenic calli were produced using the ovule culture of C. volkameriana at different immature fruit stages on EME medium supplemented with 2,4-D, BAP, and KIN.…”

Section: Discussionmentioning

confidence: 95%

Efficient protoplast isolation from ovule-derived embryogenic callus inCitrusvolkameriana

Çimen¹

2020

Turk J Agric For

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The present study reports on the isolation of viable protoplast from ovule-derived embryogenic calli of Volkameriana (Citrus volkameriana L.), which is a rootstock in high demand for lemon production.Ovules of C. volkameriana isolated at 3 different immature fruit stages, comprising4, 8, and 12 weeks after anthesis (WAA),were cultured on5 different media in order to produce embryogenic callus lines as a source material for protoplast isolation. EME medium (MT basal medium + 0.5 gL–1 malt extract), with the addition of phytohormones [kinetin (KIN), 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzylaminopurine (BAP)] at different concentrations, were tested for callogenesis. According to 2-way ANOVA, significant effects were determined as a result of the immature fruit stage and type of culture media (P ≤ 0.01) on the callogenesis and embryogenic callus induction frequency. First, callus formation was recorded after 4 WAA on medium comprising EME + 2,4-D (1.0 mg L–1) + BAP (0.5 mg L–1). Callus induction frequency was the highest (90.00%) in the same culture medium when the ovules wereculturedat8 WAA. In addition, culturing the ovules isolated from 12 WAA immature fruits of C. volkameriana resulted in the highest indirect somatic embryogenesis (24%). Embryogenic callus initiation was the highest (25.56%) using EME + KIN (1.0 mg L–1) and ovules cultured at 8 WAA (14%) resulted in the highest embryogenic callus formation. Effects of different enzyme concentrations on the efficiency of protoplast isolation were calculated using the hemocytometer cell counting method. The combination of 2% cellulase and 0.2% pectinase gave the highest numbers of protoplasts, at 12.33 × 105protoplast/mL. Embryogenic callus lines obtained by culturing ovules of C. volkameriana yielded high-quality protoplasts after isolation and could be useful as a protoplast source for further somatic hybridization studies.

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“…Friable callus has been proven to be crucial for the establishment of cell suspension cultures, which played an important role in successful somatic hybridization (Davey et al, 2005) and genetic transformation (Chumakov and Moiseeva, 2012) in many plants. In this study, the compact callus or semi-compact callus was separated from the cultures carefully and were cutted into appropriate pieces with a diameter of 3-5 mm with a sharp-pointed forceps and a scalpel blade, and then placed on the tested media, T2, T5, T8 and T9.…”

Section: Callus Induction From the Regenerated Compact/semicompact Callusmentioning

confidence: 99%

<i>In vitro</i> Induction and Proliferation of Callus from Immature Cotyledons and Embryos of Juglans regia cv. 'Xiangling'

Cai

Wang

Cao³

2013

Not Bot Hort Agrobot Cluj

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Callus holds great potential for biotechnology applications in plants. In this study, several experiments were performed to establish a protocol for in vitro induction of good quality callus by using immature cotyledons and embryos of Juglans regia cv. â€˜Xianglingâ€™ as the explants. DKW basal media supplemented with different concentrations of plant growth regulators were applied to determine the induction rate, texture and color of the recovered callus. Results showed that the optimum culture medium for the callus induction from these explants was a DKW basal medium consisting of 1 mg/L BA, 2 mg/L KT 250 mg/L glutamine, 500 mg/L casein hydrolysate, 200 mg/L Vc, 50 g/L sucrose and 2.8 g/L Gelrite (i.e., the T8 medium). The average rate of callus induction on the medium reached 80.7% for the immature cotyledons and 66.7% for the immature embryos, and all the callus was characterized by a friable, creamy or yellow appearance. In vitro proliferation of callus was not very successful as a result of the fact that the friable callus with creamy or yellow color grew very slowly, whereas the fast-growing callus was easy to turn brown and died eventually. Even though further investigation will be required to overcome the problem of browning and assess the regeneration ability of the callus, this approach for the production of friable callus may have valuable applications for walnut and other woody trees.

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Technologies of Agrobacterium plant transformation In planta

Cited by 16 publications

References 47 publications

Establishment of in planta transformation protocol of tomato (Solanum lycopersicum L.) through antiporter gene for improved salinity tolerance

Establishment of in planta transformation protocol of tomato (Solanum lycopersicum L.) through antiporter gene for improved salinity tolerance

Efficient protoplast isolation from ovule-derived embryogenic callus inCitrusvolkameriana

<i>In vitro</i> Induction and Proliferation of Callus from Immature Cotyledons and Embryos of Juglans regia cv. 'Xiangling'

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