scholarly journals Constitutive Overexpression of the Plasma Membrane Na+/H+ Antiporter for Conferring Salinity Tolerance in Rice

2016 ◽  
Vol 25 (2) ◽  
pp. 257-272 ◽  
Author(s):  
Farida Yasmin ◽  
Sudip Biswas ◽  
GM Nurnabi Azad Jewel ◽  
Sabrina M Elias ◽  
Zeba I Seraj
Keyword(s):  
Tissue Culture ◽  
Plasma Membrane ◽  
Indica Rice ◽  
Rice Variety ◽  
Segregation Ratio ◽  
Transformation Method ◽  
Cellular Level ◽  
Salt Tolerant ◽  
In Planta ◽  
Rice Transformation

At the cellular level, the Salt Overly Sensitive (SOS) signaling pathway comprising SOS3, SOS2, and SOS1 has been proposed to mediate cellular signaling under salt stress to maintain ion (Na+) homeostasis. In this regulatory pathway, both OsSOS1 encoding plasma membrane and OsNHX1 encoding vacuolar Na+/H+ antiporters are regulated by SOS3?SOS2 protein kinase complex. In the present study, the rice variety BRRI dhan28 - which is popular with farmers and high yielding, but salt sensitive, was transformed with the OsSOS1 gene isolated from salt tolerant Pokkali rice and driven by the constitutive promoter, CaMV35S. The construct was transformed through a tissue culture-independent Agrobacteriummediated in planta transformation method that circumvents the problems associated with tissue culture-based indica rice transformation methods. Integration of the foreign genes (OsSOS1) into the genome of transgenic plants was confirmed by gene-specific PCR and Southern blot analysis. The level of transgene expression (SOS1) was also quantified by semi-quantitative RT PCR and real time PCR. Genetic segregation ratio for T1 progenies was calculated and found to follow the Mendelian law of inheritance in case of positive transformants. The transformants were shown to be salt tolerant compared to wild type in molecular analysis as well as physiological screening. Future work will involve transformation of both the OsSOS1 and OsNHX1 genes together; with the expectation for enhancing the tolerance level compared to currently available transgenic rice.Plant Tissue Cult. & Biotech. 25(2): 257-272, 2015 (December)

scholarly journals Whole Genome Sequencing and Analysis of Godawee, a Salt Tolerant Indica Rice Variety

2017 ◽  
Vol 05 (01) ◽  
Author(s):  
Sanjeewa Singhabahu ◽  
Chathura Wijesinghe ◽  
Dilini Gunawardana ◽  
Muditha D Senarath Yapa ◽  
Madushani Kannangara ◽  
...  
Keyword(s):  
Whole Genome Sequencing ◽  
Genome Sequencing ◽  
Indica Rice ◽  
Rice Variety ◽  
Whole Genome ◽  
Salt Tolerant

scholarly journals Tissue culture-based Agrobacterium-mediated and in planta transformation methods

2017 ◽  
Vol 53 (No. 4) ◽  
pp. 133-143 ◽  
Author(s):  
M. Niazian ◽  
S.A. Sadat Noori ◽  
P. Galuszka ◽  
S.M.M. Mortazavian
Keyword(s):  
Tissue Culture ◽  
Transformation Method ◽  
Gene Transformation ◽  
In Planta Transformation ◽  
In Planta ◽  
Agrobacterium Mediated Transformation ◽  
Advantages And Disadvantages ◽  
Culture Independent ◽  
Transformation Methods ◽  
Mutant Lines

Gene transformation can be done in direct and indirect (Agrobacterium-mediated) ways. The most efficient method of gene transformation to date is Agrobacterium-mediated method. The main problem of Agrobacterium-method is that some plant species and mutant lines are recalcitrant to regeneration. Requirements for sterile conditions for plant regeneration are another problem of Agrobacterium-mediated transformation. Development of genotype-independent gene transformation method is of great interest in many plants. Some tissue culture-independent Agrobacterium-mediated gene transformation methods are reported in individual plants and crops. Generally, these methods are called in planta gene transformation. In planta transformation methods are free from somaclonal variation and easier, quicker, and simpler than tissue culture-based transformation methods. Vacuum infiltration, injection of Agrobacterium culture to plant tissues, pollen-tube pathway, floral dip and floral spray are the main methods of in planta transformation. Each of these methods has its own advantages and disadvantages. Simplicity and reliability are the primary reasons for the popularity of the in planta methods. These methods are much quicker than regular tissue culture-based Agrobacterium-mediated gene transformation and success can be achieved by non-experts. In the present review, we highlight all methods of in planta transformation comparing them with regular tissue culture-based Agrobacterium-mediated transformation methods and then recently successful transformations using these methods are presented.

Mapping of QTLs Controlling Na+, K+ and CI− Ion Concentrations in Salt Tolerant Indica Rice Variety CSR27

2009 ◽  
Vol 18 (2) ◽  
pp. 139-150 ◽  
Author(s):  
M. H. M. Ammar ◽  
Awadhesh Pandit ◽  
R. K. Singh ◽  
S. Sameena ◽  
M. S. Chauhan ◽  
...  
Keyword(s):  
Indica Rice ◽  
Rice Variety ◽  
Salt Tolerant ◽  
Ion Concentrations

scholarly journals An Apical Meristem-Targeted in planta Transformation Method for the Development of Transgenics in Flax (Linum usitatissimum): Optimization and Validation

2021 ◽  
Vol 11 ◽  
Author(s):  
Karthik Kesiraju ◽  
Shaily Tyagi ◽  
Soumyadeep Mukherjee ◽  
Rhitu Rai ◽  
Nagendra K. Singh ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Apical Meristem ◽  
Transformation Method ◽  
In Planta ◽  
Specific Pcr ◽  
Putative Transformants ◽  
Somaclonal Variations ◽  
Promising Tool ◽  
Culture Independent ◽  
Screening Efficiency

Efficient regeneration of explants devoid of intrinsic somaclonal variations is a cardinal step in plant tissue culture, thus, a vital component of transgenic technology. However, recalcitrance of economically important crops to tissue culture-based organogenesis ensues a setback in the use of transgenesis in the genetic engineering of crop plants. The present study developed an optimized, genotype-independent, nonconventional tissue culture-independent in planta strategy for the genetic transformation of flax/linseed. This apical meristem-targeted in planta transformation protocol will accelerate value addition in the dual purpose industrially important but recalcitrant fiber crop flax/linseed. The study delineated optimization of Agrobacterium tumefaciens-mediated transformation and stable T-DNA (pCambia2301:GUS:nptII) integration in flax. It established successful use of a stringent soilrite-based screening in the presence of 30 mg/L kanamycin for the identification of putative transformants. The amenability, authenticity, and reproducibility of soilrite-based kanamycin screening were further verified at the molecular level by GUS histochemical analysis of T0 seedlings, GUS and nptII gene-specific PCR, genomic Southern hybridization for stable integration of T-DNA, and expression analysis of transgenes by sqRT-PCR. This method resulted in a screening efficiency of 6.05% in the presence of kanamycin, indicating amenability of in planta flax transformation. The strategy can be a promising tool for the successful development of transgenics in flax.

scholarly journals In-planta Agrobacterium-mediated Stable hva1 and EPSPS Integration into Potato var. Agria Genome

2021 ◽  
Author(s):  
Darush Choobineh ◽  
nafiseh mahdi nezhad ◽  
Ali Niazi Niazi ◽  
Baratali Fakheri ◽  
Abbasali Emamjomeh
Keyword(s):  
Tissue Culture ◽  
Transgene Expression ◽  
Direct Injection ◽  
Crop Improvement ◽  
Constitutive Expression ◽  
Transformation Method ◽  
Bacterial Suspension ◽  
Potato Seed ◽  
In Planta ◽  
Culture Independent

Abstract BackgroundAfter wheat, maize, and rice, potato is not only an important food crop but also a substantial source of income throughout the world. Developing a practical and effective transformation method for cultivars that are recalcitrant in tissue culture is vital. Hva1 encodes the protein of the LEA III superfamily that involves in reactions to abiotic stresses, which holds considerable potential for use as molecular tools for genetic crop improvement toward stress tolerance.ResultsHere, a protocol has been designed for an Agrobacterium-mediated transient transformation in tissue culture-independent conditions in-planta. The protocol establishes for hva1 and EPSPS transformations by direct injection of the bacterial suspension into the potato tuber sprout to encode resistance to cold and against glyphosate herbicide. A two-stage selection was involved using 1% and 2% Glyphosate to eliminate the chimeric and non-transformed plants. Ultimately, the protocol enabled confirmation of gene integration into the plant, transgene expression of the gene and transgene expression, which was made possible by competitive PCR reaction, RT-PCR, and ELISA, respectively. In this research, the transformation efficiencies acquired in potatoes (up to 46%) were higher than those reported using conventional Agrobacterium-mediated approaches in previous studies. ConclusionsThe constitutive expression of the integrated T-DNA neither slowed down the growth rate nor affected potato tuberization significantly. The hva1 gene was expressed successfully leading to the accumulation of the hva1 protein in transgene-generated tubers. This study is the first report on a successful transformation of potato in-planta whereby Agrobacterium can be directed at potato seed sprouts through injection.

In Planta transformation for conferring salt tolerance to a tissue-culture unresponsive indica rice (Oryza sativa L.) cultivar

2017 ◽  
Vol 54 (2) ◽  
pp. 154-165 ◽  
Author(s):  
Tasnim Ahmed ◽  
Sudip Biswas ◽  
Sabrina M. Elias ◽  
M. Sazzadur Rahman ◽  
Narendra Tuteja ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Oryza Sativa ◽  
Salt Tolerance ◽  
Oryza Sativa L ◽  
Indica Rice ◽  
In Planta Transformation ◽  
In Planta

scholarly journals OsRuvBL1a DNA Helicase Boost Salinity and Drought Tolerance in Transgenic Indica Rice Raised by In-Planta Transformation

2021 ◽  
Author(s):  
Shabnam K Saifi ◽  
Nishat Passricha ◽  
Renu Tuteja ◽  
Manoj Nath ◽  
Sarvajeet Singh Gill ◽  
...  
Keyword(s):  
Abiotic Stress ◽  
Stress Tolerance ◽  
Indica Rice ◽  
Functional Characterization ◽  
Rice Genome ◽  
Transformation Method ◽  
Dna Helicase ◽  
Model Systems ◽  
In Planta ◽  
Transgenic Lines

Abstract RuvBL, is a member of SF6 superfamily of helicases and is conserved among the various model systems. Recently rice homolog of RuvBL has been biochemically characterized for its ATPase and DNA helicase activities, however its involvement in stress is not been studied yet. This study reports the detailed functional characterization of RuvBL homolog of Oryza sativa, under abiotic stress through transgenic approach. An improved Agrobacterium-mediated in-planta transformation method was developed in indica rice to generate the transgenic lines and study was focused on optimization of factors to achieve maximum transformation efficiency. Overexpressing OsRuvBL1a transgenics showed enhanced tolerance under in vivo salinity stress as compared to WT plants. The physiological and biochemical analysis of the OsRuvBL1a transgenic lines showed better performance under salinity and drought stresses. Several stress responsive interacting partners of OsRuvBL1a were identified using Y2H method. Working mechanism for boosting the stress tolerance by OsRuvBL1a has been proposed in this study. This integration of OsRuvBL1a gene in rice genome using in-planta transformation method helped us to achieve the abiotic stress tolerant smart crop. This study is the first direct evidence to show the novel function of RuvBL in boosting abiotic stress tolerance in plants.

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