In planta transformation method for T-DNA transfer in orchids

2014 ◽  
Author(s):  
Endang Semiarti ◽  
Aziz Purwantoro ◽  
Ixora S. Mercuriani ◽  
Anida M. Anggriasari ◽  
Seonghoe Jang ◽  
...  
Keyword(s):  
Transformation Method ◽  
Dna Transfer ◽  
In Planta Transformation ◽  
In Planta

scholarly journals IN-PLANTA TRANSFORMATION METHOD MEDIATED WITH Agrobacterium tumefaciens FOR T-DNA TRANSFER IN TABLE GRAPE (Vitis vinifera L.)

2019 ◽  
Vol 6 (2) ◽  
pp. 95
Author(s):  
Rindang Dwiyani ◽  
Hestin Yuswanti ◽  
Yuyun Fitriani ◽  
Bambang Sugiharto
Keyword(s):  
Transformation Method ◽  
Table Grape ◽  
Dna Transfer ◽  
Plant Genome ◽  
Nptii Gene ◽  
In Planta Transformation ◽  
In Planta ◽  
Simple Method ◽  
Leafy Shoots ◽  
Sweeping Method

The aim of the research is to investigate a simple method of in planta transformation method for T-DNA transfer in table grape.  The T-DNA harbored the S0SPS1 gene under the control of promoter of the 35S CaMV from the Cauliflower Mosaic Virus and contained the NPTII gene, a kanamycin-resistant gene as a selectable marker for transformant selection.  Six-month plants originated from cuttings were used as target plants.  We explored two methods of in planta transformation, namely ”dipping” and “sweeping”.  For both methods, the leaves of the target plants were removed and those of shoots without leaves were used as the target of  transformation.  In the “dipping method”, those shoots were dipped with the agrobacterial suspension for 60 seconds.  However, for the “sweeping method”, the scars (the spots where leaves were removed) were swept with agrobacterial suspension using a cotton bud.  Those treated non-leafy-shoots (from both methods) then were grown to be leafy shoots.  Those leafy shoots then were cut and transplanted into the soil and grown to be a whole plant. The leaves of those plants then were taken as samples for DNA extraction and PCR using primers of NPTII gene (Forward: 5’-GTCATCTCACCTTCCTCCTGCC-3’; Reverse: 5’ GTCGCTTGGTCGGTCATTTCG-3’) with expected amplified band of 550 bp. We found that only the “sweeping method” plants amplified the 550 bp bands, while those of the “dipping method”  did not.  We suggest that the T-DNA was successfully integrated into the genome of plants treated with the “sweeping method” but not with the “dipping method”.  Leaf sugar content (oBrix) of PCR-positive vines was higher than those of the wild-type vines, ensuring the integration of the T-DNA into the plant genome.

Development of simple and efficient in Planta transformation method for wheat (Triticum aestivum L.) using Agrobacterium tumefaciens

2006 ◽  
Vol 102 (3) ◽  
pp. 162-170 ◽  
Author(s):  
Putu Supartana ◽  
Tsutomu Shimizu ◽  
Masahiro Nogawa ◽  
Hidenari Shioiri ◽  
Tadashi Nakajima ◽  
...  
Keyword(s):  
Triticum Aestivum ◽  
Agrobacterium Tumefaciens ◽  
Triticum Aestivum L ◽  
Transformation Method ◽  
In Planta Transformation ◽  
In Planta

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.

A simple and efficient in planta transformation method for pommelo ( Citrus maxima ) using Agrobacterium tumefaciens

2017 ◽  
Vol 214 ◽  
pp. 174-179 ◽  
Author(s):  
Yong-yan Zhang ◽  
Dong-min Zhang ◽  
Yun Zhong ◽  
Xiao-jun Chang ◽  
Min-lun Hu ◽  
...  
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Transformation Method ◽  
In Planta Transformation ◽  
In Planta ◽  
Citrus Maxima

Development of simple and efficient in planta transformation method for rice (Oryza sativa L.) using Agrobacterium tumefaciens

2005 ◽  
Vol 100 (4) ◽  
pp. 391-397 ◽  
Author(s):  
Putu Supartana ◽  
Tsutomu Shimizu ◽  
Hidenari Shioiri ◽  
Masahiro Nogawa ◽  
Masayuki Nozue ◽  
...  
Keyword(s):  
Oryza Sativa ◽  
Agrobacterium Tumefaciens ◽  
Oryza Sativa L ◽  
Transformation Method ◽  
In Planta Transformation ◽  
In Planta

A rapid, novel and high-throughput identification of putative bell pepper transformants generated through in planta transformation approach

2011 ◽  
Vol 129 (4) ◽  
pp. 898-903 ◽  
Author(s):  
Arthikala Manoj Kumar ◽  
Kalpana N. Reddy ◽  
M. Manjulatha ◽  
Elizabeth S. Arellano ◽  
Rohini Sreevathsa ◽  
...  
Keyword(s):  
High Throughput ◽  
Bell Pepper ◽  
In Planta Transformation ◽  
In Planta ◽  
Transformation Approach

scholarly journals Optimization of Agrobacterium-mediated transformation conditions of prospective genotypes of winter bread wheat by in planta method

2021 ◽  
Vol 28 ◽  
pp. 66-71
Author(s):  
O. V. Dubrovna ◽  
L. V. Slivka
Keyword(s):  
Transgenic Plants ◽  
Bread Wheat ◽  
Optical Density ◽  
Dna Transfer ◽  
Plant Genome ◽  
In Planta ◽  
Agrobacterium Mediated Transformation ◽  
The Third ◽  
Winter Bread Wheat ◽  
Inoculation Medium

Aim. Optimization of conditions for genetic transformation of new promising genotypes of winter bread wheat (T. aestivum L.) by in planta method. Methods. Agrobacterium-mediated transformation by in planta method using the strain AGL0 and vector construct pBi2E. Results. The influence of air temperature, optical density of cells of agrobacterial suspension, inoculation day and composition of inoculation medium on the frequency of obtaining transgenic plants of new winter wheat genotypes was studied. The dependence of the frequency obtaining of transgenic plants from environmental conditions, in particular temperature, has been established. It was found that the temperature regime of 20-22°C provided the largest number (4.8%) of wheat transformants, and when the temperature is reduced to 16-18°C there is a decrease in the efficiency of T-DNA transfer into the plant genome and the lowest frequency of transformation (0.7%). Conclusions. The largest number of transformants was obtained using a inoculation medium without sucrose, the optical density of cells of the agrobacterial suspension of 0.6 op.od. and inoculation on the third day after castration of ears. Keywords: T. aestivum, Agrobacterium-mediated transformation in planta, optimization of conditions.

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