Genetic Transformation Via Biolistic Gene Gun Method and Regeneration of Common Bean

2013 ◽  
Vol 17 (4) ◽  
pp. 143-158
Author(s):  
E. A. Eissa
Keyword(s):  
Genetic Transformation ◽  
Common Bean ◽  
Gene Gun

scholarly journals Genetic Transformation ofMetroxylon sagu(Rottb.) Cultures viaAgrobacterium-Mediated and Particle Bombardment

2014 ◽  
Vol 2014 ◽  
pp. 1-9 ◽  
Author(s):  
Evra Raunie Ibrahim ◽  
Md. Anowar Hossain ◽  
Hairul Azman Roslan
Keyword(s):  
Genetic Transformation ◽  
Starch Content ◽  
Pcr Amplification ◽  
Culture Method ◽  
Helium Pressure ◽  
Gene Gun ◽  
Dot Blot ◽  
Suitable Material ◽  
Perennial Plant ◽  
Sago Palm

Sago palm (Metroxylon sagu) is a perennial plant native to Southeast Asia and exploited mainly for the starch content in its trunk. Genetic improvement of sago palm is extremely slow when compared to other annual starch crops. Urgent attention is needed to improve the sago palm planting material and can be achieved through nonconventional methods. We have previously developed a tissue culture method for sago palm, which is used to provide the planting materials and to develop a genetic transformation procedure. Here, we report the genetic transformation of sago embryonic callus derived from suspension culture usingAgrobacterium tumefaciensand gene gun systems. The transformed embryoids cells were selected against Basta (concentration 10 to 30 mg/L). Evidence of foreign genes integration and function of thebarandgusgenes were verified via gene specific PCR amplification, gus staining, and dot blot analysis. This study showed that the embryogenic callus was the most suitable material for transformation as compared to the fine callus, embryoid stage, and initiated shoots. The gene gun transformation showed higher transformation efficiency than the ones transformed usingAgrobacteriumwhen targets were bombarded once or twice using 280 psi of helium pressure at 6 to 8 cm distance.

A low-pressure gene gun for genetic transformation of maize (Zea mays L.)

2008 ◽  
Vol 2 (4) ◽  
pp. 267-270 ◽  
Author(s):  
Chien-Yuan Kao ◽  
Shin-Hui Huang ◽  
Chiu-Mei Lin
Keyword(s):  
Zea Mays ◽  
Genetic Transformation ◽  
Zea Mays L ◽  
Low Pressure ◽  
Gene Gun

scholarly journals Agrobacterium tumefaciens-mediated transformation of common bean (Phaseolus vulgaris) var. Brunca

2019 ◽  
Vol 67 (2SUPL) ◽  
pp. S83-S94 ◽  
Author(s):  
Laura-Yesenia Solís-Ramos ◽  
Julio-César Ortiz-Pavón ◽  
Antonio Andrade-Torres ◽  
Romano Porras-Murillo ◽  
Arturo Brenes-Angulo ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Phaseolus Vulgaris ◽  
Genetic Transformation ◽  
Common Bean ◽  
In Vitro Regeneration ◽  
Marker Genes ◽  
Transformation Protocol ◽  
Glucuronidase Activity ◽  
Rich Media

Common bean is a crop recalcitrant to in vitro regeneration and therefore it lacks an efficient transformation protocol that can be reproduced using A. tumefaciens. The main goal of this study was to establish a protocol for A. tumefaciens mediated transformation of Phaseolus vulgaris var. Brunca by marker genes (gusA and nptII) together with the gene for trehalose-6-phosphate synthase from Saccharomyces cerevisiae (TPS1) used in other species to increase tolerance to abiotic stress. The β-glucuronidase activity was detected in 45 % of the LBA4404 ElectroMAX® pCAMBIA1301 infected explants. Transformed explants regenerated new shoots after four to five months period in a kanamycin rich media. Surviving plants were evaluated by PCR and presented an 0.5 % efficiency of transformation. The established protocol for genetic transformation of common bean has two additional advantages with respect to previous reports: (1) it allows for obtaining transformed regenerants and (2) the genetic transformation was stable for the selective gene.

scholarly journals (283) Biolistic Genetic Transformation of Carica papaya L. using the Helios™ Gene Gun

HortScience ◽  
2006 ◽  
Vol 41 (4) ◽  
pp. 1053A-1053
Author(s):  
Salvador Guzmán-González ◽  
Pedro Valadez-Ramírez ◽  
Rosa-Edith Robles-Berber ◽  
Laura Silva-Rosales ◽  
José-Luis Cabrera-Ponce
Keyword(s):  
Genetic Transformation ◽  
Particle Bombardment ◽  
Carica Papaya ◽  
Resistance Mechanism ◽  
35S Promoter ◽  
Helium Pressure ◽  
Gene Gun ◽  
Gold Particle Size ◽  
Key Factor ◽  
Gus Transient Expression

Biolistic genetic transformation of plants with viral genes is a method for controlling plant virus diseases; however, optimization of the particle bombardment parameters according to the transformation system is a key factor for an appropiate transgene expression and, therefore, a stronger resistance mechanism in transgenic plants. In order to optimize biolistic parameters, somatic papaya (Carica papaya L.) cv. Maradol embryo masses were bombarded with the CAMBIA 1301 plasmid construction that contains the coat protein gene (CP) of the papaya ringspot virus isolate of Colima, Mexico, driven by the double constitutively CaMV 35S promoter and flanked for the GUS and hygromycin (hpt) resistance genes. Particle bombardment protocol was carried out using the Helios™ Gene Gun device (BioRad) and the manufacturer's instruction manual. Helium pressure (50, 100, and 150 psi) and gold particle size (0.6, 1.0, and 1.6 μm) were evaluated. Five days after bombardment, somatic embryo clusters were used for GUS transient expression and, during 2 months, were selected into 50, 75, and 150 mg·L-1 hygromycin-containing media to its later CP-PCR detection. Results showed that 50 psi and 1.0 μm were the two optimal values for the assayed analyses. This is the first report of genetic transformation of papaya using the Helios™ Gene Gun device as a new tool compared to conventional PDS-1000/He.

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