scholarly journals Factors Affecting Agrobacterium-mediated Transformation of Common Bean (Phaseolus vulgaris L.)

HortScience ◽  
1996 ◽  
Vol 31 (4) ◽  
pp. 616f-616
Author(s):  
Zhanyuan Zhang ◽  
A. Mitra ◽  
D.P. Coyne
Keyword(s):  
Shoot Organogenesis ◽  
Transformation Rate ◽  
Bacterial Strains ◽  
Phaseolus Vulgaris L ◽  
Reporter System ◽  
Breeding Lines ◽  
Factors Affecting ◽  
Host Interactions ◽  
High Selection ◽  
Direct Shoot Organogenesis

Factors influencing Agrobacterium–mediated DNA transfer of P. vulgaris were examined using an intron-containing β-glucuronidase (GUS) gene as a reporter system. Tissue culture procedures used were based on direct shoot organogenesis. Two A. tumefaciens strains, A2760 and EHA105, were used with more emphasis on the former due to its overall higher transformation rate. Ten bean entries including breeding lines and cultivars from both Meso-American and Andean origins were compared for compatibility with the two bacterial strains under different pre- and coculture conditions. Pinto `Othello' was extensively used in testing different transformation conditions. Factors found to have significant effects on transformation rate included Agrobacterium-host interactions, explant maturity, preculture and cocultivation conditions, as well as selection schemes, based on transient expression. Some factors, such as the effect of explant maturity and dark preconditioning of explants on gene transfer, have not been reported before. The best transformation conditions included the use of susceptible genotypes and mature explants, preconditioning of explants in darkness, followed by a maximum cocultivation period in the presence of cytokinin, and the use of high selection pressure.

scholarly journals Factors Affecting Agrobacterium-mediated Transformation of Common Bean

1997 ◽  
Vol 122 (3) ◽  
pp. 300-305 ◽  
Author(s):  
Zhanyuan Zhang ◽  
Dermot P. Coyne ◽  
Amitava Mitra
Keyword(s):  
Common Bean ◽  
Filter Paper ◽  
Blue Staining ◽  
Transformation Rate ◽  
Gus Gene ◽  
Reporter System ◽  
Factors Affecting ◽  
Host Interactions ◽  
Optimized Protocol ◽  
High Concentrations

Factors influencing Agrobacterium tumefaciens-mediated transformation of common beans (Phaseolus vulgaris L.) were examined using an intron-containing β-glucuronidase (GUS) gene as a reporter system to develop a repeatable transformation protocol. Tissue culture procedures used were based on direct shoot organogenesis. Two A. tumefaciens strains—A2760 and EHA105—were used, with emphasis on the former due to its overall higher infection rate. Eleven common-bean genotypes were compared for susceptibility to strain A2760 or EHA105. The pinto bean `Othello' was used extensively in testing different transformation conditions. Factors significantly affecting transformation rate were Agrobacterium × host interactions, explant maturity, preculture and cocultivation conditions, and selection schemes, based on transient GUS gene expression. The best transformation conditions were the use of susceptible genotypes and explants derived from mature seeds, preconditioning of explants in a medium containing 20 μmol of benzyladenine (BA) in darkness or on a filter paper, dipping explants in high concentrations of Agrobacterium cell suspension (OD650 = 0.8-1.0) followed by a long-term (6-day) cocultivation period on a semisolid agar medium in the presence of cytokinin or 3-day cocultivation on a moistened filter paper, and the use of lethal levels of selective agents. About 4% of explants, or 14% of regenerated shoots or buds, were putatively transgenic, as indicated by GUS blue staining throughout the entire shoot or bud, after explants were transformed with Agrobacterium strain A2760 using an optimized protocol.

The potential to propagate coconut clones through direct shoot organogenesis: A review

2021 ◽  
Vol 289 ◽  
pp. 110400
Author(s):  
Eveline Y.Y. Kong ◽  
Julianne Biddle ◽  
Mike Foale ◽  
Bart Panis ◽  
Stephen W. Adkins
Keyword(s):  
Shoot Organogenesis ◽  
Direct Shoot Organogenesis ◽  
Direct Shoot

scholarly journals Direct shoot organogenesis from petiole and leaf discs of Withania somnifera (L.) Dunal

2010 ◽  
Vol 9 (44) ◽  
pp. 7453-7461 ◽  
Author(s):  
Kumar Ghimire Bimal ◽  
Soo Seong Eun ◽  
Hye Kim Eun ◽  
Lamsal Kabir ◽  
Yeon Yu Chang ◽  
...  
Keyword(s):  
Withania Somnifera ◽  
Shoot Organogenesis ◽  
Leaf Discs ◽  
Direct Shoot Organogenesis ◽  
Direct Shoot

scholarly journals Direct shoot organogenesis and Agrobacterium tumefaciens mediated transformation of Solanum trilobatum L.

2016 ◽  
Vol 40 ◽  
pp. 866-877 ◽  
Author(s):  
Jayabalan SHILPHA ◽  
Manoharan JAYASHRE ◽  
Muthiah JOE VIRGIN LARGIA ◽  
Manikandan RAMESH
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Shoot Organogenesis ◽  
Solanum Trilobatum ◽  
Direct Shoot Organogenesis ◽  
Direct Shoot

scholarly journals Detecting bean golden yellow mosaic virus in bean breeding lines and in the common legume weed Macroptilium lathyroides in Puerto Rico.

1969 ◽  
Vol 85 (3-4) ◽  
pp. 165-176
Author(s):  
Lydia I. Rivera-Vargas ◽  
Vilmaris Bracero-Acosta ◽  
James S. Beaver ◽  
Dan E. Purcifull ◽  
Jane E. Polston ◽  
...  
Keyword(s):  
Puerto Rico ◽  
Mosaic Virus ◽  
Yellow Mosaic Virus ◽  
Phaseolus Vulgaris L ◽  
Breeding Lines ◽  
Golden Yellow ◽  
Enzymelinked Immunosorbent Assay ◽  
The Common ◽  
Polymerase Chain ◽  
Macroptilium Lathyroides

Bean golden yellow mosaic virus (BGYMV) is a geminivirus transmitted by whiteflies (Genus: Bemisia). This virus causes significant fosses in common bean (Phaseolus vulgaris L.). Serological techniques such as enzymelinked immunosorbent assay (ELISA) have been widely used for detection of viruses. We evaluated existing monoclonal antibodies (3F7,2G5 and 5C5) for the detection of BGYMV isolates in bean fines in Puerto Rico. Monoclonal antibody 3F7 was the most effective in detecting the virus in tissues of line DOR 364 and susceptible cuftivars Top Crop and Quest. However, it was not effective in the detection of BGYMV in lines of DOR 303, which showed typical symptoms. Sampfes from Macroptilium lathyroides, a weed that might be a possible reservoir of the virus, were also tested for viraf infection. ELISA tests were inconclusive for detection of geminiviruses in M. lathyroides. Polymerase Chain Reaction (PCR) was also used to complement BGYMV diagnosis in M. lathyroides and in bean lines that showed symptoms but were negative for the ELfSA test. Two sets of primers, specific for Begomovirus such as BGYMV, were used in PCR experiments. Using PCR, we were able to detect the virus in the line DOR 303 and in M. lathyroides tissues.

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