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- 저자 : Vasudevan Venkatachalam (검색결과 3 건)
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Vasudevan Venkatachalam,Sathish Dorairaj,Ajithan Chandrasekaran,Sathish Selvam,Manickavasagam Markandan 한국식물생명공학회 2021 Plant biotechnology reports Vol.15 No.4
The production of transgenic watermelon through Agrobacterium tumefaciens-mediated transformation with in vitro regen- eration system is a time-consuming, labor-intensive and genotype-dependent process. To acquire large number of transgenic watermelons in a shorter period of time, the half-seed explants were infected with Agrobacterium strain EHA 105 harboring pCAMBIA1301 with bar and gus genes. The factors affecting in planta transformation efficiency, such as co-cultivation duration, acetosyringone concentration, sonication duration and vacuum infiltration were assessed in the present study. The half-seed explants were sonicated for 3 min and 2 min vacuum infiltrated in Agrobacterium suspension and co-cultivated for 3 days with 100 µM acetosyringone showed maximum transformation efficiency. The transformed watermelon plants were selected against BASTA® and GUS, PCR, Southern hybridization analysis confirmed the transgene integration. The amenability of this established protocol was analyzed on four genotypes, in which the response of all genotypes was posi- tive, whereas Arka manik showed the higher transformation efficiency of 17%. The transformation protocol developed in the present study is efficient, economical and expeditious without the taking part of any tissue culture phases and produce a large number of transgenic lines within a short period of 56 days.
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Selvam Sathish,Venkatachalam Vasudevan,Sivabalan Karthik,Gadamchetty Pavan,Markandan Manickavasagam 한국식물생명공학회 2019 Plant biotechnology reports Vol.13 No.6
Hybanthus enneaspermus is an important source for L-Dopa production. This study reports elicited L-Dopa production in cell suspension cultures of Hybanthus enneaspermus. Cell suspension culture was established using green friable calli from leaf explants cultured on MS medium containing 2.0 mg/l 2,4-D. Effects of different elicitors such as SA, YE, MeJA and AgNO3 on biomass accumulation and L-Dopa content were studied. Among the elicitor tested SA treated culture produced highest biomass and L-Dopa according to their exposure time and concentration. Maximum biomass of 15.5 ± 0.16 g FW, 4.05 ± 0.18 g DW and L-Dopa production of 8.88 mg/g DW were observed at 150 μM concentration of SA. This was 9.25-fold higher compared to that of the unelicited control culture. The results obtained in this study clearly show that the elicita-tion strategy is a promising method for biosynthesis of L-Dopa production by cell suspension cultures of H. enneaspermus.
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Factors affecting Agrobacterium-mediated transformation in Hybanthus enneaspermus (L.) F. Muell
Ganeshan Sivanandhan,Chinnathambi Arunachalam,Venkatachalam Vasudevan,Gnanajothi Kapildev,Ali Alharbi Sulaiman,Natesan Selvaraj,Andy Ganapathi,임용표 한국식물생명공학회 2016 Plant biotechnology reports Vol.10 No.2
Agrobacterium tumefaciens-mediated transformation system was established for Hybanthus enneaspermus using leaf explants with the strain LBA4404 harbouring pCAMBIA 2301 carrying the nptII and gusA genes. Sensitivity of leaf explants to kanamycin was standardized (100 mg/l) for screening the transgenic plants. Transformation parameters (OD, virulence inducer, infection time, co-cultivation period, bactericidal antibiotics, etc.) influencing the gene transfer and integration were assessed in the present investigation. Fourteen-day precultured explants were subjected with Agrobacterium strain LBA4404. Optimized parameters such as culture density of 0.5 OD600, infection time of 6 min, AS concentration of 150 lM with 3 days co-cultivation revealed maximum transformation efficiency based on GUS expression assay. The presence of gusA in transgenics was confirmed by polymerase chain reaction and Southern blotting analysis. The present transformation experiment yielded 20 shoots/explant with higher transformation efficiency (28 %). The protocol could be used to introduce genes for trait improvement as well as for altering metabolic pathway for secondary metabolites production.
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