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Research articles

ScienceAsia 46 (2020): 412-419 |doi: 10.2306/scienceasia1513-1874.2020.052


Application of sonication- and vacuum infiltration-assisted Agrobacterium-mediated transformation of rice embryo (Oryza sativa L.)


Touchkanin Jongjitvimola, Kawee Sujipulib, Sittichai Urtgama,*

 
ABSTRACT:     Rice transgenics could be performed based on Agrobacterium-mediated transformation which could improve the rice breeding programs to settle for diseases, insect pests and the climate changes. In this study, the new strategy of combination of sonication-assisted Agrobacterium-mediated transformation (SAAT) and vacuum infiltration-assisted Agrobacterium-mediated transformation (VIAAT) system for producing transgenic rice was developed. This research demonstrated an Agrobacterium-mediated transformation protocol for whole rice seeds by employing both sonication and vacuum infiltration. Two Agrobacterium tumefaciens strains AGL1 and EHA105, carrying a binary vector pCAMBIA1301 (with a gusA-intron) or pCAMBIA1304 (with a gusA), were used to examine for embryo transformation efficiency of the three rice grain cultivars, RD41, RD47 and PSL2. The results show that (1) SAAT for 10 min was proved to be the best for increasing microwound regions on the embryo surface; (2) EHA105-pCAMBIA1304 gave higher transformation efficiency of dehusked seeds (87.5 ± 7.5%) than AGL1-pCAMBIA1304 (37.5 ± 2.5%); (3) the combination of SAAT for 10 min followed by another 10 min of VIAAT gave the transient GUS expression in coleoptile (18%) and primary leaf (2%) of husked seedling; and (4) the use of transformation protocol with EHA105-pCAMBIA1301 showed that the rice cultivar PSL2 gave significantly higher transformation efficiency than RD41 and RD47. This work thus suggests that genetic transformation in plants could be achieved by SAAT and VIAAT techniques without tissue culture process.

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a Biology Program, Faculty of Science and Technology, Pibulsongkram Rajabhat University, Phitsanulok 65000 Thailand
b Department of Agricultural Science, Faculty of Agriculture, Natural Resources and Environment, Naresuan University, Phitsanulok 65000 Thailand

* Corresponding author, E-mail: sittichai.u@psru.ac.th

Received 5 Dec 2019, Accepted 6 Jun 2020