PJB-2019-634
Genetic engineering of sugarcane with the rice tonoplast H+-PPase (OVP2) gene to improve sucrose content and salt tolerance
Mohammad Sayyar Khan, Sayed Usman Ali Shah, Mazhar Ullah, Muhammad Zaheer Ahmad, Waqar Ahmad and Asad Jan
Abstract
The present study describes successful transformation of sugarcane (Saccharum officinarum), CP-77/400 variety with the rice vacuolar H+-PPase (OVP2) gene. Transgenic and control plants were then tested for biochemical and salt tolerance evaluation. First, sugarcane calli of the selected variety were induced on callus induction media as optimized previously. Various concentrations of acetosyringone and cefatoxime were used along with different co-cultivation periods to obtain maximum transformation efficiency. Maximum transformation efficiency (25 %) was achieved with 60 µM. Different concentrations of cefatoxime were used for washing the calli after co-cultivation to remove excess bacteria. Out of several concentrations, 400 mg/l resulted in higher transformation frequency (26%). Highest transformation frequency of 21 % was achieved when co-cultivation period was extended to four days. Selection of transformants was achieved with various levels of hygromycine as selective antibiotic. The hygromycine concentration of 50 mg/l resulted in higher transformation frequency of 20%. The transgenic shoots were then confirmed through PCR and RT-PCR analysis. It was anticipated that constitutive expression of the OVP2 gene might accelerate the function of the vacuole localized sucrose antiporters resulting higher sucrose accumulation in the vacuole. No significant differences were found for soluble sugar contents between the non-transgenic control and the transgenic lines except T5 line that showed high sucrose content than that of the control. Transgenic and control plants were subjected to salt stress (NaCl) at concentrations of 0 mM and 100 mM. Some of the transgenic lines exhibited high chlorophyll and proline contents than that of the control plants at 100 mM stress. These results showed that expression of the OVP2 gene improved total soluble sugar content and salt tolerance in transgenic lines
To Cite this article:
Khan, M.S., S.U.A. Shah, M. Ullah, M.Z. Ahmad, W. Ahmad and A. Jan. 2021. Genetic engineering of sugarcane with the rice tonoplast H+-PPase (OVP2) gene to improve sucrose content and salt tolerance. Pak. J. Bot., 53(3): DOI: http://dx.doi.org/10.30848/PJB2021-3(36)
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