AUTHOR=Zhao Huimin  , Jia Yaqi , Cao Yanting , Wang Yucheng 

TITLE=Improving T-DNA Transfer to Tamarix hispida by Adding Chemical Compounds During Agrobacterium tumefaciens Culture

JOURNAL=Frontiers in Plant Science

VOLUME=11

YEAR=2020

URL=https://www.frontiersin.org/journals/plant-science/articles/10.3389/fpls.2020.501358

DOI=10.3389/fpls.2020.501358

ISSN=1664-462X

ABSTRACT=<p><italic>Agrobacterium tumefaciens</italic>-mediated gene transfer is the most commonly used method for plant genetic engineering. However, during the period of <italic>A. tumefaciens</italic> culture, the effects of <italic>Agrobacterium</italic> culture before inoculation on genetic transformation are poorly understood. In the present study, we investigated the factors that affect the genetic transformation efficiency during <italic>Agrobacterium</italic> culture using <italic>Tamarix hispida</italic> as transgenic plant material. <italic>Agrobacterium</italic> treatment with spermidine (Spe), azacitidine (5-AzaC), dithiothreitol (DTT), or acetosyringone (AS) alone all significantly improved the efficiency of T-DNA transfer. Treatment with 5-AzaC reduced DNA methylation in <italic>Agrobacterium</italic> to induce the expression of virulence (<italic>vir</italic>) family genes, including <italic>vir</italic>A, <italic>vir</italic>B1, <italic>vir</italic>C1, <italic>vir</italic>D2, <italic>vir</italic>D4 <italic>vir</italic>E2, and <italic>vir</italic>G. Spe treatment significantly induced the expression of all the studied genes, including <italic>vir</italic>A, <italic>vir</italic>B1, <italic>vir</italic>C1, <italic>vir</italic>D1, <italic>vir</italic>D2, <italic>vir</italic>D4, <italic>vir</italic>E2, and <italic>vir</italic>G. DTT treatment decreased reactive oxygen species accumulation. AS treatment activated the expression of the genes <italic>vir</italic>A, <italic>vir</italic>B1, <italic>vir</italic>C1, <italic>vir</italic>D1, <italic>vir</italic>D2, <italic>vir</italic>D4 and <italic>vir</italic>G. All these effects resulted in increased T-DNA transfer. Additionally, combined Spe, 5-AzaC, DTT, and AS treatment improve <italic>Agrobacterium</italic> infection to a greater extent compared with their use alone, increasing T-DNA transfer by more than 8-fold relative to no treatment. Therefore, to improve genetic transformation, pretreatment of <italic>Agrobacterium</italic> during the culture period is important for improving genetic transformation efficiency.</p>