Improvement of soybean transformation via Agrobacterium tumefaciens methods involving α-aminooxyacetic acid and sonication treatments enlightened by gene expression profile analysis

Abstract

Antagonists and sonication treatment relieved the structural barriers of Agrobacterium entering into cells; hindered signal perception and transmission; alleviated defense responses and increased cell susceptibility to Agrobacterium infection. Soybean gene expression analysis was performed to elucidate the general response of soybean plant to Agrobacterium at an early stage of infection. Agrobacterium infection stimulated the PAMPs-triggered immunity (BRI1, BAK1, BZR1, FLS2 and EFR) and effector-triggered immunity (RPM1, RPS2, RPS5, RIN4, and PBS1); up-regulated the transcript factors (WRKY25, WRKY29, MEKK1P, MKK4/5P and MYC2) in MAPK pathway; strengthened the biosynthesis of flavonoid and isoflavonoid in the second metabolism; finally led to a fierce defense response of soybean to Agrobacterium infection and thereby lower transformation efficiency. To overcome it, antagonist α-aminooxyacetic acid (AOA) and sonication treatment along with Agrobacterium infection were applied. This novel method dramatically decreased the expression of genes coding for F3'H, HCT, β-glucosidase and IF7GT, etc., which are important for isoflavone biosynthesis or the interconversion of aglycones and glycon; genes coding for peroxidase, FLS2, PBS1 and transcription factor MYC2, etc., which are important components in plant-pathogen interaction; and genes coding for GPAT and α-L-fucosidase, which are important in polyesters formation in cell membrane and the degradation of fucose-containing glycoproteins and glycolipids on the external surface of cell membrane, respectively. This analysis implied that AOA and sonication treatment not only relieved the structural membrane barriers of Agrobacterium entering into cells, but also hindered the perception of 'invasion' signal on cell membrane and intercellular signal transmission, thus effectively alleviated the defense responses and increased the cell susceptibility to Agrobacterium infection. All these factors benefit the transformation process; other measures should also be further explored to improve soybean transformation.

A giant NLR gene confers broad-spectrum resistance to Phytophthora sojae in soybean

“…Ideally, Rps11 genomic DNA would be transformed into soybean with its own promoter to best reveal the resistance conferred by Rps11 . However, given the large size of Rps11 from the TSR to the TTS (27.7 kb), the unclear boundary of its promoter region upstream of the TSR, as well as anticipated difficulty in transforming such a large segment in soybean—one of the most difficult crops to transform even for DNA fragments as small as a few kb 12 —we decided to use the promoter of an Arabidopsis ubiquitin gene, AtUbi3 (p AtUbi3 ), to drive Rps11 CDS (CDS- Rps11 ) expression (see “Methods”). The p AtUbi 3::CDS- Rps11 construct was introduced into an elite soybean variety 93Y21, which is susceptible to P. sojae races 25, 31, and OH12108-06-03.…”

Transcriptome analysis of the immature inflorescences of foxtail millet treated with Sq-1 for inducing male sterility

“…In the present study, 16 DEGs referred to other crops to verify their homogous genes' expression between the SQ-1-treated and control plants were used. These genes included DFR2A (Sachiko et al 2013), ABCB1 (Cecchetti et al 2015), NEF1 (Ariizumi et al 2004), Ms1 (Yang et al 2007 which are involved in 'anther' development along with other genes such as GH3 (Feng et al 2015), FD (Kaur et al 2021), MKK4/5P (Zhang et al 2016). Gene expression was measured by qRT-PCR using gene-specific primers.…”

Transcriptome Profiling of Barley Cultivar Hua 30 MDEC in Response to Agrobacterium Infection

“…In soybean, genes involved in this metabolic process are also upregulated. But these genes' expression was reduced when a-aminooxyacetic acid (AOA, transformation promotion factor) was added in tissue culture following transformation [22]. This suggested that the upregulation of phenylpropanoid biosynthesis pathway may negatively affect the Agrobacterium infection.…”