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Comparison of Soybean Transformation Efficiency and Plant Factors Affecting Transformation during the Agrobacterium Infection Process.

Jia Y, Yao X, Zhao M, Zhao Q, Du Y, Yu C, Xie F - Int J Mol Sci (2015)

Bottom Line: The objective of this study is to evaluate the plant factors related to transformation in cotyledonary nodes during the Agrobacterium infection process.High-efficiency genotypes had low methyl jasmonate (MeJA) content, polyphenol oxidase (PPO) and peroxidase (POD) activity, and relatively lower expression of soybean OPR3, PPO1 and PRX71.GA and ZR were positive plant factors for Agrobacterium-mediated soybean transformation by facilitating germination and growth, and increasing the number of cells in DNA synthesis cycle, respectively; MeJA, PPO, POD and ABA were negative plant factors by inducing defence reactions and repressing germination and growth, respectively.

View Article: PubMed Central - PubMed

Affiliation: Soybean Research Institute, Shenyang Agricultural University, Shenyang 110866, China. jiayuyinggood@163.com.

ABSTRACT
The susceptibility of soybean genotype to Agrobacterium infection is a key factor for the high level of genetic transformation efficiency. The objective of this study is to evaluate the plant factors related to transformation in cotyledonary nodes during the Agrobacterium infection process. This study selected three genotypes (Williams 82, Shennong 9 and Bert) with high transformation efficiency, which presented better susceptibility to Agrobacterium infection, and three low transformation efficiency genotypes (General, Liaodou 16 and Kottman), which showed a relatively weak susceptibility. Gibberellin (GA) levels and soybean GA20ox2 and CYP707A2 transcripts of high-efficiency genotypes increased and were higher than those of low-efficiency genotypes; however, the opposite performance was shown in abscisic acid (ABA). Higher zeatin riboside (ZR) content and DNA quantity, and relatively higher expression of soybean IPT5, CYCD3 and CYCA3 were obtained in high-efficiency genotypes. High-efficiency genotypes had low methyl jasmonate (MeJA) content, polyphenol oxidase (PPO) and peroxidase (POD) activity, and relatively lower expression of soybean OPR3, PPO1 and PRX71. GA and ZR were positive plant factors for Agrobacterium-mediated soybean transformation by facilitating germination and growth, and increasing the number of cells in DNA synthesis cycle, respectively; MeJA, PPO, POD and ABA were negative plant factors by inducing defence reactions and repressing germination and growth, respectively.

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Comparison of polyphenol oxidase (PPO) and peroxidase (POD) activity, and the expression of encoding genes between different genotypes. The values represent means ± SD based on three biological replications. (A) PPO activity in cotyledonary nodes after 0 h of infection, and after one day, three days, and five days of co-cultivation; (B) qRT-PCR analysis of the gene GmPPO1 that encodes PPO; (C) POD activity; and (D) qRT-PCR analysis of the gene GmPRX71 that encodes POD.
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ijms-16-18522-f008: Comparison of polyphenol oxidase (PPO) and peroxidase (POD) activity, and the expression of encoding genes between different genotypes. The values represent means ± SD based on three biological replications. (A) PPO activity in cotyledonary nodes after 0 h of infection, and after one day, three days, and five days of co-cultivation; (B) qRT-PCR analysis of the gene GmPPO1 that encodes PPO; (C) POD activity; and (D) qRT-PCR analysis of the gene GmPRX71 that encodes POD.

Mentions: PPO and POD are protective enzymes that respond to various stresses. In this study, PPO and POD activity and their encoding gene PPO1 [47] and PRX71 [48] were investigated. The results showed a striking increase of PPO and POD activity occurred during the co-cultivation period (Figure 8A,C). Moreover, PPO and POD activity in General, Liaodou 16 and Kottman was significantly higher than those in Williams 82, Shennong 9 and Bert at 3 DAC and 5 DAC. Expression of soybean homologue of PPO1 and PRX71 increased more significantly in General, Liaodou 16 and Kottman (Figure 8B,D), which followed a similar pattern with the variation of PPO and POD activity, respectively.


Comparison of Soybean Transformation Efficiency and Plant Factors Affecting Transformation during the Agrobacterium Infection Process.

Jia Y, Yao X, Zhao M, Zhao Q, Du Y, Yu C, Xie F - Int J Mol Sci (2015)

Comparison of polyphenol oxidase (PPO) and peroxidase (POD) activity, and the expression of encoding genes between different genotypes. The values represent means ± SD based on three biological replications. (A) PPO activity in cotyledonary nodes after 0 h of infection, and after one day, three days, and five days of co-cultivation; (B) qRT-PCR analysis of the gene GmPPO1 that encodes PPO; (C) POD activity; and (D) qRT-PCR analysis of the gene GmPRX71 that encodes POD.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4581258&req=5

ijms-16-18522-f008: Comparison of polyphenol oxidase (PPO) and peroxidase (POD) activity, and the expression of encoding genes between different genotypes. The values represent means ± SD based on three biological replications. (A) PPO activity in cotyledonary nodes after 0 h of infection, and after one day, three days, and five days of co-cultivation; (B) qRT-PCR analysis of the gene GmPPO1 that encodes PPO; (C) POD activity; and (D) qRT-PCR analysis of the gene GmPRX71 that encodes POD.
Mentions: PPO and POD are protective enzymes that respond to various stresses. In this study, PPO and POD activity and their encoding gene PPO1 [47] and PRX71 [48] were investigated. The results showed a striking increase of PPO and POD activity occurred during the co-cultivation period (Figure 8A,C). Moreover, PPO and POD activity in General, Liaodou 16 and Kottman was significantly higher than those in Williams 82, Shennong 9 and Bert at 3 DAC and 5 DAC. Expression of soybean homologue of PPO1 and PRX71 increased more significantly in General, Liaodou 16 and Kottman (Figure 8B,D), which followed a similar pattern with the variation of PPO and POD activity, respectively.

Bottom Line: The objective of this study is to evaluate the plant factors related to transformation in cotyledonary nodes during the Agrobacterium infection process.High-efficiency genotypes had low methyl jasmonate (MeJA) content, polyphenol oxidase (PPO) and peroxidase (POD) activity, and relatively lower expression of soybean OPR3, PPO1 and PRX71.GA and ZR were positive plant factors for Agrobacterium-mediated soybean transformation by facilitating germination and growth, and increasing the number of cells in DNA synthesis cycle, respectively; MeJA, PPO, POD and ABA were negative plant factors by inducing defence reactions and repressing germination and growth, respectively.

View Article: PubMed Central - PubMed

Affiliation: Soybean Research Institute, Shenyang Agricultural University, Shenyang 110866, China. jiayuyinggood@163.com.

ABSTRACT
The susceptibility of soybean genotype to Agrobacterium infection is a key factor for the high level of genetic transformation efficiency. The objective of this study is to evaluate the plant factors related to transformation in cotyledonary nodes during the Agrobacterium infection process. This study selected three genotypes (Williams 82, Shennong 9 and Bert) with high transformation efficiency, which presented better susceptibility to Agrobacterium infection, and three low transformation efficiency genotypes (General, Liaodou 16 and Kottman), which showed a relatively weak susceptibility. Gibberellin (GA) levels and soybean GA20ox2 and CYP707A2 transcripts of high-efficiency genotypes increased and were higher than those of low-efficiency genotypes; however, the opposite performance was shown in abscisic acid (ABA). Higher zeatin riboside (ZR) content and DNA quantity, and relatively higher expression of soybean IPT5, CYCD3 and CYCA3 were obtained in high-efficiency genotypes. High-efficiency genotypes had low methyl jasmonate (MeJA) content, polyphenol oxidase (PPO) and peroxidase (POD) activity, and relatively lower expression of soybean OPR3, PPO1 and PRX71. GA and ZR were positive plant factors for Agrobacterium-mediated soybean transformation by facilitating germination and growth, and increasing the number of cells in DNA synthesis cycle, respectively; MeJA, PPO, POD and ABA were negative plant factors by inducing defence reactions and repressing germination and growth, respectively.

No MeSH data available.


Related in: MedlinePlus