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A novel α/β-hydrolase gene IbMas enhances salt tolerance in transgenic sweetpotato.

Liu D, Wang L, Zhai H, Song X, He S, Liu Q - PLoS ONE (2014)

Bottom Line: Shangshu 19) plants exhibited significantly higher salt tolerance compared with the wild-type.Proline content was significantly increased, whereas malonaldehyde content was significantly decreased in the transgenic plants.H2O2 was also found to be significantly less accumulated in the transgenic plants than in the wild-type.

View Article: PubMed Central - PubMed

Affiliation: Beijing Key Laboratory of Crop Genetic Improvement/Laboratory of Crop Heterosis and Utilization, Ministry of Education, China Agricultural University, Beijing, China.

ABSTRACT
Salt stress is one of the major environmental stresses in agriculture worldwide and affects crop productivity and quality. The development of crops with elevated levels of salt tolerance is therefore highly desirable. In the present study, a novel maspardin gene, named IbMas, was isolated from salt-tolerant sweetpotato (Ipomoea batatas (L.) Lam.) line ND98. IbMas contains maspardin domain and belongs to α/β-hydrolase superfamily. Expression of IbMas was up-regulated in sweetpotato under salt stress and ABA treatment. The IbMas-overexpressing sweetpotato (cv. Shangshu 19) plants exhibited significantly higher salt tolerance compared with the wild-type. Proline content was significantly increased, whereas malonaldehyde content was significantly decreased in the transgenic plants. The activities of superoxide dismutase (SOD) and photosynthesis were significantly enhanced in the transgenic plants. H2O2 was also found to be significantly less accumulated in the transgenic plants than in the wild-type. Overexpression of IbMas up-regulated the salt stress responsive genes, including pyrroline-5-carboxylate synthase, pyrroline-5-carboxylate reductase, SOD, psbA and phosphoribulokinase genes, under salt stress. These findings suggest that overexpression of IbMas enhances salt tolerance of the transgenic sweetpotato plants by regulating osmotic balance, protecting membrane integrity and photosynthesis and increasing reactive oxygen species scavenging capacity.

No MeSH data available.


Related in: MedlinePlus

Production of transgenic sweetpotato plants overexpressing the IbMas gene.(A) Embryogenic suspension cultures rapidly proliferating in MS medium containing 2.0 mg L−1 2,4-D. (B) PPT-resistant calluses formed on MS medium with 2.0 mg L−1 2,4-D, 100 mg L−1 Carb and 0.8 mg L−1 PPT after 8 weeks of selection. (C) Regeneration of plantlets from PPT-resistant calluses on MS medium with 1.0 mg L−1 ABA, 100 mg L−1 Carb and 0.8 mg L−1 PPT. (D) PCR analysis of transgenic plants. Lane M: DL2000 DNA marker; Lane W: water as negative control; Lane P: plasmid pCAMBIA3301-IbMas as positive control; Lane WT: wild-type as negative control; Lanes L39, L52, L55, L99, L101, L102 and L109: transgenic plants. (E) and (F) WT and transgenic plants grown in a greenhouse, respectively. (G) and (H) WT and transgenic plants grown in a field, respectively. (I) and (J) Storage roots of WT and transgenic plants, respectively.
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pone-0115128-g004: Production of transgenic sweetpotato plants overexpressing the IbMas gene.(A) Embryogenic suspension cultures rapidly proliferating in MS medium containing 2.0 mg L−1 2,4-D. (B) PPT-resistant calluses formed on MS medium with 2.0 mg L−1 2,4-D, 100 mg L−1 Carb and 0.8 mg L−1 PPT after 8 weeks of selection. (C) Regeneration of plantlets from PPT-resistant calluses on MS medium with 1.0 mg L−1 ABA, 100 mg L−1 Carb and 0.8 mg L−1 PPT. (D) PCR analysis of transgenic plants. Lane M: DL2000 DNA marker; Lane W: water as negative control; Lane P: plasmid pCAMBIA3301-IbMas as positive control; Lane WT: wild-type as negative control; Lanes L39, L52, L55, L99, L101, L102 and L109: transgenic plants. (E) and (F) WT and transgenic plants grown in a greenhouse, respectively. (G) and (H) WT and transgenic plants grown in a field, respectively. (I) and (J) Storage roots of WT and transgenic plants, respectively.

Mentions: A total of 1000 cell aggregates of sweetpotato cv. Shangshu 19 (Fig. 4A) cocultivated with A. tumefaciens strain EHA 105 were cultured on the selective medium with 2.0 mg L−1 2,4-dichlorophenoxyacetic acid (2,4-D), 100 mg L−1 carbenicillin (Carb) and 0.8 mg L−1 PPT. Eight weeks after selection, 72 PPT-resistant embryogenic calluses were produced from them (Fig. 4B) and transferred to MS medium with 1.0 mg L−1 ABA, 100 mg L−1 Carb and 0.8 mg L−1 PPT. After 5 to 6 weeks, 61 of them formed somatic embryos which further germinated into plantlets on the same medium (Fig. 4C). These plantlets developed into whole plants on the basal medium. A total of 173 putatively transgenic plants, named L1, L2, …, L173, respectively, were obtained in the present study.


A novel α/β-hydrolase gene IbMas enhances salt tolerance in transgenic sweetpotato.

Liu D, Wang L, Zhai H, Song X, He S, Liu Q - PLoS ONE (2014)

Production of transgenic sweetpotato plants overexpressing the IbMas gene.(A) Embryogenic suspension cultures rapidly proliferating in MS medium containing 2.0 mg L−1 2,4-D. (B) PPT-resistant calluses formed on MS medium with 2.0 mg L−1 2,4-D, 100 mg L−1 Carb and 0.8 mg L−1 PPT after 8 weeks of selection. (C) Regeneration of plantlets from PPT-resistant calluses on MS medium with 1.0 mg L−1 ABA, 100 mg L−1 Carb and 0.8 mg L−1 PPT. (D) PCR analysis of transgenic plants. Lane M: DL2000 DNA marker; Lane W: water as negative control; Lane P: plasmid pCAMBIA3301-IbMas as positive control; Lane WT: wild-type as negative control; Lanes L39, L52, L55, L99, L101, L102 and L109: transgenic plants. (E) and (F) WT and transgenic plants grown in a greenhouse, respectively. (G) and (H) WT and transgenic plants grown in a field, respectively. (I) and (J) Storage roots of WT and transgenic plants, respectively.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4264881&req=5

pone-0115128-g004: Production of transgenic sweetpotato plants overexpressing the IbMas gene.(A) Embryogenic suspension cultures rapidly proliferating in MS medium containing 2.0 mg L−1 2,4-D. (B) PPT-resistant calluses formed on MS medium with 2.0 mg L−1 2,4-D, 100 mg L−1 Carb and 0.8 mg L−1 PPT after 8 weeks of selection. (C) Regeneration of plantlets from PPT-resistant calluses on MS medium with 1.0 mg L−1 ABA, 100 mg L−1 Carb and 0.8 mg L−1 PPT. (D) PCR analysis of transgenic plants. Lane M: DL2000 DNA marker; Lane W: water as negative control; Lane P: plasmid pCAMBIA3301-IbMas as positive control; Lane WT: wild-type as negative control; Lanes L39, L52, L55, L99, L101, L102 and L109: transgenic plants. (E) and (F) WT and transgenic plants grown in a greenhouse, respectively. (G) and (H) WT and transgenic plants grown in a field, respectively. (I) and (J) Storage roots of WT and transgenic plants, respectively.
Mentions: A total of 1000 cell aggregates of sweetpotato cv. Shangshu 19 (Fig. 4A) cocultivated with A. tumefaciens strain EHA 105 were cultured on the selective medium with 2.0 mg L−1 2,4-dichlorophenoxyacetic acid (2,4-D), 100 mg L−1 carbenicillin (Carb) and 0.8 mg L−1 PPT. Eight weeks after selection, 72 PPT-resistant embryogenic calluses were produced from them (Fig. 4B) and transferred to MS medium with 1.0 mg L−1 ABA, 100 mg L−1 Carb and 0.8 mg L−1 PPT. After 5 to 6 weeks, 61 of them formed somatic embryos which further germinated into plantlets on the same medium (Fig. 4C). These plantlets developed into whole plants on the basal medium. A total of 173 putatively transgenic plants, named L1, L2, …, L173, respectively, were obtained in the present study.

Bottom Line: Shangshu 19) plants exhibited significantly higher salt tolerance compared with the wild-type.Proline content was significantly increased, whereas malonaldehyde content was significantly decreased in the transgenic plants.H2O2 was also found to be significantly less accumulated in the transgenic plants than in the wild-type.

View Article: PubMed Central - PubMed

Affiliation: Beijing Key Laboratory of Crop Genetic Improvement/Laboratory of Crop Heterosis and Utilization, Ministry of Education, China Agricultural University, Beijing, China.

ABSTRACT
Salt stress is one of the major environmental stresses in agriculture worldwide and affects crop productivity and quality. The development of crops with elevated levels of salt tolerance is therefore highly desirable. In the present study, a novel maspardin gene, named IbMas, was isolated from salt-tolerant sweetpotato (Ipomoea batatas (L.) Lam.) line ND98. IbMas contains maspardin domain and belongs to α/β-hydrolase superfamily. Expression of IbMas was up-regulated in sweetpotato under salt stress and ABA treatment. The IbMas-overexpressing sweetpotato (cv. Shangshu 19) plants exhibited significantly higher salt tolerance compared with the wild-type. Proline content was significantly increased, whereas malonaldehyde content was significantly decreased in the transgenic plants. The activities of superoxide dismutase (SOD) and photosynthesis were significantly enhanced in the transgenic plants. H2O2 was also found to be significantly less accumulated in the transgenic plants than in the wild-type. Overexpression of IbMas up-regulated the salt stress responsive genes, including pyrroline-5-carboxylate synthase, pyrroline-5-carboxylate reductase, SOD, psbA and phosphoribulokinase genes, under salt stress. These findings suggest that overexpression of IbMas enhances salt tolerance of the transgenic sweetpotato plants by regulating osmotic balance, protecting membrane integrity and photosynthesis and increasing reactive oxygen species scavenging capacity.

No MeSH data available.


Related in: MedlinePlus