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Expression of Pennisetum glaucum Eukaryotic Translational Initiation Factor 4A ( PgeIF4A ) Confers Improved Drought, Salinity, and Oxidative Stress Tolerance in Groundnut

View Article: PubMed Central - PubMed

ABSTRACT

Eukaryotic translational initiation factor 4A belong to family of helicases, involved in multifunctional activities during stress and non-stress conditions. The eIF4A gene was isolated and cloned from semi-arid cereal crop of Pennisetum glaucum. In present study, the PgeIF4A gene was expressed under the regulation of stress inducible Arabidopsis rd29A promoter in groundnut (cv JL-24) with bar as a selectable marker. The de-embryonated cotyledons were infected with Agrobacterium tumefaciens (LBA4404) carrying rd29A:PgeIF4A construct and generated high frequency of multiple shoots in phosphinothricin medium. Twenty- four T0 plants showed integration of both nos-bar and rd29A-PgeIF4A gene cassettes in genome with expected amplification products of 429 and 654 bps, respectively. Transgene copy number integration was observed in five T0 transgenic plants through Southern blot analysis. Predicted Mendelian ratio of segregation (3:1) was noted in transgenic plants at T1 generation. The T2 homozygous lines (L1-5, L8-2, and L16-2) expressing PgeIF4A gene were exhibited superior growth performance with respect to phenotypic parameters like shoot length, tap root length, and lateral root formation under simulated drought and salinity stresses compared to the wild type. In addition, the chlorophyll retention was found to be higher in these plants compared to the control plants. The quantitative real time—PCR results confirmed higher expression of PgeIF4A gene in L1-5, L8-3, and L16-2 plants imposed with drought/salt stress. Further, the salt stress tolerance was associated with increase in oxidative stress markers, such as superoxide dismutase accumulation, reactive oxygen species scavenging, and membrane stability in transgenic plants. Taken together our results confirmed that the PgeIF4A gene expressing transgenic groundnut plants exhibited better adaptation to stress conditions.

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Characterization of eIF4A groundnut transgenic plants under simulated drought stress conditions. (A) Growth performance of PgeIF4A expressing transgenic plants (T2) and control (C) on plain MS medium. (B) Graphical representation of growth performance transgenic plants (T2) and control (C) on 200 and 300 mM mannitol. (C) Relative shoot length in cm. (D) Tap root length in cm. (E) Number of lateral roots. (F) Chlorophyll retention of transgenic and control plants grown under 0, 200, and 300 mM mannitol medium. All the experiments performed in triplicates and data represented as mean (n = three biological triplicates) using two way ANOVA with LSD and P < 0.005. Different alphabets indicated the significant differences between the treatments. Similar letters denotes non significant.
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Figure 5: Characterization of eIF4A groundnut transgenic plants under simulated drought stress conditions. (A) Growth performance of PgeIF4A expressing transgenic plants (T2) and control (C) on plain MS medium. (B) Graphical representation of growth performance transgenic plants (T2) and control (C) on 200 and 300 mM mannitol. (C) Relative shoot length in cm. (D) Tap root length in cm. (E) Number of lateral roots. (F) Chlorophyll retention of transgenic and control plants grown under 0, 200, and 300 mM mannitol medium. All the experiments performed in triplicates and data represented as mean (n = three biological triplicates) using two way ANOVA with LSD and P < 0.005. Different alphabets indicated the significant differences between the treatments. Similar letters denotes non significant.

Mentions: The transformed plants germinated normally without growth penalty like control plants (Figures 5A,B). There was a significant difference in the seedling growth of the wild type and transgenic lines under stress conditions. The shoot length and tap root length of transgenics exhibited higher growth than the wild type plants. Interestingly, the tap root system of transgenics treated with mannitol developed better than the control plants grown under normal conditions at both concentrations of mannitol (200 and 300 mM; Figures 5C,D; Tables 3, 4). The lateral root formation was completely inhibited in control, whereas these roots enormously proliferated under both 200 and 300 mM mannitol concentrations in case of transgenic lines (Figure 5E; Tables 3, 4).


Expression of Pennisetum glaucum Eukaryotic Translational Initiation Factor 4A ( PgeIF4A ) Confers Improved Drought, Salinity, and Oxidative Stress Tolerance in Groundnut
Characterization of eIF4A groundnut transgenic plants under simulated drought stress conditions. (A) Growth performance of PgeIF4A expressing transgenic plants (T2) and control (C) on plain MS medium. (B) Graphical representation of growth performance transgenic plants (T2) and control (C) on 200 and 300 mM mannitol. (C) Relative shoot length in cm. (D) Tap root length in cm. (E) Number of lateral roots. (F) Chlorophyll retention of transgenic and control plants grown under 0, 200, and 300 mM mannitol medium. All the experiments performed in triplicates and data represented as mean (n = three biological triplicates) using two way ANOVA with LSD and P < 0.005. Different alphabets indicated the significant differences between the treatments. Similar letters denotes non significant.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5383670&req=5

Figure 5: Characterization of eIF4A groundnut transgenic plants under simulated drought stress conditions. (A) Growth performance of PgeIF4A expressing transgenic plants (T2) and control (C) on plain MS medium. (B) Graphical representation of growth performance transgenic plants (T2) and control (C) on 200 and 300 mM mannitol. (C) Relative shoot length in cm. (D) Tap root length in cm. (E) Number of lateral roots. (F) Chlorophyll retention of transgenic and control plants grown under 0, 200, and 300 mM mannitol medium. All the experiments performed in triplicates and data represented as mean (n = three biological triplicates) using two way ANOVA with LSD and P < 0.005. Different alphabets indicated the significant differences between the treatments. Similar letters denotes non significant.
Mentions: The transformed plants germinated normally without growth penalty like control plants (Figures 5A,B). There was a significant difference in the seedling growth of the wild type and transgenic lines under stress conditions. The shoot length and tap root length of transgenics exhibited higher growth than the wild type plants. Interestingly, the tap root system of transgenics treated with mannitol developed better than the control plants grown under normal conditions at both concentrations of mannitol (200 and 300 mM; Figures 5C,D; Tables 3, 4). The lateral root formation was completely inhibited in control, whereas these roots enormously proliferated under both 200 and 300 mM mannitol concentrations in case of transgenic lines (Figure 5E; Tables 3, 4).

View Article: PubMed Central - PubMed

ABSTRACT

Eukaryotic translational initiation factor 4A belong to family of helicases, involved in multifunctional activities during stress and non-stress conditions. The eIF4A gene was isolated and cloned from semi-arid cereal crop of Pennisetum glaucum. In present study, the PgeIF4A gene was expressed under the regulation of stress inducible Arabidopsis rd29A promoter in groundnut (cv JL-24) with bar as a selectable marker. The de-embryonated cotyledons were infected with Agrobacterium tumefaciens (LBA4404) carrying rd29A:PgeIF4A construct and generated high frequency of multiple shoots in phosphinothricin medium. Twenty- four T0 plants showed integration of both nos-bar and rd29A-PgeIF4A gene cassettes in genome with expected amplification products of 429 and 654 bps, respectively. Transgene copy number integration was observed in five T0 transgenic plants through Southern blot analysis. Predicted Mendelian ratio of segregation (3:1) was noted in transgenic plants at T1 generation. The T2 homozygous lines (L1-5, L8-2, and L16-2) expressing PgeIF4A gene were exhibited superior growth performance with respect to phenotypic parameters like shoot length, tap root length, and lateral root formation under simulated drought and salinity stresses compared to the wild type. In addition, the chlorophyll retention was found to be higher in these plants compared to the control plants. The quantitative real time&mdash;PCR results confirmed higher expression of PgeIF4A gene in L1-5, L8-3, and L16-2 plants imposed with drought/salt stress. Further, the salt stress tolerance was associated with increase in oxidative stress markers, such as superoxide dismutase accumulation, reactive oxygen species scavenging, and membrane stability in transgenic plants. Taken together our results confirmed that the PgeIF4A gene expressing transgenic groundnut plants exhibited better adaptation to stress conditions.

No MeSH data available.


Related in: MedlinePlus