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Physiological and proteomic responses of diploid and tetraploid black locust (Robinia pseudoacacia L.) subjected to salt stress.

Wang Z, Wang M, Liu L, Meng F - Int J Mol Sci (2013)

Bottom Line: The results showed that diploid (2×) plants suffered from greater negative effects than those of tetraploid (4×) plants.In addition, salt stress resulted in distorted chloroplasts, swollen thylakoid membranes, accumulation of plastoglobules, and increased starch grains in 2× compared to those in 4×.Collectively, our results suggest that the plants showed significantly different responses to salt stress based on ploidy level of the plant.

View Article: PubMed Central - PubMed

Affiliation: College of Life Science, Northeast Forestry University, Harbin 150040, China. mfj19751@163.com.

ABSTRACT
Tetraploid black locust (Robinia pseudoacacia L.) is adaptable to salt stress. Here, we compared morphological, physiological, ultrastructural, and proteomic traits of leaves in tetraploid black locust and its diploid relatives under salt stress. The results showed that diploid (2×) plants suffered from greater negative effects than those of tetraploid (4×) plants. After salt treatment, plant growth was inhibited, photosynthesis was reduced, reactive oxygen species, malondialdehyde content, and relative electrolyte leakage increased, and defense-related enzyme activities decreased in 2× compared to those in 4×. In addition, salt stress resulted in distorted chloroplasts, swollen thylakoid membranes, accumulation of plastoglobules, and increased starch grains in 2× compared to those in 4×. However, 4× developed diverse responses under salt stress. A comparative proteomic analysis revealed that 41 and 37 proteins were differentially expressed in 2× and 4×, respectively. These proteins were mainly involved in photosynthesis, stress and defense, energy, metabolism, transcription/translation, and transportation. Distinct patterns of protein changes between 2× and 4× were analyzed. Collectively, our results suggest that the plants showed significantly different responses to salt stress based on ploidy level of the plant. The 4× possessed a better salt protection mechanism than that of 2×, suggesting salt tolerance in the polyploid plant.

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Coomassie Brilliant Blue (CBB)-stained two-dimensional electrophoresis gels of proteins from Robinia pseudoacacia diploid (2×) leaves. Proteins were separated on a 13 cm IPG strip (pH 4–7 linear gradient) using isoelectric focusing, followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis on a 12.5% gel. (A) 1-day NaCl-treated leaves; (B) 5-day NaCl-treated leaves; (C) 10-day NaCl-treated leaves. Blue and red numbers indicate proteins that increased and decreased between control and stressed samples, respectively. The proteins are listed in Table 1.
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f9-ijms-14-20299: Coomassie Brilliant Blue (CBB)-stained two-dimensional electrophoresis gels of proteins from Robinia pseudoacacia diploid (2×) leaves. Proteins were separated on a 13 cm IPG strip (pH 4–7 linear gradient) using isoelectric focusing, followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis on a 12.5% gel. (A) 1-day NaCl-treated leaves; (B) 5-day NaCl-treated leaves; (C) 10-day NaCl-treated leaves. Blue and red numbers indicate proteins that increased and decreased between control and stressed samples, respectively. The proteins are listed in Table 1.

Mentions: To investigate the changes in protein profiles under salt stress, total protein from the leaves of control and salt-treated plants was extracted and analyzed by 2-DE. Approximately 800 protein spots were detected in 2× and 4× plants, respectively. Master 2-D gel maps are shown in Figures 9 and 10. All protein spots were quantitatively analyzed, but only the protein spots that showed two-fold or more expression (p < 0.05) changes under the salt treatments were submitted for protein identification. All 41 protein spots from 2× gels and 37 protein spots from 4× gels were detected as differentially expressed spots (Figures 9 and 10; Tables 1 and 2). Twenty-eight proteins were up-regulated in 2× plants. Of which 13 proteins increased gradually in abundance in salt-treated plants, whereas 13 proteins increased in expression after 5 days of salt treatment and then decreased after 10 day of salt treatment. Two proteins decreased and then increased under salt stress compared to that in controls. Twenty-five proteins were up-regulated in 4× plants. Among them, 21 proteins increased gradually compared to controls and only four increased and then decreased with salt treatment.


Physiological and proteomic responses of diploid and tetraploid black locust (Robinia pseudoacacia L.) subjected to salt stress.

Wang Z, Wang M, Liu L, Meng F - Int J Mol Sci (2013)

Coomassie Brilliant Blue (CBB)-stained two-dimensional electrophoresis gels of proteins from Robinia pseudoacacia diploid (2×) leaves. Proteins were separated on a 13 cm IPG strip (pH 4–7 linear gradient) using isoelectric focusing, followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis on a 12.5% gel. (A) 1-day NaCl-treated leaves; (B) 5-day NaCl-treated leaves; (C) 10-day NaCl-treated leaves. Blue and red numbers indicate proteins that increased and decreased between control and stressed samples, respectively. The proteins are listed in Table 1.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3821616&req=5

f9-ijms-14-20299: Coomassie Brilliant Blue (CBB)-stained two-dimensional electrophoresis gels of proteins from Robinia pseudoacacia diploid (2×) leaves. Proteins were separated on a 13 cm IPG strip (pH 4–7 linear gradient) using isoelectric focusing, followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis on a 12.5% gel. (A) 1-day NaCl-treated leaves; (B) 5-day NaCl-treated leaves; (C) 10-day NaCl-treated leaves. Blue and red numbers indicate proteins that increased and decreased between control and stressed samples, respectively. The proteins are listed in Table 1.
Mentions: To investigate the changes in protein profiles under salt stress, total protein from the leaves of control and salt-treated plants was extracted and analyzed by 2-DE. Approximately 800 protein spots were detected in 2× and 4× plants, respectively. Master 2-D gel maps are shown in Figures 9 and 10. All protein spots were quantitatively analyzed, but only the protein spots that showed two-fold or more expression (p < 0.05) changes under the salt treatments were submitted for protein identification. All 41 protein spots from 2× gels and 37 protein spots from 4× gels were detected as differentially expressed spots (Figures 9 and 10; Tables 1 and 2). Twenty-eight proteins were up-regulated in 2× plants. Of which 13 proteins increased gradually in abundance in salt-treated plants, whereas 13 proteins increased in expression after 5 days of salt treatment and then decreased after 10 day of salt treatment. Two proteins decreased and then increased under salt stress compared to that in controls. Twenty-five proteins were up-regulated in 4× plants. Among them, 21 proteins increased gradually compared to controls and only four increased and then decreased with salt treatment.

Bottom Line: The results showed that diploid (2×) plants suffered from greater negative effects than those of tetraploid (4×) plants.In addition, salt stress resulted in distorted chloroplasts, swollen thylakoid membranes, accumulation of plastoglobules, and increased starch grains in 2× compared to those in 4×.Collectively, our results suggest that the plants showed significantly different responses to salt stress based on ploidy level of the plant.

View Article: PubMed Central - PubMed

Affiliation: College of Life Science, Northeast Forestry University, Harbin 150040, China. mfj19751@163.com.

ABSTRACT
Tetraploid black locust (Robinia pseudoacacia L.) is adaptable to salt stress. Here, we compared morphological, physiological, ultrastructural, and proteomic traits of leaves in tetraploid black locust and its diploid relatives under salt stress. The results showed that diploid (2×) plants suffered from greater negative effects than those of tetraploid (4×) plants. After salt treatment, plant growth was inhibited, photosynthesis was reduced, reactive oxygen species, malondialdehyde content, and relative electrolyte leakage increased, and defense-related enzyme activities decreased in 2× compared to those in 4×. In addition, salt stress resulted in distorted chloroplasts, swollen thylakoid membranes, accumulation of plastoglobules, and increased starch grains in 2× compared to those in 4×. However, 4× developed diverse responses under salt stress. A comparative proteomic analysis revealed that 41 and 37 proteins were differentially expressed in 2× and 4×, respectively. These proteins were mainly involved in photosynthesis, stress and defense, energy, metabolism, transcription/translation, and transportation. Distinct patterns of protein changes between 2× and 4× were analyzed. Collectively, our results suggest that the plants showed significantly different responses to salt stress based on ploidy level of the plant. The 4× possessed a better salt protection mechanism than that of 2×, suggesting salt tolerance in the polyploid plant.

Show MeSH
Related in: MedlinePlus