Field Supervisory Test of DREB-Transgenic Populus: Salt Tolerance, Long-Term Gene Stability and Horizontal Gene Transfer
Abstract
:1. Introduction
2. Materials and Methods
2.1. Characterization of Transformed Plants and Experimental Field Design
2.2. Materials
2.3. Screening for Kanamycin-Resistant Organisms
2.4. DNA Isolation and PCR Analyses
2.5. Total RNA Extraction and Real-Time Quantitative PCR (qRT-PCR)
2.6. Transgenic Populus Tissue Culture
3. Results and Discussion
3.1. Stability of the Foreign Gene after a Seven-Year Field Test
3.2. Expression of AhDREB1 mRNA after Three and Seven Years
3.3. Salt-Tolerant Phenotype of the Transgenic Plants
NaCl concentration (%) | Incubation Time (Days) | Leaf Color | Differentiation Rate (%) | ||
---|---|---|---|---|---|
Transgenic Populus | Controls | Transgenic Populus | Controls | ||
0.2 | 21 | Green | Green | 2.3 | 2.2 |
28 | Green | Green | 3.6 | 3.3 | |
35 | Green | Green | 4.5 | 4.3 | |
0.3 | 21 | Green | Green | 2.4 | 2.2 |
28 | Green | Green | 3.3 | 3.5 | |
35 | Green | Green | 4.6 | 4.2 | |
0.4 | 21 | Green | Green | 2.5 | 1.9 |
28 | Green | Green | 3.0 | 2.3 | |
35 | Green | Yellow/green | 4.3 | 2.7 | |
0.5 | 21 | Green | Green | 2.2 | 0.7 |
28 | Green | Green | 3.2 | 0.9 | |
35 | Green | Yellow | 4.1 | 1.1 | |
0.6 | 21 | Green | Green | 2.1 | - |
28 | Green | Yellow/green | 2.8 | - | |
35 | Yellow/green | Yellow | 3.1 | - | |
0.7 | 21 | Green | Yellow | 1.6 | - |
28 | Yellow/green | Yellow | 1.9 | - | |
35 | Yellow/green | Withered | 2.1 | - | |
0.8 | 21 | Yellow/green | Yellow | 0.6 | - |
28 | Yellow/green | Withered | 1.0 | - | |
35 | Yellow | Dead | 0.7 | - | |
0.9 | 21 | Yellow/green | Withered | 0.2 | - |
28 | Yellow | Withered | 0.4 | - | |
35 | Yellow | Dead | 0.2 | - | |
1.0 | 21 | Yellow/green | Withered | 0.2 | - |
28 | Yellow | Dead | 0.2 | - | |
35 | Withered | Dead | 0.1 | - |
NaCl concentration (%) | Incubation Time (Days) | Leaf Color | Root Number | ||
---|---|---|---|---|---|
Transgenic Populus | Controls | Transgenic Populus | Controls | ||
0.2 | 21 | Green | Green | 1.5 | 1.6 |
28 | Green | Green | 2.4 | 2.3 | |
35 | Green | Green | 3.0 | 2.8 | |
42 | Green | Green | 3.2 | 3.0 | |
0.3 | 21 | Green | Green | 1.7 | 1.5 |
28 | Green | Green | 2.5 | 2.5 | |
35 | Green | Green | 2.7 | 2.9 | |
42 | Green | Green | 3.1 | 3.2 | |
0.4 | 21 | Green | Green | 1.4 | 1.1 |
28 | Green | Green | 2.0 | 1.2 | |
35 | Green | Yellow/green | 2.4 | 1.4 | |
42 | Green | Yellow/green | 2.8 | 1.4 | |
0.5 | 21 | Green | Green | 0.7 | 0.4 |
28 | Green | Green | 1.5 | 0.4 | |
35 | Green | Yellow/green | 1.5 | 0.5 | |
42 | Yellow/green | Yellow/green | 2.1 | 0.5 | |
0.6 | 21 | Green | Green | 0.6 | 0.2 |
28 | Yellow/green | Yellow/green | 0.8 | 0.1 | |
35 | Yellow/green | Yellow | 0.9 | 0.1 | |
42 | Yellow | Withered | 0.9 | 0.1 | |
0.7 | 21 | Green | Yellow | 0.2 | - |
28 | Yellow/green | Yellow | 0.2 | - | |
35 | Yellow/green | Withered | 0.2 | - | |
42 | Yellow | Withered | 0.2 | - | |
0.8 | 21 | Yellow/green | Yellow | 0.2 | - |
28 | Yellow/green | Withered | 0.0 | - | |
35 | Yellow | Dead | 0.1 | - | |
42 | Withered | Dead | 0.1 | - | |
0.9 | 21 | Yellow/green | Withered | 0.1 | - |
28 | Yellow | Withered | - | - | |
35 | Yellow | Dead | - | - | |
42 | Withered | Dead | - | - | |
1.0 | 21 | Yellow | Withered | - | - |
28 | Yellow | Dead | - | - | |
35 | Withered | Dead | - | - | |
42 | Withered | Dead | - | - |
3.4. PCR Amplification of the NptII Gene in the Genomic DNA of Kanamycin-Resistant Microorganisms
3.5. Discussion
4. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Lu, N.; Wei, B.; Sun, Y.; Liu, X.; Chen, S.; Zhang, W.; Zhang, Y.; Li, Y. Field Supervisory Test of DREB-Transgenic Populus: Salt Tolerance, Long-Term Gene Stability and Horizontal Gene Transfer. Forests 2014, 5, 1106-1121. https://doi.org/10.3390/f5051106
Lu N, Wei B, Sun Y, Liu X, Chen S, Zhang W, Zhang Y, Li Y. Field Supervisory Test of DREB-Transgenic Populus: Salt Tolerance, Long-Term Gene Stability and Horizontal Gene Transfer. Forests. 2014; 5(5):1106-1121. https://doi.org/10.3390/f5051106
Chicago/Turabian StyleLu, Nan, Bing Wei, Yuhan Sun, Xin Liu, Shouyi Chen, Wanke Zhang, Yingzhi Zhang, and Yun Li. 2014. "Field Supervisory Test of DREB-Transgenic Populus: Salt Tolerance, Long-Term Gene Stability and Horizontal Gene Transfer" Forests 5, no. 5: 1106-1121. https://doi.org/10.3390/f5051106