Department of Horticultural Biotechnology, Kyunghee University1
Drought stress is a crucial environmental factor determining crop survival and productivity. The goal of this study was to clearly identify a new drought stress-tolerance gene in Brassica rapa. From KBGP-24K microarray data with the B. rapa ssp. pekinensis inbred line ‘Chiifu’ under drought stress treatment, a gene which was named BrDSR (B. rapa Drought Stress Resistance) was chosen among 738 drought-responsive unigenes. BrDSR function has yet to be determined, but its expression was induced over 6-fold by drought. To characterize BrDSR, the gene was isolated from B. rapa inbred line ‘CT001’ and found to contain a 438-bp open reading frame encoding a 145 amino acid protein. The full-length cDNA of BrDSR was used to construct an over-expression vector, ‘pSL100’. Tobacco transformation was then conducted to analyze whether the BrDSR gene can increase drought tolerance in plants. The BrDSR expression level in T1 transgenic tobacco plants selected via PCR and DNA blot analyses was up to 2.6-fold higher than non-transgenic tobacco. Analysis of phenotype clearly showed that BrDSR-expressing tobacco plants exhibited more tolerance than wild type under 10 d drought stress. Taking all of these findings together, we expect that BrDSR functions effectively in plant growth and survival of drought stress conditions.
1. Agarwal, P.K., P. Agarwal, M.K. Reddy, and S.K. Sopory. 2006. Role of DREB transcription factors in abiotic and biotic stress tolerance in plants. Plant Cell Rpt. 25:1263-1274.
2. Ahmed, N.U., J.I. Park, H.J. Jung, M.S. Seo, T.S. Kumar, I.H. Lee, and I.S. Nou. 2012. Identification and characterization of stress resistance related genes of Brassica rapa. Biotechnol. Lett. 34:979-987.
3. Candat, A., G. Paszkiewicz, M. Neveu, R. Gautier, D.C. Logan, M.H. Avelange-Macherel, and D. Macherel. 2014. The ubiquitous distribution of late embryogenesis abundant proteins across cell compartments in Arabidopsis offers tailored protection against abiotic stress. Plant Cell 26:3148-3166.
4. Downton, W.J.S., B.R. Loveys, and W.J.R. Grant. 1988. Stomatal closure fully accounts for the inhibition of photosynthesis by abscisic acid. New Phytol. 108:263-266.
5. Dulai, S., I. Molnar, J. Pronay, A. Csernak, R. Tarnai, and M. Molnár-Láng. 2006. Effects of drought on photosynthetic parameters and heat stability of PSII in wheat and in Aegilops species originating from dry habitats. Acta Biol. Szeged 50:11-17.
6. Fei, H., E. Tsang, and A.J. Cutler. 2007. Gene expression during seed maturation in Brassica napus in relation to the induction of secondary dormancy. Genomics 89:419-428.
7. Hara, M., S. Uchida, T. Murata, and H. Wätzig. 2014. Efficient purification of cryoprotective dehydrin protein from the radish (Raphanus sativus) taproot. Eur. Food Res. Technol. 239: 339-345.
8. Hundertmark, M. and D.K. Hincha. 2008. LEA (Late Embryogenesis Abundant) proteins and their encoding genes in Arabidopsis thaliana. BMC Genomics 9:118
9. Karimi, M., A. Depicker, and P. Hilson. 2007. Recombinational cloning with plant gateway vectors. Plant Physiol. 145:1144-1154.
10. Kauser, R., H. Athar, and M. Ashraf. 2006. Chlorophyll fluorescence: a potential indicator for assessment of water stress tolerance in canola (Brassica napus L.). Pak. J. Bot. 38:1501-1509.
11. Lee, M.K., H.S. Kim, S.H. Kim, and Y.D. Park. 2004. Analysis of T-DNA integration patterns in transgenic tobacco plants. J. Plant Biol. 47:179-186.
12. Lee, S.C., M.H. Lim, J.A. Kim, S.I. Lee, J.S. Kim, M. Jin, S.J. Kwon, J.H. Mun, Y.K. Kim, H.U. Kim, Y. Hur, and B.S. Park. 2008. Transcriptome analysis in Brassica rapa under the abiotic stresses using Brassica 24K oligo microarray. Mol. Cells 26:595-605.
13. Li, J., X.Q. Wang, M.B. Watson, and S.M. Assmann. 2000. Regulation of abscisic acid-induced stomatal closure and anion channels by guard cell AAPK kinase. Science 287:300-303.
14. Luhua, S., A. Hegie, N. Suzuki, E. Shulaev, X. Luo, D. Cenariu, V. Ma, S. Kao, J. Lim, M.B. Gunay, T. Oosumi, S.C. Lee, J. Harper, J. Cushman, M. Gollery, T. Girke, J. Bailey-Serres, R.A. Stevenson, J.K. Zhu, and R. Mittler. 2013. Linking genes of unknown function with abiotic stress responses by high- throughput phenotype screening. Physiol. Plant. 148:322-333.
15. Massacci, A., S.M. Nabiev, L. Pietrosanti, S.K. Nematov, T.N. Chernikova, K. Thor, and J. Leipner. 2008. Response of photosynthetic apparatus of cotton (Gossypium hirsutum) to the onset of drought stress under field conditions studied by gas-exchange analysis and chlorophyll fluorescence imaging. Plant Physiol. Biochem. 46:189-195.
16. Olvera-Carrillo, Y. J.L. Reyes, and A.A. Covarrubias. 2011. Late embryogenesis abundant proteins. Plant Signal. Behav. 6:586-589.
17. Oono, Y., M. Seki, M. Satou, K. Iida, K. Akiyama, T. Sakurai, M. Fujita, K. Yamaguchi-Shinozaki, and K. Shinozaki. 2006. Monitoring expression profiles of Arabidopsis genes during cold acclimation and deacclimation using DNA microarrays. Funct. Integr. Genomics 6:212-234.
18. Qin, L.X., Y. Li, D.D. Li, W.L. Xu, Y. Zheng, and X.B. Li. 2014. Arabidopsis drought-induced protein Di19-3 participates in plant response to drought and high salinity stresses. Plant Mol. Biol. 86:609-625.
19. Rabbani, M.A., K. Maruyama, H. Abe, M.A. Khan, K. Katsura, Y. Ito, K. Yoshiwara, M. Seki, K. Shinozaki, and K. Yamaguchi- Shinozaki. 2003. Monitoring expression profiles of rice genes under cold, drought, and high-salinity stresses and abscisic acid application using cDNA microarray and RNA gel-blot analyses. Plant Physiol. 133:1755-1767.
20. Rahbarian, R., R. Khavari-Nejad, A. Ganjeali, A. Bagheri, and F. Najafi. 2011. Drought stress effects on photosynthesis, chlorophyll fluorescence and water relations in tolerant and susceptible chickpea (Cicer arietinum L.) Genotypes. Acta Biol. Crac. Ser. Bot. 53:47-56.
21. RDA (Rural Development Administration). 2007. Culture plan of principal horticultural crops (Suwon, South Korea: RDA).
22. Seki, M., T. Umezawa, K. Urano, and K. Shinozaki. 2007. Regulatory metabolic networks in drought stress responses. Curr. Opin. Plant Biol. 10:296-302.
23. Shi, H., T. Ye, Y. Wang, and Z. Chan. 2013. Arabidopsis ALTERED MERISTEM PROGRAM 1 negatively modulates plant responses to abscisic acid and dehydration stress. Plant Physiol. Biochem. 67:209-216.
24. Soeda, Y., M.C. Konings, O. Vorst, A.M. van Houwelingen, G.M. Stoopen, C.A. Maliepaard, J. Kodde, R.J. Bino, S.P. Groot, and A.H. van der Geest. 2005. Gene expression programs during Brassica oleracea seed maturation, osmopriming, and germination are indicators of progression of the germination process and the stress tolerance level. Plant Physiol. 137:354-368.
25. Vaseva, I.I., B.S. Grigorova, L.P. Simova-Stoilova, K.N. Demirevska, and U. Feller. 2010. Abscisic acid and late embryogenesis abundant protein profile changes in winter wheat under progressive drought stress. Plant Biol. 12:698-707.
26. Velasco-Conde, T., I. Yakovlev, J.P. Majada, I. Aranda, and Ø. Johnsen. 2012. Dehydrins in maritime pine (Pinus pinaster) and their expression related to drought stress response. Tree Genet. Genomes 8:957-973.
27. Verbruggen, N. and C. Hermans. 2008. Proline accumulation in plants: a review. Amino Acids 35:753-759.
28. Wang, W., B. Vinocur, and A. Altman. 2003. Plant responses to drought, salinity and extreme temperatures: towards genetic engineering for stress tolerance. Planta 218:1-14.
29. Wang, X., H. Wang, J. Wang, R. Sun, J. Wu, S. Liu, Y. Bai, J.H. Mun, I. Bancroft, F. Cheng, S. Huang, X. Li, W. Hua, J. Wang, X. Wang, M. Freeling, J.C. Pires, A.H. Paterson, B. Chalhoub, B. Wang, A. Hayward, A.G. Sharpe, B.S. Park, B. Weisshaar, B. Liu, B. Li, B. Liu, C. Tong, C. Song, C. Duran, C. Peng, C. Geng, C. Koh, C. Lin, D. Edwards, D. Mu, D. Shen, E. Soumpourou, F. Li, F. Fraser, G. Conant, G. Lassalle, G.J. King, G. Bonnema, H. Tang, H. Wang, H. Belcram, H. Zhou, H. Hirakawa, H. Abe, H. Guo, H. Wang, H. Jin, I.A. Parkin, J. Batley, J.S. Kim, J. Just, J. Li, J. Xu, J. Deng, J.A. Kim, J. Li, J. Yu, J. Meng, J. Wang, J. Min, J. Poulain, J. Wang, K. Hatakeyama, K. Wu, L. Wang, L. Fang, M. Trick, M.G. Links, M. Zhao, M. Jin, N. Ramchiary, N. Drou, P.J. Berkman, Q. Cai, Q. Huang, R. Li, S. Tabata, S. Cheng, S. Zhang, S. Zhang, S. Huang, S. Sato, S. Sun, S.J. Kwon, S.R. Choi, T.H. Lee, W. Fan, X. Zhao, X. Tan, X. Xu, Y. Wang, Y. Qiu, Y. Yin, Y. Li, Y. Du, Y. Liao, Y. Lim, Y. Narusaka, Y. Wang, Z. Wang, Z. Li, Z. Wang, Z. Xiong, and Z. Zhang. 2011. The genome of the mesopolyploid crop species Brassica rapa. Nat. Genet. 43:1035-1039.
30. Warde-Farley, D., S.L. Donaldson, O. Comes, K. Zuberi, R. Badrawi, P. Chao, M. Franz, C. Grouios, F. Kazi, C.T. Lopes, A. Maitland, S. Mostafavi, J. Montojo, Q. Shao, G. Wright, G.D. Bader, and Q. Morris. 2010. The GeneMANIA prediction server: biological network integration for gene prioritization and predicting gene function. Nucleic Acids Res. W214-220.
31. Winter, D., B. Vinegar, H. Nahal, R. Ammar, G.V. Wilson, and N.J. Provart. 2007. An “electronic fluorescent pictograph” browser for exploring and analyzing large-scale biological data sets. PLoS ONE 2:e718.
32. Xiao, Q.S., X.K. Zhang, B.B. Xu, Y. Cheng, P.Y Zheng, and G.Y. Lu. 2012. Cloning and expression pattern of AnnBn1 gene in Brassica napus. Chin. J. Oil Crop Sci. 34:123-128.
33. Xin, Z., A. Mandaokar, J. Chen, R.L. Last, and J. Browse. 2007. Arabidopsis ESK1 encodes a novel regulator of freezing tolerance. Plant J. 49:786-799.
34. Yamada, M., H. Morishita, K. Urano, N. Shiozaki, K. Yamaguchi- Shinozaki, K. Shinozaki, and Y. Yoshiba. 2005. Effects of free proline accumulation in petunias under drought stress. J. Exp. Bot. 56:1975-1981.
35. Yang, M., Q. Yang, T. Fu, and Y. Zhou. 2011. Overexpression of the Brassica napus BnLAS gene in Arabidopsis affects plant development and increases drought tolerance. Plant Cell Rpt. 30:373-388.
36. Yang, Y., M. He, Z. Zhu, S. Li, Y. Xu, C. Zhang, S.D. Singer, and Y. Wang. 2012. Identification of the dehydrin gene family from grapevine species and analysis of their responsiveness to various forms of abiotic and biotic stress. BMC Plant Biol. 12:140.
37. Yin, H., S. Li, X. Zhao, Y. Du, and X. Ma. 2006. cDNA microarray analysis of gene expression in Brassica napus treated with oligochitosan elicitor. Plant Physiol. Biochem. 44:910-916.
38. Yu, J.G., G.H. Lee, S.C. Lee, and Y.D. Park. 2014. Gene expression and phenotypic analyses of transgenic Chinese cabbage over- expressing the cold tolerance gene, BrCSR. Hort. Environ. Biotechnol. 55:415-422.