DOI: 10.3724/SP.J.1006.2019.84120

Acta Agronomica Sinica (作物学报) 2019/45:5 PP.693-704

Analysis of drought resistance and DNA methylation level of resynthesized Brassica napus

Brassica napus, as one of the important resources of edible plant oil and forage protein, is a polyploid species with great economic value. However, it is sensitive to drought stress throughout whole lifecycle due to short domestication history and narrow genetic background. Thus, it is a main purpose to breed B. napus cultivar with high yield and drought resistance. In the present study, we compared the drought resistance among S1-S4 generations of resynthesized B. napus and diploid parents under different time periods of 15% PEG-6000 treatment. The different drought tolerance levels were assessed based on phenotype observation, leaf physiological indexes (MDA, soluble protein, SOD and POD). Accompanying with water content analysis, we found the drought tolerance showed a trend of B. oleracea > Bn-S3 > Bn-S4 > Bn-S1 > Bn-S2 > B. rapa. Under drought stress, POD and SOD activities in Bn-S3 and Bn-S4 were higher than these in other plants tested, and MDA content was decreased, indicating that Bn-S3 and Bn-S4 have better ability in clearing ROS, and defending from peroxidation damage. On the basis of HPLC analysis, the methylation level in all materials was the highest under drought stress of 12 h. And the methylation level in B. rapa was higher than that in others, that in Bn-S1 and Bn-S4 was between that in parents, while that in Bn-S2 and Bn-S3 was lower than that in parents. Methylation sensitive amplification polymorphism analysis also revealed multiple changes in methylation and demethylation level of resynthesized B. napus under drought stress, indicating methylation changes might be involved in plant drought tolerance.

Key words:resynthesized Brassica napus,B. rapa,B. oleracea,drought stress,DNA methylation

ReleaseDate:2019-11-05 15:30:00

[1] 朱健康, 倪建平. 植物非生物胁迫信号转导及应答. 中国稻米, 2016, 22(6):52-60. Zhu J K, Ni J P. Abiotic stress signaling and responses in plants. Chin Rice, 2016, 22(6):52-60(in Chinese with English abstract).

[2] 王汉中, 殷艳. 我国油料产业形势分析与发展对策建议. 中国油料作物学报, 2014, 36:414-421. Wang H Z, Yin Y. Analysis and strategy for oil crop industry in China. Chin J Oil Crop Sci, 2014, 36:414-421(in Chinese with English abstract).

[3] Zhu M, Assmann S M. Metabolic signatures in response to abscisic acid (ABA) treatment in Brassica napus guard cells revealed by metabolomics. Sci Rep, 2017, 7:12875-12890.

[4] Tardieu F. Plant response to environmental conditions:assessing potential production, water demand, and negative effects of water deficit. Front Physiol, 2013, 4:17-27.

[5] 纪瑞鹏, 车宇胜, 朱永宁, 梁涛, 冯锐, 于文颖, 张玉书. 干旱对东北春玉米生长发育和产量的影响. 应用生态学报, 2012, 23:3021-3026. Ji R P, Che Y S, Zhu Y N, Liang T, Feng R, Yu W Y, Zhang Y S. Impacts of drought stress on the growth and development and grain yield of spring maize in northeast China. Chin J Appl Ecol, 2012, 23:3021-3026(in Chinese with English abstract).

[6] Luo L J. Breeding for water-saving and drought-resistance rice (WDR) in China. J Exp Bot, 2010, 61:3509-3517.

[7] Fang Y J, Xiong L Z. General mechanisms of drought response and their application in drought resistance improvement in plants. Cell Mol Life Sci, 2015, 72:673-689.

[8] 王娅玲, 李维峰. 干旱胁迫对植物生长及其生理的影响概述. 南方农业, 2015, 9(6):37. Wang Y L, Li W F. Summary of effects of drought stress on plant growth and physiology. South Agric, 2015, 9(6):37(in Chinese).

[9] Enjalbert J N, Zheng S S, Johnson J J, Mullen J L, Byrne P F, McKay J K. Brassicaceae germplasm diversity for agronomic and seed quality traits under drought stress. Ind Crops Prod, 2013, 47:176-185.

[10] 焦靖芝, 谢伶俐, 李小龙, 田志宏, 许本波. 甘蓝型油菜耐旱机理研究进展. 湖北农业科学, 2015, 54:773-777. Jiao J Z, Xie L L, Li X L, Tian Z H, Xu B B. Drought tolerance in rapeseed (Brassica napus L.) oil. Hubei Agric Sci, 2015, 54:773-777(in Chinese with English abstract).

[11] Mailer R J, Cornish P S. Effects of water stress on glucosinolate and oil concentrations in the seeds of rape (Brassica napus L.) and turnip rape (Brassica rapa L. var. silvestris[Lam.] Briggs). Aust J Exp Agric, 1987, 27:707-711.

[12] Tesfamariam E H, Annandale J G, Steyn J M. Water stress effects on winter canola growth and yield. Agron J, 2010, 102:658-666.

[13] Ma Q, Turner D W. Osmotic adjustment segregates with and is positively related to seed yield in F3 lines of crosses between Brassica napus and B. juncea subjected to water deficit. Aust J Exp Agric, 2006, 46:1621-1627.

[14] Danquah A, Zelicourt A D, Colcombet J, Hirt H. The role of ABA and MAPK signaling pathways in plant abiotic stress responses. Biotechnol Adv, 2014, 32:40-52.

[15] Balestrini S, Vartanian N. Rhizogenic activity during water stress-induced senescence in Brassica napus var. oleifera. Physiol Veg, 1983, 21:269-277.

[16] Jaradat M R, Feurtado J A, Huang D Q, Lu Y Q, Cutler A J. Multiple roles of the transcription factor AtMYBR1/AtMYB44 in ABA signaling, stress responses, and leaf senescence. BMC Plant Biol, 2013, 13:192-210.

[17] Szabados L, Savouré A. Proline:a multifunctional amino acid. Trends Plant Sci, 2010, 15:89-97.

[18] Di F F, Jian H J, Wang T Y, Chen X P, Ding Y R, Du H, Lu K, Li J N, Liu LZ. Genome-wide analysis of the PYL gene family and identification of PYL genes that respond to abiotic stress in Brassica napus. Genes, 2018, 9:156-173.

[19] Luo Q X, Peng M, Zhang X L, Lei P, Ji X M, Wahsoon C, Meng F J, Sun G Y. Comparative mitochondrial proteomic, physiological, biochemical and ultrastructural profiling reveal factors underpinning salt tolerance in tetraploid black locust (Robinia pseudoacacia L.). BMC Genomics, 2017, 18:648-670.

[20] Fomeju B F, Falentin C, Lassalle G, Manzanares-Dauleux M J, Régine D. Comparative genomic analysis of duplicated homoeologous regions involved in the resistance of Brassica napus to stem canker. Front Plant Sci, 2015, 6:772-785.

[21] Ramsey J. Polyploidy and ecological adaptation in wild yarrow. Proc Natl Acad Sci USA, 2011, 108:7096-7101.

[22] Allario T, Brumos J, Colmenero-Flores J M, Iglesias D J, Pina J A, Navarro L, Talon M, Ollitrault P, Morillon R. Tetraploid Rangpur lime rootstock increases drought tolerance via enhanced constitutive root abscisic acid production. Plant Cell Environ, 2013, 36:856-868.

[23] Eliášová A, Münzbergová Z. Factors influencing distribution and local coexistence of diploids and tetraploids of Vicia cracca:inferences from a common garden experiment. J Plant Res, 2017, 130:677-687.

[24] Manzaneda A J, Rey P J, Bastida J M, Weiss-Lehman C, Raskin E, Mitchell-Olds T. Environmental aridity is associated with cytotype segregation and polyploidy occurrence in Brachypodium distachyon (Poaceae). New Phytol, 2012, 193:797-805.

[25] Novikova P Y, Hohmann N, Van de Peer Y. Polyploid Arabidopsis species originated around recent glaciation maxima. Curr Opin Plant Biol, 2018, 42:8-15.

[26] Xu Y, Xu H, Wu X, Fang X, Wang J. Genetic changes following hybridization and genome doubling in synthetic Brassica napus. Biochem Genet, 2012, 50:616-624.

[27] Golicz A A, Bayer P E, Barker G C, Edger P P, Kim H, Martinez P A, Chan C K K, Severn-Ellis A, McCombie W R, Parkin I A P, Paterson A H, Pires J C, Sharpe A G, Tang H B, Teakle G R, Town C D, Batley J, Edwards D. The pangenome of an agronomically important crop plant Brassica oleracea. Nat Commun, 2016, 7:13390-13397.

[28] Gabur I, Chawla H S, Liu X W, Kumar V, Faure S, Tiedemann A, Jestin C, Dryzska E, Volkmann S, Breuer F, Delourme R, Snowdon R, Obermeier C. Finding invisible quantitative trait loci with missing data. Plant Biotechnol J, 2018, doi:10.1111/pbi.12942.

[29] 李勤菲, 陈致富, 刘瑶, 梅家琴, 钱伟. 六倍体(AnAnCnCnCoCo)与白菜型油菜杂交可交配性及后代菌核病抗性. 中国农业科学, 2017, 50:123-130. Li Q F, Chen Z F, Liu Y, Mei J Q, Qian W. Crossability and Sclerotinia resistance among hybrids between hexaploid (AnAnCnCnCoCo) and Brassica rapa. Sci Agric Sin, 2017, 50:123-130(in Chinese with English abstract).

[30] Ran L P, Fang T T, Rong H, Jiang J J, Fang Y J, Wang Y P. Analysis of cytosine methylation in early generations of resynthesized Brassica napus. J Integr Agric, 2016, 15:1228-123.

[31] Barrs H D, Weatherley P E. A re-examination of the relative turgidity technique for estimating water deficits in leaves. Aust J Biol Sci, 1968, 15:413-428.

[32] Hodges D M, Delong J M, Forney C F, Prangel R K. Improving the thiobarbituric acid-reactive-substances assay for estimating lipid peroxidation in plant tissues containing anthocyanin and other interfering compounds. Planta, 1999, 207:604-611.

[33] Snyder J C, Desborough S L. Rapid estimation of potato tuber total protein content with coomassie brilliant blue G-250. Theor Appl Genet, 1978, 52:135-139.

[34] Giannopolitis C N, Ries S K. Superoxide dismutases:Ⅱ. Purification and quantitative relationship with water-soluble protein in seedlings (of corn, peas, and oats). Plant Physiol, 1977, 59:315-318.

[35] Xiong L Z, Xu C G, Saghai Maroof M A, Zhang Q F. Patterns of cytosine methylation in an elite rice hybrid and its parental lines, detected by a methylation-sensitive amplification polymorphism technique. Mol Gen Genet, 1999, 261:439-446.

[36] 谢涛, 戎浩, 蒋金金, 孔月琴, 冉丽萍, 吴健, 王幼平. 人工合成甘蓝型油菜及其亲本的甲基化变异模式分析. 作物学报, 2016, 42:513-524. Xie T, Rong H, Jiang J J, Kong Y Q, Ran L P, Wu J, Wang Y P. Analysis of DNA methylation patterns in resynthesized Brassica napus and diploid parents. Acta Agron Sin, 2016, 42:513-524(in Chinese with English abstract).

[37] Valliyodan B, Nguyen H T. Understanding regulatory networks and engineering for enhanced drought tolerance in plants. Curr Opin Plant Biol, 2006, 9:189-195.

[38] Qaderi M M, Kurepin L V, Reid D M. Growth and physiological responses of canola (Brassica napus) to three components of global climate change:temperature, carbon dioxide and drought. Physiol Plant, 2006, 128:710-721.

[39] Joshi R, Wani S H, Singh B, Bohra A, Dar Z A, Lone A A, Pareek A, Singla-Pareek Sneh L. Transcription factors and plants response to drought stress:current understanding and future directions. Front Plant Sci, 2016, 7:1029-1043.

[40] Nagahatenna D S K, Langridge P, Whitford R. Tetrapyrrole-based drought stress signalling. Plant Biotechnol J, 2015, 13:447-459.

[41] Shao H B, Chu L Y, Jaleel C A, Manivannan P, Panneerselvam R, Shao M A. Understanding water deficit stress-induced changes in the basic metabolism of higher plants-biotechnologically and sustainably improving agriculture and the ecoenvironment in arid regions of the globe. Crit Rev Biotechnol, 2009, 29:131-151.

[42] Liu D, Pei Z F, Naeem M S, Ming D F, Liu H B, Khan F, Zhou W J. 5-Aminolevulinic acid activates antioxidative defence system and seedling growth in Brassica napus L. under water-deficit stress. J Agron Crop Sci, 2011, 197:284-295.

[43] Shafiq S, Akram N A, Ashraf M, Arshad A. Synergistic effects of drought and ascorbic acid on growth, mineral nutrients and oxidative defense system in canola (Brassica napus L.) plants. Acta Physiol Plant, 2014, 36:1539-1553.

[44] Mittler R, Vanderauwera S, Gollery M, Gollery M, Breusegem F V. Reactive oxygen gene network of plants. Trends Plant Sci, 2004, 9:490-498.

[45] Ahmad J, Bashir H, Bagheri R, Baig A, Al-Huqail A, Ibrahim M M, Qureshi M I. Drought and salinity induced changes in ecophysiology and proteomic profile of Parthenium hysterophorus. PLoS One, 2017, 12:e0185118.

[46] Akram N A, Iqbal M, Muhammad A, Ashraf M, Al-Qurainy F, Shafiq S. Aminolevulinic acid and nitric oxide regulate oxidative defense and secondary metabolisms in canola (Brassica napus L.) under drought stress. Protoplasma, 2018, 255:163-174.

[47] Shafiq S, Akram N A, Ashraf M. Does exogenously-applied trehalose alter oxidative defense system in the edible part of radish (Raphanus sativus L.) under water-deficit conditions? Sci Hortic, 2015, 185:68-75.

[48] Latif M, Akram N A, Ashraf M. Regulation of some biochemical attributes in drought-stressed cauliflower (Brassica oleracea L.) by seed pretreatment with ascorbic acid. Sci Hortic, 2016, 91:129-137.

[49] Caverzan A, Passaia G, Rosa S B, Ribeiro C W, Lazzarotto F, Margis-Pinheiro M. Plant responses to stresses:role of ascorbate peroxidase in the antioxidant protection. Genet Mol Biol, 2012, 35:1011-1019.

[50] Ren J, Sun L N, Zhang Q Y, Song X S. Drought tolerance is correlated with the activity of antioxidant enzymes in Cerasus humilis seedlings. Biomed Res Int, 2016, 2016:2038-2047.

[51] Xia L J, Yang L Q, Sun N L, Li J, Fang Y J, Wang Y P. Physiological and antioxidant enzyme gene expression analysis reveals the improved tolerance to drought stress of the somatic hybrid offspring of Brassica napus and Sinapis alba, at vegetative stage. Acta Physiol Planta, 2016, 38:1-10.

[52] Wang T, Huang D Y, Chen B Y, Mao N N, Qiao Y S, Ji M X. Differential expression of photosynthesis-related genes in pentaploid interspecific hybrid and its decaploid of Fragaria spp. Genes Genomics, 2018, 40:321-331.

[53] Jiao J, Wu J, Lyu Z, Sun C, Gao L, Yan X, Cui L, Tang Z, Yan B, Jia Y. Methylation-sensitive amplified polymorphism-based genome-wide analysis of cytosine methylation profiles in Nicotiana tabacum cultivars. Genet Mol Res, 2015, 14:15177-15187.

[54] Fang L, Gong H, Hu Y, Liu C X, Zhou B L, Huang T, Wang Y K, Chen S Q, Fang D D, Du X M, Chen H, Chen J D, Wang S, Wang Q, Wan Q, Liu B L, Pan M Q, Chang L J, Wu H T, Mei G F, Xiang D, Li X H, Cai C P, Zhu X F, Chen Z J, Han B, Chen X Y, Guo W Z, Zhang T Z, Huang X H. Genomic insights into divergence and dual domestication of cultivated allotetraploid cottons. Genome Biol, 2017, 18:33-45.

[55] Song Q X, Zhang T Z, Stelly D M, Chen Z J. Epigenomic and functional analyses reveal roles of epialleles in the loss of photoperiod sensitivity during domestication of allotetraploid cottons. Genome Biol, 2017, 18:99-112.