Comparative Transcriptomic Analysis of Brassica napus Near-isogenic Line Resistant to Sclerotinia sclerotirium at Flowering Stage

doi:10.7668/hbnxb.20190761

Abstract

Abstract: A transcriptome comparisons of two genotypes of Brassica napus with different resistant ability against Sclerotinia sclerotirium, Xiangyou 15 with high resistance and its near-isogenic line 98C40NL with high susceptibility, were conducted by RNA-Seq in order to study the molecular mechanism of the interaction between B. napus and S. Sclerotirium and find out genes relevant with resistance and susceptibility respectively. The differentially expressed genes before inoculation with S.Sclerotirium,and 12,24 h after infection were analysed, followed by a qRT-PCR verification of gene expression. Totally, 34.16 G Clean bases and 78 582 genes were obtained through RNA-Seq. The results showed that 1 296 genes were down regulated while 1 495 genes were up regulated in Xiangyou 15 compared to 98C40NL. 54 genes expressed differentially in all of the three time points, 18 of which were down regulated and 27 genes were up regulated. The differentially expressed genes enriched in pathways related to oxidation-reduction, plant hormone signal transduction, calcium signal transduction,phenylpropane metabolism and synthesis of secondary metabolites. qRT-PCR results revealed that the genes with abundant expression in high resistant cultivar including BnaA03g09060D, BnaA01g26640D, BnaA03g09090D, BnaA04g12120D, and BnaA01g26920D could be related to resistance. However, genes BnaA06g10010D, BnaA09g16180D, BnaAnng19580D, BnaC01g40400D, and BnaC06g05340D might be connected with susceptibility, showing high expression in 98C40NL. The results can provide clue for the mechanism analysis of interactions between B. napus and S.sclerotirium and provide candidate genes for screening related resistance and receptor genes.

Key words: Brassica napus, Sclerotinia sclerotirium, Transcriptome, Differentially expressed genes, Function

CLC Number:

S432.4

ZHANG Qiuping, WEN Li, ZHANG Zhenqian, WANG Feng, GUAN Chunyun. Comparative Transcriptomic Analysis of Brassica napus Near-isogenic Line Resistant to Sclerotinia sclerotirium at Flowering Stage[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(3): 168-177. doi: 10.7668/hbnxb.20190761.

/ / Recommend / Download Citations

URL: http://www.hbnxb.net/EN/10.7668/hbnxb.20190761

http://www.hbnxb.net/EN/Y2020/V35/I3/168

References

[1] 王汉中. 以新需求为导向的油菜产业发展战略[J]. 中国油料作物学报, 2018, 40(5):613-617.doi:10.7505/j.issn.1007-9084.2018.05.001.
Wang H Z.New demand oriented oilseed rape industry developing strategy[J]. Chinese Journal of Oil Crop Sciences, 2018, 40(5):613-617.
[2] Food and agriculture organization of the United Nations Production, Crops. FAOSTAT[DB/OL]. 2017-9-26.http://faostat3.fao.org/browse/Q/QC/E.
[3] Liang X F, Rollins J A. Mechanisms of broad host range necrotrophic pathogenesis in Sclerotinia sclerotiorum[J]. Phytopathology, 2018, 108(10):1128-1140. doi:10.1094/PHYTO-06-18-0197-RVW.
[4] 官春云, 李方球, 陈社员. 双低油菜湘油15号对菌核病抗性研究简报[J]. 作物研究, 2001,15(3):33. doi:10.3969/j.issn.1001-5280.2001.03.012.
Guan C Y, Li F Q, Chen S Y. Brief report of characterization of stem rot resistance in the double-low rapeseed cultivar Xiangyou 15[J]. Crop Research,2001,15(3):33.
[5] 齐绍武,官春云,刘春林.甘蓝型油菜品系一些酶的活性与抗菌核病的关系[J].作物学报,2004,30(3):270-273. doi:10.3321/j.issn:0496-3490.2004.03.015.
Qi S W, Guan C Y, Liu C L. Relationship between some enzyme activity and resistance to Sclerotinia sclerotiorum of rapeseed cultivars[J]. Acta Agronomica Sinica,2004, 30(3):270-273.
[6] Wen L, Tan T L, Shu J B,Chen Y, Liu Y, Yang Z F, Zhang Q P, Yin M Z, Tao J, Guan C Y. Using proteomic analysis to find the proteins involved in resistance against Sclerotinia sclerotiorum in adult Brassica napus[J]. European Journal of Plant Pathology, 2013,137(3):505-523. doi:10.1007/s10658-013-0262-z.
[7] Wang Z R, Wan L L, Xin Q, Chen Y, Zhang X H, Dong F M, Hong D F, Yang G S. Overexpression of OsPGIP2 confers Sclerotinia sclerotiorum resistance in Brassica napus through increased activation of defense mechanisms[J]. Journal of Experimental Botany, 2018,69(12):3141-3155. doi:10.1093/jxb/ery138.
[8] 贾承国,向珣,王思周,汪俏梅.茉莉酸类化合物在植物防卫反应中的作用[J]. 细胞生物学杂志,2006,28(1):57-60. doi:10.3969/j.issn.1674-7666.2006.01.013.
Jia C G, Xiang X,Wang S Z,Wang Q M.The role of jasmonate in plant defense response[J]. Chinese Journal of Cell Bio1ogy, 2006,28(1):57-60.
[9] 刘庆霞,李梦莎,国静. 茉莉酸生物合成的调控及其信号通路[J]. 植物生理学报,2012,48(9):837-844.doi:10.13592/j.cnki.ppj.2012.09.013.
Liu Q X, Li M S,Guo J.Regulation of jasmonic acid biosynthesis and jasmonic acid signaling pathway[J]. Plant Physiology Journal, 2012, 48(9):837-844.
[10] Wu J, Zhao Q, Yang Q Y, Liu H, Li Q Y, Yi X Q, Cheng Y, Guo L, Fan C C, Zhou Y M. Comparative transcriptomic analysis uncovers the complex genetic network for resistance to Sclerotinia sclerotiorum in Brassica napus[J]. Sci Rep,2016(6):19007. doi:10.1038/srep19007.
[11] Wang Z, Tan X L, Zhang Z Y, Gu S L, Li G Y, Shi H F. Defense to Sclerotinia sclerotiorum in oilseed rape is associated with the sequential activations of salicylic acid signaling and jasmonic acid signaling[J]. Plant Sci, 2012, 184(9):75-82. doi:10.1016/j.plantsci.2011.12.013.
[12] Liu F, Li X X, Wang M R, Wen J, Yi B, Shen J X, Ma C Z, Fu T D, Tu J X.Interactions of WRKY15 and WRKY33 transcription factors and their roles in the resistance of oilseed rape to Sclerotinia infection[J]. Plant Biotechnol J, 2018, 16(4):911-925. doi:10.1111/pbi.12838.
[13] 郝向阳, 孙雪丽, 王天池, 吕科良,赖钟雄,程春振. 植物PAL基因及其编码蛋白的特征与功能研究进展[J]. 热带作物学报, 2018, 39(7):1452-1461. doi:10.3969/j.issn.1000-2561.2018.07.028.
Hao X Y,Sun X L, Wang T C, Lü K L, Lai Z X, Cheng C Z.Characteristics and functions of plant phenylalanine ammonia lyase genes and the encoded proteins[J]. Chinese Journal of Tropical Crops, 2018, 39(7):1452-1461.
[14] Erdemoglu N,Ozkan S,Tosun F. Alkaloid profile and antimicrobial activity of Lupinus angustifolius L. alkaloid extract[J]. Phytochemistry Reviews, 2007,6(1):197-201. doi:10.1007/s11101-006-9055-8.
[15] Erdemoglu N,Ozkan S,Duran A,Tosun F. GC-MS analysis and antimicrobial activity of alkaloid extract from Genista vuralii[J]. Pharmaceutical Biology, 2009, 47(1):81-85. doi:10.1080/13880200802448674.
[16] Zarinpanjeh N, Motallebi M, Zamani M R, Ziaei M.Enhanced resistance to Sclerotinia sclerotiorum in Brassica napus by co-expression of defensin and chimeric chitinase genes[J]. Journal of Applied Genetics, 2016, 57(4):417-425. doi:10.1007/s13353-016-0340-y.
[17] Wippich C,Wink M. Biological properties of alkaloids. Influence of quinolizidine alkaloids and gramine on the germination and development of powdery mildew, Erysiphe graminis f. sp. Hordei[J]. Experientia,1985,41(11):1477-1479. doi:10.1007/bf01950046.
[18] Zhao W M, Wolfender J L,Hostettmann K,Xu R S, Qin G W. Antifungal alkaloids and limonoid derivatives from Dictamnus dasycarpus[J]. Phytochemistry,1998,47(1):7-11. doi:10.1016/s0031-9422(97)00541-4.
[19] Cheng S H, Willmann M R, Chen H C, Sheen J. Calcium signaling through protein kinases. The Arabidopsis calcium-dependent protein kinase gene family[J]. Plant Physiolog, 2002, 129(2):469-485. doi:10.1104/pp.005645.
[20] Harmon A C, Gribskov M, Harper J f, Harmon A C, Gribskov M, Harper J F. CDPKs a kinase for every Ca ²⁺ signal?[J]. Trends in Plant Science, 2000,5(4):154-159. doi:10.1016/S1360-1385(00)01577-6.
[21] Yamniuk A P, Vogel H J. Structural investigation into the differential target enzyme regulation displayed by plant calmodulin isoforms[J]. Biochemistry, 2005, 44(8):3101-3111. doi:10.1021/bi047770y.
[22] Lee S H, Johnson J D, Walsh M P, Van Lierop J E, Sutherland C, Xu A,Snedden W A, Kosk-Kosicka D, Fromm H, Narayanan N, Cho M J. Differential regulation of Ca²⁺/calmodulin-dependent enzymes by plant calmodulin isoforms and free Ca²⁺ concentration[J]. Biochem Journal, 2000, 350(1):299-306. doi:10.1042/0264-6021:3500299.
[23] Batistic O, Kudla J. Integration and channeling of calcium signaling through the CBL calcium sensor/CIPK protein kinase network[J]. Planta, 2004, 219(6):915-924.doi:10.1007/s00425-004-1333-3.
[24] Batisti O, Kudla J. Analysis of calcium signaling pathways in plants[J]. Biochimica et Biophysica Acta (BBA)-General Subjects, 2012, 1820(8):1283-1293. doi:10.1016/j.bbagen.2011.10.012.
[25] 吕琳慧, 徐幼平, 任至玄,康冬,王继鹏,蔡新忠. Ca²⁺信号通路对本氏烟叶位介导的核盘菌抗性的影响[J]. 浙江大学学报(农业与生命科学版), 2014, 40(6):605-610. doi:10.3785/j.issn.1008-9209.2014.03.131.
Lü L H, Xu Y P, Ren Z X, Kang D, Wang J P, Cai X Z. Effect of Ca²⁺ signaling pathway on leaf position-associated resistance to Sclerotinia sclerotiorumin in Nicotiana benthamiana[J]. Journal of Zhejiang University(Agriculture and Life Sciences),2014, 40(6):605-610.
[26] 张秋平, 文李, 王峰, 廖志强,李慧,刘睿洋,官春云.油菜钙依赖蛋白激酶BnCDPK1 的克隆和表达分析[J]. 植物遗传资源学报, 2014(6):1320-1326. doi:10.13430/j.cnki.jpgr.2014.06.021.
Zhang Q P, Wen L, Wang F, Liao Z Q, Li H, Liu R Y, Guan C Y. Molecular cloning and expression analysis of calcium-dependent protein kinase BnCDPK1 in Brassica napus[J]. Journal of Plant Genetic Resources, 2014(6):1320-1326.
[27] Ding Y J, Mei J Q, Chai Y R, Yu Y, Shao C G, Wu Q N, Disi J O, Li Y H, Wan H F, Qian W.Simultaneous transcriptome analysis of host and pathogen highlights the interaction between Brassica oleracea and Sclerotinia sclerotiorum[J]. Phytopathology, 2019,109(4):542-550. doi:10.1094/PHYTO-06-18-0204-R.

Please choose a citation manager

Content to export