黄萎病菌侵染下陆地棉Dirigent-like蛋白基因表达差异分析

doi:10.3864/j.issn.0578-1752.2014.22.001

Abstract

Abstract: 【Objective】The objective of this study is to explore the Dirigent-like protein gene family and to figure out polymorphism of genes and expression differences between resistant and susceptible varieties of Upland cotton (Gossypium hirsutum) in responding to infection by Verticillium dahliae.【Method】Dirigent-like protein genes were acquired using Illumina Hiseq 2000 RNA-Seq transcriptional analysis. Multiple sequence alignment of encoding proteins was carried using online sequence analysis softwares and bioinformatics analysis tools for phylogenetic analysis. Quantitative real-time PCR with gene-specific primers was employed to identify the expression differences between resistant variety ‘Ji 79’ and susceptible variety ‘TM-1’, at 0, 1, 8, 24, 48, 72 h, and 96 h after inoculation. The quantitative reactions were performed on the ABI 7500 Real-Time PCR System with 7500 System Software (Applied Biosystems, USA) using the SYBR Green Master Mix. The comparative 2-ΔΔCt method of quantification was used with cotton GhACTIN as the reference.【Result】Totally 12 members of this gene family were found in Gossypium hirsutum, and phylogenetic analysis demonstrated 3 distinct subfamilies and 4 relatively conservative domains existed. The members of Dirigent-like protein genes were located on chromosomes 2, 3, 6, 7, 9, 10 and 13 in Gossypium raimondii, and some located in clusters on the same chromosome of D5 genome. Differential expression analysis indicated that dynamic response existed in different varieties. There were 6 members (GhDIR4, GhDIR6, GhDIR7, GhDIR9, GhDIR10 and GhDIR11) up-regulated (RQ≥2) in resistant variety Ji 79 after 1h inoculation, and RQ of GhDIR9 reached about 6.5; and 4 members (GhDIR1, GhDIR4, GhDIR7 and GhDIR11) up-regulated (RQ≥1) after 8h inoculation, only GhDIR4 up-regulated (RQ≥2) after 24h inoculation, GhDIR1, GhDIR4, GhDIR7 and GhDIR11 up-regulated (RQ≥2) after 48h inoculation; only GhDIR9 up-regulated (RQ=6.4) after 72h inoculation, up to 96 h inoculation, only GhDIR4 up-regulated (RQ≥2). GhDIR7 showed up-regulated (RQ≥2) in susceptible variety TM-1 after 1h inoculation. GhDIR7 and GhDIR10 up-regulated, and RQ of GhDIR7 increased to 325 after 8h inoculation. GhDIR4, GhDIR5, GhDIR6, GhDIR7, GhDIR9, GhDIR10 up-regulated significantly after 24h or 48h inoculation，and the RQ of GhDIR7, GhDIR9, GhDIR10 were more than 10. GhDIR4 and GhDIR7 still increased after 72h inoculation. Up to 96 h inoculation, GhDIR5, GhDIR6, GhDIR7, GhDIR9, GhDIR10 and GhDIR11 were highly expressed. So the results showed that the expression of GhDIR in response to infection by V. dahlia in resistant variety Ji 79 was earlier than that in susceptible variety TM-1. The expression of GhDIR members in susceptible variety became stronger after 8h vaccination by V. dahlia and continuously went up after 96h inoculation. Significant up-regulation of GhDIR members GhDIR4, as well as GhDIR7, GhDIR9 and GhDIR11, were observed in resistant variety, GhDIR7 and 3 other members (GhDIR5, GhDIR6 and GhDIR10) expressed in susceptible variety. And nucleotide polymorphism of 6 GhDIR genes (GhDIR1, GhDIR3, GhDIR5, GhDIR8, GhDIR10 and GhDIR12) was detected between resistant variety and susceptible variety. 【Conclusion】Dirigent-like protein plays an important role in Upland cotton in response to V. dahlia.

Key words: cotton, Dirigent-like protein gene, Verticillium dahliae, expression differences

GUO Bao-sheng, SHI Gong-yao, WANG Kai-hui, LIU Su-en, ZHAO Cun-peng, WANG Zhao-xiao, GENG Jun-yi, HUA Jin-ping. Expression Differences of Dirigent-Like Protein Genes in Upland Cotton Responsed to Infection by Verticillium dahliae[J].Scientia Agricultura Sinica, 2014, 47(22): 4349-4359.

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References

[1] 朱荷琴, 冯自力, 尹志新, 李志芳, 赵丽红, 师勇强, 杨家荣. 我国棉花黄萎病菌致病力分化及ISSR指纹分析. 植物病理学报, 2012, 42(3): 225-235.

Zhu H Q, Feng Z L, Yin Z X, Li Z F, Zhao L H, Shi Y Q, Yang J R. Pathogenicity differentiation and ISSR fingerprint analysis of cotton Verticillium dahliae in China. Acta Phytopathologica Sinica, 2012, 42(3): 225-235. (in Chinese)

[2] Cai Y F, He X H, Mo J C, Sun Q, Yang J P, Liu J G. Molecular research and genetic engineering of resistance to Verticillium wilt in cotton: A review. African Journal of Biotechnology, 2009, 8: 7363-7372.

[3] Xu L, Zhu L F, Tu L L, Guo X, Long L, Sun L, Gao W, Zhang X L. Differential gene expression in cotton defence response to Verticillium dahliae by SSH. Journal of Phytopathology, 2011, 159(9): 606-615.

[4] Zuo K, Wang J, Wu W, Chai Y, Sun X, Tang K. Identification and characterization of differentially expressed ESTs of Gossypium barbadense infected by Verticillium dahliae with suppression subtractive hybridization. Molecular Biology, 2005, 39(2): 191-199.

[5] Xu L, Zhu L F, Tu L L, Liu L L, Yuan D J, Jin L, Long L, Zhang X L. Lignin metabolism has a central role in the resistance of cotton to the wilt fungus Verticillium dahliaeas revealed by RNA-Seq-dependent transcriptional analysis and histochemistry. Journal of Experimental Botany, 2011, 62(15): 5607-5621.

[6] Burlat V, Kwon M, Davin L B, Lewis N G. Dirigent proteins and dirigent sites in lignifying tissues. Phytochemistry, 2001, 57: 883-897.

[7] Johansson C I, Saddler J N, Beatson R P. Characterization of the polyphenolics related to the colour of western red cedar (Thuja plicata Donn) heartwood. Holzforschung, 2000, 54: 246-254.

[8] Davin L B, Lewis N G. Dirigent proteins and dirigent sites explain the mystery of specificity of radical precursor coupling in lignan and lignin biosynthesis. Plant Physiology, 2000, 123: 453-461.

[9] Zhu L F, Zhang X L, Tu L L, Zeng F, Nie Y, Guo X P. Isolation and characterization of two novel dirigent-like genes highly expressed in cotton (G. barbadense) during inoculation of Verticillium daliae. Journal of Plant Pathology, 2007, 89(1): 25-29.

[10] 赵付安, 房卫平, 杨晓杰, 谢德意, 李武, 唐中杰. 陆地棉Dirigent-like基因(GhDIR)的克隆与分析. 华北农学报, 2011, 26(5): 29-33.

Zhao F A, Fang W P, Yang X J, Xie D Y, Li W, Tang Z J. Cloning and analysis of upland cotton (Gossypium hirsutum) dirigent-like gene (GhDIR). Acta Agriculturae Boreali-Sinica, 2011, 26(5): 29-33. (in Chinese).

[11] Shi H Y, Liu Z H, Zhu L, Zhang C J, Chen Y, Zhou Y, Li F G, Li X B. Overexpression of cotton (Gossypium hirsutum) dirigent1 gene enhances lignification that blocks the spread of Verticillium dahliae. Acta Biochimica et Biophysica Sinica, 2012, 44(7): 555-564.

[12] Li Q, Chen J F, Xiao Y, Di P, Zhang L, Chen W S. The dirigent multigene family in Isatis indigotica: Gene discovery and differential transcript abundance. BMC Genomics, 2014; 15(1): 388.

[13] Wall P K. Comparison of next generation sequencing technologies for transcriptome characterization. BMC Genomics, 2009, 10: 347.

[14] 郭宝生, 刘存敬, 刘素恩, 郭娴, 张建宏, 崔瑞敏, 王兆晓, 耿军义, 张香云. 棉花种间杂交渐渗系抗黄萎病性状遗传分析. 华北农学报, 2008, 23(S): 240-243.

Guo B S, Liu C J, Liu S E, Guo X, Zhang J F, Cui R M, Wang Z X, Geng J Y, Zhang X Y. Analysis on the inheritance of Verticillium wilt resistance of introgressed line from interspecific hybridization in cotton. Acta Agriculturae Boreali-Sinica, 2008, 23(S): 240-243. (in Chinese)

[15] 胡根海, 喻树迅. 利用改良的CTAB法提取棉花叶片总RNA. 棉花学报, 2007, 19(1): 69-70.

Hu G H, Yu S X. Extraction of high quality total RNA in cotton leaf with improved CTAB method. Cotton Science, 2007, 19(1): 69-70. (in Chinese)

[16] Ralph S, Park J Y, Bohlmann J, Mansfield S D. Dirigent proteins in conifer defense: Gene discovery, phylogeny, and differential wound-and insect-induced expression of a family of DIR and DIR-like genes in spruce (Picea spp.). Plant Molecular Biology, 2006, 60(1): 21-40.

[17] Ralph S G, Jancsik S, Bohlmann J. Dirigent proteins in conifer defense II: Extended gene discovery, phylogeny, and constitutive and stress-induced gene expression in spruce (Picea spp.). Phytochemistry, 2007, 68(14): 1975-1991.

[18] Fristensky B, Riggleman R C, Wagoner W, Hadwiger L A. Gene expression in susceptible and disease resistant interactions of peas induced with Fusarium solanipathogens and chitosan. Physiology Plant Pathology, 1985, 27: 15-28.

[19] Riggleman R C, Fristensky B, Hadwiger L A. The disease resistance response in pea is associated with increased levels of specific mRNAs. Plant Molecular Biology, 1985, 4: 81-86.

[20] Grlach J, Volrath S, Knauf-Beiter G, Hengy G, Beckhove U, Kogel K H, Oostendorp M, Staub T, Ward E, Kessmann H, Ryals J. Benzothiadiazole, a novel class of inducers of systemic acquired resistance, activates gene expression and disease resistance in wheat. The Plant Cell, 1996, 8: 629-643.

[21] 文婷, 李朋波, 姜淑琴, 蒲剑锋, 王玉美, 刘文欣, 华金平. 嫁接对棉花抗黄萎病的影响. 中国农业科学, 2011, 44(24): 5130-5136.

Wen T, Li P B, Jiang S Q, Pu J F, Wang Y M, Liu W X, Hua J P. Study on cotton resistance to Verticillium wilt by grafting. Scientia Agricultura Sinica, 2011, 44(24): 5130-5136. (in Chinese)

[22] Wang K B, Wang Z W, Li F G, Wang J, Song G, Yue Z, Cong L, Shang H, Zhu S, Zou C, Li Q, Yuan Y, Lu C, Wei H, Gou C, Zheng Z, Yin Y, Zhang X, Liu K, Wang B, Song C, Shi N, Kohel R J, Percy RG, Yu J Z, Zhu Y X, Wang J, Yu S. The draft genome of a diploid cotton Gossypium raimondii. Nature Genetics, 2012, 44: 1098-1103.

[23] 葛海燕, 汪业春, 郭旺珍, 张天真. 陆地棉抗黄萎病性状的遗传及分子标记研究. 棉花学报, 2008, 20(1): 19-22.

Ge H Y, Wang Y C, Guo W Z, Zhang T Z. Inheritance and molecular tagging of resistance against Verticillium wilt in upland cotton. Cotton Science, 2008, 20(1):19-22. (in Chinese)

[24] 蒋锋, 赵君, 周雷, 郭旺珍, 张天真. 陆地棉抗黄萎病基因的分子标记定位. 中国科学C辑: 生命科学, 2009, 39(9): 849-861.

Jiang F, Zhao J, Zhou L, Guo W Z, Zhang T Z. QTL mapping for resistance to Verticillium wilt in upland cotton. Science in China Series C: Life Sciences, 2009, 39(9): 849-861. (in Chinese)

[25] 杨昶, 郭旺珍, 张天真. 陆地棉抗黄萎病、纤维品质和产量等农艺性状的QTL定位. 分子植物育种, 2007, 5(6): 797-805.

Yang C, Guo W Z, Zhang T Z. QTL mapping for resistance to Verticillium wilt, fiber quality and yield traits in upland cotton (Gossypium hirsutum L.). Molecular Plant Breeding, 2007, 5(6): 797-805. (in Chinese)

[26] 张仁善. SSR分子标记辅助聚合陆地棉抗黄萎病QTL[D]. 南京: 南京农业大学, 2010.

Zhang R S. SSR marker-assisted pyramiding of Verticillum wilt resistant QTL in upland cotton[D]. Nanjing: Agricultural University, 2010. (in Chinese)

[27] 潘家驹. 棉花育种学. 北京: 中国农业出版社, 1998: 212-249.

Pan J J. Cotton Breeding. Beijing: Chinese Agricultural Press, 1998: 212-249. (in Chinese)

[28] Wang Z W, Wang J, Yue Z, Cong L, Zhu S, Li Q, Wei H, Gou C, Zheng Z, Yin Y, Zhang X, Liu K, Wang B, Song C, Shi N, Li F G, Fan G Y, Wang K B, Sun F M, Yuan Y L, Song G L, Qin L, Ma Z Y, Lu C, Zou C S, Chen W B, Liang X M, Shang H H, Liu W Q, Shi C C, Xiao G H, Gou C Y, Ye W W, Xu X, Zhang X Y, Wei H L, Li Z F, Zhang G Y, Wang J Y, Liu K, Kohel R J, Percy R G, Yu J Z, Zhu Y X, Wang J, Yu S X. Genome sequence of the cultivated cotton Gossypium arboretum. Nature Genetics, 2014, 46(6): 567-572.

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Expression Differences of Dirigent-Like Protein Genes in Upland Cotton Responsed to Infection by Verticillium dahliae

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