Comparative study on the mRNA expression of Pinus massoniana infected by Bursaphelenchus xylophilus

  • Wanfeng Xie
  • Guanghong Liang
  • Aizhen Huang
  • Feiping ZhangEmail author
  • Wenshuo GuoEmail author
Original Paper


Pine wilt disease (PWD) is a devastating disease affecting the growth of Pinus massoniana, often leading to withering and death. To reveal the changes involved during disease progression, we investigated the mRNA expression profile of P. massoniana infested by Bursaphelenchus xylophilus. The infestation resulted in the downregulation of genes involved in interactions with pathogenic pathways such as disease resistance gene, CC-NBS-LRR resistance-like protein, and the gene encoding a putative nematode resistance protein. Increased infestation pressure (number of nematodes inoculated) caused a continuous decline in the gene expression of stem samples. An infestation of P. massoniana also resulted in a pathway enrichment of genes involved in phenylpropanoid metabolism and flavonoid biosynthesis, which in turn reduced the levels of total phenols and total flavonoids. A downregulation of auxin responsive family protein was observed in infested samples, which resulted in a suppression of plant growth. Thus, upon B. xylophilus infestation, a downregulation of genes associated with the recognition of pathogens, PWD resistance, and growth regulation was observed in P. massoniana, together with a decrease in the levels of phytoalexin-like secondary substances, all of which resulted in withering and ultimately death of P. massoniana.


Auxin/IAA Bursaphelenchus xylophilus Pinus massoniana Resistance gene Phytoalexin 

Supplementary material

11676_2018_824_MOESM1_ESM.docx (2.4 mb)
Supplementary material 1 (DOCX 2501 kb)


  1. Cai D, Kleine M, Kifle S, Harloff HJ, Sandal NN, Marcker KA, Klein-Lankhorst RM, Salentijn EMJ, Lange W, Stiekema WJ, Wyss U, Grundler FMW, Jung C (1997) Positional cloning of a gene for nematode resistance in sugar beet. Science 275:832–834CrossRefGoogle Scholar
  2. Dangl JL, Jones JD (2001) Plant pathogens and integrated defence responses to infection. Nature 411:826–833CrossRefGoogle Scholar
  3. Ellis J, Dodds P, Pryor T (2000) Structure, function and evolution of plant disease resistance genes. Curr Opin Plant Biol 3:278–284CrossRefGoogle Scholar
  4. Fukuda K (1997) Physiological process of the symptom development and resistance mechanism in pine wilt disease. J For Res 2:171–181CrossRefGoogle Scholar
  5. Futai K (2013) Pine wood nematode, Bursaphelenchus xylophilus. Annu Rev Phytopathol 51:61–83CrossRefGoogle Scholar
  6. Gao L, Tu ZJ, Millett BP, Bradeen JM (2013) Insights into organ-specific pathogen defense responses in plants: RNA-seq analysis of potato tuber-Phytophthora infestans interactions. BMC Genom 14:340CrossRefGoogle Scholar
  7. Grabherr MG, Haas BJ, Yassour M, Levin JZ, Thompson DA, Amit I, Adiconis X, Fan L, Raychowdhury R, Zeng Q (2011) Full-length transcriptome assembly from RNA-Seq data without a reference genome. Nat Biotechnol 29:644–652CrossRefGoogle Scholar
  8. Hanin M, Brini F, Ebel C, Toda Y, Takeda S, Masmoudi K (2011) Plant dehydrins and stress tolerance: versatile proteins for complex mechanisms. Plant Signal Behav 6:1503–1509CrossRefGoogle Scholar
  9. Hirao T, Fukatsu E, Watanabe A (2012) Characterization of resistance to pine wood nematode infection in Pinus thunbergii using suppression subtractive hybridization. BMC Plant Biol 12:13CrossRefGoogle Scholar
  10. Jones JD, Dangl JL (2006) The plant immune system. Nature 444:323–329CrossRefGoogle Scholar
  11. Kanehisa M, Araki M, Goto S, Hattori M, Hirakawa M, Itoh M, Katayama T, Kawashima S, Okuda S, Tokimatsu T (2008) KEGG for linking genomes to life and the environment. Nucleic Acids Res 36:480–484CrossRefGoogle Scholar
  12. Kortekamp A (2006) Expression analysis of defence-related genes in grapevine leaves after inoculation with a host and a non-host pathogen. Plant Physiol Biochem 44:58–67CrossRefGoogle Scholar
  13. Li G, Asiegbu FO (2004) Induction of Pinus sylvestris PsACRE, a homology of Avr9/Cf-9 rapidly elicited defense-related gene following infection with root rot fungus Heterobasidion annosum. Plant Sci 167:535–540CrossRefGoogle Scholar
  14. Liu Y, Wang L, Xing X, Sun L, Pan J, Kong X, Zhang M, Li D (2013) ZmLEA3, a multifunctional group 3 LEA protein from maize (Zea mays L.), is involved in biotic and abiotic stresses. Plant Cell Physiol 54:944–959CrossRefGoogle Scholar
  15. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408CrossRefGoogle Scholar
  16. McLean MD, Hoover GJ, Bancroft B, Makhmoudova A, Clark SM, Welacky T, Simmonds DH, Shelp BJ (2007) Identification of the full-length Hs1 pro1 coding sequence and preliminary evaluation of soybean cyst nematode resistance in soybean transformed with Hs1 pro1 cDNA. Botany 85:437–441Google Scholar
  17. Mortazavi A, Williams BA, McCue K, Schaeffer L, Wold B (2008) Mapping and quantifying mammalian transcriptomes by RNA-Seq. Nat Med 5:621–628Google Scholar
  18. Park S, Han K-H (2003) An auxin-repressed gene (RpARP) from black locust (Robinia pseudoacacia) is posttranscriptionally regulated and negatively associated with shoot elongation. Tree Physiol 23:815–823CrossRefGoogle Scholar
  19. Petit P, Granier T, d’Estaintot BL, Manigand C, Bathany K, Schmitter JM, Lauvergeat V, Hamdi S, Gallois B (2007) Crystal structure of grape dihydroflavonol 4-reductase, a key enzyme in flavonoid biosynthesis. J Mol Biol 368:1345–1357CrossRefGoogle Scholar
  20. Tan S, Wu S (2012) Genome wide analysis of nucleotide-binding site disease resistance genes in Brachypodium distachyon. Comp Funct Genom 3:418208Google Scholar
  21. Tronchet M, Balague C, Kroj T, Jouanin L, Roby D (2010) Cinnamyl alcohol dehydrogenases-C and D, key enzymes in lignin biosynthesis, play an essential role in disease resistance in Arabidopsis. Mol Plant Pathol 11:83–92CrossRefGoogle Scholar
  22. Westermann AJ, Gorski SA, Vogel J (2012) Dual RNA-seq of pathogen and host. Nat Rev Microbiol 10:618–630CrossRefGoogle Scholar
  23. Xie W, Huang A, Li H, Feng L, Zhang F, Guo W (2017) Identification and comparative analysis of microRNAs in Pinus massoniana infected by Bursaphelenchus xylophilus. Plant Growth Regul 83:223–232CrossRefGoogle Scholar
  24. Xu L, Liu ZY, Zhang K, Lu Q, Liang J, Zhang XY (2013) Characterization of the Pinus massoniana transcriptional response to Bursaphelenchus xylophilus infection using suppression subtractive hybridization. Int J Mol Sci 14:11356–11375CrossRefGoogle Scholar
  25. Zhao BG (2008) Pine wilt disease in China. In: Zhao BG, Futai K, Sutherland JR, Takeuchi Y (eds) Pine wilt disease. Springer, Tokyo, pp 18–25CrossRefGoogle Scholar
  26. Zheng HY, Xu M, Xu FY, Ye JR (2015) A comparative proteomics analysis of Pinus massoniana inoculated with Bursaphelenchus xylophilus. Pak J Bot 47:1271–1280Google Scholar

Copyright information

© Northeast Forestry University and Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Jinshan CollegeFujian Agriculture and Forestry UniversityFuzhouChina
  2. 2.Institute of Forestry Protection, Forestry CollegeFujian Agriculture and Forestry UniversityFuzhouChina

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