Screening for differential expression of genes for resistance to Sitodiplosis mosellana in bread wheat via BSR-seq analysis

  • Zhiming Hao
  • Miaomiao Geng
  • Yanran Hao
  • Yue Zhang
  • Lijing Zhang
  • Shumin Wen
  • Ruihui WangEmail author
  • Guiru LiuEmail author
Original Article


Key message

Five putative candidate genes for OWBM resistance in Chinese winter wheat ‘Jimai 24’ were identified via BSR-seq and differential expression analyses.


Orange wheat blossom midge (OWBM), Sitodiplosis mosellana, is one of the most serious threats to wheat production worldwide. Conventional gene mapping methods to identify genes require significant amounts of financial support and time. Here, bulked segregant RNA-seq (BSR-seq) was applied to profile candidate genes and develop associated markers for OWBM resistance. Previously, we identified a major QTL (QSm.hebau-4A) for OWBM resistance on the long arm of chromosome 4A. In this study, we aimed at screening differentially expressed resistance genes associated with this QTL. Twelve differentially expressed genes (DEGs) were obtained based on BSR-seq and differential expression analyses. Among them, four were confirmed to be associated with OWBM resistance via quantitative reverse transcription PCR, using an additional set of wheat samples subjected to OWBM invasion. One SPI-like gene and one Malectin-like gene were revealed by gene annotation, respectively. Sequencing results confirmed that the four DEGs and the SPI gene had SNP polymorphisms between wheat parents. All these five resistance-related genes for OWBM were located in the same genomic region with QSm.hebau-4A. Furthermore, six new markers developed based on sequences of the five genes were also mapped in the same genomic region using genetic population. These five genes may be the candidate genes for OWBM resistance in Chinese wheat ‘Jimai 24’ and should be the targets for further positional isolation.


Bread wheat (Triticum aestivum L.) Orange wheat blossom midge (Sitodiplosis mosellana Géhin) Bulked segregant analysis RNA-seq Differentially expressed gene SNP (single nucleotide polymorphism) 



We are grateful to Dr. Minglin Lang (University of Chinese Academy of Sciences), Dr. Jianjun Zhao (Hebei Agricultural University) and Dr. Jingmei Mu (Northwest Agriculture & Forestry University) for their critical reading and editing of the manuscript; Dr. Guangyao Zhao (Institute of Crop Sciences, Chinese Academy of Agricultural Sciences) for his advices in methods of genomic analysis; Prof. Xinming Yang (Institute of Crop Sciences, Chinese Academy of Agricultural Sciences) for his supplying of wheat accessions; and Ms. Manyuan Ma and Mr. Liangfeng Zou (Total Genomics Solution (Shenzhen) Co., Ltd.) for their contributions to transcriptome data analysis.

Author’s contribution

ZH and RW designed the study. LZ, MG, SW, and GL provided advices to write the paper. SW, GL, and RW performed RIL population construction. YH, GL, and RW performed wheat invasion treatment and analyzed resistance index. YH, ZH, LZ, and YZ performed experimental material collection for RNA-seq and qRT-PCR. ZH, YH, and LZ performed resistant evaluation.


This work was supported by the National Key Research and Development Program of China (2016YFD0100102).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical standards

The authors note that this study was performed and reported in accordance with the ethical standards of scientific conduct.

Supplementary material

122_2019_3419_MOESM1_ESM.pdf (121 kb)
Supplementary file1 (PDF 120 kb)
122_2019_3419_MOESM2_ESM.pdf (734 kb)
Supplementary file2 (PDF 733 kb)
122_2019_3419_MOESM3_ESM.pdf (299 kb)
Supplementary file3 (PDF 298 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.North China Key Laboratory for Crop Germplasm Resources of Education Ministry, College of AgronomyHebei Agricultural UniversityBaodingPeople’s Republic of China

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