转录组分析不同砧木对嫁接烤烟抗青枯病的影响

doi:10.7668/hbnxb.20190870

摘要/Abstract

摘要： 为研究不同砧木对烟草嫁接苗抗病性的影响，明确抗病材料云烟87（Y87）作为砧木提高接穗红大金元（HD）对青枯病抗性的机理，采用劈接加生料带缠绕法以易感青枯病品种HD作为接穗，以HD和Y87作为砧木，分别构建HD/HD和HD/Y87嫁接烤烟苗，然后测量HD/HD和HD/Y87保护性酶苯丙氨酸解氨酶（PAL）活性的变化，最后利用高通量测序技术和Illumina HiSeqTM 4000测序平台，提取5株HD/HD或HD/Y87 mRNA，反转录构建总cDNA文库，测序，过滤低质量Reads后，用TopHat2比对参考基因组，用Cufflinks和DESeq软件获得基因表达量和差异表达基因。结果发现，不同取样时间HD/Y87 PAL活性均显著高于HD/HD，4个样品分别得到48 414 474~48 697 874条Clean Reads和38 272~40 938条基因，但HD/Y87发生选择性剪切事件数高于HD/HD；比较HD/Y87和HD/HD两组间基因表达量，得到3 904条差异基因，其中3 096条基因上调，808条基因下调；在上调基因中有参与木质素合成的3条编码苯丙氨酸解氨酶基因，5条Myb家族转录因子基因，5条编码多酚氧化酶基因；还有2条病程相关蛋白基因和3条相应的ERF转录因子基因等。这些结果表明，Y87作为砧木能够提高接穗HD PAL活性和多种抗性基因（包含木质素、多酚及病程相关蛋白）表达量，从而提高接穗红大金元对青枯病等病害抗性。

关键词: 烟草, 嫁接, 转录组, 苯丙氨酸解氨酶, 多酚氧化酶, 病程相关蛋白

Abstract: In order to study the effect of different rootstocks on the disease resistance of tobacco grafted seedlings, and to clarify the mechanism of Yunyan 87 (Y87) as rootstock to improve the resistance of scion Hongdajinyuan (HD) to bacterial wilt, this study used split grafting and raw material band winding method to construct HD/HD grafted and HD/Y87 flue-cured tobacco seedlings respectively, using HD as the scion and Y87 as the rootstock. The PAL activity of HD/HD and HD/Y87 protective enzymes was measured. Finally, mRNA of 5 HD/HD or HD/Y87 were extracted and reverse transcribed to construct a total cDNA library by high-throughput sequencing technology and Illumina HiSeqTM 4000 sequencing platform. After sequencing and filtering low-quality Reads, the reference genome was compared with TopHat2, and the genes expression amount and differentially expressed genes were obtained by Cufflinks and DESeq software Heterogene expression. The results showed that the PAL enzyme activity of HD/Y87 was significantly higher than that of HD/HD at different sampling times, 48 414 474-48 697 874 Clean Reads and 38 272-40 938 genes were obtained from four samples, respectively, and the number of selective shear events of HD/Y87 was higher than HD/HD. After comparing the gene expression between HD/Y87 and HD/HD, 3 904 differential genes were obtained, 3 096 genes were up-regulated, 808 genes were down regulated; among the up-regulated genes, there were 3 genes ecoding phenylalanine ammonia lyases, 5 genes ecoding Myb family transcription factors, and 5 genes ecoding polyphenol oxidase involved in lignin synthesis, it also had 2 disease-related proteins and 3 genes ecoding ERF transcription factors. These results showed that Y87 as rootstock could improve the activity of HD PAL and gene expression of various resistance genes (including PAL, PPO and disease related protein) in scion, so as to improve the resistance of HD to bacterial wilt and other diseases.

Key words: Tobacco, Grafting, Transcriptome, Phenylalanine ammonia lyase, Polyphenol oxidase, Disease-related proteins

中图分类号:

S604
Q78

谢冰, 詹小旭, 罗延宏, 刘斌祥, 孔凡磊, 袁继超. 转录组分析不同砧木对嫁接烤烟抗青枯病的影响[J]. 华北农学报, 2020, 35(4): 177-186. doi: 10.7668/hbnxb.20190870.

XIE Bing, ZHAN Xiaoxu, LUO Yanhong, LIU Binxiang, KONG Fanlei, YUAN Jichao. Transcriptome Analysis of the Effect of Different Rootstocks on the Resistance of Grafted Flue-cured Tobacco to Bacterial Wilt[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(4): 177-186. doi: 10.7668/hbnxb.20190870.

http://www.hbnxb.net/CN/Y2020/V35/I4/177

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