Identification of Ramie Genes in Response to Pratylenchus coffeae Infection Challenge by Digital Gene Expression Analysis.
Identification of Ramie Genes in Response to Pratylenchus coffeae Infection Challenge by Digital Gene Expression Analysis.
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DOI：
10.3390/ijms160921989
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发表时间：
2015-09-11
影响因子：
5.6
通讯作者：
Zhu A
中科院分区：
文献类型：
--
作者：
Yu Y;Zeng L;Yan Z;Liu T;Sun K;Zhu T;Zhu A
由咖啡短体线虫（Pratylenchuscoffeae）引起的根腐病严重危害苎麻的生长和产量。苎麻对咖啡疫霉感染的防御机制知之甚少，这阻碍了通过育种计划提高抗性的努力。本研究利用Illumina测序技术对苎麻抗病品种青大叶的转录组进行了分析。产生了约4630万个干净的对端（PE）读段，并组装成40，826个平均长度为830 bp的单基因。对对照根（CK）和受咖啡疫霉感染的根（CH）进行数字基因表达（DGE）分析，并鉴定差异表达基因（DEG）。分别对CK和CH cDNA文库中的约1016和807万个cDNA读段进行了测序。两个文库中共有137个基因表现出不同的转录本丰度。其中，117和20个DEG在咖啡疫霉感染的苎麻中分别表达上调和下调。通过qRT-PCR确定的15个候选基因的表达模式证实了DGE分析的结果。感病和抗病苎麻品种接种咖啡疫霉后，8个防御相关基因的时程表达谱存在差异。在咖啡疫霉侵染过程中，蛋白酶抑制剂、病程相关蛋白（PRs）和转录因子在抗感苎麻中的差异表达表明，半胱氨酸蛋白酶抑制剂可能在线虫抗性中起重要作用。
Root lesion disease, caused by Pratylenchus coffeae, seriously impairs the growth and yield of ramie, an important natural fiber crop. The ramie defense mechanism against P. coffeae infection is poorly understood, which hinders efforts to improve resistance via breeding programs. In this study, the transcriptome of the resistant ramie cultivar Qingdaye was characterized using Illumina sequence technology. About 46.3 million clean pair end (PE) reads were generated and assembled into 40,826 unigenes with a mean length of 830 bp. Digital gene expression (DGE) analysis was performed on both the control roots (CK) and P. coffeae-challenged roots (CH), and the differentially expressed genes (DEGs) were identified. Approximately 10.16 and 8.07 million cDNA reads in the CK and CH cDNA libraries were sequenced, respectively. A total of 137 genes exhibited different transcript abundances between the two libraries. Among them, the expressions of 117 and 20 DEGs were up- and down-regulated in P. coffeae-challenged ramie, respectively. The expression patterns of 15 candidate genes determined by qRT-PCR confirmed the results of DGE analysis. Time-course expression profiles of eight defense-related genes in susceptible and resistant ramie cultivars were different after P. coffeae inoculation. The differential expression of protease inhibitors, pathogenesis-related proteins (PRs), and transcription factors in resistant and susceptible ramie during P. coffeae infection indicated that cystatin likely plays an important role in nematode resistance.