cDNA-AFLP analysis reveals differential gene expression in incompatible interaction between infected non-heading Chinese cabbage and Hyaloperonospora parasitica.
cDNA-AFLP analysis reveals differential gene expression in incompatible interaction between infected non-heading Chinese cabbage and Hyaloperonospora parasitica.
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cDNA-AFLP分析揭示了受感染的不结球大白菜和寄生大膜孢菌之间不相容相互作用中的差异基因表达
DOI：
10.1038/hortres.2016.34
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发表时间：
2016
影响因子：
8.7
通讯作者：
Hu CM
中科院分区：
文献类型：
--
作者：
Xiao D;Liu ST;Wei YP;Zhou DY;Hou XL;Li Y;Hu CM
不结球白菜（Brassica rapa ssp.）chinensis）是世界上主要的绿色叶菜类蔬菜之一，在我国具有重要的经济价值。寄生透明霜霉是引起大白菜霜霉病的主要病原真菌，严重影响大白菜的生产。本研究的目的是鉴定不结球白菜与H.利用互补DNA扩增片段长度多态性（cDNA-AFLP）技术对寄生虫进行了研究。我们在抗病品系‘苏州青’中获得了129个可靠的差异转录衍生片段（TDFs）。其中，121个上调的TDFs在接种后24-48 h（h.p.i.）显示表达高峰。进一步选择15个基因进行cDNA-AFLP表达模式的验证，使用定量逆转录PCR。结果证实了cDNA-AFLP分析的13个基因（86.7%）的表达模式发生了改变。在15个TDFs中，我们确定了4个与真菌抗性相关的TDFs。此外，对抗性品系‘苏州青’和感病品系‘矮脚黄’的4个TDFs的比较分析表明，TDF 14（BcLIK1_A01）的转录水平在48 h.抗病系比感病系增加25.1倍。类似地，其他三种基因TDF 42（BcCAT3_A07）、TDF 75（BcAAE3_A06）和TDF 88（BcAMT2_A05）的转录水平在24、48和24 h. p. i达到峰值。分别增加了25.1倍、100倍和15.8倍。结果表明，抗病基因在抗病系中的转录水平高于感病系，这可能是抗病基因对病原菌侵染的抗性。本研究有助于阐明侵染过程的分子基础和鉴定感病品种抗病性改良的候选基因。
Non-heading Chinese cabbage (Brassica rapa ssp. chinensis) is one of the main green leafy vegetables in the world, especially in China, with significant economic value. Hyaloperonospora parasitica is a fungal pathogen responsible for causing downy mildew disease in Chinese cabbage, which greatly affects its production. The objective of this study was to identify transcriptionally regulated genes during incompatible interactions between non-heading Chinese cabbage and H. parasitica using complementary DNA-amplified fragment length polymorphism (cDNA-AFLP). We obtained 129 reliable differential transcript-derived fragments (TDFs) in a resistant line ‘Suzhou Qing’. Among them, 121 upregulated TDFs displayed an expression peak at 24–48 h post inoculation (h.p.i.). Fifteen genes were further selected for validation of cDNA-AFLP expression patterns using quantitative reverse transcription PCR. Results confirmed the altered expression patterns of 13 genes (86.7%) revealed by the cDNA-AFLP. We identified four TDFs related to fungal resistance among the 15 TDFs. Furthermore, comparative analysis of four TDFs between resistant line ‘Suzhou Qing’ and susceptible line ‘Aijiao Huang’ showed that transcript levels of TDF14 (BcLIK1_A01) peaked at 48 h.p.i. and 25.1-fold increased in the resistant line compared with the susceptible line. Similarly, transcript levels of the other three genes, TDF42 (BcCAT3_A07), TDF75 (BcAAE3_A06) and TDF88 (BcAMT2_A05) peaked at 24, 48 and 24 h.p.i. with 25.1-, 100- and 15.8-fold increases, respectively. The results suggested that the resistance genes tended to transcribe at higher levels in the resistance line than in the susceptible line, which may provide resistance against pathogen infections. The present study might facilitate elucidating the molecular basis of the infection process and identifying candidate genes for resistance improvement of susceptible cultivars.