摘要
[目的]本文旨在研究大白菜ADF7和ADF10基因对真菌侵染的响应。[方法]从拟南芥基因组数据库中找到全部的At ADF7和At ADF10基因,通过与大白菜基因组数据库序列比对分析,得到对应的基因序列。利用生物信息学分析其特性,通过半定量和定量分析2个基因在受到真菌侵染的大白菜中的表达情况。[结果]系统进化树分析表明,大白菜ADF7基因与其他近似植物的ADF7基因有较高的同源性。对大白菜ADF7和ADF10编码的蛋白进行二级结构分析,它们具有相近的相对分子质量和等电点,都没有信号肽。采用半定量检测大白菜受灰霉菌侵染前后ADF基因表达特点,结果显示:ADF7和ADF10在未受侵染的根、茎、叶中几乎不表达,在灰霉菌侵染的叶中表达量最高,茎次之,根中表达不明显。实时荧光定量PCR(qRTPCR)分析进一步验证了基因的组织特异性表达,病原菌侵染大白菜后,ADF7和ADF10在根、茎、叶中均呈现双波表达特点且表达趋势一致,在侵染12和48 h后均出现表达高峰值且在‘新烟杂3号’的表达量明显高于‘包头莲’。[结论]ADF7和ADF10在根、茎、叶中具有组织差异表达特性,并且在抗病品种中的表达较高。这为进一步研究生物胁迫情况下肌动蛋白的聚合和解聚机制奠定了理论基础。
[Objectives]The response of ADF7 and ADF10 genes to fungal infection was researched in Chinese cabbage. [Methods]At ADF7 and At ADF10 gene were found from Arabidopsis thaliana genome database,and the corresponding gene sequences were got from cabbage genome database by sequence alignment analyze. Using bioinformatics we analyzed their features. Semi-quantitative and quantitative analysis of the two genes expression was analyzed in Chinese cabbage after fungal infection. [Results]Sequence analysis and similarity comparison showed that ADF7 had a high homology between Chinese cabbage and other similar species. The secondary structure of ADF7 and ADF10 genes were analyzed in this study,they had similar molecular weight and isoelectric point,and no signal peptide existed. Semi-quantitative detection of the expression characteristics of ADF genes by Botrytis cinerea infection indicated that ADF7 and AFD10 genes had almost no expression in uninfected root,stem and leaf of Chinese cabbage. However,the two genes had the highest expression in leaf with Botrytis cinerea treatment,and the second was stem. They had an unconspicuous expression in root. The expression characteristics of ADF7 and AFD10 genes after Botrytis cinerea infection using real-time quantitative PCR（ qRT-PCR） showed a double waves characteristic expression and further validated the tissue specific expression. Both genes had the first expression peak at12 h after infection and then reached the highest expression levels at 48 h,showing a similar up-regulated trend expression in the entire treatment. They had similar expression in root,stem,and leaf with mock inoculation,however,they had the highest expression in leaf,therewith stem and had an unobvious expression in root. All results showed ADF genes had a certain relationship with the resistance to fungal infection and the expression level of‘Xinyanza 3＇was higher than that of‘Baotoulian＇. [Conclusions]The two genes had a different expression among the root,stem and leaf after Botrytis cinerea infection and they all had a higher expression on the resistant variety than the sensitive variety. It laid a theoretical foundation for further study of the mechanism on the process of regulating actin polymerization and depolymerization under the biological stress conditions.
出处
《南京农业大学学报》
CAS
CSCD
北大核心
2015年第5期742-747,共6页
Journal of Nanjing Agricultural University
基金
国家自然科学基金重点项目(31272173)
山东省自然科学基金项目(ZR2012cq019)
关键词
大白菜
肌动蛋白解聚因子
灰霉菌
实时定量PCR
Chinese cabbage
actin depolymerizing factor
Botrytis cinerea
real-time quantitative PCR（qRT-PCR）
