Gene differential expression of Phytophthora sojae during sporangium formation process
Gene differential expression of Phytophthora sojae during sporangium formation process
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DOI：
10.5897/ajmr12.729
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发表时间：
2012-10
影响因子：
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通讯作者：
Wen Jing-zhi;Li Yan;L. Yonggang;Yang Mingxiu;Shilova Na;L. Wei-ting
中科院分区：
文献类型：
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作者：
Wen Jing-zhi;Li Yan;L. Yonggang;Yang Mingxiu;Shilova Na;L. Wei-ting
采用差异显示反转录聚合酶链反应（DDRT-PCR）技术，研究了大豆疫霉菌（Phytophthora sojae）孢子囊形成早期（0 ~ 7 h）基因的差异表达。从78对引物组合中筛选出30对引物组合，得到90条稳定的差异带。通过反向北方杂交，共得到39条阳性差异片段，测序后得到36条差异片段序列。利用BLASTX软件对大豆疫霉基因组数据库进行同源性检索和分析。结果表明，23个片段与大豆疫霉基因组数据库中的蛋白质序列具有显著的相似性，其中15个片段与已知的参与代谢、细胞过程和信号转导过程的功能蛋白相匹配。对冗余度大于5的4个片段进行数据挖掘和生物信息学分析，结果表明，序列AR 15 -258在疫霉菌丝体中有很高的表达量，可能与菌丝感染或饥饿有关; AR 10 -263和GR 1 -304仅在疫霉中发现，可能是该属特有表达。对大豆疫霉孢子囊形成的重要调控基因的分析和研究，为进一步研究控制大豆疫霉生长发育的关键基因奠定了基础。关键词：大豆疫霉，孢子囊形成，mRNA差异显示，差异片段。
The gene differential expression of Phytophthora sojae during early stage (0 to 7 h) of sporangium formation was studied by differential display reverse transcription polymerase chain reaction (DDRT-PCR) in the present study. Thirty out of 78 primer combinations were selected and 90 stable differential bands were obtained. Thirty-nine positive differential fragments were found in the experiments by using the reverse northern blot, in which 36 sequences of differential fragments were obtained after sequencing. Homology search and analysis were tested by BLASTX in the P. sojae genome database. The results showed that 23 fragments had significant similarities with proteins in P. sojaegenome database, 15 of which matched with the known functional proteins involved in metabolism, cellular processes, and signal transduction processes. Data mining and bioinformatics analysis on 4 fragments which redundancy was greater than 5 showed that sequence AR15-258 had a very high expression level in Phytophthora mycelium, which may be related to mycelium infection or hunger; AR10-263 and GR1-304 were only found in Phytophthora, which might be unique expression in this genus. The analysis and research on important regulatory genes controlling sporangium formation of P. sojae was a foundation for studying of key genes controlling growth and development of P. sojae. Key words: Phytophthora sojae, sporangium formation, mRNA differential display, differential fragments.