摘要
利用Affymetrix小麦基因芯片研究了小麦成熟胚在MS+2,4-D(2mgL-1)培养基上脱分化过程种基因表达变化,用NCBI、DATF和DRTF等生物信息学相关网站对基因表达信息进行处理,并针对生长素相关基因的变化情况进行分析。结果表明,有80个生长素相关基因在小麦成熟胚脱分化过程中的2、6、12、24和72h等不同时间点,至少发生了一次有意义的表达变化,其中41个在整个过程中上调,29个下调;10个在不同时点分别表现上调或下调。这些基因涉及到生长素的运输、响应、诱导、合成和降解等多个生物学过程。对BG906698、BQ281752、CD454626、CD864552、CD938626、BJ233383和AY543630基因的半定量RT-PCR验证结果表明,它们的表达变化与基因芯片的结果基本一致。质膜H+-ATPase基因(AY543630)在2h时间点即有较高强度表达,然后下降,24h降至最低水平,表明该基因在小麦成熟胚脱分化的启动中可能有重要作用。
The quality of dedifferentiation determines the capacity of callus regeneration. Mature embryo is considered to be of the most potential for genetic transformation, but its transformation efficiency is still lower now. To elucidate the mechanism of mature wheat embryo dedifferentiation, we studied the expression profile of genes in dedifferentiation using Affymetrix microarray technique in mature embryos of wheat cultivar Yumai 18 cultured on MS medium supplemented with 2 mg L^-1 of 2,4-dichlorophenolxyacetic acid （2,4-D） at different time points of 2, 6, 12, 24, and 72 h. Using online tools at NCBI, DATF, and DRTF websites, 80 auxin-related genes changed at least at one time point were detected with the 41 up-regulated and 29 down-regulated during the whole period. The remained 10 genes showed up-regulation or down-regulation at different time points. Up-regulated genes were more than down-regulated genes, especially at 24 h and 72 h points. These genes were involved in seve- ral biological processes, such as transportation, response, induction, synthesis, and degeneration of auxin. Using semi-quantative RT-PCR technique, the expression changes of seven genes, coded as BG906698, BQ281752, CD454626, CD864552, CD938626, BJ233383, and AY543630, were confirmed to be consistent with the result by microarray. Gene of plasma membrane （PM） H^＋-ATPase （GenBank accession number： AY543630） expressed at a high level at the 2 h time point, and then decreased gradually till the minimum at the 24 h time point. This result implied that PM H^＋-ATPase gene might be a trigger in dedifferentiation of mature embryo in wheat.
出处
《作物学报》
CAS
CSCD
北大核心
2009年第10期1798-1805,共8页
Acta Agronomica Sinica
基金
国家转基因植物研究与产业化专项基金(JY03-B-19-2)
河南省杰出人才创新基金(022100090)资助
