《植物生理学报》 2012, 48(9): 901-908

利用巴西橡胶树寡核苷酸芯片筛选死皮相关基因

李德军^1,*, 覃碧¹, 邓治¹, 刘向红^1,2

1中国热带农业科学院橡胶研究所, 农业部橡胶树生物学与遗传资源利用重点实验室, 海南儋州571737; 2海南大学农学院,

通信作者：李德军；E-mail: djli.rricatas@gmail.com；Tel: 0898-23301174

摘　要：

以健康和死皮巴西橡胶树品系热研7-33-97树皮为实验材料, 利用定制橡胶树寡核苷酸芯片筛选橡胶树死皮相关基 因。在橡胶树寡核苷酸芯片包含的566个基因中, 死皮与健康树树皮差异表达倍数在2倍或2倍以上的有56个, 占筛选转录 本总数的9.9%。在56个死皮相关基因中, 死皮树中上调表达基因有3个, 下调表达基因有53个。这些死皮相关基因共涉及8 个功能分类, “抗性及防御反应”所占比例最高, 接下来是“蛋白质合成、加工及转运”和“代谢和能量”, 以上三类功能基因占66.07%。此外, 死皮相关基因还涉及“细胞结构、生长及分化”、“细胞信号转导”、“转录相关”、“橡胶生物合成”和“未知 功能”。为验证芯片结果的可信性, 随机选取18个基因进行RT-PCR分析, 结果表明被检测基因表达模式均与芯片结果完全 一致。本研究鉴定并分析了死皮相关基因, 为进一步揭示橡胶树死皮发生机制奠定了基础。

关键词：巴西橡胶树; 树皮; 死皮分子机制; 死皮相关基因; 寡核苷酸芯片

收稿：2012-06-11   修定：2012-06-28

资助：海南省自然科学基金(310072)、国家自然科学基金(30960310)和中国热带农业科学院橡胶研究所基本科研业务费专项(1630022011024和RC201204)。

Identifying the Genes Related to Tapping Panel Dryness by Hevea brasiliensisOligonucleotide Microarrays

LI De-Jun^1,*, QIN Bi¹, DENG Zhi¹, LIU Xiang-Hong^1,2

¹Key Laboratory of Biology and Genetic Resources of Rubber Tree, Ministry of Agriculture, Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou, Hainan 571737, China; ²College of Agriculture, Hainan University, Haikou 570228, China

Corresponding author: LI De-Jun; E-mail: djli.rricatas@gmail.com; Tel: 0898-23301174

Abstract:

With the barks of healthy and tapping panel dryness (TPD) rubber tree clone (RY7-33-97) as experimental materials, the genes associated with TPD were identified by custom-designed oligonucleotide microarrays. Among 566 transcripts related to TPD of the oligonucleotide microarrays, 56 transcripts (about 9.9%) indicated 2-fold or higher expression differences between healthy and TPD rubber trees. Compared with healthy rubber tree, 3 and 53 genes were up- and down-regulated in TPD rubber tree, respectively. The TPD-related genes identified in the study were involved in 8 functional classifications. Of them, “stress/defense response” was the largest classification, followed by “protein synthesis, process and transport”, and “metabolism and energy”, and these three classifications mentioned above represented 66.07%. In addition, the TPD-related genes were associated with “cell structure, growth and development”, “cell signal transduction”, “transcription-related”, “rubber biosynthesis”, and “unknown function”. To validate the microarray results, 18 genes randomly selected were analyzed with RT-PCR. The results showed that the expression patterns of all the selected genes were consistent with ones from microarray analyses. The genes related to TPD were identified by oligonucleotide microarrays, which laid foundations for elucidating the molecular mechanism underlying TPD in rubber tree.

Key words: Hevea brasiliensis; bark; molecular mechanism involved in tapping panel dryness (TPD); TPD-related genes; oligonucleotide microarrays