利用oligo芯片技术鉴定橡胶树死皮相关基因  

Identification of Genes Associated with Tapping Panel Dryness in Hevea brasiliensis Using Oligonucleotide Microarrays

作　　者：覃碧[1] 刘向红[1,2] 邓治[1] 李德军[1] 

机构地区：[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571101 [2]海南大学农学院,海南海口570228

出　　处：《热带作物学报》2012年第2期296-301,共6页Chinese Journal of Tropical Crops

基　　金：国家自然科学基金“利用分子生物学方法研究橡胶树死皮发生分子机制”(No.30960310);海南省自然科学基金“橡胶树死皮相关基因芯片开发与应用”(No.310072);中国热带农业科学院橡胶研究所基本科研业务费专项“橡胶树死皮发生关键基因HbTCTP功能研究”(No.1630022011024)

摘　　要：橡胶树死皮(Tapping Panel Dryness,TPD)是天然橡胶单产提高的主要限制因子之一,给橡胶种植业带来严重危害。本研究利用定制橡胶树oligo芯片,通过提取死皮和健康橡胶树胶乳总RNA,标记并反转录成cDNA后与橡胶树oligo芯片进行杂交鉴定TPD相关基因,利用RT-PCR技术对芯片结果进行验证。对芯片数据分析结果显示,以2倍标准在死皮与健康橡胶树间筛选到26个差异表达基因,其功能涉及橡胶生物合成、细胞程序性死亡、细胞抗性及防御反应、活性氧代谢、DNA甲基化、泛素-蛋白酶体、信号传导、蛋白质合成、加工及转运等调控途径。进一步的RT-PCR验证结果表明,随机选取12个基因的表达模式均与芯片结果一致。本研究为大规模鉴定TPD相关基因和揭示TPD发生分子机制奠定了基础。Tapping panel dryness（TPD）is one of the main limiting factors for increasing the yield of natural rubber,and causes serious economic loss to the rubber production.In this research,the genes associated with TPD were identified by oligonucleotide microarrays hybridized with the latex from TPD and healthy rubber trees,and validated with RT-PCR method.The results from microarray analyses showed that 26 genes were differentially expressed between the TPD and healthy rubber trees.The 26 differentially expressed genes were classified into the following functional categories：rubber biosynthesis,programmed cell death,stress/defense response,metabolism of reactive oxygen species,DNA methylation,ubiquitin-proteasome pathway,cellular communication and signaling,and protein synthesis,process and transport,etc.In addition,the expression patterns of 12 differentially expressed genes randomly selected were further validated by RT-PCR analyses.This research laid foundations for identifying genes related to TPD on large scale and revealing the molecular mechanism of TPD in rubber tree.

关 键 词：橡胶树 死皮 oligo芯片 TPD相关基因 RT-PCR 

分 类 号：S794.1[农业科学—林木遗传育种]