利用cDNA-AFLP技术分析‘窄冠黑白杨’杂种优势的差异表达基因  

cDNA-AFLP analysis of the differentially expressed genes of heterosis in Populus ’Zhaiguanheibaiyang’

作　　者：侯丽丽 姚培娟[1] 李际红[1] 李景涛 李柱[3] HOU Lili;YAO Peijuan;LI Jihong;LI Jingtao;LI Zhu(College of Forestry,Shandong Agricultural University/Mountain Tai Forest Ecosystem Research Station of State Forestry and Grassland Administration,State Forestry and Grassland Administration Key Laboratory of Silviculture in Downstream Areas of the Yellow River,Taian,Shangdong 271018,China;Shandong Forest Seedling Station,Jinan 250014,China;Shandong Agricultural University Scientific Research Department,Taian,Shangdong 271018,China)

机构地区：[1]山东农业大学林学院/泰山森林生态系统国家定位观测研究站/黄河下游森林培育国家林业和草原局重点实验室,山东泰安271018 [2]山东省林木种苗和花卉站,济南250014 [3]山东农业大学科研处,山东泰安271018

出　　处：《植物生理学报》2020年第12期2695-2704,共10页Plant Physiology Journal

基　　金：转基因生物新品种培育重大专项(2018ZX08020002);山东省自然基金项目(ZR2013CM020);山东省农业良种项目(201496)。

摘　　要：本研究以‘窄冠黑白杨’(Populus ‘Zhaiguanheibaiyang’)及其亲本为材料,通过cDNA扩增片段长度多态性分析技术(cDNA-AFLP)构建扩增图谱。从96对扩增引物中筛选出35对可用于条带分析的扩增引物,获得761个修饰位点, 5种基因差异表达模式。回收cDNA-AFLP图谱中分离的19条差异表达的转录衍生片段(transcript-derived fragments, TDFs)进行同源性比对分析,发现其中14条TDFs与杨树基因库中的已知基因同源,如丝氨酸/苏氨酸蛋白激酶(STPK)、泛素激活酶(E1)、糖基转移酶(UDP)、ARGOS-like蛋白基因等,在植物生长代谢活动、信号转导调控等途径中具有重要作用。利用实时荧光定量PCR技术检测差异表达TDFs在杂种及父本中芽、叶、茎等不同组织的表达量,发现在叶中表达量有明显差异,这可能与杂种优势密切相关。结果与cDNA-AFLP图谱一致,说明研究结果稳定可靠,为后续杨树杂种优势的研究提供新的见解。In this study, the Populus ’Zhaiguanheibaiyang’ and its parents were used as experimental materials, and the amplification map was constructed by cDNA amplified fragment length polymorphism(cDNA-AFLP) in this study. Thirty-five pairs of primers were selected from 96 pairs of primers that can be used for strip analysis, 761 modified loci and 5 patterns of gene differential expression were obtained. Nineteen differentially expressed transcriptional derived fragments(TDFs) isolated from cDNA-AFLP map were recovered for homology analysis, 14 TDFs were compared to known genes in the poplar gene pool, such as serine/threonine protein kinase(STPK), ubiquitin activating enzyme(E1), glycosyl transferase(UDP), ARGOS-like protein gene and other genes, which play an important role in plant metabolism, signal transduction, regulate cell proliferation, differentiation, organ size, etc. The fluorescent quantitative PCR was used to detect the expression levels of differentially expressed TDFs in buds, leaves and stems, in the male parent and hybrids, and it was found that there were significant differences in the expression levels in leaves, which may be closely related to heterosis. The results were consistent with the cDNA-AFLP map, indicating that the results were stable and reliable, and providing new insights for the subsequent studies on poplar heterosis.

关 键 词：‘窄冠黑白杨’ 杂种优势 CDNA-AFLP 差异基因 

分 类 号：S792.11[农业科学—林木遗传育种]