中国蔬菜
中國蔬菜
중국소채
China Vegetables
2015年
11期
19-27
,共9页
蒲全明%高启国%张林成%施松梅%刘晓欢%张莹%毕云龙%任雪松%刘豫东%朱利泉
蒲全明%高啟國%張林成%施鬆梅%劉曉歡%張瑩%畢雲龍%任雪鬆%劉豫東%硃利泉
포전명%고계국%장림성%시송매%류효환%장형%필운룡%임설송%류예동%주리천
结球甘蓝%叶片卷曲%生长素相关基因%基因克隆%表达分析
結毬甘藍%葉片捲麯%生長素相關基因%基因剋隆%錶達分析
결구감람%협편권곡%생장소상관기인%기인극륭%표체분석
Brassica oleracea L. var.capitata L.%Leaf curl%Auxin-related genes%Gene cloning%Expression analysis
生长素在结球甘蓝球叶向上、向内自然卷曲的过程中发挥着重要的调控作用。为挖掘重要的影响结球甘蓝叶片卷曲的生长素相关基因，通过对结球甘蓝莲座期叶片与结球期叶片的转录组进行对比分析，筛选出6个显著差异表达的生长素相关基因，分别为调控生长素动态平衡的BoILL6、BoASA1基因，控制极性运输的BoSF21、BoPIN4基因，参与信号转导的BoARF8、BoGH3.5基因。对上述6个基因进行同源克隆，分别获得全长cDNA序列，序列分析发现6个基因均含有生长素相关的结构域。分子进化树分析发现各基因与芜菁的对应基因遗传关系最近，拟南芥次之。荧光定量PCR检测结果显示， BoILL6、BoASA1与BoSF21的表达量变化最为显著，结球期叶片相对莲座期叶片的表达量差异均高达12倍以上，变化趋势与转录组数据分析结果基本一致，说明这3个基因可能与结球甘蓝叶片的卷曲发育密切相关。
生長素在結毬甘藍毬葉嚮上、嚮內自然捲麯的過程中髮揮著重要的調控作用。為挖掘重要的影響結毬甘藍葉片捲麯的生長素相關基因，通過對結毬甘藍蓮座期葉片與結毬期葉片的轉錄組進行對比分析，篩選齣6箇顯著差異錶達的生長素相關基因，分彆為調控生長素動態平衡的BoILL6、BoASA1基因，控製極性運輸的BoSF21、BoPIN4基因，參與信號轉導的BoARF8、BoGH3.5基因。對上述6箇基因進行同源剋隆，分彆穫得全長cDNA序列，序列分析髮現6箇基因均含有生長素相關的結構域。分子進化樹分析髮現各基因與蕪菁的對應基因遺傳關繫最近，擬南芥次之。熒光定量PCR檢測結果顯示， BoILL6、BoASA1與BoSF21的錶達量變化最為顯著，結毬期葉片相對蓮座期葉片的錶達量差異均高達12倍以上，變化趨勢與轉錄組數據分析結果基本一緻，說明這3箇基因可能與結毬甘藍葉片的捲麯髮育密切相關。
생장소재결구감람구협향상、향내자연권곡적과정중발휘착중요적조공작용。위알굴중요적영향결구감람협편권곡적생장소상관기인，통과대결구감람련좌기협편여결구기협편적전록조진행대비분석，사선출6개현저차이표체적생장소상관기인，분별위조공생장소동태평형적BoILL6、BoASA1기인，공제겁성운수적BoSF21、BoPIN4기인，삼여신호전도적BoARF8、BoGH3.5기인。대상술6개기인진행동원극륭，분별획득전장cDNA서렬，서렬분석발현6개기인균함유생장소상관적결구역。분자진화수분석발현각기인여무정적대응기인유전관계최근，의남개차지。형광정량PCR검측결과현시， BoILL6、BoASA1여BoSF21적표체량변화최위현저，결구기협편상대련좌기협편적표체량차이균고체12배이상，변화추세여전록조수거분석결과기본일치，설명저3개기인가능여결구감람협편적권곡발육밀절상관。
Auxin plays an important role in regulating the process ofBrassica oleracea L. var.capitata L. leaf curl.In order to find out the auxin relevant genes involved in cabbage leaf curling,we analyzed transcriptome of cabbage leaf at rosette and heading stages and identified 6 significantly differentially expressed auxin-related genes,including regulation of homeostasis genes(BoILL6,BoASA1),controlling polar transport genes(BoSF21, BoPIN4)and involving in signal transduction genes(BoARF8,BoGH3.5).The full-length cDNA sequences of 6 genes were obtained,respectively using homologous clone method.The protein structure prediction revealed that 6 genes contained auxin-related domains.Phylogenetic analysis though MEGA 5.1 comparison displayed that these genes had the closest genetic relationship with correspondence genes ofBrassica rapa,andArabidopsis thaliana followed. The real-time Fluorescence quantitative PCR analysis suggested that the expression level of BoILL6,BoASA1andBoSF21 showed significantly different trend.The expression level of these 3 genes in leaf during heading stage were 12 folds more than that in the leaf at rosette stage.The trend was consistent with the results of transcriptome analysis.The up-regulated expression of these 3 genes in the leaf during heading stage maybe lead to enhance the auxin concentration and change the auxin polar contribution in the leaves,and finally lead to format of the leaf curl.
