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作 者:栾爱萍 贺军虎[1] 刘佳柔 张伟[2] 谢桃[2] 何业华[2] LUAN Aiping;HE Junhu;LIU Jiarou;ZHANG Wei;XIE Tao;HE Yehua(Tropic Crops Genetic and Resources Institute,Chinese Academy of Tropic Agricultural Sciences,Haikou 571101,China;College of Horticulture,South China Agricultural University,Guangzhou 510642,China)
机构地区:[1]中国热带农业科学院热带作物品种资源研究所,海口571101 [2]华南农业大学园艺学院,广州510642
出 处:《园艺学报》2020年第4期734-740,共7页Acta Horticulturae Sinica
基 金:国家自然科学基金项目(31901355);海南省重点研发计划项目(ZDYF2019109)。
摘 要:利用AFLP分子标记技术,在DNA水平分析矮凤梨(Ananas nanus)品种‘Hime Pineapple’与其短营养期突变体‘14-1’之间的差异,鉴定突变体变异的真实性,为短营养期菠萝的分子育种提供参考依据。以‘14-1’和‘Hime Pineapple’的幼叶为试材,采用改良CTAB法提取叶片基因组DNA,经64对AFLP引物组合的选择性扩增,共获得1952条清晰谱带,平均每对引物产生30.5条谱带,其中有18对检测出DNA分子多态性,获得多态性条带76条,差异片段主要集中在200~500 bp,多态性条带比例为4.54%,遗传相似系数为95.39%。76条AFLP扩增差异条带经筛选、回收、克隆、测序和同源比对分析后显示,其中2个差异条带序列与菠萝MADS-box transcription factor 6-like基因同源,表明‘14-1’和‘Hime Pineapple’之间的性状差异与遗传物质的改变相关,相应的特异条带可作为调控营养期时长或是与调控营养期时长基因相连锁的候选基因片段。该结果对后续挖掘早花关键基因,解析短营养期突变体早花变异机理具有较好的参考价值。The genetic differences between‘Hime Pineapple’(Ananas nanus)and its short-vegetativegrowth-period mutant‘14-1’were analyzed by AFLP molecular marker technology.Identifying the authenticity of mutants could provide references for the molecular breeding of short-vegetativegrowth-period pineapples.The young leaves of mutant‘14-1’and‘Hime Pineapple’were used as experimental objects and their leaf genomic DNAs were extracted by improved CTAB method.Totally 1952 clear bands were obtained by 64 pairs of AFLP selective amplification primers(each pair of primers produced 30.5 bands on average),among them,DNA molecular polymorphisms were detected in 18 pairs and 76 polymorphism bands were obtained which accounted for 4.54%.The difference fragments mainly appeared between 200-500 bp and the genetic similarity coefficient was 95.39%.After selecting,recycling,cloning,sequencing and homologous analysis of 76 amplified difference bands of AFLP,two of them were homologous to the MADS-box transcription factor 6-like gene of pineapple,which indicated that the character differences between mutant‘14-1’and‘Hime Pineapple’were related to the changes of genetic material.Corresponding specific bands could be used as candidate gene segments that regulate vegetative duration or were linked with genes that regulate vegetative duration.The results could provide good reference value for exploration of the key genes of early flowering and variation mechanism of early flowering with short-vegetative-growth-period varieties.
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