作物学报 ›› 2020, Vol. 46 ›› Issue (4): 631-642.doi: 10.3724/SP.J.1006.2020.94135
• 研究简报 • 上一篇
WANG Heng-Bo,QI Shu-Ting,CHEN Shu-Qi,GUO Jin-Long,QUE You-Xiong()
摘要:
甘蔗是世界上最重要的糖料作物之一, 由于尚未完全破译栽培种基因组, 导致SSR标记匮乏, 难以覆盖全基因组, 限制了甘蔗遗传研究的进展。本研究以栽培种R570的4660个BAC文库片段序列(累计总长为382 Mb, 预测到25,316个编码蛋白基因)组装成的一套甘蔗单倍体基因组的模板, 利用MISA (Microsatellite identification tool)软件, 发掘SSR位点; 并综合分析其与4种禾本科植物(高粱、玉米、水稻和二岁短柄草)SSR位点的分布特征; 选取50对以TG和AG重复基序的SSR引物, 分别利用4个甘蔗属材料(R570、ROC1、LA purple和SES208)和24个重要甘蔗亲本, 对SSR引物进行扩增效率验证和多态性分析。共发掘到27,241个SSR位点, 平均每个BAC片段有6.29个SSR位点, 平均密度为71.33个SSR Mb -1, 远低于高粱的平均密度(350.00个SSR Mb -1)。在重复基序中, 占比前2位的分别为单核苷酸基序(11,079个)和三核苷酸重复基序(6447个), 合计占总SSR位点数的64.33%。与甘蔗不同的是, 4种禾本科植物中的三核苷酸基序类型数量最多、占比最大。此外, 在单核苷酸重复基序中, A/T所占比例最高, 为84.8%, C/G所占比例最低, 为15.2%; 在三核苷酸重复基序中, TGT/ACA所占比例最高, 为16.04%。总之, 禾本科植物基因组富含A/T的基序。在50对SSR引物(TG基序41对和AG基序9对)的多态性验证中, 共有45对(90%)能够扩增出清晰的条带, 其中35对(70%)在4个甘蔗材料上呈现多态性。进一步利用20对多态性较高的SSR引物对24个甘蔗重要亲本材料进行分析, 共扩增到95个等位基因, 平均每对引物扩增4.75个, 验证了这些引物应用于甘蔗遗传多样性研究的可行性。本研究鉴定的甘蔗栽培种单倍体基因组SSR标记, 有效增加了甘蔗遗传研究中可用的分子标记数量, 可直接用于甘蔗群体遗传多样性分析和重要性状遗传机制的解析, 为甘蔗分子育种的深入研究奠定了基础。
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