参薯miR156和靶基因SPL特征分析

Analysis of Characteristics of miR156 and Target Gene SPL in Dioscorea alata

参薯(Dioscorea alata)在热带和亚热带广泛种植,为该地区的人提供了粮食,具有重要的经济价值。miR156及其靶基因SPL对植物的生长发育的调控起到重要的作用,为了分析参薯中miR156-SPL的调控模块,本研究对参薯miR156基因位点和对应靶基因进行预测和验证。基于参薯基因组以及转录组信息,一共得到DamiR156的8个基因位点,预测了miR156的6个靶基因,其中4个属于SPL基因家族。多序列比对分析发现8个Da-miR156的前体序列在发卡区高度保守,根据序列同源性可以分为3组。表达谱数据和RT-qPCR表明DamiR156在参薯组培再生植株的叶、茎、块茎各个组织表达量远高于田间参薯,且在块茎中表达量也高于其他组织,初步推断Da-miR156在薯块发育的过程中发挥作用。5'-RLM-RACE实验验证了DaSPL被miR156切割的位点,为开展参薯SPL基因功能分析提供了参考依据。本研究通过对参薯miR156和靶基因SPL特征研究为分析miR156-SPL调控模块在参薯薯块发育中的作用打下科学基础。

Water yam(Dioscorea alata)is widely grown in the tropics and subtropics and provides staple food for these areas,having great economic value.The miR156 genes and their targets SQUAMOSA promoter-binding protein-like(SPL)play a vital role in plant growth and development.To analyze the miR156-SPL module in water yam,the study conducted the identification of miR156 gene loci and prediction and validation of their targets.Based on water yam genome sequence and transcriptome datasets,the study obtained eight DamiR156 loci and six target genes among which four belong to SPL gene family.Multiple-sequence alignments indicated that the eight miR156 genes were highly conserved in their hairpin regions and could be clustered into three groups.Transcriptome datasets and RT-qPCR showed that the expression level of miR156 is much higher in leaves,stems and tubers of tissue-cultured regeneration yam than that in the yams grown in the field,while the expression level of which is higher in the leaves,stems and tubers of the regenerated plant were cultured by tissue culture than that in other tissues.The study preliminary inferred that DalmiR156 may play an important role in the tuber development.The study used the 5'RLM-RACE to verify the cleavage site of a target SPL gene,which provided reference basis for further gene function verification of the SPL gene in yam.The research laid a scientific foundation for the study of the miR156-SPL regulation model in water yam.