新疆野苹果幼苗叶片响应NaCl胁迫的miRNAs及靶基因筛选  

Selection of miRNAs and Target Genes in Leaves of Malus sieversii under NaCl Stress

作　　者：何晨晨 刘俐君 鲁晓燕[1] HE Chenchen;LIU Lijun;LU Xiaoyan(Key Laboratory of Cultivation Physiology and Germplasm Resources Utilization of Characteristic Fruits and Vegetables of XPCC/College of Agronomy,Shihezi University,Shihezi Xinjiang 832003,China)

机构地区：[1]石河子大学农学院/特色果蔬栽培生理与种质资源利用兵团重点实验室,新疆石河子832000

出　　处：《新疆农业科学》2021年第4期589-598,共10页Xinjiang Agricultural Sciences

基　　金：国家自然科学基金委NSFC-新疆联合基金(培育)(U1703116)。

摘　　要：【目的】筛选新疆野苹果响应NaCl胁迫相关差异miRNA及靶基因。【方法】以新疆野苹果组培苗为材料,对150 mmol/L NaCl处理6和48 h时的新疆野苹果幼苗叶片进行small RNA测序。【结果】NaCl处理6 h时3个miRNA差异表达,其中2个上调表达,1个下调表达,miRNA对应的靶基因数目为64个;NaCl处理48 h时13个miRNA差异表达,其中4个上调表达,9个下调表达,miRNA对应的靶基因数目为108个。对差异靶基因进行GO和KEGG分类注释,NaCl处理6 h时,GO主要富集包括:肌醇磷酸2激酶活性、寡糖基转移酶活性、蛋白丝氨酸/苏氨酸磷酸酶抑制剂活性、木聚糖生物合成过程等;NaCl处理48 h时,GO主要富集包括:氧氧化还原酶活性、DNA结合、质外体、次生代谢过程等。KEGG共同富集在N-glycan生物合成通路。【结论】mdm-miR168b、mdm-miR159c、mdm-miR827、mdm-miR390f、mdm-miR171i、mdm-miR399j基因在NaCl处理6和48 h时表达量存在差异,其中mdm-miR390f及其靶基因HF10976、mdm-miR171i及其靶基因HF33844、mdm-miR399j及其靶基因HF11095表达量呈负相关,3个miRNA参与了新疆野苹果对NaCl胁迫的响应过程。【Objective】To screen miRNA and target genes in response to NaCl stress in Malus sieversii.【Methods】In this study,tissue culture seedlings of Malus sieversii were used as experimental materials and small RNA sequencing was performed on leaves of Malus sieversii seedlings treated with 150 mmol/L NaCl for 6 h and 48 h.【Results】The results showed that:compared with the control group,3 miRNAs were differentially expressed when NaCl treatment lasted for 6 h,among which 2 were up-regulated and 1 was down-regulated,and the corresponding target genes of miRNA were 64.When NaCl treatment lasted for 48 h,13 miRNAs were differentially expressed,among which 4 were up-regulated,9 were down-regulated,and the corresponding target genes of miRNA were 108.The differential target genes were classified and annotated by GO and KEGG.When NaCl treatment lasted for 6 h,GO enrichment mainly included:inositol pentakisphosphate 2-kinase activity,oligosaccharyl transferase activity,tRNA wobble adenosine to inosine editing,xylan biosynthetic process,etc.When NaCl treatment lasted for 48 h,GO enrichment mainly included:oxidoreductase activity,DNA binding,Apoplast,secondary metabolic process,etc.KEGG co-accumulate in the N-glycan biosynthesis pathway.【Conclusion】qRT-PCR verified that the expression levels of mdm-mir168b,mdm-mir159c,mdm-mir827,mdm-mir390f,mdm-mir171i and mdm-mir399j were different when NaCl treatment lasted for 6 h and 48 h,and the expression levels of mdm-mir390f and its target genes HF10976,mdm-mir171i and its target genes HF33844,mdm-mir399j and its target genes HF11095 were negatively correlated.These three miRANs were involved in the response process of Malus sieversii to NaCl stress.

关 键 词：新疆野苹果 NACL胁迫 MICRORNA 

分 类 号：S661.1[农业科学—果树学] S188[农业科学—园艺学]