月季‘绿萼’花器官发育相关microRNA的鉴定及分析  

Identification of microRNA associated with flower organ development in Rosa chinensis 'Viridiflora'

作　　者：眭梦洁 晏慧君[2] 王珍珍 邱显钦[2] 蹇洪英[2] 王其刚[2] 陈敏[2] 张颢[2] 唐开学[2] Sui Meng-Jie;Yan Hui-Jun;Wang Zhen-Zhen;Qiu Xian-Qin;Jian Hong-Ying;Wang Qi-Gang;Chen Min;Zhang Hao;Tang Kai-Xue(College of Landscape and Horticulture,Yunnan Agricultural University,Kunming 650201,China;Flower Research Institute of Yunnan Academy of Agricultural Sciences,Kunming 650205,China;School of Agriculture,Yunnan University,Kunming 650091,China)

机构地区：[1]云南农业大学园林园艺学院,昆明650201 [2]云南省农业科学院花卉研究所,昆明650205 [3]云南大学农学院,昆明650091

出　　处：《植物科学学报》2019年第1期37-46,共10页Plant Science Journal

基　　金：国家自然科学基金项目(31660579;31872144);云南省学术技术带头人后备人才项目(2017HB067)~~

摘　　要：利用高通量测序技术,构建了中国古老月季‘绿萼’(Rosa chinensis ‘Viridiflora’)和‘月月粉’(R.chinensis‘Old Blush’)花蕾期的microRNA(miRNA)文库,并对其进行了测序和序列分析。结果显示,在‘绿萼’文库中,鉴定到已知的miRNA成熟体39个,miRNA前体42个;预测到新的miRNA成熟体56个,前体57个。在‘月月粉’文库中,鉴定到已知RNA成熟体39个,已知miRNA前体40个;预测到新的miRNA成熟体53个,前体57个。与‘月月粉’相比,‘绿萼’中差异表达的miRNA有31个,其中17个上调、14个下调。荧光定量PCR实验结果表明,miR156、miR398和miR535在2种月季的花蕾期表达上调,而miR167、miR172和miR396表达下调。进一步检测miR172和miR156在2种月季不同花器官中的表达差异,发现miR172在‘绿萼’的花瓣、雌、雄蕊中表达显著下调,提示miR172可能通过负调控其靶基因RcAP2的表达,在‘绿萼’花器官发育过程中起重要作用。In this study,two independent microRNA(miRNA)libraries of Rosa chinensis‘Viridiflora’and‘Old Blush’at the flower bud development stage were constructed and sequenced with Illumina sequencing.In total,39 known miRNAs and 42 known pre-miRNAs were discovered,and 56 novel miRNAs and 57 novel pre-miRNAs were predicted in the‘Viridiflora’library.In total,39 known miRNAs and 40 known pre-miRNAs were identified,and 53 novel miRNAs and 57 novel pre-miRNAs were predicted in the‘Old Blush’library.Compared with‘Old Blush’,there were 31 differentially expressed miRNAs in‘Viridiflora’,with 17 up-regulated and 14 down-regulated.The RT-qPCR results showed that the expressions of miR156,miR398,and miR535 were up-regulated,whereas those of miR167,miR172,and miR396 were down-regulated in‘Viridiflora',identical to those obtained by sequencing.The expressions of miR172 and miR156 in different floral organs of the two rose species were detected by RT-qPCR.We found that miR172 was down-regulated in the petals,pistils,and stamens of‘Viridiflora’.Furthermore,RcAP2 was previously reported to be up-regulated in relative flower organs,suggesting that miR172 may negatively regulate the expression of its target gene RcAP2.Thus,miR172 may play an important role in the development of floral organs of‘Viridiflora’.

关 键 词：月季品种'绿萼’ 花器官发育 MiRNA高通量测序 荧光定量PCR 

分 类 号：Q943.2[生物学—植物学]