摘要
该研究采用高通量测序技术Illumina HiSeq 2000,对小花草玉梅花器官进行转录组测序,挖掘参与其花发育相关的基因。测序结果得到54 513 822个序列读取片段(reads),包含7 826 726 115bp的碱基序列信息。将测序数据进行序列组装后,获得43 767个单基因簇(unigenes),平均长度为926bp。转录物注释结果显示,28 130条unigenes有同源比对信息。在43 767条unigenes中检测到5 015个SSR位点,且SSR不同重复基序类型中,出现频率最高的是AG/CT,其次是AAG/CTT和ACC/GGT。通过转录因子分析,进一步筛选得到了12个与花发育紧密相关的MADS基因,分别为FUL1,FUL2,AP3-1,AP3-2,AP3-3,PI1,PI2,AG1,AG2,SEP1,SEP3和AGL6。实时荧光定量PCR分析表明,与小花草玉梅正常花相比,全白、绿白相间、五瓣变异、全绿变异花中的FUL1、SEP1,SEP3和AGL6基因均显著上调表达,而12个基因在极端变异花中的表达水平与正常花的差异均不明显。定量结果经主成分分析显示,AGL6、SEP3、FUL1、PI2及SEP1的表达量均为小花草玉梅花形态建成的主要指标。研究结果在一定程度上丰富了小花草玉梅的基因信息,为后续研究其花器官变异的分子机制提供了基础数据。
The transcriptome of Anemone rivularis var.flore-minore was sequenced by Illumina HiSeq 2000 platform to find flower development gene.The result indicated that 54 513 822 sequence reads containing 7 826 726 115 bp nucleotide sequence information were obtained.A total of 43 767 unigenes were formed by initial sequence splicing,with an average read length of 926 bp.Annotation analysis of unigene indicated that 28 130 unigenes had homolog in public protein database.Total 5 015 unigenes included SSRs and the highest frequency was AG/CT,followed by AAG/CTT and ACC/GGT.Through the analysis of transcription factors,12 MIKC-type MADS genes closely related to flower development were selected,and they were FUL 1,FUL 2,AP3-1,AP3-2,AP3-3,PI 1,PI 2,AG 1,AG 2,SEP 1,SEP 3 and AGL 6.Quantitative PCR analysis showed that the expression of FUL 1,SEP 1,SEP 3 and AGL 6 were significant up regulate in white,green-white,five-flap and green plants than that in normal flower of A.rivularis var.flore-minore,and the expression of 12 genes had no significant difference in normal and extreme variation plants.The expression of AGL 6,SEP 3,FUL 1,PI 2 and SEP 1 were the main indexes of flower morphogenesis based on Principal Component Analysis(PCA).These results greatly enriched genetic information of A.rivularis var.flore-minore and provided basic data for the study of mechanism on diversity of floral organ variants in A.rivularis var.flore-minore in the future.
作者
戎利勤
李晓冬
刘虎岐
RONG Liqin;LI Xiaodong;LIU Huqi(College of Life Science,Northwest A&F University,Yangling,Shaanxi 712100,China)
出处
《西北植物学报》
CAS
CSCD
北大核心
2018年第8期1437-1442,共6页
Acta Botanica Boreali-Occidentalia Sinica
基金
国家中医药管理局2017年中医药公共卫生服务补助专项"全国中药资源普查项目"(财社[2017]66号)
关键词
小花草玉梅
转录组
高通量测序
花发育基因
Anemone rivularis var.flore-minore
transcriptome
high-throughput sequencing technology
flower development gene
