CAJ | 학술논문

类黄酮途径中, TT3编码的4-二氢黄铜醇还原酶是参与原花色素和花青素合成的关键酶。为了明确该基因可能的上游调控网络,利用黄籽母本 GH06和黑籽父本 ZY821构建的遗传图谱,以 BnTT3基因在高世代重组自交系群体中随机选取的94个株系花后40 d种子的表达量作为性状,采用复合区间作图法进行eQTL分析。结果共检测到5个表达量相关的eQTL,分别位于A03、A08、A09和C01染色体,单个eQTL解释表型变异的5.22%~24.05%。A09染色体上存在2个主效 eQTL,单个 eQTL 分别解释24.05%和16.55%的表型变异,分别位于标记 KS10260~KBrB019I24.15和B055B21-5~KS30880之间,微效eQTL分布于A03、A08和C01染色体上。A09染色体上的2个主效eQTL区间(包含200 kb侧翼序列)与拟南芥、白菜、甘蓝和芸薹族近缘物种基因组同源区段具有很好的共线性关系。基因注释结果表明检测到的eQTL均为trans-QTL,2个主效eQTL区段共包含78个基因,包括MYB51、MYB52和bZIP5转录因子,可能为 BnTT3基因的上游直接调控因子,对这些基因功能的深入分析将有助于阐明甘蓝型油菜黄籽性状形成的分子调控机制,为黄籽候选基因的克隆筛选奠定基础。
류황동도경중, TT3편마적4-이경황동순환원매시삼여원화색소화화청소합성적관건매。위료명학해기인가능적상유조공망락,이용황자모본 GH06화흑자부본 ZY821구건적유전도보,이 BnTT3기인재고세대중조자교계군체중수궤선취적94개주계화후40 d충자적표체량작위성상,채용복합구간작도법진행eQTL분석。결과공검측도5개표체량상관적eQTL,분별위우A03、A08、A09화C01염색체,단개eQTL해석표형변이적5.22%~24.05%。A09염색체상존재2개주효 eQTL,단개 eQTL 분별해석24.05%화16.55%적표형변이,분별위우표기 KS10260~KBrB019I24.15화B055B21-5~KS30880지간,미효eQTL분포우A03、A08화C01염색체상。A09염색체상적2개주효eQTL구간(포함200 kb측익서렬)여의남개、백채、감람화예대족근연물충기인조동원구단구유흔호적공선성관계。기인주석결과표명검측도적eQTL균위trans-QTL,2개주효eQTL구단공포함78개기인,포괄MYB51、MYB52화bZIP5전록인자,가능위 BnTT3기인적상유직접조공인자,대저사기인공능적심입분석장유조우천명감람형유채황자성상형성적분자조공궤제,위황자후선기인적극륭사선전정기출。
In flavonoid biosynthesis pathway, the key enzyme dihydroflavonol 4-reductase (DFR) involved in the proanthocya-nidin and anthocyanin biosynthesis pathway is encoded byTRANSPARENT TESTA3 (TT3) gene. The objective of this research was to identify the upstream regulatory networks ofBnTT3 using the composite interval mapping method (CIM). Hence, we per-formed an eQTL analysis for the transcript-level variation ofBnTT3 gene in seeds at 40 days after flower (DAF) in 94 recombi-nant inbred lines (RILs) derived from a cross between the yellow-seeded female parent GH06 and black-seeded male parent ZY821. Five eQTLs for expression levels ofBnTT3 were detected on four different chromosomes (A03, A08, A09, and C01) inB. napus, accounting for 5.22% to 24.05% of phenotypic variation. Two major eQTLs were found and located among the markers KS10260–KBrB019I24.15 and B055B21-5–KS30880 of chromosome A09, with explained 24.05% and 16.55% of phenotypic variation, respectively. Three minor eQTLs were also detected to be located on chromosomes A03, A08, and C01. The flanking sequences with 200 kb of two major eQTLs on chromosome A09 ofB. napus showed well synteny to those ofA. thaliana,Bras-sica rapa,Brassica oleracea and other Brassiceae relatives. Furthermore, the annotation results showed that they belong to the trans-QTL, containing 78 genes in the two trans-QTL regions. Some transcription factors (MYB51,MYB52, andbZIP5) might be upstream regulatory factors associated with transcriptional regulation ofBnTT3. Therefore, further study about these genes func-tion will be helpful to elucidate the molecular mechanism of the seed coat colour formation, as well as lay the foundation for se-lecting candidate genes of seed coat colour in B. napus.