Somyong Suthasinee, Poopear Supannee, Jomchai Nukoon, Uthaipaisanwong Pichahpuk, Ruang-Areerate Panthita, Sangsrakru Duangjai, Sonthirod Chutima, Ukoskit Kittipat, Tragoonrung Somvong, Tangphatsornruang Sithichoke
National Center for Genetic Engineering and Biotechnology (BIOTEC), 113 Thailand Science Park, Phaholyothin Road, Klong Nueng, Klong Luang, Pathum Thani, 12120, Thailand,
Funct Integr Genomics. 2015 May;15(3):349-62. doi: 10.1007/s10142-014-0423-y. Epub 2014 Dec 12.
Sex ratio (SR), the ratio of female inflorescences to total inflorescences, is one of the main yield components of oil palm (Elaeis guineensis Jacq). The SR quantitative trait locus (QTL) was recently identified on linkage (LG) 8 with a phenotype variance explained (PVE) of 11.3 %. The use of both genetic and physical mapping is one strategy for uncovering the genetic basis of the traits. Here, we report the construction of bacterial artificial chromosome (BAC) and fosmid libraries, and their use for physical mapping in oil palm. Combined, the libraries consist of more than 200,000 clones, representing 6.35 genome equivalents. Physical mapping at the SR locus was implemented by incorporating the published oil palm genome sequence and positive BAC/fosmid clones as identified by colony PCR screening. Based on the previously published sequences, the interval (about 184 kb) was comprised of 19 contigs of the known sequences (117 kb, 64 %). After, combining the 454 pyrosequences of 15 positive clones and the previously published sequences, the known sequences were revealed to cover about 82 % of the interval (150 kb), and were used for identifying the new markers by designing 35 gene-based and 23 simple sequence repeat (SSR)-amplified primers. As a result, a putative aldo-keto reductase gene (named EgAKR1) was revealed to be a promising candidate for sex ratio determination, via controlling female inflorescence number (11 % of PVE). This was predicted from the two newly identified polymorphic marker loci (mEgSSRsr8-21LB and mEgAKR1-9) designing from EgAKR1. The functions of AKR gene families in other plant species and our promoter analysis suggested that EgAKR1 may contribute to the sex ratio through abiotic stress responsiveness.
性别比(SR),即雌花序与总花序的比例,是油棕(Elaeis guineensis Jacq)的主要产量构成要素之一。最近在连锁群(LG)8上鉴定出了SR数量性状位点(QTL),其表型变异解释率(PVE)为11.3%。综合运用遗传图谱和物理图谱是揭示性状遗传基础的一种策略。在此,我们报告了细菌人工染色体(BAC)和fosmid文库的构建及其在油棕物理图谱构建中的应用。这些文库合起来包含超过200,000个克隆,代表6.35个基因组当量。通过整合已发表的油棕基因组序列以及经菌落PCR筛选鉴定出的阳性BAC/fosmid克隆,在SR位点进行了物理图谱构建。基于先前发表的序列,该区间(约184 kb)由19个已知序列的重叠群组成(约117 kb,占64%)。之后,将15个阳性克隆的454焦磷酸测序结果与先前发表的序列相结合,发现已知序列覆盖了该区间约82%(约150 kb)的区域,并通过设计35个基于基因的引物和23个简单序列重复(SSR)扩增引物用于鉴定新标记。结果表明,一个假定的醛酮还原酶基因(命名为EgAKR1)通过控制雌花序数量(PVE的11%),有望成为性别比决定的候选基因。这是根据从EgAKR1设计的两个新鉴定的多态性标记位点(mEgSSRsr8 - 21LB和mEgAKR1 - 9)预测出来的。其他植物物种中AKR基因家族的功能以及我们的启动子分析表明,EgAKR1可能通过非生物胁迫响应来影响性别比。