光周期介导藜麦育性相关Cq_AGL基因的克隆与功能分析

藜麦是苋科藜属作物，因对逆境有较强抗性和籽粒具优越营养与保健价值而被全球广泛种植。与拟南芥清晰的开花调控与配子体发育途径相比，藜麦光周期诱导成花及配子发育的分子机制仍不清楚。为阐明光周期影响藜麦育性的分子机制，研究组前期以短日藜麦W25（长日不结籽）和光周期不敏感W19（短日和长日均结籽）为材料，利用转录组测序分析W25和W19不同光周期诱导后基因表达与植株相态和育性变化的相关性，筛选到3个与育性调控相关的AG相似基因（Cq_AGL-5、6、7）。据此，本项目以三代转录组测序进行候选Cq_AGL的全长CDS和可能的外显子选择拼接分析；qRT-PCR分析验证候选基因在不同器官中的表达；进行全长CDS的克隆及编码蛋白的亚细胞定位；藜麦中进行病毒诱导的基因沉默及拟南芥野生型和agl突变中表达后植株形态变化分析来确定其基因的功能。研究为阐明光周期调控藜麦育性分子机制及分子育种等提供参考。

Chenopodium quinoa is a crop of the Amaranthaceae, which is world widely cultivated because of its strong resistance to abiotic stress and its superior nutritional and health care value in seeds. Compared with the clear flowering regulation and gametophyte development pathway of Arabidopsis thaliana, the molecular mechanism of quinoa photoperiod-induced flower formation and gametophyte development is still unclear. To elucidate the molecular mechanism of photoperiod on the flowering and fertility of quinoa, both quinoa strains W25, which is short day plant inducing seedless development in long-day, and W19, which is photoperiod-insensitive plant inducing seed development in both short and long day, were selected as studying materials. After analyzing the correlation between genes expression, which were determined by RNS-seq, and plant phenotypes and fertility changes after induction of different photoperiods on W25 and W19, three C.quinoa agamous-like genes (Cq_AGL-5, 6, 7) which were predicted to correlated to fertility regulation of quinoa. The proposal intends to confirm the gene function of Cq_AGL-5, 6, 7 by (1) Their full-length CDS of and their possible exon alternative splicing were analyzed by three-generation RNA-seq. (2) Analysis of the candidate genes expression in different organs were done by qRT-PCR. (3) All full length CDSs were cloned by RT-PCR and subcellular localization of encoding proteins were analyzed by using both bioinformatics prediction and GFP fusion expression. (4) Plant fertility changes were determined in both quinoa after virus-induced gene silencing (VIGS) and in both wild and agl mutated Arabidopsis thaliana after expression of candidate genes respectively. The research provides a basis for elucidating the molecular mechanism of fertility regulation by photoperiod inducing and molecular breeding in quinoa.