■大麻二酚(CBD),作为大麻植物的重要治疗特性,主要产于花器官。生长素响应因子(ARFs)在花卉发育和次生代谢产物产生中起着至关重要的作用。然而,ARF基因家族在大麻中的具体作用尚不清楚.
■在这项研究中,使用在线网站和生物信息学对CsARF基因进行了各种生物信息学分析,利用实时定量PCR技术研究CsARF基因家族在不同大麻品种不同组织中的表达模式,并在烟叶中进行了亚细胞定位分析。
■在这项研究中,鉴定了22个CsARF基因,发现它们在大麻基因组的9条染色体上分布不均。系统发育分析表明,ARF蛋白分为4个亚组。重复分析确定了一对节段/全基因组重复的CsARF,和三对串联复制的CsARF。共线性分析显示,两个CsARF基因,CsARF4和CsARF19在水稻和大豆中都是直系同源的。此外,亚细胞定位分析显示CsARF2定位于细胞核内。组织特异性表达分析显示,六个基因在大麻雄花中高表达,在这些基因中,进一步发现3个基因在雄花的不同发育阶段高表达。同时,两个品种“H8”和“Y7”中CsARF基因的表达水平与CBD含量之间的相关性分析表明,CsARF13的表达水平与CBD含量呈负相关,6个基因的表达水平与CBD含量呈正相关。此外,大多数CsARF基因对IAA治疗有反应。
■我们的研究为进一步研究CsARFs在大麻中的功能奠定了基础,并为在大麻生产中培育具有高CBD产量的品种提供候选基因。
UNASSIGNED: Cannabidiol (CBD), as an important therapeutic property of the cannabis plants, is mainly produced in the flower organs. Auxin response factors (ARFs) are play a crucial role in flower development and secondary metabolite production. However, the specific roles of ARF gene family in cannabis remain unknown.
UNASSIGNED: In this study, various bioinformatics analysis of CsARF genes were conducted using online website and bioinformatics, quantitative real time PCR technology was used to investigate the expression patterns of the CsARF gene family in different tissues of different cannabis varieties, and subcellular localization analysis was performed in tobacco leaf.
UNASSIGNED: In this study, 22 CsARF genes were identified and found to be unevenly distributed across 9 chromosomes of the cannabis genome. Phylogenetic analysis revealed that the ARF proteins were divided into 4 subgroups. Duplication analysis identified one pair of segmental/whole-genome duplicated CsARF, and three pairs of tandemly duplicated CsARF. Collinearity analysis revealed that two CsARF genes, CsARF4 and CsARF19, were orthologous in both rice and soybean. Furthermore, subcellular localization analysis showed that CsARF2 was localized in the nucleus. Tissue-specific expression analysis revealed that six genes were highly expressed in cannabis male flowers, and among these genes, 3 genes were further found to be highly expressed at different developmental stages of male flowers. Meanwhile, correlation analysis between the expression level of CsARF genes and CBD content in two cultivars \'H8\' and \'Y7\' showed that the expression level of CsARF13 was negatively correlated with CBD content, while the expression levels of six genes were positively correlated with CBD content. In addition, most of CsARF genes were responsive to IAA treatment.
UNASSIGNED: Our study laid a foundation for the further studies of CsARFs function in cannabis, and provides candidate genes for breeding varieties with high CBD yield in cannabis production.