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作 者:李朝闯 马关鹏 谢婷[1,3] 陈娇[1] 王志敏[1] 宋明[1] 汤青林[1]
机构地区:[1]西南大学园艺园林学院,南方山地园艺学教育部重点实验室,重庆市蔬菜学重点实验室,重庆400715 [2]贵州省园艺研究所,贵阳550006 [3]毕节市农业技术推广站,贵州毕节551700
出 处:《园艺学报》2017年第3期463-474,共12页Acta Horticulturae Sinica
基 金:国家重点基础研究发展计划(‘973’)项目(2012CB113900);国家自然科学基金项目(31000908);中央高校基本科研业务费专项(XDJK2012B020)
摘 要:为阐明芥菜开花抑制因子AGL18与开花整合子SOC1间的互作调控机制,在芥菜‘QJ’的开花期克隆了AGL18-1,幼苗期克隆了AGL18-2和AGL18-3,它们分别编码257、257和258个氨基酸,为AGL18家族的3个成员。序列比对表明,芥菜AGL18家族成员与十字花科芜菁和油菜同源性均高达90%。酵母双杂交和BiFC试验表明:芥菜AGL18-1、AGL18-2和AGL18-3蛋白与SOC1不会发生蛋白相互作用。酵母单杂交和Dual-Glo~ Luciferase试验表明:AGL18-1、AGL18-2和AGL18-3中仅有花期AGL18-1蛋白与SOC1启动子间存在互作。为进一步筛选AGL18-1/SOC1的互作区域,分别截取了AGL18-1蛋白的M域和IKC域,发现仅M域与SOC1启动子存在相互作用,说明M域是介导花期AGL18-1蛋白与SOC1启动子互作的关键区域。这为深入研究AGL18与SOC1互作的分子机制及其对开花时间的调控奠定了基础。AGAMOUS-LIKE 18 (AGL18) is a key regulatory factor in flowering time control of Brassica juncea. In order to clarify the regulatory mechanism of AGL18 protein interact with SOC 1 in flowering pathways, AGL18-1 gene from reproductive shoots as well as AGL18-2 and AGL18-3 from vegetative shoots in 'QJ' germplasm ofBrassicajuneea, were cloned respectively. AGL18-1, A GL18-2 and AGL18-3 were members of AGL18 family and respectively encoded 257, 257 and 258 amino acids. Sequences analysis showed that they were more than 90% similar to Brassiea rapa and Brassica napus. Both Yeast two hybrid and BiFC showed that SOC 1 protein could not interact with AGL 18-1, AGL 18-2 or AGL18-3. Yeast one hybrid and Dual-Glo~ Luciferase assays indicated that SOC1 promoter could interact with AGL18-1 but not AGL18-2 and AGL18-3. To screen the interaction regions of AGL18-1/SOC1, the M-domain and IKC-domain truncations of AGL18-1 protein were separated and found that only the M-domain truncation could interact with SOCI promoter. It suggested that M-domain was the key region that mediated the interaction between AGL18-1 and SOC1 promoter. The work provided valuable information for in-depth studies on the molecular mechanisms of AGL18 protein with SOC1 and the flowering time in Brassicajuncea.
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