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机构地区:[1]河北农业大学,河北保定071001 [2]深圳职业技术学院,广东深圳518000
出 处:《热带作物学报》2016年第6期1147-1154,共8页Chinese Journal of Tropical Crops
基 金:国家自然基金(No.31572162);深圳市科技项目(No.JCYJ20130331151531198)
摘 要:低温引起切花月季花朵过度重瓣化导致畸形在生产上亟需解决。以‘芬得拉’(Rosa hybrida‘Vendela’)为试材,以RT-PCR研究与月季花器官发育密切相关基因Rh AP1,Rh AP2,Rh AP3,Rh TM6,Rh PI,Rh AG,Rh SHP,Rh WUS在不同温度不同花芽分化时期表达量。发现过度重瓣化形成关键时期,只有Rh AG表达水平相对较高,且低温下表达水平明显低于常温,相比其它基因变化最明显。本课题组推测Rh AG在低温导致的过度重瓣化中可能起着主要调控作用。为深入研究,以RT-PCR结合RACE克隆到Rh AG全长。其全长4 942 bp,包含7个内含子、8个外显子,开放阅读框747 bp,编码248个氨基酸,N端含保守MADS-box、K-box结构域,属MADS-box家族。洋葱表皮亚细胞定位发现其位于细胞核内,属核蛋白。上述结果对明确低温导致月季花朵过度重瓣化分子机制提供了理论依据。The excessive double flowers in rose caused by low temperature,resulting to abnormal flowers needs prompt solutions in production.To use Rosa hybrida ‘Vendela' as the material and take Rh AP1,Rh AP2,Rh AP3,Rh TM6,Rh PI,Rh AG,Rh SHP,Rh WUS which are closely related to rose floral organ development as the research objects.Their expression quantity at different temperature and different flower bud differentiation period were analyzed by RT-PCR technique.We found that the expression level of Rh AG was higher than others at the key period of excessive double flowers in rose caused by low temperature,the expression at low temperature was much lower and its change was the most obvious.It indicated that Rh AG played a major regulatory role in the excessive double flowers in rose caused by low temperature.For further research,we amplified the full length of Rh AG by RACE and RT-PCR techniques.The results showed that the cloned full-length c DNA of Rh AG was 4 942 bp,containing a 747 bp open reading frame(ORF) which encoded 248 amino acids.It had 7 introns and 8 exons.Protein alignment analysis showed that Rh AG protein contained a MADS domain and a K domain at N terminal which indicated that Rh AG was a member of MADS-box family.We used onion epidermal sub-cellular location to locate Rh AG,and found that Rh AG was located in cell nucleus,it was a nucleoprotein.
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