中国农业科学院郑州果树研究所,郑州450009
研究方向为桃种质资源评价与利用,E-mail: wangjiaolingling@163.com
王力荣,研究方向为桃种质资源与遗传育种研究,E-mail: wanglirong@caas.cn
中国农业科学院科技创新工程专项经费项目(CAAS-ASTIP-2021-ZFRI-01)
Zhengzhou Fruit Research Institute, Chinese Academy of Agricultural Sciences, Zhengzhou 450009
Foundation project: The Agricultural Science and Technology Innovation Program (CAAS-ASTIP-2021-ZFRI-01)
以红色深浅不一的红肉桃种质为材料,探讨影响其花色素苷含量的分子机理,为高效选育红色深浅不等的红肉桃品种提供理论依据。利用GUS染色测定桃果肉花色素苷重要基因PpMYB10.1的启动子活性,利用DNA-pulldown鉴定结合于PpMYB10.1启动子上的转录抑制因子,利用双荧光素酶及酵母双杂交验证转录抑制因子的功能。结果表明:(1)具有深红、红、浅红的桃果肉,其对应的PpMYB10.1表达量及花色素苷含量依次下降。(2)具有483 bp序列的PpMYB10.1启动子,其启动活性弱于缺失该序列的启动子。(3)利用该483 bp序列鉴定到的转录抑制子基因Prupe.2G302800,虽然不能直接抑制PpMYB10.1的转录,但能够结合花色素苷合成的主效基因PpBL,并抑制其转录活性,可能对降低PpMYB10.1表达具有一定功能。本研究通过483 bp缺失序列鉴定到的转录抑制子Prupe.2G302800,虽然不是红肉变浅的直接因素,但通过抑制PpBL转录活性,对于红肉桃红色变浅,可能具有一定作用。
Based on the red-flesh color peaches showing different intensities, this study attempted to decipher their formation mechanism in order to provide theoretical basis for efficient breeding of red peach varieties. The promoter activity of PpMYB10.1 was detected via GUS staining, and the candidate transcription repressors binding on its promoter were captured through DNA-pull down assay. The function of these candidate genes were determined by double luciferase and yeast two-hybrid assay. The results showed that: (1) The expression of PpMYB10.1 and anthocyanin content in flesh peaches with deep-red, red and light-red were gradually decreasing. (2) Activity of PpMYB10.1 promoter with a 483 bp deletion was weaker than that without the sequence. (3) Interestingly, we identified a candidate transcription repressor Prupe.2G302800 based on the 483bp deletion. The protein strongly interacted with PpBL, a major factor in anthocyanin synthesis and resulted in a reduction on the transcription of PpMYB10.1. Prupe.2G302800 is unlikely the direct factor modulating the red flesh of peach, whereas it might play an important role in decreasing red-flesh color by inhabiting PpBL transcription activity.
王蛟,曹珂,王玲玲,等.