新疆土桃果皮纯色形成与PpMYB10.1启动子变异的关系

doi:10.3864/j.issn.0578-1752.2025.02.009

中国农业科学 ›› 2025, Vol. 58 ›› Issue (2): 326-338.doi: 10.3864/j.issn.0578-1752.2025.02.009

新疆土桃果皮纯色形成与PpMYB10.1启动子变异的关系

郭天发¹^,²(), 吴金龙², 仇倩倩³, 马新超¹, 王力荣²(), 吴翠云¹()

¹ 塔里木大学园艺与林学学院/南疆特色果树高效优质栽培与深加工技术国家地方联合工程实验室，新疆阿拉尔 843300
² 中国农业科学院郑州果树研究所，郑州 450009
³ 阿拉尔国家农业科技园区，新疆阿拉尔 843300

收稿日期:2024-05-16 接受日期:2024-07-01 出版日期:2025-01-21 发布日期:2025-01-21
通信作者:
王力荣，E-mail：wanglirong@caas.cn。
吴翠云，E-mail：wucuiyun@taru.edu.cn
联系方式: 郭天发，E-mail：gtfzky@taru.edu.cn。
基金资助:
塔里木大学校长基金(TDZKSS202257); 兵团创新人才培养示范基地建设项目(2019CB001); 中国农业科学院科技创新工程项目(CAAS-ASTIP-ZFRI-01)

Relationship Between the Formation of Non-Red Color in the Fruit Skin of Xinjiang Local Peach Varieties and the Variation of PpMYB10.1 Promoter

GUO TianFa¹^,²(), WU JinLong², QIU QianQian³, MA XinChao¹, WANG LiRong²(), WU CuiYun¹()

¹ College of Horticulture and Forestry, Tarim University/The National and Local Joint Engineering Laboratory of High Efficiency and Superior-Quality Cultivation and Fruit Deep Processing Technology of Characteristic Fruit Trees in South Xinjiang, Aral 843300, Xinjiang
² Zhengzhou Fruit Research Institute, Chinese Academy of Agricultural Sciences, Zhengzhou 450009
³ Aral National Agricultural Science and Technology Park, Aral 843300, Xinjiang

Received:2024-05-16 Accepted:2024-07-01 Published:2025-01-21 Online:2025-01-21

摘要/Abstract

摘要：

【目的】探究新疆土桃资源果皮纯色形成的分子机理，为选育果皮纯色桃新品种提供理论依据。【方法】通过对85株新疆土桃实生后代和3个栽培品种的果皮颜色进行表型调查，利用GUS染色检测不同长度PpMYB10.1启动子活性，在PlantCARE（https://bioinformatics.psb.ugent.be/webtools/plantcare/html/）网站上预测不同材料PpMYB10.1启动子的作用元件，用PCR克隆鉴定88份材料PpMYB10.1启动子上5 243 bp转座子插入和483 bp缺失的基因型，利用双荧光素酶试验验证Pp.4G186800对PpMYB10.1的激活转录。【结果】根据果皮颜色，将85株新疆土桃实生后代分为纯色和红色两类，多数单株表现为纯色（绿色、浅黄色），果皮不积累或极少积累花色苷，与全红品种‘中桃金蜜’相比，其果皮花色苷含量和PpMYB10.1表达量明显降低。通过GUS染色检测启动子变异对其活性的影响，结果显示，当PpMYB10.1启动子上存在5 243 bp插入时，果盘蓝色变浅，表明活性减弱；而携带483 bp缺失后，果盘蓝色加深，表明启动子活性增强，启动子上插入/缺失影响了其活性；新疆土桃实生后代的PpMYB10.1启动子均不存在5 243 bp插入和483 bp缺失。克隆了土桃PpMYB10.1启动子序列，并将启动子顺式作用元件分为4类：光响应元件、植物激素响应元件、非生物胁迫和生长发育响应元件及一些未知功能的元件。由于土桃实生后代成熟较晚，鉴定了成熟期调控基因Pp.4G186800的基因型，均为0 bp/0 bp；通过双荧光素酶试验发现Pp.4G186800能够结合PpMYB10.1启动子，当PpMYB10.1启动子存在483 bp缺失时，Pp.4G186800（+9 bp）与PpMYB10.1启动子形成“强强联合”，可能促进早熟桃果皮花色苷积累。而在晚熟桃中，Pp.4G186800（-9 bp）对PpMYB10.1启动子无促进作用；反之，若PpMYB10.1启动子无483 bp缺失时，其本身活性弱，尽管Pp.4G186800对PpMYB10.1启动子有作用，但影响甚微。【结论】新疆土桃果皮纯色与PpMYB10.1启动子上的插入和缺失变异无直接关系，在早、中熟品种中，Pp.4G186800可激活PpMYB10.1的转录，有助于花色苷合成；而晚熟桃中Pp.4G186800对PpMYB10.1的作用减弱或消失。

关键词: 新疆, 土桃, 果皮, PpMYB10.1, 启动子活性, 花色苷

Abstract:

【Objective】This study aims to explore the molecular mechanisms underlying the formation of Non-red ground color in Xinjiang local peach (Prunus ferganensis and P. persica), providing a theoretical basis for the breeding of new peach varieties with Non-red skin color. 【Method】A phenotypic survey of the skin color of 85 progenies of Xinjiang peach and three cultivated varieties was conducted. The activity of various lengths of the PpMYB10.1 promoter was assessed using β-glucuronidase (GUS) staining. Predictive analysis of the cis-acting elements of the PpMYB10.1 promoter from different materials was performed using the PlantCARE (https://bioinformatics.psb.ugent.be/webtools/plantcare/html/). Genotypes with a 5 243 base pairs (bp) transposon insertion and a 483 bp deletion in the PpMYB10.1 promoter of 88 materials were identified and cloned using polymerase chain reaction (PCR). The transcriptional activation of PpMYB10.1 by Pp.4G186800 was validated through dual-luciferase assays. 【Result】Based on skin color, 85 Xinjiang peach progenies were classified into Non-red and red types. The majority exhibited Non-red colors (green or light yellow) with minimal or no anthocyanin accumulation. Compared to the fully red variety Zhongtao Jinmi, these exhibited significantly lower levels of anthocyanins and reduced expression of PpMYB10.1. Using GUS staining to assess the effects of promoter mutations on activity, the results showed that a 5 243 bp insertion in the PpMYB10.1 promoter caused the fruit discs to display a lighter blue, indicating decreased activity. Conversely, a 483 bp deletion resulted in a darker blue, suggesting increased promoter activity. No such insertions or deletions were found in the PpMYB10.1 promoter of the Xinjiang peach progenies. The promoter sequences of PpMYB10.1 were cloned, and cis-acting elements were categorized into four groups: light-responsive, plant hormone-responsive, abiotic stress and growth development-responsive, and elements of unknown function. Due to the late maturation of the Xinjiang peach progenies, all were found to have a 0 bp/0 bp genotype for the maturity-regulating gene Pp.4G186800. Dual-luciferase assays showed that Pp.4G186800 could bind to the PpMYB10.1 promoter. Notably, when the promoter had a 483 bp deletion, Pp.4G186800 (+9 bp) formed a strong interaction, potentially enhancing early maturation and anthocyanin accumulation in the peach skin. However, in late-ripening peaches, Pp.4G186800 (-9 bp) had no enhancing effect on the promoter’s activity; If the PpMYB10.1 promoter lacked the 483 bp deletion, its intrinsic activity was weak, even though Pp.4G186800 could still interact with it, the effect was minimal.【Conclusion】The presence of insertions and deletions in the PpMYB10.1 promoter is not directly related to the Non-red skin color of Xinjiang peach. In early and mid-ripening varieties, Pp.4G186800 can activate the transcription of PpMYB10.1, facilitating anthocyanin synthesis; however, its influence is reduced or absent in late-ripening peaches.

Key words: Xinjiang, local peach, fruit skin, PpMYB10.1, promoter activity, anthocyanin

郭天发, 吴金龙, 仇倩倩, 马新超, 王力荣, 吴翠云. 新疆土桃果皮纯色形成与PpMYB10.1启动子变异的关系[J]. 中国农业科学, 2025, 58(2): 326-338.

GUO TianFa, WU JinLong, QIU QianQian, MA XinChao, WANG LiRong, WU CuiYun. Relationship Between the Formation of Non-Red Color in the Fruit Skin of Xinjiang Local Peach Varieties and the Variation of PpMYB10.1 Promoter[J]. Scientia Agricultura Sinica, 2025, 58(2): 326-338.

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新疆土桃果皮纯色形成与PpMYB10.1启动子变异的关系

Relationship Between the Formation of Non-Red Color in the Fruit Skin of Xinjiang Local Peach Varieties and the Variation of PpMYB10.1 Promoter

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