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2025年7月30日 星期三
农业生物技术学报  2019, Vol. 27 Issue (4): 624-635    DOI: 10.3969/j.issn.1674-7968.2019.04.006
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
PpmMDH基因克隆及采后激素处理下的表达分析
张姗姗1, *, 王滨1, *, 李静媛2, 沈俊岭1, 马春晖1, 黄永红1, 段艳欣1*, *
1 青岛市现代农业质量与安全工程重点实验室/青岛农业大学 园艺学院,青岛 266109;
2 青岛农业大学 食品科学与工程学院,青岛 266109
Gene Cloning of PpmMDH and Expression Analysis Under Postharvest Hormone Treatments in Peach (Prunus persica)
ZHANG Shan-Shan1, *, WANG Bin1, *, LI Jing-Yuan2, SHEN Jun-Ling1, MA Chun-Hui1, HUANG Yong-Hong1, DUAN Yan-Xin1, **
1 Qingdao Key Lab of Modern Agriculture Quality and Safety Engineering/College of Horticulture, Qingdao Agricultural University, Qingdao 266109, China;
2 College of Food Science and Engineering, Qingdao Agricultural University, Qingdao 266109, China
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摘要 苹果酸脱氢酶在植物的生长发育中发挥重要作用。本研究克隆了桃(Prunus persica)苹果酸脱氢酶(malate dehydrogenase, mMDH)基因(PpmMDH),分析了其序列特征及其在硬肉和软肉桃品种不同桃组织及果实成熟前后的表达情况,并探讨了不同激素处理对桃果实中PpmMDH基因表达的影响。结果表明,克隆得到了PpmMDH基因,GenBank登陆号为KF017594,该cDNA片段长度为1 239 bp,具有1个1 020 bp的ORF,编码339个氨基酸,序列分析表明,PpmMDH编码的氨基酸序列与其他植物的mMDH蛋白有较高的相似性,系统进化分析结果表明PpmMDH与梅(Prunus mume)和甜樱桃(Pr. avium)同源关系最近。qRT-PCR结果表明,PpmMDH在雄蕊中表达量较高(P<0.05),其次是花瓣,在叶片、幼果和雌蕊中表达量较低。果实成熟软化后期,PpmMDH在硬肉桃'双久红'果实中的表达量高于软肉桃'川中岛白桃'的。脱落酸(abscisic acid, ABA)、萘乙酸(1-naphthylacetic acid, NAA)、乙烯利(ethephon, ETH)处理桃果实得出,软肉桃果实成熟软化过程中PpmMDH的表达与乙烯有拮抗作用,硬肉桃与软肉桃在果实成熟与贮藏过程中内源激素动态不同,PpmMDH表达水平与酶活动态不同。上述结果表明,PpmMDH不一定是维持果实硬度的上游基因,但是在软肉桃与硬肉桃不同材料间表现出了与生理差异同步的酶活差异。本研究结果为进一步探索PpmMDH在桃果实成熟软化中的作用提供了科学依据。
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张姗姗
王滨
李静媛
沈俊岭
马春晖
黄永红
段艳欣
关键词 PpmMDH基因克隆激素处理表达分析    
Abstract:Malate dehydrogenase plays an important role in plant growth and development. The objective of this study was to clone malate dehydrogenase (PpmMDH), a gene possibly related to fruit ripening and softening in peach (Prunus persica), investigate its sequence characteristics and analyze its expression on different peach tissues and fruits before and after ripening firm-fleshed and soft-fleshed peach varieties. The results indicated that the full-length cDNA of PpmMDH (GenBank No. KF017594) in peach was cloned. The sequence consisted of 1 239 bp with an ORF of 1 020 bp, encoding a polypeptide of 339 amino acids. Homology analysis showed that the deduced PpmMDH protein was highly homologous to other mMDH proteins from different species. Phylogenetic analysis also indicated that PpmMDH was very closely related to mMDH of plum blossom (Prunus mume)和 cherry (Pr. avium). qRT-PCR results showed that the PpmMDH expression was abundant in stamens, followed by petals and leaves, and lower in young fruits and pistil. During later stage of fruit ripening, the accumulation of PpmMDH was significantly higher in the firm-fleshed mutant 'Shuangjiuhong' than that in the soft-fleshed 'Kawanakajima Hakuto' (P<0.05). Abscisic acid (ABA), 1-naphthylacetic acid (NAA) and ethephon (ETH) treatments showed that the expression of PpmMDH was up-regulated by ABA, NAA in both cultivars, but the soft-fleshed peach variety 'Kawanakajima Hakuto' was induced in a short period of time, and the firm-fleshed peach variety 'Shuangjiuhong' was upregulated continuously. Different from ABA, NAA, the effect of ETH treatment on the induction of the gene in the soft-fleshed peach variety 'Kawanakajima Hakuto' was very weak, even negligible, but the gene was continuously upregulated in 'Shuangjiuhong', firm-fleshed peach variety. In the control group, the expression level of PpmMDH in 'Kawanakajima Hakuto' was significantly lower than that in 'Shuangjiuhong'. Infer from this, the expression of PpmMDH was antagonistic to ethylene during ripening and softening of soft-fleshed peach, the endogenous hormone dynamics of firm-fleshed peach and soft-fleshed each were different during fruit ripening and storage, and PpmMDH expression level was different from enzyme activity state. The above results indicated that PpmMDH was not necessarily the upstream gene to maintain fruit firmness, but showed the same enzyme activity difference as physiological difference between different materials of firm-fleshed and soft-fleshed peach. The present study laid a foundation for further exploring the role of PpmMDH in peach fruit ripening and softening. It also provides a scientific basis for the identification of the gene at the molecular level.
Key wordsPrunus persica    PpmMDH    Gene cloning    Hormone treatment    Expression analysis
收稿日期: 2018-07-13     
ZTFLH:  S66  
基金资助:国家重点研发计划(No.2018YFD1000200)、山东省农业良种工程项目(No. 2016LZGC008)、山东省青岛市民生科技计划项目(No. 14-2-3-38-nsh)和国家级大学生创新训练项目(No. 201510435044)
通讯作者: dyxdyx2007@163.com   
引用本文:   
张姗姗, 王滨, 李静媛, 沈俊岭, 马春晖, 黄永红, 段艳欣. 桃PpmMDH基因克隆及采后激素处理下的表达分析[J]. 农业生物技术学报, 2019, 27(4): 624-635.
ZHANG Shan-Shan, WANG Bin, LI Jing-Yuan, SHEN Jun-Ling, MA Chun-Hui, HUANG Yong-Hong, DUAN Yan-Xin. Gene Cloning of PpmMDH and Expression Analysis Under Postharvest Hormone Treatments in Peach (Prunus persica). 农业生物技术学报, 2019, 27(4): 624-635.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2019.04.006     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2019/V27/I4/624
 
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