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
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.
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