桑树脱落酸生物合成相关基因的鉴定及转录表达分析

doi:10.3864/j.issn.0578-1752.2015.05.19

Abstract

Abstract: 【Objective】 9-cisepoxycarotenoid dioxygenase (NCED), ABA-aldehyde oxidase(AAO), Zeaxanthin epoxidase (ZEP) are the key enzymes involved in biosynthesis of abscisic acid (ABA) via indirect pathway. The aim of this study was to analyze the transcriptional expression of ABA biosynthesis related genes and detect the content of ABA during Jialing 40 (Morus atropurpurea Roxb.) fruit development. The effects of ABA and its synthesis inhibitors on fruit development were also explored. This work will lay a foundation for further studying the role of ABA on mulberry (Morus alba L.) fruit maturity and senescence. 【Method】In this study, the sequences of six putative ABA biosynthesis related genes were obtained from Morus notabilis genome database and its deduced amino acid sequences were analyzed by using bioinformatics tools. Total RNA was extracted from mulberry fruit using RNAiso Plus (TaKaRa) and reverse transcribed to synthesize cDNA. The relative transcriptional expression of ABA biosynthesis related genes in different developmental stages and treated mulberry fruits was investigated by using qRT-PCR. The content of ABA was determined by using HPLC.【Result】Six ABA biosynthesis related genes were isolated from mulberry, including one AAO gene, two ZEP genes and three NCED genes. Multiple-sequence alignment results showed that MnNCEDs amino acids are relatively conserved in higher plants. Cluster analysis revealed that MnNCED1-3 are closely related with dicotyledons plants and farther with monocotyledons plants. Transcription analysis indicated that the expression of ABA biosynthesis related genes was higher in leaves and the lowest expression was detected in roots. Particularly, the expression level of MnNCED2 and MnNCED3 was higher in different plant tissues compared to other genes. The concentration of ABA was strongly increased after the conversion stage of fruit. Exogenous ABA significantly promoted fruit maturity, whereas fluridone remarkably inhibited fruit ripening compared with the negative control treated with distilled water. The results of real-time PCR showed that the expression abundance of MnNCEDs was higher in the middle and latter periods than the early stage. Again, ABA biosynthesis related genes have shown different expression patterns under the same treatment. The expression level of ABA biosynthesis related genes was up-regulated in the fourth day after ABA treatment while down-regulated in the first day after fluridone treatment, unlikely the expression level of MnNCED2 was down-regulated after both ABA and fluridone treatments. 【Conclusion】 In this study six putative ABA biosynthetic genes in M. notabilis genome were obtained. The expression levels of MnNCEDs were higher in middle and later stages than early fruit developmental stage, which is in coincidence with the trend of ABA content during fruit development. Thus, MnNCEDs may play important roles in ABA biosynthesis. Exogenous ABA significantly promoted fruit maturity, while fluridone markedly inhibited fruit ripening.

Key words: mulberry, ABA, NCED, AAO, ZEP, transcriptional expression analysis, fluridone

ZHU Pan-pan, LIU Chang-ying, ZHAO Ai-chun, PEI Rui-chao, LI Jun, WANG Xiao-hong, LI Zhen-gang, WANG Xi-ling, LU Cheng, YU Mao-de. Characterization and Transcriptional Expression Analysis of ABA Biosynthesis Related Genes from Mulberry (Morus alba L.)[J].Scientia Agricultura Sinica, 2015, 48(5): 1011-1022.

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Characterization and Transcriptional Expression Analysis of ABA Biosynthesis Related Genes from Mulberry (Morus alba L.)

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