紫花苜蓿不同发育时期次生壁合成调控的转录组分析

doi:10.3864/j.issn.0578-1752.2018.11.003

Abstract

Abstract: 【Objective】The aim of the study was to investigate the gene network regulation dynamics and expression pattern of secondary cell wall synthesis and to identify the key candidate genes and transcription factors in regulatory network for the understanding of molecular mechanism of the secondary cell wall thickening in alfalfa.【Method】The stem samples collected at vegetative stage (S1), flower bud stage (S2), early flower stage (S3) and late flower stage (S4) were used to determine the main substance contents in the secondary cell wall of the ‘Zhongmu No.1’ cultivar of alfalfa using a near-infrared-reflectance spectroscopy. The high-throughput transcriptome sequencing of these tissues were performed using an Illumina HiSeq^TM 2500 platform. The genome of Medicago truncatula was used as a reference to align the sequences and construct the Unigenes. The gene expression level was calculated in the term of Fragments Per Kilobase of transcript per Million fragments (FPKM). The thresholds to judge the significance of gene expression differences between three adjacent growth periods (S2 vs S1, S3 vs S2 and S4 vs S3) were set as fold change ≥2 or≤0.5 for up or down regulation with a false discovery rate (FDR) ≤ 0.001. The Gene Ontology database and KEGG Pathway database were used to annotate the functions and metabolic pathways of differentially expressed genes. 【Result】A total of 41 734 Unigenes were identified at different developmental stages of alfalfa in this study, of which 27 differentially expressed genes (DEGs) were annotated with functions closely related to cellulose and lignin biosynthesis. The expression levels of these DEGs gradually increased along with the accumulation of cellulose and lignin over the developmental stages. The early flower stage was found to be a transitional period at which the contents of cellulose and lignin in the secondary cell wall and expression of synthetic genes were all increased significantly. For instance, the expression of cellulose synthase genes of MTR_2g016630 (Ces) and MTR_7g103590 (Ces A1) were significantly increased at the early flower stage, the expression of lignin biosynthesis genes of MTR_1g064090 (PAL1), MTR_1g111240 (C4H) and MTR_2g104960 (CCR) were significantly up-regulated more than 10 times at the early or late flower stage compared to the vegetative stage. A total of 27 transcription factors were differentially expressed at different developmental stages of alfalfa which were related to plant growth and development, of which 18 belonged to NAC and MYB transcription factor family while the rest were of WRKY, BHLH, ERF and C3H transcription factor family.【Conclusion】We acquired the gene expression profiling data of the stem at four developmental stages of the ‘Zhongmu No. 1’ cultivar of alfalfa. A total of 54 DEGs were identified, of which 24 were up-regulated while 30 were downregulated. These genes were postulated to be involved in the synthesis and regulation of the secondary cell wall in alfalfa.

Key words: Medicago sativa;developmental stage, secondary cell wall, transcriptome analysis, transcription factor

LIU XiQiang, ZHANG Han, GONG Pan, GONG WenLong, WANG Zan. Transcriptome Analysis of Secondary Cell Wall Synthesis Regulation at Different Developmental Stages in Alfalfa (Medicago sativa L.)[J].Scientia Agricultura Sinica, 2018, 51(11): 2049-2059.

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Transcriptome Analysis of Secondary Cell Wall Synthesis Regulation at Different Developmental Stages in Alfalfa (Medicago sativa L.)

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