Sequencing of anthocyanin synthesis-related enzyme genes and screening of reference genes in leaves of four dominant subtropical forest tree species
Sequencing of anthocyanin synthesis-related enzyme genes and screening of reference genes in leaves of four dominant subtropical forest tree species
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亚热带四种优势森林树种叶片花青素合成相关酶基因测序及内参基因筛选
DOI：
10.1016/j.gene.2019.144024
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发表时间：
2019
期刊：
影响因子：
3.5
通讯作者：
Peng ChangLian
中科院分区：
文献类型：
--
作者：
Yu ZhengChao;Zhang Peng;Lin Wei;Zheng XiaoTing;Cai MinLing;Peng ChangLian
亚热带森林优势种的幼叶一般积累一定浓度的花青素，花青素合成相关酶基因表达水平的变化对研究亚热带森林幼叶花青素的光保护作用有重要影响。花色素苷合成相关酶基因序列的测定和合适参比基因的选择为分析花色素苷的功能特性提供了基础。在这项研究中，四个优势亚热带森林物种（即，以木荷（Schima superba）、栗实（Castanopsisfissa）、厚壳桂（Cryptocarya concinna）、厚壳桂（Acmena acuminatissima）为试材。为了获得正确的花色素苷相关酶的核苷酸序列，对各优势种的CHS、DFR和ANS进行了测序和比较。为了筛选出在不同发育时期和不同光照条件下叶片最稳定的参考基因，采用实时荧光定量PCR（qRT-PCR）技术对18 S、Actin、GAPDH、TUB、EF 1和UBQ等6个参考基因的表达水平进行了研究，并利用GeNorm和Normandise软件对参考基因的稳定性进行了分析。结果表明，Actin在木荷、木霉、木acuminatissima和C.而GAPDH在C.裂。最后，对花色素苷合成基因CHS、DFRandANS的表达水平进行分析，发现其表达水平与花色素苷在叶片中的积累趋势一致。本研究对亚热带森林优势树种花青苷合成相关酶基因表达的研究具有重要的理论和实践意义，揭示了花青苷对强光环境下幼叶的光保护作用。
The young leaves generally accumulate a certain concentration anthocyanins in the dominant species of the subtropical forest, and the changes of anthocyanin synthesis-related enzyme genes expression levels had an important effect on the study photoprotection of anthocyanins in the young leaves of subtropical forests. The determination of anthocyanin synthesis-related enzyme gene sequences and the selection of appropriate reference genes provide a basis for analyzing the functional properties of anthocyanins. In this study, four dominant subtropical forest species (i.e.,Schima superba,Castanopsisfissa,Acmena acuminatissima,Cryptocarya concinna) were taken as materials. To obtain the correct nucleotide sequences of anthocyanin-related enzymes, the nucleotide sequences of CHS, DFR andANSin each dominant species were obtained by sequencing and comparison. Then, to select the most stable reference genes for leaves at different developmental stages and different light conditions, the expression levels of six reference genes, including18S,Actin, GAPDH,TUB,EF1andUBQ, were studied by real-time fluorescent quantitative PCR (qRT-PCR), and reference gene stability was analyzed by GeNorm and NormFinder software. The results showed that the expression level ofActinwas the most stable inS. superba,A. acuminatissimaandC. concinna, and the expression level ofGAPDHwas the most stable inC. fissa. Finally, the expression levels of the anthocyanin synthesis genesCHS,DFRandANSwere analyzed and found to be consistent with the accumulation trend of anthocyanins in leaves. This study has important theoretical and practical significance for future research into the expression of anthocyanin synthesis-related enzyme genes in the dominant tree species in subtropical forests and reveals that anthocyanin has a photoprotective effect for young leaves in high-light environments.