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作 者:李遥 何彦岑 林宇环 万婷 李蒙 陈智勇[1,2] Li Yao;HeYancen;LinYuhuan;Wan Ting;Li Meng;Chen Zhiyong(College of Bioscience and Biotechnology,Hunan Agricultural University,Changsha,410128;Miscanthus Ecological Application Technology,Hunan Engineering Laboratory,Hunan Agricultural University,Changsha,410128)
机构地区:[1]湖南农业大学生物科学技术学院,长沙410128 [2]湖南农业大学,芒属植物生态应用技术湖南省工程实验室,长沙410128
出 处:《分子植物育种》2021年第13期4378-4385,共8页Molecular Plant Breeding
基 金:国家自然科学基金面上项目(31471557);国家自然科学基金青年项目(32000260);湖湘高层次人才聚集工程(2019RS1051)共同资助。
摘 要:于PacBio SMRT单分子实时测序技术进行3代转录组测序,辅以2代测序,构建出一个完整的,准确的南荻Unigene库。通过与近源物种比对后,拼接出南荻(Miscanthus lutarioriparius)纤维素合成酶基因(cellulose synthase gene)CesA4、CesA7和CesA9的序列,并将其命名为MlCesA4、MlCesA7、MlCesA9。使用生物信息学分析软件构建出蛋白质系统进化树,对氨基酸翻译后修饰的磷酸化位点进行预测和分析,对蛋白质的保守结构域进行预测和分析。结果显示,MlCesA4、MlCesA7和MlCesA9基因序列长度分别为2980、3310、3208 bp与高粱和玉米的亲缘关系最近;南荻纤维素合成酶(MlCesA)MlCesA4和MlCesA9有43个磷酸化位点,MlCesA7有48个磷酸化位点;Ml CesA4是不稳定蛋白,具有6个跨膜结构域,其中4个在膜外,3个在膜内,MlCesA7和MlCesA9是稳定蛋白质,具有8个跨膜结构域,其中5个在膜外,4个在膜内。三者都是亲水性蛋白,二级结构均以α-螺旋和无规则卷曲为主。Based on 3 generations transcriptome sequencing technology PacBio SMRT single-molecule real-time sequencing method,supplemented with 2 generation sequencing method,a complete and accurate Unigene library was constructed.After comparing with closely related species,sequences of cellulose synthase genes CesA4,CesA7 and CesA9 of Miscanthus lutarioriparius were spliced and named as MlCesA4,MlCesA7 and MlCesA9.Bioinformatics analysis software was used to construct phylogenetic trees of proteins,to predict and analyze the phosphorylation sites of post-translational modifications of amino acids,and to predict and analyze conserved domains of proteins.The analysis results showed that the sequence lengths of MlCesA4,MlCesA 7,MlCesA 9 genes were 2980,3310,and 3208 bp,respectively,which were closely related to Sorghum bicolor and Zea mays.Cellu lose synthase of Miscanthus lutarioriparius(MlCesA)MlCesA4 and MlCesA9 have 43 phosphorylation sites,and Ml CesA7 has 48 phosphorylation sites.MlCesA4 is an unstable protein with 6 transmembrane domains,4 domains are outside the membrane and 3 are inside the membrane.Ml CesA7 and MlCesA9 are stable proteins,both of which have 8 transmembrane domains,5 domains are outside the membrane and 4 are inside the membrane.The three are all hydrophilic proteins,and the secondary structure is dominated byα-helix and random coils.
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