摘要
MADS-box家族基因编码的转录因子,在植物、动物及微生物的发育过程中起着重要调控作用,本研究从马尾松幼苗转录组测序结果中筛选到1条具有完整开放阅读框的MADS-box基因序列,设计特异性引物并克隆获得该基因,通过生物信息学方法分析该基因的序列特征、编码蛋白的理化特性、结构域以及进化关系等,并应用荧光定量PCR技术分析该基因在马尾松幼苗不同组织中的表达活性。结果表明,克隆得到的MADS-box基因开放阅读框为672 bp,命名为Pm MADS4。Pm MADS4编码223个氨基酸,含有MADS保守域和K-box次级保守域。蛋白质理论分子量为24854.15 kD,理论等电点为7.83;其蛋白质结构主要由大量的α-螺旋(58.74%)和大量随机卷曲(31.84%)构成;亚细胞预测该基因主要在细胞核(43.5%)、线粒体(17.4%)、高尔基体(13.0%)和细胞质(13.0%)中发挥生物学作用。系统进化分析显示,Pm MADS4属于MADS-box家族基因中MIKC型的GMADS分支。组织特异性表达分析发现,Pm MADS4在马尾松幼苗的顶芽和茎中高表达,表达量分别是根部的239倍和123倍;在根部、初生叶和针叶中的表达量相对较低,表明Pm MADS4可能广泛参与马尾松幼苗地上部分分生组织的发育过程。本研究结果为Pm MADS4基因在马尾松幼苗生长发育调控方面的深入研究提供了数据基础。
The transcription factors encoded by MADS-box family genes are essential to development of plants,animals and microorganisms.In this study,a MADS-box gene with a complete open reading frame(ORF)was screened from the transcriptome results of Pinus massoniana seedlings.Specific primers including start and end codes were used to clone the ORF sequence.Furthermore,sequence characteristics of the gene,physicochemical characteristics,structural domains and evolutionary relationship of the encoded protein were analyzed by bioinformatics methods.And the expressions of the gene in different tissues of P.massoniana seedlings were detected by qPCR.The results showed that ORF of the gene was 672 bp and named as Pm MADS4.The Pm MADS4 encoded223 amino acids and contained the MADS-box conserved and K-box secondary conserved domain.The theoretical molecular weight of the protein was 24854.15 kD,and the theoretical isoelectric point was 7.83.The protein structure was composed ofα-helix(58.74%)and coils(31.84%).The results of subcellular localization analysis indicated that the proteins were mainly in the nucleus(43.5%).And they also took effect in mitochondria(17.4%),Golgi apparatus(13.0%)and cytoplasm(13.0%).Phylogenetic analysis showed that PmMADS4 belongs to the GMADS clade of MIKC type in MADS-box family genes.Moreover,expressions of PmMADS4 in the shoot apex and stems of the seedlings were 239 and 123 times higher than that in root,respectively.However,the expressions were relatively low in the cataphyll and needles,which indicated PmMADS4 might be widely involved in the development of shoot meristem of P.massoniana seedlings.This study provides data foundation for further research on the regulation and development of seeding growth in P.massoniana.
作者
石长双
朱小坤
涂晶晶
吴峰
Shi Changshuang;Zhu Xiaokun;Tu Jingjing;Wu Feng(Institute for Forest Resources&Environment of Guizhou,Guizhou University,Guiyang,550025)
出处
《分子植物育种》
CAS
CSCD
北大核心
2020年第3期847-852,共6页
Molecular Plant Breeding
基金
国家重点研发计划课题(2017YFD0600302)
国家自然科学基金(31660201)
贵州省科技计划项目(黔科合LH字[2016]7447)和(黔科合平台人才[2019]5102)共同资助。
作者简介
通信作者:吴峰,wufengyjy@163.com。
