齐明,何贵平,周建革,程亚平,徐肇友.杉木生长性状的杂种优势转录组分析[J].林业科学研究,2019,32(3):113-120
杉木生长性状的杂种优势转录组分析
Transcriptome Analysis of Heterosis of Growth Traits in Chinese Fir
投稿时间:2018-03-25  修订日期:2019-02-11
DOI:10.13275/j.cnki.lykxyj.2019.03.015
中文关键词:  杉木  生长  杂种优势  转录组测序  分子机理
英文关键词:Chinese fir  growth  Cunninghamia lanceolata  heterosis  transcriptome sequencing  molecular mechanism
基金项目:浙江省"十三五"林木新品种育种专项(2016C02056-5);中国林科院中央级公益性科研院所基本科研业务费专项资金资助子项(CAFYBB2017ZA001-1-2)
作者单位E-mail
齐明 中国林业科学研究院亚热带林业研究所, 浙江省林木育种技术研究重点实验室, 浙江 杭州 311400  
何贵平 中国林业科学研究院亚热带林业研究所, 浙江省林木育种技术研究重点实验室, 浙江 杭州 311400 guipinghe@126.com 
周建革 浙江省遂昌县林业技术推广总站, 浙江 遂昌 323300  
程亚平 浙江省庆元县庆元林场, 浙江 庆元 323805  
徐肇友 浙江省龙泉市林业科学研究院, 浙江 龙泉 323700  
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中文摘要:
      [目的]研究杉木杂种F1的基因表达模式,从基因表达水平揭示杂种优势形成分子机理,为杉木杂种深度开发利用提供参考依据。[方法]采用Illumina HiSeq4000高通量测序技术对不同生长势的杉木杂种(龙15×1339)HF1(3个超亲杂种)、LF2(3个低亲杂种)及其亲本进行转录组测序和差异比较。利用无参转录组分析,先将测序reads进行de novo拼接,组装获得unigenes和transcripts,然后进行功能注释、差异表达等项目分析。[结果]12个样本,转录组测序共产生Clean reads5.8E+08条,总拼接长度49 803 726 pb,BLASTX分析,比对结果产生80 171个基因。韦恩图揭示基因在上下代间的传递模式有5种:(1)双亲可表达但杂种中不表达(双亲共沉默型);(2)只在双亲之一中表达,不在杂种中表达(亲本特异表达型);(3)只在杂种中表达,不在双亲中表达(杂种特异表达型);(4)在杂种和一个亲本中表达(单亲表达一致型);(5)在双亲和杂种中都表达。在HF1VSP1比较组中,筛选出236个不同差异表达的基因;在HF1VSP2比较组中,筛选出505个差异表达基因。在LF2VSP1中,筛选出1 483个差异表达基因;在LF2VSP2中筛选出了2 335个差异表达基因。从亲代和子代不同样本组间,各挑选出100个左右差异表达极显著的基因,采用其表达量进行热聚类分析,得到不同的聚类块,聚类块内的基因表达量在父母本间是互补的,说明杉木杂种优势分子机理是超显性;聚类块有大小之别,则说明杉木有些性状是寡基因控制,有些性状是多基因控制;杉木生长的超亲优势是龙15中下调而在超亲子代中上调的这14个基因决定的。[结论]杉木杂种优势的分子机理是超显性,杉木超亲杂种高生产力与14个上调基因有关,环境显著刺激了这14个基因的上调表达,从而促进生长优势的产生。
英文摘要:
      [Objective] To study the changes of gene expression patterns in F1 hybrids of Chinese fir, analyze the transcriptomic profiling and to identify the associative genes related to heterosis, which presented a useful reference for the deep development and utilization of Chinese fir (Cunninghamia lanceolata).[Method] Using the latest second-generation hybrids (Long 15×1339), HF1 (3 super-parent hybrids), LF2 (3 low-parent hybrids) and parents (P1 and P2), all sample groups were compared by transcriptome sequencing. Since Chinese fir transcriptome sequencing is lack of reference genome, so it is needed to do the sequencing reads first for de novo splice to get unigenes and transcripts. Subsequently, the functional annotation of transcripts, differential expression and other bioinformatic analysis approaches were adopted.[Result] In a total of 12 samples, 5.8E+08 clean reads were generated by transcriptome sequencing, and the total length was 49 803 726 pb. BLASTX analysis is performed on clean reads in six databases (Nr, Swiss-prot, KOG, KEGG, Pfam, GO), through sequence alignment which results in 80 171 unigenes. The Venn diagram analysis revealed 5 transmission modes of DEGs from the parents to the progenies:1. parental expression but not expressed in hybrid (biparent silent type); 2. expression in one parent only, not in hybrids (parental specific expression); 3. only expressed in hybrids, not expressed in parents (hybrid specific expression); 4. expression in hybrid and one parent (single expression consistent); 5. expression in both parents and hybrids. 236 unigenes with differentially expressed (DEGs) were identified in HF1VSP1 comparison group, in HF1VSP2, LF2VSP1, and LF2VSP2, 1 483, 505, and 2 335DEGs were revealed respectively. 100 significant different unigenes among parents and progenies are screened out, and the heatmap clustering analysis was carried out. The results revealed the molecular mechanism of the heterosis of Chinese fir was over-dominant. The sizes of clustering block were different, indicating that some traits were controlled by oligogenes and some by polygenes. The superparental dominance of Chinese fir growth was determined by the 14 genes down-regulated in long15 and up-regulated in the superparent progeny.[Conclusion] It is concluded that the molecular mechanisms of Chinese fir heterosis are over-dominant. The high productivity of Chinese fir superparental offsprings is related to 14 up-regulated genes. Environment significantly stimulates the up-regulated expression of these 14 genes, thus promotes the generation of growth advantages.
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