Expression of Genes in New Sprouts of Cunninghamia lanceolata Grown Under Dark and Light Conditions
Expression of Genes in New Sprouts of Cunninghamia lanceolata Grown Under Dark and Light Conditions
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暗光条件下杉木新芽基因的表达
DOI：
10.1007/s00344-019-09996-9
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发表时间：
2020
影响因子：
4.8
通讯作者：
Wan Yinglang
中科院分区：
文献类型：
--
作者：
Wang Qiaojun;Liu Shuai;Li Xinyu;Wu Hongyang;Shan Xiaoyi;Wan Yinglang
杉木子叶在黑暗中呈现绿色，是由于功能性的暗操作原叶绿素还原酶（DPOR）和光依赖原叶绿素还原酶（LPOR）共存所致。在这项研究中，我们发现LPOR的POR基因和DPOR三个亚基的ChlB，ChlL和ChlN基因在叶绿素生物合成受到抑制的黑暗生长的新芽中强烈下调。我们从黄化（黑暗生长）和绿色（光照生长）芽中提取总RNA，测序并注释。共发现70，499个unigenes，其中47，410个被注释。其中，3263差异表达基因之间的黑暗和光照生长的样品进行了鉴定。根据京都基因百科全书和基因组数据库，五个最受影响的代谢途径是类黄酮的生物合成，光合作用天线蛋白，光合作用，角质，枯草和蜡的生物合成和芪类化合物，二芳基庚烷类化合物，姜酚的生物合成。根据获得的cDNA序列，我们进一步利用定量PCR分析了18个核基因和11个叶绿体基因的相对表达水平。转录组和定量PCR结果表明，黑暗处理可以显着阻断所有已鉴定的基因的表达相关的光合电子流的蛋白质复合物在新芽。暗处理杉木新梢通过阻断DPOR和LPOR编码基因的表达而引起黄化。与光合作用相关的核基因组和质体基因组基因在黄化芽中均显著下调。这一结果为理解不同光合器官叶绿体发育的不同机制提供了新的线索。
Chinese fir (Cunninghamia lanceolata) cotyledons are green in darkness due to the coexistence of functional dark-operative protochlorophyllide reductase (DPOR) and light-dependent protochlorophyllide reductase (LPOR). In this study, we found that thePORgene for LPOR and theChlB,ChlL,andChlNgenes for three DPOR subunits were strongly downregulated in dark-grown new sprouts in which chlorophyll biosynthesis was inhibited. We extracted, sequenced, and annotated the total RNA from etiolated (dark-grown) and green (light-grown) sprouts. A total of 70,499 unigenes were found, of which 47,410 were annotated. Among them, 3263 differentially expressed genes were identified between dark- and light-grown samples. According to the Kyoto Encyclopedia of Genes and Genomes databases, the five most affected metabolic pathways were flavonoid biosynthesis, photosynthesis-antenna proteins, photosynthesis, cutin, suberine and wax biosynthesis and stilbenoid, diarylheptanoid, and gingerol biosynthesis. Based on the acquired cDNA sequences, we applied quantitative PCR to further analyze the relative expression levels of 18 nuclear genes and 11 chloroplast genes related to photophosphorylation. Both transcriptomic and quantitative PCR results indicated that dark treatment can significantly block the expression of all of the identified genes related to the protein complexes of photosynthetic electron flow in new sprouts. Dark treatment of new sprouts of Chinese fir caused etiolation via blockage of the expression of DPOR- and LPOR-encoding genes. Both nuclear and plastid genomic genes related to photosynthesis were significantly downregulated in etiolated sprouts. The results provide new clues to understanding the different mechanisms of chloroplast development in different photosynthetic organs.