杨树性别决定基因对杨树叶片叶绿素含量的影响及相关差异表达基因分析
Effects of poplar sex-determining gene on chlorophyll contents of poplar leaves and analysis of the relevant differentially expressed genes
[目的]在植物界中,雌雄异株植物除了生殖器官表现出的二态性,在一些非生殖器官上也会观察到明显的次生性别二态性。这种次生性别二态性在植物的生长发育、形态特征和代谢物质积累等方面都可能产生重要的影响,尤其在种群水平上。本研究旨在通过在分子水平上调控杨树性别决定基因的表达,探究植物性别决定基因是否对植物非生殖器官的次生性别二态性产生影响。[方法]通过在杨树雄株无性系"84K"(Populus.Alba×P.tremula var.glandulosa)中过表达和在杨树雌株无性系"南林895"(P.deltoides×P.euramericana)中敲除杨树性别决定基因,对抗性植株进行分子检测之后,分别获得杨树性别决定基因的过表达株系和基因敲除系。对获得的阳性转基因株系进行了表型观察和相关指标测定,并对过表达植株开展了基因差异表达分析。[结果]本研究共获得了91 个"南林895"过表达株系和139 个"84K"基因敲除系。在过表达株系中,普遍观察到了植株叶片黄化的现象。对叶色差异显著株系的叶绿素a、b、(a+b)的含量测定显示,过表达株系叶绿素a、叶绿素b、总叶绿素(a+b)的含量显著低于野生型,其中叶绿素b含量下降最为显著。相比之下,基因编辑植株没有出现黄化的现象,其叶色与野生型没有差异。对过表达株系开展的基因差异表达分析结果显示,过表达系与野生型植株叶片共有 899 个差异表达基因。将这 899 个差异表达基因进行了GO分类与KEGG通路分析显示,类黄酮生物合成途径显著富集,也是差异表达基因数量最多的通路。[结论]杨树性别决定基因过表达可能影响了类黄酮生物合成途径基因的表达,从而影响了过表达株系叶片叶绿素含量,最终导致叶片黄化现象。这一发现对植物性别决定基因影响植物非生殖器官的次生性别二态性提供了直接证据。同时,这项研究也为植物育种和遗传改良提供了新的思路和理论基础。
[Objective]Except for the dimorphism in the reproduction organs,dioecious plants exhibited secondary sexual dimorphism in some non-reproductive characteristics.This secondary sexual dimorphism can have significant impacts on plant growth,development,morphological traits,and accumulation of metabolic substances,particularly at the population level.This study aims to investigate whether plant sex-determining gene affects the secondary sexual characteristics by manipulating the expression of poplar sex determining gene at molecular level.[Method]To established the over-expression and knockout lines of the poplar sex determining gene,we over-expressed the poplar sex determining gene in a male poplar"84K"(Populus.Alba×P.tremula var.glandulosa)and edited the poplar sex determining gene in a female poplar"Nanlin 895"(P.deltoides×P.euramericana).For the positive transgenic lines,we observed the phenotypic changes and measured the relevant physiological parameters.Subsequently,we analyzed the relevant differentially expressed genes(DEGs)in over-expression lines.[Result]In this study,a total of 139 over-expression and 91 knockout lines were obtained.For the over-expression lines,an obvious phenotypic change of leaf yellowing was observed.Measurement of the chlorophyll a,b,and(a+b)contents showed that all these parameter were significantly lower in the over-expression than in the knockout lines,with chlorophyll b decreased the most.By contrast,no leaf yellowing was observed with the knockout lines,which exhibited the similar leaves color as that of the wild type plants.Analyzing the DEGs in the over-expression lines against the wild type plants revealed a total of 899 DEGs,and the most abundant DEGs were found to involve in the flavonoid biosynthesis pathway base on GO clustering and KEGG mapping.[Conclusion]The poplar sex determining gene negatively affected chlorophyll contents,which might occur through its effect on the expression of genes relevant to the flavonoid biosynthesis pathway,leading to leaf yellowing.The findings of this study provided direct evidence that plant sex determining gene affected the secondary sexual dimorphism of the non-reproductive characteristics.Moreover,this study provides new insights and theoretical foundations for plant breeding and genetic improvement.
陆静;尹佟明;
南京林业大学 南京市 210037;南京林业大学 南京市 210037;
雌雄异株植物 性别决定基因 次生性别二态性 叶绿素含量 差异表达基因
dioecious plant sex determining gene secondary sexual dimorphism chlorophyll content differentially expressed gene.
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