Divergent DNA methylation contributes to duplicated gene evolution and chilling response in tea plants

The tea plant (Camellia sinensis) is a thermophilic cash crop and contains a highly duplicated and repeat-rich genome. It is still unclear how DNA methylation regulates the evolution of duplicated genes and chilling stress in tea plants. We therefore generated a single-base-resolution DNA methylation map of tea plants under chilling stress. We found that, compared with other plants, the tea plant genome is highly methylated in all three sequence contexts, including CG, CHG and CHH (where H = A, T, or C), which is further proven to be correlated with its repeat content and genome size. We show that DNA methylation in the gene body negatively regulates the gene expression of tea plants, whereas non-CG methylation in the flanking region enables a positive regulation of gene expression. We demonstrate that transposable element-mediated methylation dynamics significantly drives the expression divergence of duplicated genes in tea plants. The DNA methylation and expression divergence of duplicated genes in the tea plant increases with evolutionary age and selective pressure. Moreover, we detect thousands of differentially methylated genes, some of which are functionally associated with chilling stress. We also experimentally reveal that DNA methyltransferase genes of tea plants are significantly downregulated, whereas demethylase genes are upregulated at the initial stage of chilling stress, which is in line with the significant loss of DNA methylation of three well-known cold-responsive genes at their promoter and gene body regions. Overall, our findings underscore the importance of DNA methylation regulation and offer new insights into duplicated gene evolution and chilling tolerance in tea plants.     

Gene expression remodelling and immune response during adaptive divergence in an African cichlid fish

Variation in gene expression contributes to ecological speciation by facilitating population persistence in novel environments. Likewise, immune responses can be of relevance in speciation driven by adaptation to different environments. Previous studies examining gene expression differences between recently diverged ecotypes have often relied on only one pair of populations, targeted the expression of only a subset of genes or used wild‐caught individuals. Here, we investigated the contribution of habitat‐specific parasites and symbionts and the underlying immunological abilities of ecotype hosts to adaptive divergence in lake–river population pairs of the cichlid fish Astatotilapia burtoni. To shed light on the role of phenotypic plasticity in adaptive divergence, we compared parasite and microbiota communities, immune response, and gene expression patterns of fish from natural habitats and a lake‐like pond set‐up. In all investigated population pairs, lake fish were more heavily parasitized than river fish, in terms of both parasite taxon composition and infection abundance. The innate immune response in the wild was higher in lake than in river populations and was elevated in a river population exposed to lake parasites in the pond set‐up. Environmental differences between lake and river habitat and their distinct parasite communities have shaped differential gene expression, involving genes functioning in osmoregulation and immune response. Most changes in gene expression between lake and river samples in the wild and in the pond set‐up were based on a plastic response. Finally, gene expression and bacterial communities of wild‐caught individuals and individuals acclimatized to lake‐like pond conditions showed shifts underlying adaptive phenotypic plasticity.     

滇龙胆GrHDR基因的克隆与表达分析

龙胆苦苷(gentiopicroside)是中药龙胆中的主要药效成分,属于萜类化合物的衍生物。1-羟基-2-甲基-2-(E)-丁烯基-4-二磷酸还原酶[1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate reductase,HDR]是萜类物质合成途径中的关键酶。为探讨不同光照条件下滇龙胆HDR(GrHDR)基因的表达与龙胆苦苷含量之间的关系,该文以滇龙胆叶片cDNA为模板,采用PCR和TA克隆技术获得GrHDR基因序列,对该序列进行生物信息学分析和表达分析,并采用高效液相色谱法测定龙胆苦苷含量,对该基因表达与龙胆苦苷含量进行比较。结果表明:(1)GrHDR基因(GenBank登录号: KJ917165.1)全长1 398 bp,编码465个氨基酸,推定GrHDR蛋白是亲水且稳定的,相对分子质量是52 281.25 Da,理论等电点是5.32;(2)该蛋白属于LYTB蛋白家族,可能定位于叶绿体上,无信号肽,二级结构主要由α-螺旋(45.16%)、β-转角(6.24%)、无规卷曲(33.98%)、延伸链(14.62%)构成;(3)GrHDR蛋白序列与同属植物秦艽的HDR蛋白相似性最高(95.71%);(4)实时荧光定量PCR结果显示GrHDR基因在滇龙胆中的表达量为根 > 叶 > 茎,而在10%、30%、100%全光光照条件下各组织的表达量有很大差异;(5)高效液相色谱法结果显示,不同光照条件下龙胆苦苷含量一致,均为根 > 叶 > 茎,其中100%全光光照下,药用部位根中龙胆苦苷含量达到7.141%,约是30%、10%全光光照条件的两倍,但该结果与同一光照条件下GrHDR基因表达规律不完全一致。该研究为阐述HDR基因功能及其与龙胆苦苷含量的关系提供参考。     

毛竹APX基因系统进化与表达分析

抗坏血酸过氧化物酶(aseorbate peroxidase, APX)是植物活性氧代谢中重要的抗氧化酶之一,尤其是叶绿体中清除H₂O₂的关键酶,也是维生素C代谢的主要酶类。该文基于生物信息学方法,利用毛竹的基因组及转录组数据鉴定毛竹中的APX基因家族成员,并对其编码的蛋白基本理化性质、基因结构、启动子元件、系统进化及共线性关系、重复串联基因、GO注释及表达模式进行综合分析,共鉴定出21种编码APX的基因。结果表明:(1)PeAPX基因家族成员多为不稳定疏水蛋白,基因结构、基序及结构域相对较为保守,大多数APX基因具有高度保守的内含子模式。(2)系统进化关系显示毛竹APX基因与水稻APX基因有着较高的同源性关系,PeAPX具有较高的进化保守性。(3)Ka/Ks分析表明PeAPX基因都经历了纯化选择压力,此外在每个APX基因的启动子序列中发现有许多与应激反应和植物激素相关的顺式作用元件,结合表达量分析,表明毛竹APX基因在毛竹生长发育中起着正向促进作用。该研究为进一步了解毛竹APX基因家族基本功能及其抗氧化机制提供了一定的参考,为毛竹APX基因功能的深层次鉴定提供了重要依据。