蜡梅比较基因组学及木兰类植物系统进化分析

基于蜡梅（Chimonanthus praecox）及相关植物物种的基因组数据,进行基因家族聚类,估算物种分化时间,并开展有关物种的共线性分析,计算不同类别代表性物种内旁系同源及物种间的直系同源同义替换率（synonymous substitution rate,Ks）,同时基于核苷酸和氨基酸序列的串联法及并联法等多种策略,进行木兰类植物系统进化分析。结果表明,蜡梅科（Calycanthaceae）植物共享了两次全基因组复制（whole-genome duplication,WGD）事件,樟目（Laurales）的共同祖先经历了二倍化事件,木兰类植物未共享WGD事件。长枝吸引（long branch attraction,LBA）会影响系统发育的准确性,消除LBA可使用1以及0和1相位点构建系统发育树。以往对木兰类植物系统发育位置的研究结果不一致,可能是由于系统发育树的构建方法不同、不完全谱系分选（incomplete lineage sorting, ILS）的存在、分类单元取样的局限性,以及木兰类植物在进化早期的快速分化等原因。在充分考虑多种可能影响木兰类植物进化位置多种因素的基础...     

全基因组复制事件的绝对定年揭示莲座蕨属植物的迟滞演化

全基因组复制在维管植物的物种形成过程中普遍存在, 被认为是物种适应极端环境的重要机制之一。确定全基因组复制事件的发生时间对理解生物的适应性演化具有重要意义。然而, 在维管植物, 特别是蕨类植物中, 全基因组复制事件的发生时间及其演化意义仍知之甚少。本研究以蕨类植物重要基部类群——福建莲座蕨(Angiopteris fokiensis)为例, 基于不同采样点(广东、广西、上海)的3个转录组学数据, 利用同义替换率(Ks)和绝对定年的方法分析全基因组复制事件的发生时间和物种单位时间内的分子演化速率, 并对事件发生后保留下的基因进行基因功能注释和富集分析。结果表明, 福建莲座蕨在159‒165 Mya发生了一次全基因组复制事件, 该复制事件优先保留的基因主要与营养代谢、信号传导、适应调节和组织结构生长相关。另外, 福建莲座蕨的分子演化速率为1.66 × 10^‒9 (同义替换/位点/年), 是除裸子植物外, 陆生植物中已知演化速率最缓慢的类群。综合以上研究结果, 我们推测福建莲座蕨全基因组复制的发生可能与裸子植物繁盛、核心被子植物集中兴起或托阿尔阶灭绝事件有关。而复制后显著保留基因可能促进了莲座蕨属(Angiopteris)植物的遗传和形态创新, 从而帮助其快速适应环境的剧烈变化。进一步对该类群植物演化速率缓慢的原因进行讨论, 推测莲座蕨属缓慢的演化速率可能与其本身世代周期长、基因组较大及其生长环境稳定有关。本研究通过分析福建莲座蕨的全基因组复制历史和复制基因的保留模式, 推测全基因组复制事件对促进演化速率较慢的植物适应极端环境变化具有重要意义, 可为理解其他陆生植物的适应性演化提供更多启发。     

转录组测序揭示翼盖蕨(Didymochlaena trancatula)的全基因组复制历史

全基因组复制在动植物中普遍存在, 被认为是促进物种进化的重要动力之一。作为蕨类植物的单种科物种, 翼盖蕨(Didymochlaena trancatula)是真水龙骨类I的基部类群, 在蕨类中具有独特的演化地位。本研究基于高通量测序, 通过同义替换率(Ks)分析、相对定年分析揭示翼盖蕨的全基因组复制发生情况。Ks分析表明, 翼盖蕨至少经历了两次全基因组复制事件, 其中一次发生于59-62 million years ago (Mya), 另一次发生于90-94 Mya, 这两次全基因组复制事件分别和白垩纪第三纪的Cretaceous-Tertiary (C-T)大灭绝事件以及翼盖蕨的物种分化时间相吻合。进一步对两次全基因组复制保留的基因进行功能注释和富集分析, 结果显示与转录及代谢调控相关的基因优势被保留。翼盖蕨的全基因组复制事件可能促进了该物种的分化及其对极端环境的适应性。     

四川都江堰迁地保育的42种杜鹃属植物开花–展叶物候节律

目前有研究表明被子植物类群中单一的开花或展叶物候模式可能具有遗传性,但两类物候事件的节律关系是否受遗传因素控制,各种节律模式的形成与演化机制及其与生物系统演化的关系仍不清楚。作者对四川都江堰市迁地保育的2个海拔点的杜鹃属(Rhododendron)4亚属4组13亚组42种植物的开花–展叶节律模式进行了5年的观察研究。作者假定杜鹃属的开花–展叶节律模式为进化遗传特性,包括先花后叶(F→L)、先花后叶部分重叠(F+L)、叶期包花期(L∈F)、先叶后花部分重叠(L+F)和先叶后花(L→F)等5种类型。同一地点不同年份以及不同地点同一物种的相关物候观察结果均显示,尽管它们5年的物候发生时间随年度和观察点海拔不同而有波动,但物候发生次序和开花–展叶节律模式均未见随年份而变化,即每个物种只对应一种节律模式,支持了上述假设。作者进一步提出了开花–展叶节律模式与物种的系统演化存在进化程度上的差异和序列关系的假设。不同节律模式的物种其区系地理以及不同海拔和分类群中的物种数量分布特征表明:(1)进化程度不同的节律模式分别对应于古老的中国–日本森林植物亚区和年轻的中国–喜玛拉雅森林植物亚区物种,较原始的节律模式倾向于出现在中高山海拔区域,并与杜鹃属可能起源山地的一定海拔区域构成关联,而进化程度较高的节律模式倾向于出现在更高或更低的海拔地段;(2)节律模式的进化程度与有关亚属、组、亚组的系统位置和进化程度有一定关联;(3)节律模式的演化与生活型、叶性进化同步,环境胁迫、生长期长度变化、光资源竞争等都可能是节律模式演化的外部动力。研究还表明,在气候波动条件下物候事件间的发生次序比单一物候事件的时间属性更加稳定。     

气候生态位进化与物种多样化的关系——以泛热带植物番荔枝科为例

热带地区较高的物种多样性与其气候条件有关,但气候如何影响热带地区物种的多样化却未有定论。为了解气候对热带植物多样性的影响,本研究以泛热带植物番荔枝科为研究对象,利用系统发育比较分析方法,计算了该科植物气候生态位的进化速率,并与该科的净多样化速率进行了相关性分析。结果显示:(1)番荔枝科气候生态位的进化速率较低,但新近分化类群的进化速率相对较高;(2)气候生态位下界(低温与低降水)的进化速率较上界(高温与高降水)快;(3)净多样化速率与气候生态位的进化速率,尤其是温度生态位的进化速率紧密相关。本研究揭示了番荔枝科植物气候生态位的进化对其物种多样性形成的重要作用,对该科在当前气候变化下的保护具有参考意义。     

被子植物成花诱导和性别决定机制的研究进展

开花植物具有多样性的生殖系统,其中单性花的形成是促进异交、避免自交衰退、保持遗传多样性的重要途径。单性花物种分布于被子植物不同进化分支上的事实表明,物种的雌雄异花性可能是通过不同的机制进化形成的。本文从花发育、性染色体、植物激素和环境因素四个方面,阐述了被子植物性别分化调控机制的研究进展。     

云南被子植物菊类分支的系统发育多样性及其分布格局

全球气候变化与人为活动等因素导致的生物多样性丧失,引起了全球各界对生物多样性保护的高度关注。传统生物多样性保护主要对物种、特有种、受威胁物种的种类组成及其分布模式开展研究,忽视了进化历史在生物多样性保护中的作用。云南是全球生物多样性热点地区的交汇区,生物多样性的保护历来受到广泛关注,为了更好地探讨云南生物多样性的保护措施,该研究以云南被子植物菊类分支物种为研究对象,基于物种间的演化关系,结合其地理分布,从进化历史的角度探讨物种、特有种、受威胁物种的种类组成及系统发育组成的分布格局,并整合自然保护地的空间分布,识别生物多样性的重点保护区域。结果表明：云南被子植物菊类分支的物种、特有种及受威胁物种的物种密度与系统发育多样性均显著正相关;通过零模型分析发现,由南向北标准化系统发育多样性逐渐降低;云南南部、东南部、西北部是云南被子植物菊类分支的重点保护区域,加强这些区域的保护,将最大化地保护生物多样性的进化历史和进化潜能。由此可见,融合进化历史信息的植物多样性格局分析不仅有助于更加深入地理解植物多样性的形成与演变,也为生物多样性保护策略的制定提供更多的思路。     

染色体数据的挖掘及其在植物多样性进化研究中的利用

多倍化(或全基因组加倍)是植物物种形成的重要途径,现存的被子植物可能都发生过一次甚至多次多倍化事件。多倍化传统的定义是染色体数目相对于祖先类群呈整倍性增加。其中最常用的研究方法是核型分析,核型能够提供物种的基本细胞学参数,包括染色体数目、倍性水平、核型不对称性、核型变异系数等。目前核型研究的趋势表现出从物种基本核型参数分析逐渐演化到多类群、多学科交叉融合的特点:一方面植物核型分析从种群、物种、科属的类群到生命之树,探讨染色体核型在各支系的进化特征、趋势以及驱动植物系统进化的细胞学机制;另一方面探讨和分析区域或生态系统植物区系的染色体谱或倍性等细胞学特征,可以探究区域地质环境变化或生态环境对染色体倍性等的影响,或通过区域染色体谱的构建,分析区域植物区系的形成和进化历史。因而,植物核型研究为系统发育、分子系统进化、生命之树以及植物区系地理的起源和演化研究提供了新思路。越来越多的新方法、新手段在植物核型分析与多倍化研究中得到运用,从而揭示了植物类群或植物区系的染色体进化以及细胞地理特征。今后植物细胞学研究趋势会向多学科交叉融合,整合各研究领域证据,从不同水平角度综合分析植物核型多样性形成的原因及意义,从而更加全面地认识和理解植物物种多样化与物种形成原因。     

植物CO基因研究进展

CO(constans)是植物开花时间光周期调控途径中的一个重要基因.目前从拟南芥、水稻、油菜、马铃薯等多个物种中都已经克隆到CO同源基因.CO基因在不同物种中具有保守的锌指结构和核定位区域,但是不同植物中的作用机理并不完全相同.序列分析表明该基因在被子植物与裸子植物之间、双子叶植物与单子叶植物之间以及不同科、属的植物之间均有明显分化,说明CO基因可能在植物进化中起到了重要作用.本文综述了近年来有关植物CO基因的研究进展,并对其在物种中的进化进行分析,为CO基因进一步研究提供参考.     

横断山区六种八居群鼠尾草属植物的核型分析

鼠尾草属(Salvia)是唇形科(Lamiaceae)最大的属,属下多种为民间常用草药,亦有供观赏的种类。为探究横断山区物种在细胞学水平的进化方式,讨论形态分类学与分子系统学之间的分类关系,该研究通过广泛收集染色体文献资料,采用植物常规压片法对采集自横断山地区6种8居群鼠尾草属植物进行核型分析,并构建了中国地区分布的鼠尾草属植物叶绿体系统发育树。统计结果表明:(1)全世界范围内报道了约23%的鼠尾草属植物染色体数据,其中分布在中国地区的鼠尾草属植物染色体报道率为32.10%,分布在横断山地区的鼠尾草属植物报道率为40.54%,(2)鼠尾草属植物染色体基数以x=8和x=11为主,分布在中国地区的鼠尾草属植物染色体基数均为x=8。实验结果表明:(1)西藏鼠尾草(S. wardii)核型数据为首次报道。(2)雪山鼠尾草(S. evansiana)首次在云南德钦地区发现二倍体居群。将细胞学数据结合叶绿体进化树开展染色体进化关联分析,论证多倍化可能不是鼠尾草属物种适应高海拔环境的主要机制,表明多倍体不是该属物种形成的主要进化途径而是以二倍体水平为主,推测染色体组的加倍可能是物种在形态学与分子系统学上分类关系不一致的原因之一。该研究丰富了横断山区鼠尾草属植物的染色体核型数据,结合区域分子系统树探讨染色体特征的进化关系,为今后深入研究该属物种的核型进化做出了探索,为开展祖先物种染色体基数推演分析补充了基础数据。     

Whole genome duplication of intra- and inter-chromosomes in the tomato genome

Whole genome duplication(WGD)events have been proven to occur in the evolutionary history of most angiosperms.Tomato is considered a model species of the Solanaceae family.In this study,we describe the details of the evolutionary process of the tomato genome by detecting collinearity blocks and dating the WGD events on the tree of life by combining two different methods:synonymous substitution rates(Ks)and phylogenetic trees.In total,593 collinearity blocks were discovered out of 12 pseudo-chromosomes constructed. It was evident that chromosome 2 had experienced an intra-chromosomal duplication event.Major inter-chromosomal duplication occurred among all the pseudo-chromosome.We calculated the Ks value of these collinearity blocks.Two peaks of Ks distribution were found,corresponding to two WGD events occurring approximately 36-82 million years ago(MYA)and 148-205 MYA.Additionally, the results of phylogenetic trees suggested that the more recent WGD event may have occurred after the divergence of the rosidasterid clade,but before the major diversification in Solanaceae.The older WGD event was shown to have occurred before the divergence of the rosid-asterid clade and after the divergence of rice-Arabidopsis(monocot-dicot).     

Recurrent genome duplication events likely contributed to both the ancient and recent rise of ferns

Ferns, the second largest group of vascular plants, originated ～400 mil ion years ago(Mya). They became dominant in the ancient Earth landscape before the angiosperms and are stil important in current ecosystems.Many ferns have exceptional y high chromosome numbers,possibly resulting from whole-genome duplications(WGDs).However, WGDs have not been investigated molecularly across fern diversity. Here we detected and dated fern WGDs using a phylogenomic approach and by calculating synonymous substitution rates(Ks). We also investigated a possible correlation between proposed WGDs and shifts in species diversification rates. We identified 19 WGDs: three ancient events along the fern phylogenetic backbone that are shared by 66%–97% of extant ferns, with additional lineage-specific WGDs for eight orders, providing strongevidence for recurring genome duplications across fern evolutionary history. We also observed similar Ks peak values for more than half of these WGDs, with multiple WGDs occurring close to the Cretaceous(～145–66 Mya). Despite the repeated WGD events, the biodiversity of ferns declined during the Cretaceous, implying that other factors probably contributed to the floristic turnover from ferns to angiosperms. This study provides molecular evidence for recurring WGDs in ferns and offers important clues to the genomic evolutionary history of ferns.     

ABSOLUTE DIVERSIFICATION RATES IN ANGIOSPERM CLADES

Abstract The extraordinary contemporary species richness and ecological predominance of flowering plants (angiosperms) are even more remarkable when considering the relatively recent onset of their evolutionary diversification. We examine the evolutionary diversification of angiosperms and the observed differential distribution of species in angiosperm clades by estimating the rate of diversification for angiosperms as a whole and for a large set of angiosperm clades. We also identify angiosperm clades with a standing diversity that is either much higher or lower than expected, given the estimated background diversification rate. Recognition of angiosperm clades, the phylogenetic relationships among them, and their taxonomic composition are based on an empirical compilation of primary phylogenetic studies. By making an integrative and critical use of the paleobotanical record, we obtain reasonably secure approximations for the age of a large set of angiosperm clades. Diversification was modeled as a stochastic, time‐homogeneous birth‐and‐death process that depends on the diversification rate (r) and the relative extinction rate (∈). A statistical analysis of the birth and death process was then used to obtain 95% confidence intervals for the expected number of species through time in a clade that diversifies at a rate equal to that of angiosperms as a whole. Confidence intervals were obtained for stem group and for crown group ages in the absence of extinction (∈= 0.0) and under a high relative extinction rate (∈= 0.9). The standing diversity of angiosperm clades was then compared to expected species diversity according to the background rate of diversification, and, depending on their placement with respect to the calculated confidence intervals, exceedingly species‐rich or exceedingly species‐poor clades were identified. The rate of diversification for angiosperms as a whole ranges from 0.077 (∈= 0.9) to 0.089 (∈= 0.0) net speciation events per million years. Ten clades fall above the confidence intervals of expected species diversity, and 13 clades were found to be unexpectedly species poor. The phylogenetic distribution of clades with an exceedingly high number of species suggests that traits that confer high rates of diversification evolved independently in different instances and do not characterize the angiosperms as a whole.     

Evolutionary Dynamics of Microsatellite Distribution in Plants: Insight from the Comparison of Sequenced Brassica,Arabidopsis and Other Angiosperm Species

Despite their ubiquity and functional importance, microsatellites have been largely ignored in comparative genomics, mostly due to the lack of genomic information. In the current study, microsatellite distribution was characterized and compared in the whole genomes and both the coding and non-coding DNA sequences of the sequenced Brassica, Arabidopsis and other angiosperm species to investigate their evolutionary dynamics in plants. The variation in the microsatellite frequencies of these angiosperm species was much smaller than those for their microsatellite numbers and genome sizes, suggesting that microsatellite frequency may be relatively stable in plants. The microsatellite frequencies of these angiosperm species were significantly negatively correlated with both their genome sizes and transposable elements contents. The pattern of microsatellite distribution may differ according to the different genomic regions (such as coding and non-coding sequences). The observed differences in many important microsatellite characteristics (especially the distribution with respect to motif length, type and repeat number) of these angiosperm species were generally accordant with their phylogenetic distance, which suggested that the evolutionary dynamics of microsatellite distribution may be generally consistent with plant divergence/evolution. Importantly, by comparing these microsatellite characteristics (especially the distribution with respect to motif type) the angiosperm species (aside from a few species) all clustered into two obviously different groups that were largely represented by monocots and dicots, suggesting a complex and generally dichotomous evolutionary pattern of microsatellite distribution in angiosperms. Polyploidy may lead to a slight increase in microsatellite frequency in the coding sequences and a significant decrease in microsatellite frequency in the whole genome/non-coding sequences, but have little effect on the microsatellite distribution with respect to motif length, type and repeat number. Interestingly, several microsatellite characteristics seemed to be constant in plant evolution, which can be well explained by the general biological rules.     

Unraveling the Distribution and Evolution of miR156 targeted SPLs in Plants by Phylogenetic Analysis

Squamosa promoter binding protein like genes (SPLs) are critical during plant development and mostly regulated by miR156. However, little is known about phylogenetic distribution and evolutionary patterns of miR156 targeted SPLs. In this study, 183 SPLs from nine genome sequenced species representing algae, bryophytes, lycophyte, monocots, and eudicots were computationally analyzed. Our results showed that miR156 responsive elements (MREs) on SPLs were present in land plants but absent from unicellular green algae. Phylogenetic analysis revealed that miR156 targeted SPLs only distributed in group II not group I of land plants, suggesting they originated from a common ancestor. In addition, group II were further divided into seven subgroups (IIa IIg) and miR156 targeted SPLs distributed in some specific members of SPLs from six subgroups except subgroup IId. Such distribution pattern was well elucidated by gene structure evolution of miR156 targeted SPLs based on the correlation of phylogenetic classification and gene structure. They could suffer from the exon loss events combined with MREs loss during evolution. Moreover, gene duplication contributed to the abundance of miR156 targeted SPLs, which had significantly increased after angiosperms and lower plants split. With Arabidopsis as the model species, we found segmental and tandem gene duplications predominated during miR156 targeted SPLs expansion. Taken together, these results provide better insights in understanding the function diversity and evolution of miR156 targeted SPLs in plants.     

miR156靶基因SPL在植物中的系统分布和进化模式分析

Squamosa promoterbinding proteinlike genes （SPLs）在植物发育过程中具有重要作用。很多SPLs被miR156调节,然而,对于它们在植物中的系统分布和进化模式还知之甚少。本文对9个测序物种（藻类,苔藓,石松,单子叶和双子叶植物）的183个SPLs进行了生物信息学分析。结果表明miR156应答元件（MREs）仅在陆生植物SPLs中发现,藻类中不存在。系统进化分析显示陆生植物SPLs分为两大分支：group I和group II。 MiR156靶基因仅分布于group II,表明它们有着共同的祖先。Group II进一步分为7个亚支（IIaIIg）,miR156靶基因分布在除IId外的其余6个亚支的特定SPLs。系统分类与基因结构的相关性反映了SPL靶基因结构上的变化。在进化过程中,它们可能发生外显子的丢失且伴随MRE的丢失。另外,基因重复对SPL靶基因的丰度变化影响很大,尤其是被子植物与低等植物分歧后它们数量明显增加。以拟南芥为模式植物分析发现串联重复和片段重复是SPL靶基因扩张的主要机制。     

Size is not everything: rates of genome size evolution,not C-value,correlate with speciation in angiosperms

Angiosperms represent one of the key examples of evolutionary success, and their diversity dwarfs other land plants; this success has been linked, in part, to genome size and phenomena such as whole genome duplication events. However, while angiosperms exhibit a remarkable breadth of genome size, evidence linking overall genome size to diversity is equivocal, at best. Here, we show that the rates of speciation and genome size evolution are tightly correlated across land plants, and angiosperms show the highest rates for both, whereas very slow rates are seen in their comparatively species-poor sister group, the gymnosperms. No evidence is found linking overall genome size and rates of speciation. Within angiosperms, both the monocots and eudicots show the highest rates of speciation and genome size evolution, and these data suggest a potential explanation for the megadiversity of angiosperms. It is difficult to associate high rates of diversification with different types of polyploidy, but it is likely that high rates of evolution correlate with a smaller genome size after genome duplications. The diversity of angiosperms may, in part, be due to an ability to increase evolvability by benefiting from whole genome duplications, transposable elements and general genome plasticity.     

Networks of gene sharing among 329 proteobacterial genomes reveal differences in lateral gene transfer frequency at different phylogenetic depths

Lateral gene transfer (LGT) is an important mechanism of natural variation among prokaryotes. Over the full course of evolution, most or all of the genes resident in a given prokaryotic genome have been affected by LGT, yet the frequency of LGT can vary greatly across genes and across prokaryotic groups. The proteobacteria are among the most diverse of prokaryotic taxa. The prevalence of LGT in their genome evolution calls for the application of network-based methods instead of tree-based methods to investigate the relationships among these species. Here, we report networks that capture both vertical and horizontal components of evolutionary history among 1,207,272 proteins distributed across 329 sequenced proteobacterial genomes. The network of shared proteins reveals modularity structure that does not correspond to current classification schemes. On the basis of shared protein-coding genes, the five classes of proteobacteria fall into two main modules, one including the alpha-, delta-, and epsilonproteobacteria and the other including beta- and gammaproteobacteria. The first module is stable over different protein identity thresholds. The second shows more plasticity with regard to the sequence conservation of proteins sampled, with the gammaproteobacteria showing the most chameleon-like evolutionary characteristics within the present sample. Using a minimal lateral network approach, we compared LGT rates at different phylogenetic depths. In general, gene evolution by LGT within proteobacteria is very common. At least one LGT event was inferred to have occurred in at least 75% of the protein families. The average LGT rate at the species and class depth is about one LGT event per protein family, the rate doubling at the phylum level to an average of two LGT events per protein family. Hence, our results indicate that the rate of gene acquisition per protein family is similar at the level of species (by recombination) and at the level of classes (by LGT). The frequency of LGT per genome strongly depends on the species lifestyle, with endosymbionts showing far lower LGT frequencies than free-living species. Moreover, the nature of the transferred genes suggests that gene transfer in proteobacteria is frequently mediated by conjugation.     

Transfer of chloroplast genomic DNA to mitochondrial genome occurred at least 300 MYA

With the completion of the first gymnosperm mitochondrial genome (mtDNA) from Cycas taitungensis and the availability of more mtDNA taxa in the past 5 years, we have conducted a systematic analysis of DNA transfer from chloroplast genomes (cpDNAs) to mtDNAs (mtpts) in 11 plants, including 2 algae, 1 liverwort, 1 moss, 1 gymnosperm, 3 monocots, and 3 eudicots. By using shared gene order and boundaries between different mtpts as the criterion, the timing of cpDNA transfer during plant evolution was estimated from the phylogenetic tree reconstructed independently from concatenated protein-coding genes of 11 available mtDNAs. Several interesting findings emerged. First, frequent DNA transfer from cpDNA to mtDNA occurred at least as far back as the common ancestor of extant gymnosperms and angiosperms, about 300 MYA. The oldest mtpt is trnV(uac)-trnM(cau)-atpE-atpB-rbcL. Three other mtpts--psaA-psaB, rps19-trnH(gug)-rpl2-rpl23, and psbE-psbF--were dated to the common ancestor of extant angiosperms, at least 150 MYA. However, all protein-coding genes of mtpts have degenerated since their first transfer. Therefore, mtpts contribute nothing to the functioning of mtDNA but junk sequences. We discovered that the cpDNA transfers have occurred randomly at any positions of the cpDNAs. We provide strong evidence that the cp-derived tRNA-trnM(cau) is the only mtpt (1 out of 3 cp-derived tRNA shared by seed plants) truly transferred from cpDNA to mtDNA since the time of the common ancestor of extant gymnosperms and angiosperms. Our observations support the proposition of Richly and Leister (2004) that "primary insertions of organellar DNAs are large and then diverge and fragment over evolutionary time."