植物LTR反转录转座子的研究进展

反转录转座子是真核生物基因组中普遍存在的一类可移动的遗传因子,它们以RNA为媒介,在基因组中不断自我复制。在高等植物中,反转录转座子是基因组的重要成分之一。反转录转座子可以分为5大类型,其中以长末端重复（LTR）类型报道较多。LTR类型由于其首尾具有长末端重复序列,内部含有PBS、PPT、GAG和POL开放阅读框、TSD等结构,可以采用生物信息学软件进行预测。LTR反转录转座子的活性受到自身甲基化和环境因素的影响,DNA甲基化抑制反转录转座子转座,而外界环境的刺激能够激活转座子,从而影响插入位点周边基因的表达。同时由于LTR反转录转座子在植物中普遍存在,丰富的拷贝数以及多态性为新型分子标记（RBIP、SSAP、IRAP、REMAP）的开发提供了良好的素材。该文对近年来国内外有关植物反转录转座子的类型、结构特征、 LTR反转录转座子的活性及其影响因素、 LTR反转录转座子的预测以及标记开发等方面的研究进展进行综述。     

植物LTR类反转录转座子序列分析识别方法

LTR类反转录转座子(Long terminal repeat retrotransponson)是真核生物中的一类重要转座元件,具有分布广泛、异质性高等特点,在真核生物基因组进化中起着重要作用,现广泛应用于植物的基因功能分析和遗传多样性研究等方面。LTR类反转录转座子的序列识别是其应用的前提条件,因此对LTR类反转录转座子的序列鉴定和分析方法的研究具有重要的理论意义和实际应用价值。LTR类反转录转座子序列的生物信息学分析软件按原理可大致分为序列比对分析和相关序列保守区域识别鉴定两类。比对软件如BLAST、DNAstar等,是一种序列相似性搜索程序,通过与已知的反转录转座子序列比对后的序列相似性来判断未知序列是否是反转录转座子序列,但这类软件不能直接获得具体的LTR等特征序列的相关信息,不能对反转录转座子序列的全长进行识别。识别鉴定软件按原理可分为从头算起法、比较基因组法、同源搜索法和结构基础法4种,如LTR-Finder等基于从头算起法的识别鉴定软件,可对LTR类反转录转座子全序列进行较准确地预测和注释,RepeatMasker等基于同源搜索法的软件,通过与数据库中的序列的相似性比对后发现可能存在的LTR类反转录转座子。文章对不同的LTR类反转录转座子预测方法进行了比较和分析,在此基础上归纳总结出一套分析LTR类反转录转座子序列的操作流程,旨在为LTR类反转录转座子序列的分析提供参考。     

四倍体海岛棉LTR反转录转座子的数量与分布

棉花是重要的纤维作物,在四个栽培棉种中,海岛棉纤维品质最优,了解LTR反转录转座子的数量与分布,可以促进海岛棉基因组的研究。通过综合不同方法挖掘海岛棉基因组中的LTR反转录转座子序列,并进行家族归类和数据分析。结果表明海岛棉A亚组和D亚组共有的LTR反转录转座子家族占全部家族的95%,LTR反转录转座子的Copia超家族和Gypsy超家族的分布特征有明显的不同,但相同超家族在相同亚组染色体上则表现出相似的特征。LTR反转录转座子周边基因的GO注释主要富集在结合活性、催化活性和代谢过程等方面。研究结果揭示了LTR反转录转座子在海岛棉染色体上的分布特征及其周边基因的功能富集。     

植物基因组中的非LTR反转录转座子SINEs和LINEs

反转录转座子是基因组进化的推动者之一。分为LTR和非LTR两种类型。前者是真核基因组的主要组分,结构和转座方式与逆转录病毒类似。后者是最初发现于动物基因组新近发现在植物基因组中也广泛存在的新型重复序列,包括LINEs(long interspersed nuclear elements)和SINEs(short interspersed nuclear elements)两个亚型。它们大多因自身或受宿主基因组的调控而失去转座活性。其转座机理目前还不十分清楚,推测LINEs可以自主转座,SINEs依赖其他转座子被动转座。种系分析认为LINEs可能是最古老的反转录转座子,SINEs的起源未知。文章对以上内容进行了归纳和讨论。     

白菜和甘蓝基因组转座子表达及其对基因调控的潜在影响

转座子或转座元件是大多数真核生物基因组的主要组成成分。甘蓝(Brassica oleracea)基因组比白菜(B. rapa)大主要是转座子的扩增差异造成的。然而, 这两个芸薹属近缘物种转座子表达水平以及对基因的调控和功能的影响目前还不清楚。文章对白菜和甘蓝叶、根、茎3个器官的转录组数据进行了初步分析。结果显示, 转座子的表达量很低, 转录组reads中有1%来自转座子的转录本; 转座子的表达存在器官差异, 且不同类别和家族的转座子表达量相差很大, 相同类别和同一家族的转座子在白菜和甘蓝基因组中的表达活性也不相同。进一步鉴定到转录读出的LTR反转座子, 其与下游基因距离小于2 kb的有41个, 小于100 bp的有9个, 这些LTR的转录读出很可能通过正义或反义的转录本激活或干扰下游基因的表达。同时, 具有转录读出的intact LTR比solo LTR具有更强的读出活性。通过深入分析转座子的插入位点发现, 白菜基因组中转座子插入基因内部的频率比甘蓝基因组中的高; 与反转座子相比, DNA转座子更偏向于插入或保留在基因的内含子当中。这些结果为认识转座子对其他蛋白编码基因的影响提供了基础。     

毛竹LTR反转录转座子PHRE6的克隆与转录活性分析

长末端重复序列(long terminal repeat,LTR)反转录转座子是真核生物基因组中普遍存在的一类可移动的DNA序列,因两端具有长末端重复序列而得名。大多数LTR反转录转座子能够感受外界环境的变化,具有转录激活特性和转座激活特性。该研究从毛竹(Phyllostachys edulis,Ph.edulis)基因组中克隆出一条完整的LTR反转录转座子,命名为PHRE6(Phyllostachys edulisretrotransposons 6),该转座子全长为5 620 bp,具有GAG和POL保守结构域。通过荧光定量PCR检测了PHRE6在DNA甲基化抑制剂和不同胁迫处理(包括辐照、高温、低温、高盐)的毛竹实生苗中转录水平的变化,结果表明,在DNA甲基化抑制剂处理后和高温(42°C)、低温(16°C、4°C)、高盐(100 mmol/L、200 mmol/L、300 mmol/L NaCl溶液)胁迫处理下PHRE6表达水平均有显著提高。以上结果说明,PHRE6是一个具有转录活性的反转录转座子,可能参与毛竹逆境响应过程。     

植物基因组中微型反向重复转座元件研究进展

微型反向重复转座元件(miniature inverted repeat transposable element,MITE)是一类特殊的转座元件,在结构上与有缺失的DNA转座子相似,但具有反转录转座子高拷贝数的特点.MITE时常与基因相伴,对基因调控可能起重要作用,因此,MITE正逐渐成为基因和基因组进化及生物多样性研究的一种重要工具.本文综述了植物基因组MITE的结构、分类、活性及其应用研究进展.     

美洲大蠊基因组重复序列分析

重复序列是真核生物基因组的重要组成部分。一些重复序列,如自主型的逆转录转座子LINE,在昆虫的系统进化和遗传多样性研究方面得到了广泛的应用。de novo从头预测和基于同源比对预测相结合的方法被用来搜索美洲大蠊Periplaneta americana基因组,共鉴定出大约占全基因组62%的重复序列。研究发现,散在重复序列中,DNA转座子占美洲大蠊基因组的16.18%;逆转座元件中LINE最多,占基因组的13.64%,SINE和LTR分别占基因组的3.52%和1.32%。LINEs中的Bov Bs亚家族在所有转座子亚家族中比例最高(约6.73%)。美洲大蠊与德国小蠊Blattella germanica相比,除LTR外,其他类型的转座子占基因组的比例均高于德国小蠊。通过分析逆转录转座子反转录酶完整度、氨基酸序列相似度及遗传距离,从美洲大蠊基因组中鉴定出一类BovBs:RTE-1_PAm。BovBs的反转录酶氨基酸序列的系统树表明,美洲大蠊与内华达古白蚁Zootermopsis nevadensis的进化关系比与其同属蜚蠊科Blattidae的德国小蠊的关系更近。昆虫中BovBs的进化关系与传统核基因进化关系的不同,表明转座子的进化相对宿主基因的进化具有一定的独立性。     

酵母Ty1和Ty3转座调控机制研究进展

LTR(Long Terminal Repetition, LTR)反转录转座子广泛存在于真核生物界,是逆转录病毒的进化祖先。LTR反转录转座子有两个古老的家族,Ty1/Copia和Ty3/Gypsy。目前关于LTR反转录转座子转座机制及调控机制研究最透彻的是来源于酵母的两个活性转座子Ty1和Ty3。全面综述了Ty1和Ty3的分子生物学机制相关的最新研究进展。系统总结了Ty1和Ty3的结构特征及转座特性,归纳了Ty1和Ty3与宿主共生的调控机制,为进一步了解酵母LTR反转录转座子相关转座调控机制提供参考。     

Young but not relatively old retrotransposons are preferentially located in gene‐rich euchromatic regions in tomato (Solanum lycopersicum) plants

Long terminal repeat (LTR) retrotransposons are the major DNA components of flowering plants. They are generally enriched in pericentromeric heterochromatin regions of their host genomes, which could result from the preferential insertion of LTR retrotransposons and the low effectiveness of purifying selection in these regions. To estimate the relative importance of the actions of these two factors on their distribution pattern, the LTR retrotransposons in Solanum lycopersicum (tomato) plants were characterized at the genome level, and then the distribution of young elements was compared with that of relatively old elements. The current data show that old elements are mainly located in recombination‐suppressed heterochromatin regions, and that young elements are preferentially located in the gene‐rich euchromatic regions. Further analysis showed a negative correlation between the insertion time of LTR retrotransposons and the recombination rate. The data also showed there to be more solo LTRs in genic regions than in intergenic regions or in regions close to genes. These observations indicate that, unlike in many other plant genomes, the current LTR retrotransposons in tomatoes have a tendency to be preferentially located into euchromatic regions, probably caused by their severe suppression of activities in heterochromatic regions. These elements are apt to be maintained in heterochromatin regions, probably as a consequence of the pericentromeric effect in tomatoes. These results also indicate that local recombination rates and intensities of purifying selection in different genomic regions are largely responsible for structural variation and non‐random distribution of LTR retrotransposons in tomato plants.     

Novel clades of chromodomain-containing Gypsy LTR retrotransposons from mosses (Bryophyta)

Retrotransposons are the major component of plant genomes. Chromodomain-containing Gypsy long terminal repeat (LTR) retrotransposons are widely distributed in eukaryotes. Four distinct clades of chromodomain-containing Gypsy retroelements are known from the vascular plants: Reina, CRM, Galadriel and Tekay. At the same time, almost nothing is known about the repertoire of LTR retrotransposons in bryophyte genomes. We have combined a search of chromodomain-containing Gypsy retroelements in Physcomitrella genomic sequences and an experimental investigation of diverse moss species. The computer-based mining of the chromodomain-containing LTR retrotransposons allowed us to describe four different elements from Physcomitrella. Four novel clades were identified that are evolutionarily distinct from the chromodomain-containing Gypsy LTR retrotransposons of other plants.     

Evidence of multiple horizontal transfers of the long terminal repeat retrotransposon RIRE1 within the genus Oryza

Horizontal gene transfer, defined as the transmission of genetic material between reproductively isolated species, has been considered for a long time to be a rare phenomenon. Most well-documented cases of horizontal gene transfer have been described in prokaryotes or in animals and they often involve transposable elements. The most abundant class of transposable elements in plant genomes are the long terminal repeat (LTR) retrotransposons. Because of their propensity to increase their copy number while active, LTR retrotransposons can have a significant impact on genomics changes during evolution. In a previous study, we showed that in the wild rice species Oryza australiensis , 60% of the genome is composed of only three families of LTR retrotransposons named RIRE1 , Wallabi and Kangourou . In the present study, using both in silico and experimental approaches, we show that one of these three families, RIRE1 , has been transferred horizontally between O. australiensis and seven other reproductively isolated Oryza species. This constitutes a new case of horizontal transfer in plants.     

利用iPBS技术克隆牡丹反转录转座子LTR序列

利用iPBS方法从西北牡丹(Paeonia suffruticosa)品种红绣球和中原牡丹品种洛阳红中扩增出相应片段,经回收、克隆及测序,获得了12条来自牡丹LTR类反转录转座子的LTR序列,并用相关生物信息学软件对序列进行分析。结果表明,这些核苷酸序列表现出较高的异质性,主要表现为缺失突变,序列长度变化范围为313–894 bp,同源性从31.1%–65.8%不等。将其氨基酸序列与已登录的不同植物LTR类反转录转座子LTR氨基酸序列进行聚类分析,结果显示与某些植物相应序列具有较高的同源性,表明可能存在LTR类反转录转座子的横向传递关系。根据克隆出的LTR序列设计SSAP引物,对牡丹29个品种进行了SSAP分子标记分析,结果显示具丰富的多态性。实验验证了用iPBS技术分离牡丹LTR序列的适用性,并为牡丹种质资源评价提供了新的技术手段。     

Retrotranspositions in orthologous regions of closely related grass species

Background  

Retrotransposons are commonly occurring eukaryotic transposable elements (TEs). Among these, long terminal repeat (LTR) retrotransposons are the most abundant TEs and can comprise 50–90% of the genome in higher plants. By comparing the orthologous chromosomal regions of closely related species, the effects of TEs on the evolution of plant genomes can be studied in detail.     

Ogre elements--a distinct group of plant Ty3/gypsy-like retrotransposons

Ogre elements are a group of LTR retrotransposons recently discovered in legume plants, where they constitute almost 40% of the genome in some species. They are exceptional in their size (reaching 25 kb) and possess several specific features, including an intron within a polyprotein-coding region, and an extra open reading frame (ORF1) encoding a protein of unknown function located upstream of the gag gene. Although these features make Ogres interesting for further research, identification of additional elements from a broader range of plant taxa has been complicated by the divergence of their sequences, preventing their detection using similarity-based searches. Here we report the results of structure-based computational searches for Ogre elements in available plant genomic sequences, which proved to be more efficient and revealed occurrences of Ogres in three families of dicot plants (Leguminosae, Solanaceae and Salicaceae). In addition, a representative set of 85 elements was retrieved from a model legume species Medicago truncatula. All identified full-length elements were used for comparative analysis, which showed that in spite of only little conservation of their nucleotide sequences, their protein domains were highly conserved, including several regions within ORF1. Further, the elements shared the same functional regions, including a primer binding site complementary to tRNA_arg, a conserved motif within a polypurine tract, and a putative intron between the pro and rt/rh coding domains. These findings, together with analysis of their phylogenetic relationship to other retrotransposons based on similarities of rt domains suggest that Ogre elements from different plant taxa have a common origin and thus constitute a distinct group of Ty3/gypsy retrotransposons.