Messages from Powdery Mildew DNA:How the Interplay with a Host Moulds Pathogen Genomes

The genomes of the barley,Arabidopsis and pea powdery mildew are signif icantly larger than those of related fungi.This is due to an extraordinary expansion of retro-trasposons that are evident as repetitive elements in the sequence.The protein coding genes are fewer than expected due to an overall reduction in the size of gene families,a reduction in the number of paralogs and because of the loss of certain metabolic pathways.Many of these changes have also been observed in the genomes of other taxonomically unrelated obligate biotrophic pathogens.The only group of genes that bucks the trend of gene loss,are those encoding small secreted proteins that bear the hall marks of effectors.     

Identification and Bioinformatics Analysis of SnRK2 and CIPK Family Genes in Sorghum

Through bioinformatic data mining, 10 SnRK2 and 31 CIPK genes were identified from sorghum genome. They are unevenly distributed in the sorghum chromosomes. Most SnRK2 genes have 8 introns, while the CIPK genes have a few （no intron or less than 3 introns） or more than I0 introns. Phylogenetic analysis revealed that SnRK2 genes belong to one cluster and CIPK genes form the other independent cluster. The sorghum SnRK2s are subgrouped into three parts, and CIPK into five parts. More than half SnRK2 and CIPK genes present in homologous pairs, suggesting gene duplication may be due to the amplification of SnRK family genes. The kinase domains of SnRK2 family are highly conserved with 88.40% identity, but those of the CIPK family are less conserved with 63.72% identity. And the identity of sorghum CBLinteracting NAF domains of CIPKs is 61.66%. What＇s more, regarding to the sorghum SnRK2 and CIPK kinases, they are characterized with distinct motifs and their subcellular localization is not necessarily the same, which suggests they may be divergent in functions. Due to less conserved sequences, complex subcellular localization, and more family members, sorghum CIPK genes may play more flexible and multiple biological functions. According to the phylogenetic analysis of SnRK genes and SnRK functional studies in other plants, it is speculated that sorghum SnRK2 and CIPK genes may play important roles in stress response, growth and development.     

Differential Gene Expression Between Hybrids and Their Parents During the Four Crucial Stages of Cotton Growth and Development

The study aims to clarify the differential gene expression between cotton hybrids and their parents in order to better understand the molecular basis of cotton heterosis. The research focused on cotton heterotic and lower heterotic hybrids and their parents during the four crucial stages, which were analyzed using a differential display technique. The results indicated that there were both quantitative and qualitative differences in gene expression amongst them. The quantitative differences include over- and under-expression of parental genes and the dominant expression of highly-expressed parental genes in hybrids. In contrast, the qualitative differences are the following： （i） Bands were observed in both parents but not in the F1 hybrid （BPnF1）; （ii） bands occurred in either of the parents but not in the F1 hybrid （UPnF1）; （iii） bands presented only in the F1 hybrid but not in either of the parents （UF1nP）; and （iv） bands were detected in either of the parents and the F1 hybrid （UPF1）. Overall, the major differences of gene expression occurred in the qualitative level and four related differential patterns were observed. Furthermore, the amount of differential patterns during the flowering stage was relatively higher than those of other stages. At this juncture, both the amount of hybrid-specific expression patterns at flowering stage and the silenced expression patterns at boll-forming stage in highly heterotic hybrids were found higher than those in the lower heterotic ones. It was concluded that significant differences of gene expression in leaves were present between cotton hybrid and its parents during the whole growing stages. Hence, these differences might be responsible for the observed cotton heterosis.     

Inheritance of Powdery Mildew Resistance in Cucumber （Cucumis sativus L.） and Development of an AFLP Marker for Resistance Detection

Cucumber powdery mildew is one of the most destructive diseases of cucumber throughout the world. In the present study, inheritance of powdery mildew resistance in three crosses, and linkage of resistance with amplified fragment length polymorphism （AFLP） markers are studied to formulate efficient strategies for breeding cultivars resistant to powdery mildew. The joint analysis of multiple generations and AFLP technique has been applied in this study. The best model is the one with two major genes, additive, dominant, and epistatic effects, plus polygenes with additive, dominant, and epistatic effects （E-l-0 model）. The heritabilities of the major genes varied from 64.26% to 97.82%, and susceptibility was incompletely dominant for the two major genes in the three crosses studied. The additive effects of the two major genes and the dominant effect of the second major gene were high, and the epistatic effect of the additive-dominant between the two major genes was the highest in cross I . In cross II, the absolute value of the additive effect, dominant effect, and potential ratio of the first major gene were far higher than those of the second major gene, and the epistatic effect of the additive-additive was the highest. The genetic parameters of the two major genes in cross III were similar to those in cross II. Correlation and regression analyses showed that marker E25/M63-103 was linked to a susceptible gene controlling powdery mildew resistance. The marker could account for 19.98% of the phenotypic variation. When the marker was tested on a diverse set of 29 cucumber lines, the correlation between phenotype and genotype was not significant, which suggested cultivar specialty of gene expression or different methods of resistance to powdery mildew. The target DNA fragment was 103 bp in length, and only a small part was found to be homologous to DNA in the other species evaluated, which indicated that it was unique to the cucumber genome.     

Cloning and Molecular Identification of A Fatty Acid Desaturase 2 Gene in a and C Genome of Brassica Species

The fatty acid desaturase 2 (fad2) gene was proven to be a major locus for high oleic acid (C18:1). Brassica napus is an amphidiploid species originating from a spontaneous hybridization of Brassica rapa and Brassica oleracea. B. napus contains multiple copies in genome for most of the genes, including fad2 genes. The research cloned nine fad2 genes from 3 varieties of B. rapa and 3 varieties of B. oleracea, respectively. Alignment of the nine fad2 sequences from B. rapa and B. oleracea detect-ed 6 single nucleotide polymorphic sites, which resulted in 6 amino-acid substitutions. The nucleotide substitutions at position 743 bp in the fad2-A gene and position 947 bp in the fad2-C gene were used as 3’ end of al ele-specific primers. In use of the al-lele-specific primers to amplify fad2 gene, we could identify if the fad2 gene originated from A genome or C genome. Besides, the research found that fad2 genes in C genome are more conserved in evolutionary process than those in A genome. The fad2 expression data reported in this study revealed that fad2-A from B. rapa was not only expressed in siliques same as fad2-C from B. oleracea, but also expressed in a high level in stems. Not even the less, fad2 gene from B. napus was expressed higher in roots and flowers. Al these results provided evidences that fad2, though it was expressed differently in B. rapa and B. oleracea, but it was regulated by the same approach in B. napus.     

Characterization of the Promoter of a Homolog of Maize MADS-Box Gene m18

Maize （Zea mays L.） is one of the world’s major food crops, and often suffers from tremendous yield loss caused by abiotic stresses. The MADS-box genes are known to play versatile roles in plants, controlling plant responses to multiple abiotic stresses. However, understanding of regulation of their expressions by the conventional loss-of-function approach is very dififcult. So far, regulation of MADS-box gene expression is little known. The best approach to retrieve expression regulation of this category of genes is to characterize expression of their promoters. In this study, the promoter of a homolog （GenBank accession no. EC864166） of maize MADS-box gene m18 was cloned by way of genome-walking PCR, named Pro66. Predicative analysis indicated that Pro66 contains more than one TATA box and multiple cis-acting environmental conditions-responsive elements （ECREs）. Pro66 could drive expression of theβ-glucuronidase （GUS）-encoding gene in maize, and heterologous expression of GUS in red pepper stressed by water deifcit, salt, copper, iron deifciency, heat, cold, and grown under short and long photoperiods, echoing predicative ECREs. Conclusively, maize MADS-box gene m18 likely plays versatile functions in maize response to multiple abiotic stresses due to the promoter with multiple cis-acting elements. The complex arrangement of multiple cis-acting elements in the promoter features meticulously regulated expression of m18. The results give informative clues for heterologous utilisation of the promoters in monocot and dicot species. The copy of the ECREs and heterologous expression of the promoter in dicot species are also discussed.     

The impact of tandem duplication on gene evolution in Solanaceae species

Whole genome duplication(WGD) and tandem duplication(TD) are important modes of gene amplification and functional innovation, and they are common in plant genome evolution. We analyzed the genomes of three Solanaceae species(Solanum lycopersicum, Capsicum annuum, and Petunia inflata), which share a common distant ancestor with Vitis vinifera, Theobroma cacao, and Coffea canephora but have undergone an extra whole genome triplication(WGT) event. The analysis was used to investigate the phenomenon...     

Phylogenetics and the cohesion of bacterial genomes

Gene acquisition is an ongoing process in many bacterial genomes, contributing to adaptation and ecological diversification. Lateral gene transfer is considered the primary explanation for discordance among gene phylogenies and as an obstacle to reconstructing the tree of life. We measured the extent of phylogenetic conflict and alien-gene acquisition within quartets of sequenced genomes. Although comparisons of complete gene inventories indicate appreciable gain and loss of genes, orthologs available for phylogenetic reconstruction are consistent with a single tree.     

50 million years of genomic stasis in endosymbiotic bacteria

Comparison of two fully sequenced genomes of Buchnera aphidicola, the obligate endosymbionts of aphids, reveals the most extreme genome stability to date: no chromosome rearrangements or gene acquisitions have occurred in the past 50 to 70 million years, despite substantial sequence evolution and the inactivation and loss of individual genes. In contrast, the genomes of their closest free-living relatives, Escherichia coli and Salmonella spp., are more than 2000-fold more labile in content and gene order. The genomic stasis of B. aphidicola, likely attributable to the loss of phages, repeated sequences, and recA, indicates that B. aphidicola is no longer a source of ecological innovation for its hosts.     

Rediscovery and analysis of Phytophthora carbohydrate esterase (CE) genes revealing their evolutionary diversity

A continuous co-evolutionary arms-race between pathogens and their host plants promotes the development of pathogenic factors by microbes, including carbohydrate esterase (CE) genes to overcome the barriers in plant cell walls. Identification of CEs is essential to facilitate their functional and evolutionary investigations; however, current methods may have a limit in detecting some conserved domains, and ignore evolutionary relationships of CEs, as well as do not distinguish CEs from proteases. Here, candidate CEs were annotated using conserved functional domains, and orthologous gene detection and phylogenetic relationships were used to identify new CEs in 16 oomycete genomes, excluding genes with protease domains. In our method, 41 new putative CEs were discovered comparing to current methods, including three CE4, 14 CE5, eight CE12, five CE13, and 11 CE14. We found that significantly more CEs were identified in Phytophthora than in Hyaloperonospora and Pythium, especially CE8, CE12, and CE13 that are putatively involved in pectin degradation. The abundance of these CEs in Phytophthora may be due to a high frequency of multiple-copy genes, supporting by the phylogenetic distribution of CE13 genes, which showed five units of Phytophthora CE13 gene clusters each displaying a species tree like topology, but without any gene from Hyaloperonospora or Pythium species. Additionally, diverse proteins associated with products of CE13 genes were identified in Phytophthora strains. Our analyses provide a highly effective method for CE discovery, complementing current methods, and have the potential to advance our understanding of function and evolution of CEs.     

Comparative genomics of trypanosomatid parasitic protozoa

A comparison of gene content and genome architecture of Trypanosoma brucei, Trypanosoma cruzi, and Leishmania major, three related pathogens with different life cycles and disease pathology, revealed a conserved core proteome of about 6200 genes in large syntenic polycistronic gene clusters. Many species-specific genes, especially large surface antigen families, occur at nonsyntenic chromosome-internal and subtelomeric regions. Retroelements, structural RNAs, and gene family expansion are often associated with syntenic discontinuities that-along with gene divergence, acquisition and loss, and rearrangement within the syntenic regions-have shaped the genomes of each parasite. Contrary to recent reports, our analyses reveal no evidence that these species are descended from an ancestor that contained a photosynthetic endosymbiont.     

通过多基因组比较的方法在5种绿球蓝细菌中识别基因组岛(英文)

为探索水平基因转移在绿球蓝细菌基因组多样性中的作用,5个高光适应性绿球蓝细菌基因组被比较.先用BLASTP两两最佳匹配搜索和基因位置保守性的方法预测直系同源基因,并进一步计算每个基因组中所有基因在其他4种绿球蓝细菌基因组中直系同源基因的数量,从而达到识别外源基因的目的;而基因组岛是指由1个或几个连续外源基因组成的区域.结果表明:在5个绿球蓝细菌基因组中共识别出了16至52个不等的基因组岛,它们中的许多基因在病毒基因组或富含病毒的环境样本中都有同源基因.以上这些结果显示,在绿球蓝细菌中,病毒可能介导了基因的水平转移,从而导致了基因组的多样性.     

Metabolic diversification--independent assembly of operon-like gene clusters in different plants

Operons are clusters of unrelated genes with related functions that are a feature of prokaryotic genomes. Here, we report on an operon-like gene cluster in the plant Arabidopsis thaliana that is required for triterpene synthesis (the thalianol pathway). The clustered genes are coexpressed, as in bacterial operons. However, despite the resemblance to a bacterial operon, this gene cluster has been assembled from plant genes by gene duplication, neofunctionalization, and genome reorganization, rather than by horizontal gene transfer from bacteria. Furthermore, recent assembly of operon-like gene clusters for triterpene synthesis has occurred independently in divergent plant lineages (Arabidopsis and oat). Thus, selection pressure may act during the formation of certain plant metabolic pathways to drive gene clustering.     

Comparative genome and proteome analysis of Anopheles gambiae and Drosophila melanogaster

Comparison of the genomes and proteomes of the two diptera Anopheles gambiae and Drosophila melanogaster, which diverged about 250 million years ago, reveals considerable similarities. However, numerous differences are also observed; some of these must reflect the selection and subsequent adaptation associated with different ecologies and life strategies. Almost half of the genes in both genomes are interpreted as orthologs and show an average sequence identity of about 56%, which is slightly lower than that observed between the orthologs of the pufferfish and human (diverged about 450 million years ago). This indicates that these two insects diverged considerably faster than vertebrates. Aligned sequences reveal that orthologous genes have retained only half of their intron/exon structure, indicating that intron gains or losses have occurred at a rate of about one per gene per 125 million years. Chromosomal arms exhibit significant remnants of homology between the two species, although only 34% of the genes colocalize in small "microsyntenic" clusters, and major interarm transfers as well as intra-arm shuffling of gene order are detected.     

Widespread lateral gene transfer from intracellular bacteria to multicellular eukaryotes

Although common among bacteria, lateral gene transfer-the movement of genes between distantly related organisms-is thought to occur only rarely between bacteria and multicellular eukaryotes. However, the presence of endosymbionts, such as Wolbachia pipientis, within some eukaryotic germlines may facilitate bacterial gene transfers to eukaryotic host genomes. We therefore examined host genomes for evidence of gene transfer events from Wolbachia bacteria to their hosts. We found and confirmed transfers into the genomes of four insect and four nematode species that range from nearly the entire Wolbachia genome (>1 megabase) to short (<500 base pairs) insertions. Potential Wolbachia-to-host transfers were also detected computationally in three additional sequenced insect genomes. We also show that some of these inserted Wolbachia genes are transcribed within eukaryotic cells lacking endosymbionts. Therefore, heritable lateral gene transfer occurs into eukaryotic hosts from their prokaryote symbionts, potentially providing a mechanism for acquisition of new genes and functions.     

药敏试验与比较基因组揭示嗜水气单胞菌耐药潜力

为了揭示嗜水气单胞菌的耐药潜力,运用微量肉汤稀释法和比较基因组学方法对其进行了分析。结果显示：恩诺沙星、硫酸新霉素、氟苯尼考、盐酸多西环素对3株菌株有抑制作用,而磺胺间甲氧嘧啶对3株菌株均无抑制作用。菌株的耐药表型未与耐药基因一一对应,某些菌株虽然未注释到耐药基因,也可以表现出相关耐药表型;自测的3株菌株基因组大小差异较大,亲缘关系非近缘,但耐药基因数量基本一致;经ANI（平均核苷酸一致性）计算后,基因组比较分析发现嗜水气单胞菌基因组具有较强的可变性,即使是近缘菌株,其基因组间仍有较大差异;系统发育分析显示,不同宿主来源的菌株在进化树上呈现分散分布,近缘菌株分布在不同国家和地区;所有菌株在CARD（综合抗生素耐药性数据库）中共注释到21种耐药基因,其中7种耐药基因为所有菌株共享。造成这一现象的原因,很可能是由于嗜水气单胞菌基因组较强的可变性以及其宿主分布的广泛性,菌株间进行频繁的基因交流,促进了耐药基因在整个物种的快速扩散,所以即使菌株间基因组变化较大,但耐药基因在菌株间的变化较小。