Different expression patterns of duplicated PHANTASTICA-like genes in Lotus japonicus suggest their divergent functions during compound leaf development

INTRODUCTION The leaf organs of higher plants can be classified as simple or compound leaves. Compound leaves are found in distantly related groups, and differ from simple leaves in that each petiole bears multiple leaflets lacking auxiliary buds [1, 2]. …     

Analysis of expressed sequence tags of flower buds in Lotus japonicus.

In order to study gene expression in a reproductive organ, we constructed a cDNA library of mature flower buds in Lotus japonicus, and characterized expressed sequence tags (ESTs) of 842 clones randomly selected. The EST sequences were clustered into 718 non-redundant groups. From BLAST and FASTA search analyses of both protein and DNA databases, 58.5% of the EST groups showed significant sequence similarities to known genes. Several genes encoding these EST clones were identified as pollen-specific genes, such as pectin methylesterase, ascorbate oxidase, and polygalacturonase, and as homologous genes involved in pollen-pistil interaction. Comparison of these EST sequences with those derived from the whole plant of L. japonicus, revealed that 64.8% of EST sequences from the flower buds were not found in EST sequences of the whole plant. Taken together, the EST data from flower buds generated in this study is useful in dissecting gene expression in floral organ of L. japonicus.     

Genomic clustering of cyanogenic glucoside biosynthetic genes aids their identification in Lotus japonicus and suggests the repeated evolution of this chemical defence pathway

Cyanogenic glucosides are amino acid-derived defence compounds found in a large number of vascular plants. Their hydrolysis by specific β-glucosidases following tissue damage results in the release of hydrogen cyanide. The cyanogenesis deficient1 (cyd1) mutant of Lotus japonicus carries a partial deletion of the CYP79D3 gene, which encodes a cytochrome P450 enzyme that is responsible for the first step in cyanogenic glucoside biosynthesis. The genomic region surrounding CYP79D3 contains genes encoding the CYP736A2 protein and the UDP-glycosyltransferase UGT85K3. In combination with CYP79D3, these genes encode the enzymes that constitute the entire pathway for cyanogenic glucoside biosynthesis. The biosynthetic genes for cyanogenic glucoside biosynthesis are also co-localized in cassava (Manihot esculenta) and sorghum (Sorghum bicolor), but the three gene clusters show no other similarities. Although the individual enzymes encoded by the biosynthetic genes in these three plant species are related, they are not necessarily orthologous. The independent evolution of cyanogenic glucoside biosynthesis in several higher plant lineages by the repeated recruitment of members from similar gene families, such as the CYP79s, is a likely scenario.     

以可转化人工染色体(TAC)载体为基础的百脉根基因组文库的构建及筛选

可转化人工染色体(transformation-competentartificial chromosome,TAC)载体是具有克隆和转移大片段DNA特征的新型载体,是植物基因克隆和转化的有效工具.该研究把它用于豆科植物百脉根(Lotus japonicus)基因组文库的构建.此文库由1.8×105个克隆组成,平均插入片段大小为15kb左右,约覆盖百脉根基因组6倍.文库保存在12块96孔板中,每个孔中约含150个不同的重组克隆.用与花发育相关的同源基因Ljcen1片段为探针,筛选得到6个阳性克隆,酶切后验证这些阳性克隆,结果表明这些克隆含有同一个基因片段.此基因组文库可直接用于植物转化,为百脉根功能基因组的研究打下基础.     

Heterologous expression of flowering locus T promotes flowering but does not affect diurnal movement in the legume Lotus japonicus

Flowering locus T (FT) is known to promote flowering in response to photoperiodic conditions and has recently been shown to contribute to other phenomenon, such as diurnal stomatal movement. In legumes, FTs are classified into three subtypes, though the role of each subtype is not well defined. It has been reported that when FT of Lotus japonicus (LjFT) is heterologously expressed in Arabidopsis, LjFT functions as a mobile florigen to promote flowering, similar to Arabidopsis FT (AtFT). In this study, we expressed AtFT in L. japonicus using the SUC2 promoter and showed that heterologous expression of AtFT was able to promote flowering in the plant. We also showed that AtFT expression does not affect stomatal closing nor nyctinastic leaf movement. These findings contribute to our understanding of flower development and have potential application to breeding or plant biotechnology.     

Ecotypes of the model legume Lotus japonicus vary in their interaction phenotypes with the root-knot nematode Meloidogyne incognita

BACKGROUND AND AIMS: Knowledge of host factors affecting plant-nematode interactions is scarce. Here, relevant interaction phenotypes between a nodulating model host, Lotus japonicus, and the endoparasitic root-knot nematode Meloidogyne incognita are assessed via a genetic screen. METHODS: Within an alpha experimental design, 4-week-old replicate plants from 60 L. japonicus ecotypes were inoculated with 1000 nematodes from a single egg mass population, and evaluated for galling and nematode egg masses 6 weeks after inoculation. KEY RESULTS: Statistical analysis of data for 57 ecotypes showed that ecotype susceptibilities ranged from 3.5 to 406 galls per root, and correlated strongly (r = 0.8, P < 0.001, log scale) with nematode reproduction (ranging from 0.6 to 34.5 egg masses per root). Some ecotypes, however, showed a significant discrepancy between disease severity and nematode reproduction. Necrosis and developmental malformations were observed in other infected ecotypes. CONCLUSIONS: The first evidence is provided of significant variability in the interactions between L. japonicus and root-knot nematodes that may have further implications for the genetic dissection and characterization of host pathways involved in nematode parasitism and, possibly, in microbial symbiosis.     

Gametophytic expression of genes controlling endosperm development in maize

Summary An indirect approach was adopted to select viable mutants affecting the male gametophytic generation in maize. This approach consists of a selection of endosperm defective mutants followed by a test for gametophytic gene expression, based on the distortion from mendelian segregation and on the measurement of pollen size and pollen sterility. The material used consisted of 34 endosperm defective viable mutants introgressed in B37 genetic background. Complementation tests indicated that the mutation in the collection of mutants affected different genes controlling endosperm development. The study of the segregation in F2 revealed four classes of de (defective endosperm) mutants: (1) mutants in which the mutation does not affect either gametophytic development or function; (2) mutants in which the effect on the gametophyte affects pollen development processes; (3) mutants showing effects on both pollen development and function, and (4) mutants where only pollen tube growth rate is affected. Positive and negative interactions between pollen and style were detected by means of mixed pollination (pollen produced by de/de plants and pollen from an inbred line used as a standard and carrying genes for colored aleurone), on de/de and de/ + plants. Positive interactions were interpreted as methabolic complementation between defective pollen and normal styles.     

Evolution of nucleotide sequences and expression patterns of hydrophobin genes in the ectomycorrhizal fungus Paxillus involutus

Hydrophobins are small, secreted proteins that play important roles in the development of pathogenic and symbiotic fungi. Evolutionary mechanisms generating sequence and expression divergence among members in hydrophobin gene families are largely unknown. Seven hydrophobin (hyd) genes and one hyd pseudogene were isolated from strains of the ectomycorrhizal fungus Paxillus involutus. Sequences were analysed using phylogenetic methods. Expression profiles were inferred from microarray experiments. The hyd genes included both young (recently diverged) and old duplicates. Some young hyd genes exhibited an initial phase of enhanced sequence evolution owing to relaxed or positive selection. There was no significant association between sequence divergence and variation in expression levels. However, three hyd genes displayed a shift in the expression levels or an altered tissue specificity following duplication. The Paxillus hyd genes evolve according to the so-called birth-and-death model in which some duplicates are maintained for a long time, whereas others are inactivated through mutations. The role of subfunctionalization and/or neofunctionalization for preserving the hyd duplicates in the genome is discussed.     

Multiple ferritin subunit genes of the Pacific oyster Crassostrea gigas and their distinct expression patterns during early development

Multiple ferritin subunit genes are reported in mollusks, but they have not been systematically classified. Based on the recently published whole genome sequence, we screened out the four ferritin subunit genes (cgi-fer1–cgi-fer4) from the Pacific oyster Crassostrea gigas. The four genes were predicted to encode two non-secretory and two secretory peptides. Further phylogenetic analyses revealed two groups of non-secretory and secretory ferritin subunits in mollusks. This differs dramatically from the situation in mammals or insects, which contain only non-secretory or secretory ferritin subunits. These results emphasize the evolution of molluscan ferritin subunit genes. The expression patterns of the four genes during early development exhibited dramatic differences, indicating the functional diversity of these genes. Among them, cgi-fer2 was the only gene expressed prevalently and is thus suggested to be the major house-keeping ferritin subunit gene. The expression of the other three genes was tissue-specific beginning in the D-veliger stage. Based on their expression patterns, we inferred important functions of cgi-fer2 in ciliated tissues and of the other three genes in the digestive system. Moreover, our results indicated potentially different roles of ferritin subunit genes during larval shell formation in gastropods and bivalves, which may be helpful in understanding the molecular mechanisms that cause the different shells of gastropods and bivalves. In addition, we conducted a further semi-quantitative analysis of the four genes in four major developmental stages and five adult tissues. The results also revealed dramatically different expression patterns of the genes, which brought additional functional indications. This work may promote studies on molluscan ferritins and shed light on the evolution of ferritin subunit genes among different animal groups.