Jer-Ming Hu

Phylogenetic Relationships of Basal Papilionoid Legumes Based Upon Sequences of the Chloroplast trnL Intron

Abstract The Swartzieae, Sophoreae, Dipterygeae, and Dalbergieae are considered the most basal tribes of the subfamily Papilionoideae (Leguminosae). Nucleotide sequences from the chloroplast trnL intron for the majority of genera of these tribes were analyzed cladistically together with placeholder representatives of more derived tribes. Preliminary results indicate radical changes are necessary for papilionoid classification because Swartzieae, Sophoreae, and Dalbergieae are polyphyletic. Their constituent genera are mixed in a series of monophyletic groups, many of which have never been proposed previously, and the relationships amongst which are poorly resolved. Some of these groups, such as the genistoid and dalbergioid clades, are species-rich because they contain major papilionoid radiations. In other cases, putatively basal genera form small clades with no derived taxa included. There is weak evidence that Bobgunnia, Swartzia, Cyathostegia, Bocoa, and Ateleia (all Swartzieae) may be the sister group to all other papilionoids, and that a large clade is congruent with a 50kb inversion in the chloroplast large single copy (LSC) region. Communicating Editor: Kathleen A. Kron

LZF1, a HY5-regulated transcriptional factor, functions in Arabidopsis de-etiolation

We surveyed differential gene expression patterns during early photomorphogenesis in both wild-type and mutant Arabidopsis defective in HY5, an influential positive regulator of the responses of gene expression to a light stimulus, to identify light-responsive genes whose expression was HY5 dependent. These gene-expression data identified light-regulated zinc finger protein 1 (LZF1), a gene encoding a previously uncharacterized C2C2-CO B-box transcriptional regulator. HY5 has positive trans-activating activity toward LZF1 and binding affinity to LZF1 promoter in vivo. HY5 is needed but not sufficient for the induction of LZF1 expression. Anthocyanin content is significantly diminished in lzf1 under far red, which is the most efficient light for the induction of LZF1. The expression of PAP1/MYB75 is elevated in plants overexpressing LZF1, which leads to the hyperaccumulation of anthocyanin in transgenic Arabidopsis. The transition from etioplast to chloroplast and the accumulation of chlorophyll were notably compromised in the lzf1 mutant. We provide molecular evidence that LZF1 influences chloroplast biogenesis and function via regulating genes encoding chloroplast proteins. In the absence of HY5, mutation of LZF1 leads to further reduced light sensitivity for light-regulated inhibition of hypocotyl elongation and anthocyanin and chlorophyll accumulation. Our data indicate that LZF1 is a positive regulator functioning in Arabidopsis de-etiolation.

Phylogenetic Analysis of Nuclear Ribosomal ITS/5.8S Sequences in the Tribe Millettieae (Fabaceae): Poecilanthe-Cyclolobium, the core Millettieae, and the Callerya Group

Abstract The taxonomic composition of three principal and distantly related groups of the former tribe Millettieae, which were first identified from nuclear phytochrome and chloroplast trnK/matK sequences, was more extensively investigated with a phylogenetic analysis of nuclear ribosomal DNA ITS/5.8S sequences. The first of these groups includes the neotropical genera Poecilanthe and Cyclolobium, which are resolved as basal lineages in a clade that otherwise includes the neotropical genera Brongniartia and Harpalyce and the Australian Templetonia and Hovea. The second group includes the large millettioid genera, Millettia, Lonchocarpus, Derris, and Tephrosia, which are referred to as the “core Millettieae” group. Phylogenetic analysis of nuclear ribosomal DNA ITS/5.8S sequences reveals that Millettia is polyphyletic, and that subclades of the core Millettieae group, such as the New World Lonchocarpus or the pantropical Tephrosia and segregate genera (e.g., Chadsia and Mundulea), each form well supported monophyletic subgroups. The third lineage includes the genera Afgekia, Callerya, and Wisteria. These genera are resolved as a basal subclade in the inverted-repeat-lacking clade, which is a large legume group that includes the many well known temperate and herbaceous legumes, such as Astragalus, Medicago and Pisum, but not any other Millettieae. Communicating Editor: Jerrold I. Davis

Reassortment and Concerted Evolution in Banana Bunchy Top Virus Genomes

ABSTRACT The nanovirus Banana bunchy top virus (BBTV) has six standard components in its genome and occasionally contains components encoding additional Rep (replication initiation protein) genes. Phylogenetic network analysis of coding sequences of DNA 1 and 3 confirmed the two major groups of BBTV, a Pacific and an Asian group, but show evidence of web-like phylogenies for some genes. Phylogenetic analysis of 102 major common regions (CR-Ms) from all six components showed a possible concerted evolution within the Pacific group, which is likely due to recombination in this region. The CR-M of additional Rep genes is close to that of DNA 1 and 2. Comparison of tree topologies constructed with DNA 1 and DNA 3 coding sequences of 14 BBTV isolates showed distinct phylogenetic histories based on Kishino-Hasegawa and Shimodaira-Hasegawa tests. The results of principal component analysis of amino acid and codon usages indicate that DNA 1 and 3 have a codon bias different from that of all other genes of nanoviruses, including all currently known additional Rep genes of BBTV, which suggests a possible ancient genome reassortment event between distinctive nanoviruses.

The duplicated B-class MADS-box genes display dualistic characters in orchid floral organ identity and growth

Orchidaceae are an excellent model to examine perianth development because of their sophisticated floral architecture. In this study, we identified 24 APETALA3 (AP3)-like and 13 PISTILLA (PI)-like genes from 11 species of orchids and characterized them into four AP3- and two PI-duplicated homologs. The first duplication event in AP3 homologs occurring in the early evolutionary history of the Orchidaceae gave rise to AP3A and AP3B clades. Further duplication events resulted in four subclades, namely AP3A1, AP3A2, AP3B1 and AP3B2, during the evolution of Orchidaceae. The AP3 paralogous genes were expressed throughout inflorescence and floral bud development. From the in situ hybridization results, we noticed that the transition timings from ubiquitous to constrained expression in floral organs for both clades are different. The transition point of expression of the AP3A clade (clades 3 and 4) was at the late floral organ primordia stage. In contrast, that for the AP3B clade (clades 1 and 2) was not observed until the late inflorescence and floral bud stages. In addition, the AP3 orthologous genes revealed diverse expression patterns in various species of orchids, whereas the PI homologs were uniformly expressed in all floral whorls. AP3A2 orthologs play a noticeable role in lip formation because of their exclusive expression in the lip. Further evidence comes from the ectopic expression of AP3A2 detected in the lip-like petals extending from the lip in four sets of peloric mutants. Finally, a Homeotic Orchid Tepal (HOT) model is proposed, in which dualistic characters of duplicated B-class MADS-box genes are involved in orchid perianth development and growth.

Complete Plastid Genome Sequence of the Basal Asterid Ardisia polysticta Miq. and Comparative Analyses of Asterid Plastid Genomes

Ardisia is a basal asterid genus well known for its medicinal values and has the potential for development of novel phytopharmaceuticals. In this genus of nearly 500 species, many ornamental species are commonly grown worldwide and some have become invasive species that caused ecological problems. As there is no completed plastid genome (plastome) sequence in related taxa, we sequenced and characterized the plastome of Ardisia polysticta to find plastid markers of potential utility for phylogenetic analyses at low taxonomic levels. The complete A. polysticta plastome is 156,506 bp in length and has gene content and organization typical of most asterids and other angiosperms. We identified seven intergenic regions as potentially informative markers with resolution for interspecific relationships. Additionally, we characterized the diversity of asterid plastomes with respect to GC content, plastome organization, gene content, and repetitive sequences through comparative analyses. The results demonstrated that the genome organizations near the boundaries between inverted repeats (IRs) and single-copy regions (SCs) are polymorphic. The boundary organization found in Ardisia appears to be the most common type among asterids, while six other types are also found in various asterid lineages. In general, the repetitive sequences in genic regions tend to be more conserved, whereas those in noncoding regions are usually lineage-specific. Finally, we inferred the whole-plastome phylogeny with the available asterid sequences. With the improvement in taxon sampling of asterid orders and families, our result highlights the uncertainty of the position of Gentianales within euasterids I.

The Identification of A‐, B‐, C‐, and E‐Class MADS‐box Genes and Implications for Perianth Evolution in the Basal Eudicot Trochodendron aralioides (Trochodendraceae)

Trochodendron aralioides is the sole member of the family Trochodendraceae, which belongs to the basal eudicots, has vesselless wood, and lacks a distinct perianth. Our observations confirmed that there are numerous perianth‐like structures and that the number of these structures differs between protandrous and protogynous flowers and among the positions within an inflorescence. The epidermal cells on many floral parts of T. aralioides are papillate or conical, similar to the ones of ordinary showy petals of other species. The data in this article support the hypothesis that the perianth of Trochodendron has been secondarily lost and suggest that some aspects of petal identity, e.g., papillate cells, have been retained that might be important for pollinator attraction. We have identified 11 homologues of floral organ identity genes—two A‐class, three B‐class, two C‐class, and four E‐class homologous genes—from T. aralioides. Phylogenetic analysis shows that all of the genes arose before a major duplication of MADS‐box genes at the base of the core eudicots. Expression patterns for those floral organ identity gene homologues was determined by reverse transcriptase PCR, which showed variations that do not conform well to the current floral ABCDE model. In addition, all paralogous genes have distinct expression patterns, suggesting that they had undergone functional divergence.

Unusual Events Involved in Banana bunchy top virus Strain Evolution

Banana bunchy top virus (BBTV) can be transmitted by aphids and consists of at least six integral components (DNA-R, -U3, -S, -M, -C, and -N). Several additional replication-competent components (additional Reps) are associated with some BBTV isolates. A collected BBTV strain (TW3) that causes mild symptoms was selected to study the processes in BBTV evolution. Southern blot hybridization, polymerase chain reaction (PCR), and real-time PCR did not detect DNA-N in TW3. Real-time PCR quantification of BBTV components revealed that, except for the copy number of TW3 DNA-U3, each detected integral component of BBTV TW3 was at least two orders lower than that of the severe strains. No infection was observed in plants inoculated with aphids, which were first given acquisition access to the TW3-infected banana leaves. Recombination analysis revealed recombination between the integral component TW3 DNA-U3 and the additional Rep DNA-Y. All BBTV integral components contain a replication initiation region (stem-loop common region) that share high sequence identity. Sequence alignment revealed that TW3 DNA-R, -S, -M, and -C all have a stem-loop common region containing a characteristic 9-nucleotide deletion found only in all reported DNA-N. Our data suggest that the additional Rep DNAs can serve as sources of additional genetic diversity for integral BBTV components.

Morphology and phylogenetics of two holoparasitic plants, Balanophora japonica and Balanophora yakushimensis (Balanophoraceae), and their hosts in Taiwan and Japan

Balanophora japonica and B. yakushimensis are two putatively agamospermic taxa previously reported from southern Japan. Their inflorescences superficially represent those of B. laxiflora and B. fungosa. In this study we confirmed their presence in Taiwan by morphological and phylogenetic analysis using nuclear 18S rDNA and nrITS sequences with related taxa. B. japonica, B. yakushimensis, and B. laxiflora formed a well-supported clade that is distinct from other Balanophora. All three taxa also show considerable differences on morphological and nucleotide sequence differences, therefore the name of B. yakushimensis is retained. The results provide new insights on the intrageneric classification of Balanophora and suggest the positioning of female flowers should be down-weighted. We also successfully identify the hosts of B. japonica and B. yakushimensis by amplifying chloroplast matK sequences from the connected root tissues. The results showed that B. japonica parasitizes on Symplocos species, and that B. yakushimensis parasitizes on Distylium racemosum in Japan and Schima superba in Taiwan’s population.

The complete chloroplast genome of hemiparasitic flowering plant Schoepfia jasminodora

Abstract The complete chloroplast genome hemiparasitic plant Schoepfia jasminodora (Schoepfiaceae), was determined in this study by de novo assembly with whole-genome sequence data. The chloroplast genome is 118,743 bp in length with a typical quadripartite structure containing a pair of inverted repeats of 12,406 bp, separated by a large and a small single copy fragments of 84,168 bp and 9763 bp, respectively. The chloroplast genome contains 112 genes that consisting of 69 protein-coding, 27 tRNA, 4 rRNA and 3 pseudolized genes. All of the ndh (except for ndhA) and two tRNA (trnV-UAG and trnG-UCC) genes were found to be lost. The three pseudogenes are ndhA, ycf15 and trnL-CAA. Schoepfia represents the early stages of chloroplast genome degradation along with its transition to heterotrophy in related taxa.