Horticulture,hybrid cultivars and exotic plant invasion: a case study of Wisteria (Fabaceae)

Exotic Wisteria species are highly favoured for their horticultural qualities and have been cultivated in North America since the early 1800s. This study determines the identity, genetic diversity and hybrid status of 25 Asian Wisteria cultivars using plastid, mitochondrial and nuclear DNA data. Fifteen (60%) hybrid cultivars were identified. All of the ‘Wisteria sinensis’ cultivars sampled are hybrids with W. floribunda. Although W. sinensis and W. floribunda are recognized invasive species in the southeastern USA, the relationships of horticultural cultivars to naturalized plants was previously unknown. Haplotype analysis of nuclear data identifies four haplotypes shared between cultivated stock and naturalized populations in the southeastern USA. In addition, US invasive haplotypes are present in New Zealand‐derived cultivars although, to date, naturalized Wisteria has not been documented in New Zealand. Finally, these data are used to make recommendations to horticulturalists of select species cultivars which may be less likely to invade US landscapes. © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society, 2008, 158 , 593–601.     

紫藤脉花叶病毒cp基因在大肠杆菌中的表达及抗血清的制备

采用RT-PCR方法自紫藤脉花叶病毒北京分离物(WVMV-BJ)的基因组中分离出其CP基因,连接到原核表达载体pET22b(+)上.获得的重组子pET-WVMVCP转化大肠杆菌BL21(DE3)后,用IPTG进行诱导表达.SDS-PAGE和Western blot分析表明,cp基因在大肠杆菌中获得了高效表达,融合蛋白分子量约为34.4 kDa.将融合蛋白纯化后免疫兔子,获得了特异性较高的抗血清.微量免疫沉淀法测定该抗血清的效价为1/1024,酶联法(enzyme-linkedimmunosorbant assay,ELISA)测定的效价为1/8192.     

紫藤脉花叶病毒cp基因在大肠杆菌中的表达及抗血清的制备

采用RT-PCR方法自紫藤脉花叶病毒北京分离物(WVMV-BJ)的基因组中分离出其CP基因,连接到原核表达载体pET22b( )上。获得的重组子pET-WVMVCP转化大肠杆菌BL21(DE3)后,用IPTG进行诱导表达。SDS-PAGE和Western blot分析表明,cp基因在大肠杆菌中获得了高效表达,融合蛋白分子量约为34．4kDa。将融合蛋白纯化后免疫兔子,获得了特异性较高的抗血清。微量免疫沉淀法测定该抗血清的效价为1／1024,酶联法(enzyme-linked immunosorbant assay,ELISA)测定的效价为1／8192。     

Bi-parental cytoplasmic DNA inheritance in Wisteria (Fabaceae): evidence from a natural experiment

Cytoplasmic inheritance was investigated in interspecific hybrids of Wisteria sinensis and W. floribunda. Species-specific nuclear, mitochondrial and plastid DNA markers were identified from wild-collected plants of each species in its native range. These markers provide evidence for the bi-parental transmission of plastids in hybrid swarms of these two species in the southeastern USA. These population level molecular data corroborate previous cytological evidence of this phenomenon in Wisteria.     

不同土壤水分条件下紫藤叶片生理参数的光响应

测定了不同土壤湿度下2年生紫藤叶片光合速率（P_n）、蒸腾速率（T_r）及水分利用效率（WUE）等生理参数的光响应过程,探讨了紫藤正常生长发育所需的土壤水分和光照条件.结果表明:紫藤叶片的P_n、T_r及WUE对土壤湿度和光照强度的变化具有明显的阈值响应.维持紫藤正常生长（同时具有较高P_n和WUE）的土壤湿度范围为:体积含水量（W_v）15.3%～26.5%、相对含水量（W_r）46.4%～80.3%,最佳土壤湿度约为W_v 23.3%、W_r 70.6%.紫藤叶片对光照环境的适应性较强,在光合有效辐射强度（PAR）为600～1 600 μmol·m^-2·s^-1时,P_n和WUE具有较高水平,饱和光强在PAR为800～1 000 μmol·m^-2·s^-1.紫藤叶片光合作用非气孔限制的发生与土壤湿度与光照强度密切相关,W_v为18.4%～26.5%、W_r为55.8%～80.3%时,光合作用主要受气孔限制,光照强度的影响较小;超出此范围后,其受光照强度的影响较大,出现由气孔限制转变为非气孔限制的PAR临界值.紫藤正常生长允许的最低土壤湿度约为W_v 11.9%、W_r 36.1%,允许最高PAR约为1 000 μmol·m^-2·s^-1,是紫藤叶片光合机构受到破坏的临界点.     

Large‐scale identification of secretome glycoproteins recognized by Wisteria floribunda agglutinin: A glycoproteomic approach to biomarker discovery

One of the promising approaches to the development of cancer diagnostic systems is quantification of a specific protein carrying cancerous glycans. Potential utility of Wisteria floribunda agglutinin (WFA) for such assays has been suggested for several cancer types. To develop such diagnostic systems, identification of WFA‐recognized glycoproteins is essential. Here, we successfully identified 504 WFA‐recognized glycoproteins from the secretome of HEK293T cells. Most of the identified proteins were likely soluble or single‐pass transmembrane proteins, which may serve as specific proteins for the diagnosis using biological fluids. Our method may help to discover marker glycoproteins for various cancers generating WFA‐recognized glycans.     

Wisteria floribunda agglutinin positive glycobiomarkers: a unique lectin as a serum biomarker probe in various diseases

Introduction: Serum proteins are generally glycosylated and solubilized, and are thus present as glycoproteins. The glycan structure of glycoproteins reflects cell differentiation status; glycan structures generated by diseased cells are distinguishable from those produced by healthy cells. Proteins may therefore serve as markers of tissues that secrete them. Several strategies for the identification of novel serum biomarkers using a combination of glycoscience-based technologies have been recently proposed. The selection of lectins for use as probes for identification of altered glycan structures represents a critical step.

Areas covered: This review describes the identification of Wisteria floribunda agglutinin (WFA) as a probe that recognizes the altered glycan structure of glycoproteins secreted by diseased cells. WFA may be employed as a probe for several diseases, e.g., liver fibrosis, liver cirrhosis, prostate cancer, ovarian cancer, and IgA nephropathy. The advantage of employing WFA as a serum biomarker probe is that only very small amounts of WFA-positive glycoproteins are present in serum; therefore, WFA background in serum is very low.

Expert commentary: Based on the findings to date, several WFA-positive serum biomarkers may be measured without pre-purification of target glycoproteins, indicating their utility as serum biomarkers in patients with various diseases.     

重瓣多花紫藤(豆科),一种中国新记录园艺植物

首次报道一种中国新记录园艺植物,重瓣多花紫藤[Wisteria floribunda(Willd.) DC. f. violaceo-plena(Schneider) Rehder & Wilson].     

New pterocarpon flavonoids from Wisteria sinensis tumor

Three new pterocarpon flavonoids, wisterones B-D, together with nine isoflavone known compounds, were isolated from Wisteria sinensis Tumor. The structure of those compounds were elucidated on the basis of spectroscopic analyses, including UV, IR, MS and NMR experiments. These new compounds were investigated for their activities against anti-crop pathogenic fungi.     

Potential cytoplasmic inheritance in Wisteria sinensis and Robinia pseudoacacia (Leguminosae)

We examined pollen cells of Wisteria sinensis and Robinia pseudoacacia (Leguminosae) to determine a possible mode for cytoplasmic inheritance in these species. Epifluorescence microscopy revealed distinct mature generative cells. Mature generative cells of W. sinensis were associated with large numbers of punctuated fluorescent signals corresponding to cytoplasmic DNA aggregates, but no fluorescent signals were observed in the generative cells of R. pseudoacacia. Closer examination showed that the punctate fluorescent signals corresponded to plastid but not mitochondrial DNA. These results suggest a strong potential for paternal transmission of the plastid genome in W. sinensis. Electron microscopy confirmed the presence of plastids in the generative cells of W. sinensis and the absence of plastids in R. pseudoacacia cells due to an unequal distribution of plastids during the first pollen mitosis. Mitochondria were present and intact in the mature generative cells of both species. The lack of fluoresced mitochondrial DNA suggests a very low level of mitochondrial DNA in the cells. Immunoelectron microscopy demonstrated that the labeling of mitochondrial DNA in these cells was reduced by nearly 90% during pollen development. Such a dramatic reduction suggests an active degradation of paternal mitochondrial DNA, which may contribute greatly to the maternal inheritance of mitochondria. In short, we found that W. sinensis exhibits a strong potential for paternal transmission of plastids and that both W. sinensis and R. pseudoacacia appear to share the same mechanism for maternal mitochondrial inheritance.     

Molecular systematics and biogeography of Wisteria inferred from nucleotide sequences of nuclear and plastid genes

Previous molecular phylogenetic studies of Fabaceae indicated that species of Wisteria, an intercontinental disjunct genus between eastern Asia and eastern North America, formed a clade derived from within Callerya. However, interspecific relationships were not well resolved or supported. In this study, we used sequences of the nuclear ribosomal DNA internal transcribed spacer region and the chloroplast gene matK to examine interspecific relationships and explore implications of the phylogeny for the systematics and biogeography of Wisteria. Our results showed that Wisteria with deciduous leaves and racemose inflorescences formed a strongly supported clade derived from within the paraphyletic Callerya. Afgekia was also found to be included within Callerya. Therefore, our data support the merger ofAfgekia, Callerya, and Wisteria. The phylogenetic pattern suggested that the deciduousness in Wisteria may be a derived trait likely in response to temperate climate, and the racemose inflorescences in the Afgekia–Callerya–Wisteria clade may have evolved from panicles. Our study also provided strong support for the sister relationship of the North American and eastern Asian species of Wisteria. In the Asian clade, Wisteria brachybotrys Siebold & Zucc. of Japan was sister to the clade containing W. floribunda (Willd.) DC of Japan and Korea, and W. sinensis (Sims) Sweet of China. However, our data offered weak support for the sister relationship ofW. floribunda and W. sinensis. Our divergence time and biogeographic analyses suggested that the eastern Asian–North American disjunction in Wisteria may have occurred through a dispersal event in the middle Miocene (13.4 Mya) from the Old World to the New World across the Bering land bridge followed by vicariance in the late Miocene (6.8 Mya). This study added another example to the “out of Asia” migration for the eastern Asian–eastern North American disjunction.     

五种攀缘植物光合作用与光因子关系的初步研究

城市垂直绿化常用的攀缘植物爬山虎、美国爬山虎、紫藤、常春藤、洋常春藤的净光合速率、呼吸速率、光能利用率及光补偿点等指标进行了测定和分析．结果表明,5种攀缘植物的净光合速率日变化显示出“双峰”。第一峰值出现在10：00～12：00,第二峰值出现在14：00～16：00,“午休”现象明显;呼吸速率的日变化也表现出“双峰”,第一峰值出现在11：00～13：00,第二峰值出现在14：00～17：00;常春藤、洋常春藤、爬山虎、美国爬山虎及紫藤的光补偿点分别为5．73、5．07、9．96、6．40和18．93μmol·m^-2·s^-1;紫藤在强光下的光能利用率较高,其次为美国爬山虎,而在弱光下洋常春藤光能利用率较高,可以认为,紫藤为典型的阳性植物,爬山虎、美国爬山虎为中性植物,常春藤、洋常春藤为阴性植物。     

Terminal N-Acetylgalactosamine-Specific Leguminous Lectin from Wisteria japonica as a Probe for Human Lung Squamous Cell Carcinoma

Millettia japonica was recently reclassified into the genus Wisteria japonica based on chloroplast and nuclear DNA sequences. Because the seed of Wisteria floribunda expresses leguminous lectins with unique N-acetylgalactosamine-binding specificity, we purified lectin from Wisteria japonica seeds using ion exchange and gel filtration chromatography. Glycan microarray analysis demonstrated that unlike Wisteria floribunda and Wisteria brachybotrys lectins, which bind to both terminal N-acetylgalactosamine and galactose residues, Wisteria japonica lectin (WJA) specifically bound to both α- and β-linked terminal N-acetylgalactosamine, but not galactose residues on oligosaccharides and glycoproteins. Further, frontal affinity chromatography using more than 100 2-aminopyridine-labeled and p-nitrophenyl-derivatized oligosaccharides demonstrated that the ligands with the highest affinity for Wisteria japonica lectin were GalNAcβ1-3GlcNAc and GalNAcβ1-4GlcNAc, with K _a values of 9.5 × 10⁴ and 1.4 × 10⁵ M^-1, respectively. In addition, when binding was assessed in a variety of cell lines, Wisteria japonica lectin bound specifically to EBC-1 and HEK293 cells while other Wisteria lectins bound equally to all of the cell lines tested. Wisteria japonica lectin binding to EBC-1 and HEK293 cells was dramatically decreased in the presence of N-acetylgalactosamine, but not galactose, mannose, or N-acetylglucosamine, and was completely abrogated by β-hexosaminidase-digestion of these cells. These results clearly demonstrate that Wisteria japonica lectin binds to terminal N-acetylgalactosamine but not galactose. In addition, histochemical analysis of human squamous cell carcinoma tissue sections demonstrated that Wisteria japonica lectin specifically bound to differentiated cancer tissues but not normal tissue. This novel binding characteristic of Wisteria japonica lectin has the potential to become a powerful tool for clinical applications.     

山东临朐中中新世山旺组紫藤属(豆科)荚果化石的再观察

豆科紫藤属Wisteria约有5-6个现生种,间断分布于中国、日本和美国的温带地区,但化石记录表明,该属在新近纪可能广泛分布于捷克、荷兰、格鲁吉亚阿布哈兹、保加利亚、罗马尼亚、俄罗斯远东、日本和中国。因此,研究紫藤属化石有助于深入认识它的早期演化、分类、多样性、古生态和生物地理,其中荚果化石的分类价值和演化意义尤为显著。文中基于对产自山东临朐中中新世山旺组的山旺紫藤W.shanwangensis荚果化石的再观察,并结合紫藤属3个现生种——紫藤W.sinensis、藤萝W.villosa和多花紫藤W.floribunda的荚果发育特征,讨论这些化石的分类、演化、发育和埋藏学意义。结果进一步证明,山旺紫藤荚果化石与国产的2个现生种——紫藤和藤萝的荚果更为相似,呈倒披针形、种子较少和室间缢缩明显。比较而言,日本和美国产的紫藤属现生种——多花紫藤和美国紫藤W.frutescens的荚果呈线形、种子较多和室间缢缩不明显,而且日本中新世和上新世报道的紫藤属荚果化石与多花紫藤的荚果更为相似。然而,中国和日本报道的紫藤属荚果化石迄今都没发现被毛,这与现生种中最原始的美国紫藤的荚果相似,而与东亚紫藤属现生种密被绒毛的荚果形成显著差别。因此,中国、日本和美国的紫藤属种类可能早在中新世就已经发生了形态地理分化,而荚果无毛或许是该属演化过程中一个比较原始的性状;紫藤属现生种荚果在发育的中、后期果壁上具有与纵轴方向成锐角的倾斜纤维纹饰,它们在荚果完全成熟后导致果瓣沿缝线开裂并卷曲,卷曲的果瓣放入水中又能恢复平整。值得注意的是,山旺紫藤荚果化石果壁上也发现了类似的倾斜纤维纹饰,这表明它们在脱落保存时处在发育的中、后期,这一发育时期脱落的荚果更有可能保存为化石记录;山旺紫藤荚果化石果壁的碳质残片中还富含硅藻类,近似于远距直链藻Melosira distans和颗粒直链藻M.granulata这些浮游相的、生活在深水区的优势种。因此,山旺紫藤荚果脱离母体后可能沉积在湖水较深的地方,而且它也可能是在成熟开裂的状态下脱落,瓣片本来卷曲,被短程搬运至湖中,又在湖水的浸泡下恢复平整状态,而后经沉积物掩埋后形成化石。     

Phylogenomics and biogeography of Wisteria: Implications on plastome evolution among inverted repeat-lacking clade (IRLC) legumes

The genus Wisteria (Fabaceae) is disjunctly distributed in eastern Asian and eastern North American temperate deciduous forests, and it is widely cultivated around the world as spectacular garden plants. It is a member of inverted repeat-lacking clade (IRLC). The IRLC Species are characterized by the loss of an IR region in their plastomes, which has long been of great interest. In this research, we report whole plastome sequences from all four Wisteria species and a Wisteriopsis japonica, combining these with existing data to explore phylogenetic relationships and biogeography of Wisteria, as well as plastome evolution of IRLC species. Phylogenetic analyses recognized a clade containing Glycyrrhiza–Wisteriopsis–Wisteria as sister to the remaining genera of IRLC. North American Wisteria frutescens and the three Asian species formed reciprocal clades, and Wisteria brachybotrys was sister to Wisteria floribunda and Wisteria sinensis. Wisteria may have originated in Japan near the boundary of the Oligocene and Miocene. The disappearance of Bering Land Bridge in the late Miocene might lead to the Eastern Asian–Eastern North American disjunction of Wisteria. Allopatric speciation of Wisteria between the Japanese archipelago and the Asian continent in the Quaternary increased the species richness of eastern Asia in comparison with eastern North America. Synonymous substitution rates (d_S) of protein-coding genes in the IRLC species were around 2-fold (SC genes) or 11-fold (IR genes) higher than those of non-IRLC species. For both SC and IR genes, herbaceous legumes have around 3-fold higher d_S than woody ones. Both loss of one IR region and herbaceous habit elevated substitution rates of the plastomes.     

Diverse rhizobia associated with woody legumes Wisteria sinensis, Cercis racemosa and Amorpha fruticosa grown in the temperate zone of China

Fifty-nine bacterial isolates from root nodules of the woody legumes Wisteria sinensis, Cercis racemosa and Amorpha fruticosa grown in the central and eastern regions of China were characterized with phenotypic analysis, PCR-based 16S and 23S rRNA gene RFLP, Box PCR and 16S rRNA gene sequencing. Seven main phena were defined in numerical taxonomy, which corresponded to distinct groups within the genera Agrobacterium, Bradyrhizobium, Mesorhizobium and Rhizobium in 16S and 23S rRNA gene PCR-RFLP. The phylogenetic relationships of the 16S rRNA genes supported the grouping results of PCR-RFLP. Most of the isolates from Amorpha fruticosa were classified into two groups closely related to Mesorhizobium amorphae. Seventeen of the 21 isolates from Wisteria sinensis were identified as two groups related to Rhizobium and Agrobacterium. Six out of 10 isolates from Cercis racemosa were identified as a group related to Bradyrhizobium. Our results indicated that each of the investigated legumes nodulated mainly with one or two rhizobial groups, although isolates from different plants intermingled in some small bacterial groups. In addition, correlation between geographic origin and grouping results was found in the isolates from Amorpha fruticosa. These results revealed that the symbiotic bacteria might have been selected by both the legume hosts and the geographic factors.     

Purification and complex formation analysis of a cysteine proteinase inhibitor (cystatin) from seeds of Wisteria floribunda

Seeds of Wisteria floribunda contain several kinds of cysteine proteinase inhibitor (cystatin). We purified and characterized one of these inhibitors, named WCPI-3. The molecular weight of WCPI-3 was estimated to be 17,500 and 15,700 by gel filtration and SDS-PAGE, respectively. The isoelectric point was 5.7. WCPI-3 formed an equimolar complex with native papain and the dissociation constant was estimated to be 6.1 nM. Complex formation between WCPI-3 and Cys25-modified papain, such as S-carboxy-methylated or S-carbamoylmethylated papain, could not be observed by gel filtration or native PAGE analysis. A peptide fragment derived from WCPI-3 digested by Achromobacter proteinase (lysyl endopeptidase) had the amino acid sequence of VVAGVNYRFVLK. The VVAG sequence in this fragment corresponds to the conserved sequence QVVAG which is considered to be one of binding regions to cysteine proteinases. The amino acid sequence of the amino-terminal portion (34 residues) of WCPI-3 was highly homologous to that of oryzacystatin from rice seeds.