Epimedium species are traditional Chinese medicinal plants as well as potential groundcover and ornamental plants. In this study, genome size and genome structures of Epimedium species were investigated using flow cytometric and fluorescence in situ hybridization (FISH). The nuclear DNA content of Epimedium species ranged from 8.42 pg/2C (8230.7 Mbp) to 9.97 pg/2C (9752.8 Mbp). The pairwise nucleotide diversity (π) of the fragments of the genes for reverse transcriptase (rt) of Ty1-copia retrotransposon within a species of rt fragments ranged from 0.251 to 0.428 in 10 Epimedium species. Phylogenetic analysis of the sequences revealed four major clades with the largest subclade containing 72 sequences of relatively low nucleotide diversity. FISH indicated that Ty1-copia retrotransposons are distributed unevenly along the pachytene chromosomes of E. wushanense and E. sagittatum, mostly associated with the pericentromeric and terminal heterochromatin. The relatively low sequence heterogeneity of Ty1-copia rt sequences implies that the Epimedium genomes have experienced a few relatively large-scale proliferation events of copia elements, which could be one of the major forces resulting in the large genome size of Epimedium species.
Jianjun Chen, Lijia Li and Ying Wang
Yau-Wen Yang, Pai-Yean Tai and Ying Chen
There are two evolutionary pathways in the genus of Brassica, one is rapa/oleracea lineage and the other is nigra lineage. Based on the morphological characteristics and nuclear RAPD or RFLP markers, genus Raphanus was thought more closely related to nigra lineage than to rapa/oleracea lineage (Song et al., 1990; Thormann et al., 1994). RFLP data of both chloroplast and mitochondria revealed that Raphanus is more closely related to rapa/oleracea lineage (Palmer and Herbon, 1988; Warwick and Black, 1991; Pradhan et al., 1992). We have previously demonstrated that Raphanus sativus is more closely related to nigra lineage using nuclear intergenic spacer between 5S rDNA and internal transcribed spacer region between 18S and 25S rDNA. In this study, we analyzed DNA sequences from different regions of chloroplast and showed that Raphanus sativus was closely related to rapa/oleracea lineage than to nigra lineage. These results suggest that Raphanus sativus is a hybrid between B. rapa/oleracea and B nigra lineages as proposed by Song et al (1990). The split time between these two lineages and the divergent time of Raphanus was also determined based on these chloroplast DNA sequences.
Ying Chen, Xinlu Chen, Fei Hu, Hua Yang, Li Yue, Robert N. Trigiano and Zong-Ming (Max) Cheng
Agave species are economically important plants in tropical and subtropical desert ecosystems as ornamentals as well as potential bioenergy crops. However, their relatively long life cycles and the current lack of biotechnology tools hinder their breeding. In this study, an efficient system for micropropagation was developed for Agave americana L. by using basal stems as explants and grown on a modified Murashige and Skoog medium (MSI) or a 1/2 MSI medium supplemented with various concentrations of 6-benzylaminopurine (BA) for shoot proliferation. The highest number of shoots (18.5 shoots/explant) from basal stems was obtained on MSI supplemented with 13.32 μM BA. An efficient shoot regeneration system was also developed from leaf tissues. Combinations of auxin with cytokinin, basal media, and leaf regions were optimized for shoot induction. Adventitious shoot formation from leaf segments was induced and proliferated with combination ranging of 0.54 to 2.68 μM [α-naphthaleneacetic acid (NAA)] with 8.88 to 13.32 μM (BA), and the maximum frequency (≈69%) was obtained with 2.68 μM NAA plus 13.32 μM BA. MSI medium and the basal segment of leaf affected shoot induction. The highest rooting frequency and mean number of shoots occurred in 1/2 MSI containing with 4.92 μM indole-3-butyric acid (IBA) alone (90%, 3.4) or 1.48 μM IBA plus 1.61 μM NAA (92%, 5.2). Survival of in vitro plantlets after transfer and acclimatization to ex vitro conditions was 87%. This is the first complete protocol for micropropagation of A. americana.
Luping Qu, Ying Chen, Xiping Wang, Richard Scalzo and Jeanine M. Davis
We investigated patterns of variation in alkamides and cichoric acid accumulation in the roots and aboveground parts of Echinacea purpurea (L.) Moench. These phytochemicals were extracted from fresh plant parts with 60% ethanol and quantified by high performance liquid chromatography (HPLC) analysis. Concentrations of alkamides and cichoric acid were measured on a dry-weight basis (mg·g–1). For total alkamides, concentrations among individual plants varied from 5.02 to 27.67 (mean = 14.4%) in roots, from 0.62 to 3.42 (mean = 1.54) in nearly matured seed heads (NMSH), and 0.22 to 5.25 (mean = 0.77) in young tops (about ½ flower heads, ¼ leaves, and ¼ stems). For cichoric acid, concentrations among individual plants varied from 2.65 to 37.52 (mean = 8.95), from 2.03 to 31.58 (mean = 10.9), and from 4.79 to 38.55 (mean = 18.88) in the roots, the NMSH, and the tops, respectively. Dodeca-2E, 4E, 8Z, 10E-tetraenoic acid isobutylamide and dodeca-2E, 4E, 8Z, 10Z-tetraenoic acid isobutylamide (alkamides 8/9) accounted for only 9.5% of the total alkamides in roots, but comprised 87.9% in the NMSH, and 76.6% in the young tops. Correlations of concentrations of alkamides or cichoric acid between those of roots and those of the NMSH were not statistically significant, and either within the roots, the NMSH, and the young tops. However, a significant negative correlation was observed between the concentration of cichoric acid in the roots and in young tops, and a significant positive correlation was observed between total alkamide concentration in the roots and cichoric acid concentration in the young tops. These results may be useful in the genetic improvement of E. purpurea for medicinal use.
Hadi Susilo, Ying-Chun Peng, Shui-Cheng Lee, Yu-Chun Chen and Yao-Chien Alex Chang
Phalaenopsis is currently the world’s number one potted flower crop. It is a slow-growing plant that responds slowly to nitrogen (N) fertilization and is noted for great resilience against N deficiency. Despite the great significance of N during the cultivation of Phalaenopsis, little has been studied on the uptake and partitioning of N in this crop. The stable isotope 15N was used as a tracer to investigate the uptake and partitioning of N and the roles of organs in sink and source relationship of N partitioning during different stages in Phalaenopsis. Fertilizer labeled with 15N was applied to Phalaenopsis Sogo Yukidian ‘V3’ during the vegetative growth stage on different parts of plants. Both leaves and roots were able to take up N. Nitrogen uptake efficiency of young roots was the highest, followed by old roots, whereas that of leaves was lowest. No difference of N uptake efficiency was found between the upper and lower leaf surfaces. Movement of fertilizer N to the leaves occurred as early as 0.5 day after fertilizer application to the roots. The partitioning of N depended on organ sink strength. During the vegetative growth stage, newly grown leaves and newly formed roots were major sinks. Sink strength of leaves decreased with the increase in leaf age. Stalks and flowers were major sinks during the reproductive growth stage. Mature leaves were a major location where N was stored and could serve as a N source during the reproductive growth stage and also for new leaf growth.
Hui-Shan Chan, Hui-Ying Chu and Mei-Fang Chen
In floriculture design, “shaping” is the use of floral materials as media for expressing ideas. Common floriculture techniques include tying, pasting, winding, connecting, overlapping, and weaving. Shaping is also a key factor in the appeal of the final product. Therefore, this study recruited 149 university students to explore how their floriculture material-shaping skills are affected by factors such as creative personality traits, spatial abilities, and shaping creativity. Students were allowed to use three different leaf materials in their floriculture works: planar leaf, linear leaf, and amorphous leaf materials. Representative planar, linear, and amorphous floriculture materials used in the current study were yellow palm (Chrysalidocarpus lutescens), veitch’s screw pine (Pandanus baptistii), and tree fern (Asparagus virgatus), respectively. The average score for creativity in shaping floriculture material was (±sd) 3.26 ± 0.84 (range, 1.33–4.67). Comparisons of the three leaf materials showed that the score for shaping creativity was highest for the planar leaf material (3.70 ± 1.23), followed by the amorphous leaf material (3.18 ± 0.99) and the linear leaf material (2.91 ± 0.94). The chi-square test results indicated that creative personality traits affected the number of shaping skills used, and that spatial abilities and floriculture material-shaping creativity further enhanced skills in floriculture material-shaping. Suggestions for floriculture educators and practitioners are provided accordingly.
Joseph N. Wolukau, Xiao-Hui Zhou, Ying Li, Yong-Bin Zhang and Jin-Feng Chen
Gummy stem blight incited by the fungus Didymella bryoniae is a major disease of melons worldwide. The objectives of the present study were to critically evaluate melon (Cucumis melo L.) germplasm for resistance to D. bryoniae and to characterize the genetics of resistance in the resistant accessions. Two hundred sources of germplasm (plant introduction accessions, cultivars, breeding lines, landraces, and wild relatives) were screened against a single highly virulent isolate (IS25) of D. bryoniae in a plastic tunnel. The genetics of resistance to D. bryoniae was studied in three crosses between plant introductions 157076, 420145, and 323498, resistant parents that were fairly adapted (flowering, fruiting, powdery mildew tolerance) to Nanjing conditions, and plant introductions 268227, 136170, and NSL 30032 susceptible parents, respectively. Six populations of each cross (susceptible parent, resistant parent, F1, F2, the two reciprocal backcrosses) were analyzed for their responses to D. bryoniae. Seedlings in both studies were inoculated with a spore suspension (5 × 105 spores/mL−1) of D. bryoniae at the four to six true-leaf stages and assessed for leaf and stem damage at 7, 14, and 21 d postinoculation. Results of germplasm screening indicated most germplasms reported as resistant elsewhere were confirmed resistant under our conditions. However, some plant introductions identified as highly resistant elsewhere were susceptible under our conditions, the most interesting being plant introduction 482399. This plant introduction that was considered resistant was highly susceptible in our study. We also identified other sources of resistance not reported previously, for example, JF1; a wild Cucumis from the highlands of Kenya was rated highly resistant. Analysis of segregation of F1, F2, and backcross generations of the three crosses indicated that each of the three plant introductions carry a single dominant gene for resistance to the D. bryoniae.
Chen Chen, Meng-Ke Zhang, Kang-Di Hu, Ke-Ke Sun, Yan-Hong Li, Lan-Ying Hu, Xiao-Yan Chen, Ying Yang, Feng Yang, Jun Tang, He-Ping Liu and Hua Zhang
Aspergillus niger is a common pathogenic fungus causing postharvest rot of fruit and vegetable, whereas the knowledge on virulence factors is very limited. Superoxide dismutase [SOD (EC 126.96.36.199)] is an important metal enzyme in fungal defense against oxidative damage. Thus, we try to study whether Cu/Zn-SOD is a virulence factor in A. niger. Cu/Zn-SOD encoding gene sodC was deleted in A. niger [MA70.15 (wild type)] by homologous recombination. The deletion of sodC led to decreased SOD activity in A. niger, suggesting that sodC did contribute to full enzyme activity. ΔsodC strain showed normal mycelia growth and sporulation compared with wild type. However, sodC deletion markedly increased the cell’s sensitivity to intracellular superoxide anion generator menadione. Besides, spore germination under menadione and H2O2 stresses were significantly retarded in ΔsodC mutant compared with wild type. Further results showed that sodC deletion induced higher superoxide anion production and higher content of H2O2 and malondialdehyde (MDA) compared with wild type, supporting the role of SOD in metabolism of reactive oxygen species (ROS). Furthermore, ΔsodC mutant had a reduced virulence on chinese white pear (Pyrus bretschneideri) as lesion development by ΔsodC was significantly less than wild type. The determination of superoxide anion, H2O2, and MDA in A. niger-infected pear showed that chinese white pear infected with ΔsodC accumulated less superoxide anion, H2O2, and MDA compared with that of wild type A. niger, implying that ΔsodC induced an attenuated response in chinese white pear during fruit–pathogen interaction. Our results indicate that sodC gene contributes to the full virulence of A. niger during infection on fruit. Aspergillus niger is one of the most common species found in fungal communities. It is an important fermentation industrial strain and is also known to cause the most severe symptoms in fruit during long-term storage (Pel et al., 2007). Meanwhile, plants activate their signaling pathways to trigger defense responses to limit pathogen expansion. One of the earliest host responses after pathogen attack is oxidative burst, during which large quantities of ROS are generated by different host enzyme systems, such as glucose oxidase (Govrin and Levine, 2000). ROS such as singlet oxygen, superoxide anion, hydroxyl (OH−), and H2O2 are released to hinder the advance of pathogens (Gara et al., 2003). ROS can react with and damage cellular molecules, such as DNA, protein, and lipids, which will limit fungal propagation in the host plant (Apel and Hirt, 2004).
Ting-Ting Li, Zhi-Rong Li, Kang-Di Hu, Lan-Ying Hu, Xiao-Yan Chen, Yan-Hong Li, Ying Yang, Feng Yang and Hua Zhang
Kiwifruit (Actinidia deliciosa) is a typical climacteric fruit, and its ripening is closely associated with ethylene. In this study, we present evidence that H2S alleviated ethylene-induced ripening and senescence of kiwifruit. Kiwifruit were fumigated with ethylene released from 0.4 g·L−1 ethephon solution or H2S with 1 mm sodium hydrosulfide (NaHS) as the donor or in combination. Fumigation with ethylene was found to accelerate kiwifruit ripening and H2S treatment effectively alleviated ethylene-induced fruit softening in parallel with attenuated activity of polygalacturonase (PG) and amylase. Ethylene + H2S treatment also maintained higher levels of ascorbic acid, titratable acid, starch, soluble protein, and reducing sugar compared with ethylene group, whereas suppressed the increase in chlorophyll and carotenoid. Kiwifruit ripening and senescence under ethylene treatment was accompanied by elevation in reactive oxygen species (ROS) levels, including H2O2 and superoxide anion and malondialdehyde (MDA), but combined treatment of ethylene plus H2S alleviated oxidative stress in fruit. Furthermore, the activities of antioxidative enzymes catalase (CAT) and ascorbate peroxidase (APX) were increased by ethylene + H2S treatment in comparison with ethylene alone, whereas the activities of lipoxygenase (LOX) and polyphenol oxidase (PPO) were attenuated by H2S treatment. Further investigations showed that H2S repressed the expression of ethylene synthesis-related genes AdSAM, AdACS1, AdACS2, AdACO2, and AdACO3 and cysteine protease genes, such as AdCP1 and AdCP3. Taken together, our findings suggest that H2S alleviates kiwifruit ripening and senescence by antagonizing the effect of ethylene through reduction of oxidative stress and inhibition of ethylene synthesis pathway.
Luping Qu, Xiping Wang, Ying Chen, Richard Scalzo, Mark P. Widrlechner, Jeanine M. Davis and James F. Hancock
Seed germination patterns were studied in Echinacea purpurea (L.) Moench grouped by seed source, one group of seven lots from commercially cultivated populations and a second group of nine lots regenerated from ex situ conserved wild populations. Germination tests were conducted in a growth chamber in light (40 μmol·m–2·s–1) or darkness at 25 °C for 20 days after soaking the seeds in water for 10 minutes. Except for two seed lots from wild populations, better germination was observed for commercially cultivated populations in light (90% mean among seed lots, ranging from 82% to 95%) and in darkness (88% mean among seed lots, ranging from 82% to 97%) than for wild populations in light (56% mean among seed lots, ranging from 9% to 92%) or in darkness (37% mean among seed lots, ranging from 4% to 78%). No germination difference was measured between treatments in light and darkness in the commercially cultivated populations, but significant differences were noted for treatments among wild populations. These results suggest that repeated cycles of sowing seeds during cultivation without treatments for dormancy release resulted in reduced seed dormancy in E. purpurea.