Inhibitory effect of cinnamaldehyde on Fusarium solani and its application in postharvest preservation of sweet potato.

Root rot caused by Fusarium solani is one of major postharvest diseases limiting sweet potato production. Antifungal effect and possible mode of action of cinnamaldehyde (CA) against F. solani were investigated. CA concentration of 0.075 g/L inhibited conidial viability of F. solani. CA vapor of 0.3 g/L in air completely controlled the F. solani development in sweet potatoes during storage for 10 days at 28 °C, and protected soluble sugar and starch in the flesh from depletion by the fungus. Further results demonstrated that CA induced reduction in mitochondrial membrane potential (Δψm), ROS accumulation, and cell apoptosis characterized by DNA fragmentation in F. solani. Moreover, CA facilitated decomposition of mitochondria-specific cardiolipin (CL) into its catabolites by the catalytic action of phospholipases. Altogether, the results revealed a specific antifungal mechanism of CA against F. solani, and suggest that CA holds promise as a preservative for postharvest preservation of sweet potato.

Potassium Fertilization Stimulates Sucrose-to-Starch Conversion and Root Formation in Sweet Potato (Ipomoea batatas (L.) Lam.).

A field experiment was established to study sweet potato growth, starch dynamic accumulation, key enzymes and gene transcription in the sucrose-to-starch conversion and their relationships under six K2O rates using Ningzishu 1 (sensitive to low-K) and Xushu 32 (tolerant to low-K). The results indicated that K application significantly improved the biomass accumulation of plant and storage root, although treatments at high levels of K, i.e., 300-375 kg K2O ha-1, significantly decreased plant biomass and storage root yield. Compared with the no-K treatment, K application enhanced the biomass accumulation of plant and storage root by 3-47% and 13-45%, respectively, through promoting the biomass accumulation rate. Additionally, K application also enhanced the photosynthetic capacity of sweet potato. In this study, low stomatal conductance and net photosynthetic rate (Pn) accompanied with decreased intercellular CO2 concentration were observed in the no-K treatment at 35 DAT, indicating that Pn was reduced mainly due to stomatal limitation; at 55 DAT, reduced Pn in the no-K treatment was caused by non-stomatal factors. Compared with the no-K treatment, the content of sucrose, amylose and amylopectin decreased by 9-34%, 9-23% and 6-19%, respectively, but starch accumulation increased by 11-21% under K supply. The activities of sucrose synthetase (SuSy), adenosine-diphosphate-glucose pyrophosphorylase (AGPase), starch synthase (SSS) and the transcription of Susy, AGP, SSS34 and SSS67 were enhanced by K application and had positive relationships with starch accumulation. Therefore, K application promoted starch accumulation and storage root yield through regulating the activities and genes transcription of SuSy, AGPase and SSS in the sucrose-to-starch conversion.

Antioxidant, hepatoprotective and antifungal activities of black pepper (Piper nigrum L.) essential oil.

The essential oil extracted from the black Piper nigrum L. (BPEO) was analyzed for antioxidant, hepatoprotective and antifungal activities. BPEO is rich in total phenolics, total flavonoids and proanthocyanidins, and showed good free radicals and lipid peroxidation scavenging capacities. In a CCl4-induced liver injury mice model, the BPEO treated groups showed increases in the catalase (CAT), glutathione (GSH) and total superoxide dismutase (T-SOD) activities present in the liver and kidney, and reverses the CCl4-elevated total bilirubin (TBIL), glutamate pyruvate transaminase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (AKP) and malondialdehyde (MDA) level, which were confirmed in further analyses of kidney tissue sections. BPEO can effectively inhibit the growth of Aspergillus flavus spoilage fungus in maize. Further analyses indicated that BPEO disrupt the permeability barrier of the cell membrane and lead to mitochondrial dysfunction in A. flavus. Therefore, the current study proved BPEO's potential as hepatoprotective products and natural food preservatives.

Comparative Transcriptome and Proteome Analysis of Salt-Tolerant and Salt-Sensitive Sweet Potato and Overexpression of IbNAC7 Confers Salt Tolerance in Arabidopsis.

Salt stress is one of the major devastating factors affecting the growth and yield of almost all crops, including the crucial staple food crop sweet potato. To understand their molecular responses to salt stress, comparative transcriptome and proteome analysis of salt-tolerant cultivar Xushu 22 and salt-sensitive cultivar Xushu 32 were investigated. The results showed the two genotypes had distinct differences at the transcription level and translation level even without salt stress, while inconsistent expression between the transcriptome and proteome data was observed. A total of 16,396 differentially expressed genes (DEGs) and 727 differentially expressed proteins (DEPs) were identified. Wherein, 1,764 DEGs and 93 DEPs were specifically expressed in the tolerant genotype. Furthermore, the results revealed that the significantly upregulated genes were mainly related to the regulation of ion accumulation, stress signaling, transcriptional regulation, redox reactions, plant hormone signal transduction, and secondary metabolite accumulation, which may be involved in the response of sweet potato to salt stress and/or may determine the salt tolerance difference between the two genotypes. In addition, 1,618 differentially expressed regulatory genes were identified, including bZIP, bHLH, ERF, MYB, NAC, and WRKY. Strikingly, transgenic Arabidopsis overexpressing IbNAC7 displayed enhanced salt tolerance compared to WT plants, and higher catalase (CAT) activity, chlorophyll and proline contents, and lower malondialdehyde (MDA) content and reactive oxygen species (ROS) accumulation were detected in transgenic plants compared with that of WT under salt stress. Furthermore, RNA-seq and qRT-PCR analysis displayed that the expression of many stress-related genes was upregulated in transgenic plants. Collectively, these findings provide revealing insights into sweet potato molecular response to salt stress and underlie the complex salt tolerance mechanisms between genotypes, and IbNAC7 was shown as a promising candidate gene to enhance salt tolerance of sweet potato.

Perillaldehyde Controls Postharvest Black Rot Caused by Ceratocystis fimbriata in Sweet Potatoes.

Black rot caused by Ceratocystis fimbriata is the most damaging postharvest disease among sweet potatoes. Black rot can be controlled by synthetic fungicides, but these synthetic fungicides also have several negative effects. Perillaldehyde (PAE), a major component of the herb perilla, is an effective and eco-friendly method of controlling this disease. The antifungal activity of PAE on the mycelial growth in C. fimbriata was evaluated in vitro. Sweet potatoes at the postharvest stage were surfaced-disinfected with 75% ethanol. Artificially created wounds were inoculated with a C. fimbriata cell suspension, and then, the PAE was spontaneously volatilized inside the residual airspace of the containers at 28°C. Samples were collected at 0, 3, 6, 9, 12, 15, 18, and 21 days from each group, and the tissues around the wounds of the sweet potatoes were collected using a sterilized knife and then homogenized to determine their defense-related enzyme activity and quality parameters. In vitro assays showed that the mycelial growth of C. fimbriata was inhibited by PAE in a dose-dependent manner. An in vivo test demonstrated that 25, 50, and 100 µl/l PAE doses, when applied to sweet potatoes inoculated with C. fimbriata, could remarkable lower lesion diameter as compared to the control. Even though the storage time was prolonged, PAE vapor treatment still drastically inhibited sweet potato decay during storage at 28°C. These PAE vapor treatments also enhanced the activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), peroxidase (POD), polyphenol oxidase (PPO), and phenylalanine ammonia-lyase (PAL). These treatments remarkably decreased weight loss rates and had minor effects on other fruit quality parameters, such as anthocyanin content and vitamin C content. In our study, the results suggested that the effects of PAE on postharvest sweet potatoes may be attributed to the maintenance of enzymatic activity and fruit quality. In sum, PAE may be a promising approach to controlling C. fimbriata in sweet potatoes.

Characterization of actin and tubulin promoters from two sap-sucking pests, Nilaparvata lugens (Stål) and Nephotettix cincticeps (Uhler).

The brown planthopper, Nilaparvata lugens (N. lugens, Hemiptera: Delphacidae), and the green rice leafhopper, Nephotettix cincticeps (N. cincticeps, Hemiptera: Cicadellidae), two sap-sucking feeders, have caused many destructive agricultural disasters in East Asia, as they can bring diseases like 'hopper burn' and transmit plant viruses. Recently, continuously-cultured cell lines from both insects have been reported. However, exogenous protein expression systems have not yet been established. Here, we identified thirteen tubulin genes and three actin genes from N. lugens, and one tubulin gene and two actin genes from N. cincticeps. Furthermore, putative promoter regions of these genes were analyzed by bioinformatic approaches and 5'-RACE assay, and the promoter strength was evaluated by driving the enhanced green fluorescent protein expression in three insect cell lines, S2, Sf9, and BmN. Finally, we identified three effective promoters (Nl_αTub1 promoter, Nl_act3 promoter, and Nc_act1 promoter) among all candidates we screened. The Nc_act1 promoter showed the strongest activity, while the Nl_αTub1 promoter only worked in S2 cells. In conclusion, we identified and functionally characterized three native promoters from N. lugens and N.cincticeps, which would facilitate the establishment of exogenous protein expression systems suitable for these two insect pests.

PMRD: a curated database for genes and mutants involved in plant male reproduction.

BACKGROUND: Male reproduction is an essential biological event in the plant life cycle separating the diploid sporophyte and haploid gametophyte generations, which involves expression of approximately 20,000 genes. The control of male reproduction is also of economic importance for plant breeding and hybrid seed production. With the advent of forward and reverse genetics and genomic technologies, a large number of male reproduction-related genes have been identified. Thus it is extremely challenging for individual researchers to systematically collect, and continually update, all the available information on genes and mutants related to plant male reproduction. The aim of this study is to manually curate such gene and mutant information and provide a web-accessible resource to facilitate the effective study of plant male reproduction. DESCRIPTION: Plant Male Reproduction Database (PMRD) is a comprehensive resource for browsing and retrieving knowledge on genes and mutants related to plant male reproduction. It is based upon literature and biological databases and includes 506 male sterile genes and 484 mutants with defects of male reproduction from a variety of plant species. Based on Gene Ontology (GO) annotations and literature, information relating to a further 3697 male reproduction related genes were systematically collected and included, and using in text curation, gene expression and phenotypic information were captured from the literature. PMRD provides a web interface which allows users to easily access the curated annotations and genomic information, including full names, symbols, locations, sequences, expression patterns, functions of genes, mutant phenotypes, male sterile categories, and corresponding publications. PMRD also provides mini tools to search and browse expression patterns of genes in microarray datasets, run BLAST searches, convert gene ID and generate gene networks. In addition, a Mediawiki engine and a forum have been integrated within the database, allowing users to share their knowledge, make comments and discuss topics. CONCLUSION: PMRD provides an integrated link between genetic studies and the rapidly growing genomic information. As such this database provides a global view of plant male reproduction and thus aids advances in this important area.

Involvements of S-nitrosylation and denitrosylation in the production of polyphenols by Inonotus obliquus.

Nitric oxide (NO) has been evidenced to mediate biosynthesis of polyphenols in Inonotus obliquus. However, it remains unknown how NO regulates their biosynthesis. Here we show that higher cellular NO levels coincided with higher accumulation of S-nitrosothiols (SNO; the products of NO combined with a specific residue in glutathione or proteins) and polyphenols, and higher activity of denitrosylated S-nitrosoglutathione reductase (GSNOR) and thioredoxin reductase (TrxR). This homeostasis was breached by GSNOR or TrxR inhibitors. Inhibiting GSNOR boosted TrxR activity, but reduced SNO formation, coinciding with an enhanced production of polyphenols. Likewise, inhibiting TrxR increased GSNOR activity and SNO production, but downregulated accumulation of polyphenols. Inhibiting GSNOR or TrxR also modified the polyphenolic profiles of I. obliquus. Suppressing GSNOR-enhanced biosynthesis of phelligridins C and H, inoscavin C and methyl inoscavin B, but reduced that of phelligridin D, methyl inoscavin A, davallialactone and methyl davallialactone, the typical polyphenols in I. obliquus. Similarly, downregulating TrxR increased production of phelligridin D, methyl inoscavin A, davallialactone, and methyl davallialactone, but shrinking that of phelligridins C and H, methyl inoscavin B and inoscavin C. Thus, in I. obliquus, the state of S-nitrosylation and denitrosylation affects not only the accumulation of polyphenols, but also their metabolic profiles.

Analysis of antioxidant metabolites by solvent extraction from sclerotia of Inonotus obliquus (Chaga).

INTRODUCTION: The sclerotia of Inonotus obliquus (Chaga) are effective therapeutic agents to treat several human malignant tumours and other diseases without unacceptable toxic side-effects. OBJECTIVE: To investigate solvent effects on metabolic profiles and antioxidant activities of extracts of Chaga. METHODOLOGY: Chaga was extracted by petroleum ether, chloroform, ethyl acetate, acetone, ethanol and water. Solvent effects on metabolites in the extracts were assayed by NMR-based metabolomic analysis. Antioxidant activities were indicated as capacities for scavenging superoxide anion, DPPH and hydroxyl radicals. RESULTS: Petroleum ether and chloroform extracts contained primarily lanostane-type triterpenoids (LT), whereas the extracts of ethyl acetate, acetone and ethanol were characterised by the predominant presence of hispidin analogues and LT, and water extracts by polysaccharides and phenolic compounds. The ethyl acetate, acetone, ethanol and water extracts revealed remarkable potential for scavenging the tested radicals, while those of petroleum ether and chloroform did not. Polyphenols are the major contributors for quenching the tested free radicals, while in LT only compounds 16, 17 and 22 participated in scavenging hydroxyl radicals. CONCLUSION: Polyphenols in Chaga are the principles for quenching free radicals while polysaccharides and a few LT compounds contribute partially in scavenging DPPH and hydroxyl radicals, respectively. NMR-based metabolomic analysis is a useful method by which to correlate ¹H-NMR spectra of Chaga extracts with their antioxidant activities, and this allows the prediction of potentials for scavenging free radicals by ¹H-NMR spectroscopy.

Production of antioxidant and antitumor metabolites by submerged cultures of Inonotus obliquus cocultured with Phellinus punctatus.

While Inonotus obliquus produces a diverse range of bioactive metabolites in its natural habitats, it accumulates less in its submerged cultures. We show here that coculture of I. obliquus with Phellinus punctatus resulted in less production of mycelial biomass but an increased accumulation of phenolic compounds, melanins, and lanostane-type triterpenoids. Metabolites increased in production by coculture include phelligridin C, phelligridin H, methyl inoscavin A, inoscavin C, inoscavin B, davallialactone, methyl davallialactone, foscoparianol D, 21,24-cyclopentalanosta-3ß,21,25-triol-8-en, lanosta-7,9(11),23-triene-3ß,22,25-triol, and inotodisaccharide and melanins. Metabolites from coculture also showed an increased potential for scavenging free radicals and inhibiting the proliferation of HeLa 229 cells. Davallialactone, methyl davallialactone, and minor phenolic components are the major contributors for scavenging DPPH and hydroxyl radical in monoculture, and phelligridin C, phelligridin H, methyl inoscavin A, inoscavin C, methyl davallialactone, foscoparianol D, and inotodisaccharide are those for scavenging the tested radicals in coculture. Lanostane-type triterpenoids indicated limited roles in scavenging free radicals. Nearly all the detected metabolites correlate positively with inhibiting proliferation of HeLa 229 cells. Thus, coculture of I. obliquus with other fungi seems to be a cost-effective strategy for upregulating biosynthesis of bioactive metabolites.

Chemical diversity of biologically active metabolites in the sclerotia of Inonotus obliquus and submerged culture strategies for up-regulating their production.

Inonotus obliquus (Fr.) Pilat is a white rot fungus belonging to the family Hymenochaetaceae in the Basidiomycota. In nature, this fungus rarely forms a fruiting body but usually an irregular shape of sclerotial conk called 'Chaga'. Characteristically, I. obliquus produces massive melanins released to the surface of Chaga. As early as in the sixteenth century, Chaga was used as an effective folk medicine in Russia and Northern Europe to treat several human malicious tumors and other diseases in the absence of any unacceptable toxic side effects. Chemical investigations show that I. obliquus produces a diverse range of secondary metabolites including phenolic compounds, melanins, and lanostane-type triterpenoids. Among these are the active components for antioxidant, antitumoral, and antiviral activities and for improving human immunity against infection of pathogenic microbes. Geographically, however, this fungus is restricted to very cold habitats and grows very slowly, suggesting that Chaga is not a reliable source of these bioactive compounds. Attempts for culturing this fungus axenically all resulted in a reduced production of bioactive metabolites. This review examines the current progress in the discovery of chemical diversity of Chaga and their biological activities and the strategies to modulate the expression of desired pathways to diversify and up-regulate the production of bioactive metabolites by the fungus grown in submerged cultures for possible drug discovery.

Nitric oxide mediates the fungal-elicitor-enhanced biosynthesis of antioxidant polyphenols in submerged cultures of Inonotus obliquus.

A fungal elicitor prepared from the cell debris of the plant-pathogenic ascomycete Alternaria alternata induces multiple responses by Inonotus obliquus cells, including an increase in generation of nitric oxide (NO), activity of phenylalanine ammonia lyase (PAL) and accumulation of total mycelial phenolic compounds (TMP), but does not trigger production of oxylipins or jasmonic acid (JA). The role of NO in TMP production was investigated via the effects of the NO-specific scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPITO) and the nitric oxide synthase (NOS) inhibitor aminoguanidine (AG). TMP profiles were assayed using (1)H NMR spectroscopy combining multivariate pattern recognition strategies. Pretreatment of I. obliquus mycelia with cPITO or AG suppressed not only elicitor-enhanced NO generation and PAL activity, but also the elicitor-induced increase in TMP production. This TMP reduction by either a NO scavenger or a NOS inhibitor was reversed by exogenous addition of either a NO donor, sodium nitroprusside, or JA separately. NMR-based metabonomic analysis of TMP profiles showed that the induced TMP were hispidin analogues including inoscavins, phelligridins, davallialactone and methyldavallialactone, which possess high antioxidant activities. Thus, NO mediates an elicitor-induced increase in production of antioxidant polyphenols in I. obliquus via a signalling pathway independent of oxylipins or JA, a mechanism which differs from those in some higher plants.

[Design and application of computer-assisted software for teaching and research of population genetics].

The computer-assisted software for population genetics' teaching and research is designed with Microsoft Visual Basic 6.0. It takes all factors affecting population genetic structure into account. By way of setting various conditional parameters, calculating theoretically, simulating with computer and choosing the states of show, the system displays the results in charts and tables accurately. It also presents distribution pattern, statistical characteristics and the change of population gene frequencies and genotype frequencies in the inter-generation visually. The software is a mature system for teaching and researches on the aspects of its functions, operations and interface.

Molecular phylogeny of the domesticated silkworm, Bombyx mori, based on the sequences of mitochondrial cytochrome b genes.

Pupae from the Chinese wild mulberry silkworm, Bombyx mandarina, and 11 representative strains of the domesticated silkworm, Bombyx mori were selected for preparation of mitochondrial DNA. The 5'-end fragments of cytochrome b genes (Cytb) were generated by polymerase chain reaction products and sequenced directly. The homologous sequences of the Japanese B. mandarina and three strains of B. mori were from the GenBank database. The sequences of the 16 silkworm strains were analysed with DNASTAR software and a phylogenic tree was constructed using PHYLIP software. The result showed that: (i) The sequence divergence between the strains of B. mori and the Japanese B. mandarina was larger (5.4%-5.8%) compared with that between strains of B. mori and the Chinese B. mandarina (0.8% -1.9%). Analysis of clustering also showed that the sequences of B. mori strains and Chinese B. mandarina clustered into group (B group), while that of Japanese B. mandarina (A group) was outside this cluster. This may be evidence for the hypothesis that B. mori originated from Chinese B. mandarina. (ii) Among 14 strains of B. mori, sequence divergence was small and the most divergence was seen between strains Yanhe-1 and Chuxiong, whose sequences branched off from those of the other B. mori strains on the phylogenetic tree. From this and from historical records, we infer that the strains Yanhe-1 and Chuxiong originated independently from southwest China.

[Principle of genetic equilibrium for two gene loci].

Because linkage equilibrium is introduced by directly quoting the conclusions or imprecise mathematical reasoning in most of textbooks, many students are puzzled with the problem of linkage equilibrium when they learn population genetics. Based on the radical conditions of genetic equilibrium, the principle of linkage equilibrium condition and process, for two gene loci is introduced by precise mathematical reasoning. The article may provide reference to teachers and students in the teaching and learning of population genetics.

[A system for capturing and showing micrographs of genetics experiments].

A system for capturing and showing micrographs of genetics was designed with Microsoft Visual Basic 6.0. The system includes many functions such as capturing and editing images, typing and editing text, teaching experiments, showing images, image retrieval, database management, system maintenance and help, all of them were developed with the form of Windows. The system could collect images not only from image-grabber card in real-time but also from scanner, digital camera, clipboard and files. After utilizing the image compression technology, the images will be saved in database along with experiment instruction. With all the features referred above, the system can be used as a wonderful assistant both for the teaching of genetics experiments and for the students' learning by themselves.