Hsiao-Feng Lo

Reduced susceptibility to waterlogging together with high-light stress is related to increases in superoxide dismutase and catalase activities in sweet potato

We investigated the changes in antioxidative enzyme activities of two sweet potato cultivars under waterlogging and high-light conditions in the growth chamber. The activities of antioxidative enzymes were measured from leaf crude extract of sweet potato during the first five days of the treatments. Activities of superoxide dismutase and catalase were consistently increased in Taoyuan 1 sweet potato over time under waterlogging and high-light conditions. However, decreases in both superoxide dismutase and catalase activities were observed for cultivar Yongtsai under waterlogging and high-light conditions. Waterlogging, together with high-light intensity, impairs superoxide dismutase and catalase activities in the cultivar Yongtsai indicating its greater susceptibility to waterlogging and high-light stress. In contrast, the increase in activities of superoxide dismutase and catalase in Taoyuan 1 indicated its greater ability to detoxify reactive oxygen species during the treatment and ensured its reduced susceptibility to waterlogging and high-light stress. The activities of peroxidase may be inactivated by high-light treatment and, therefore, may not be associated with tolerance of sweet potato plants to waterlogging and high-light stress. Differences in susceptibility to waterlogging and high-light conditions in the leafy vegetable Yongtsai and storage root Taoyuan 1 are discussed.

Antioxidant activity of herbaceous plant extracts protect against hydrogenperoxide-induced DNA damage in human lymphocytes

BackgroundHerbaceous plants containing antioxidants can protect against DNA damage. Thepurpose of this study was to evaluate the antioxidant substances,antioxidant activity, and protection of DNA from oxidative damage in humanlymphocytes induced by hydrogen peroxide (H2O2). Ourmethods used acidic methanol and water extractions from six herbaceousplants, including Bidens alba (BA), Lycium chinense (LC),Mentha arvensis (MA), Plantago asiatica (PA),Houttuynia cordata (HC), and Centella asiatica(CA).MethodsAntioxidant compounds such as flavonol and polyphenol were analyzed.Antioxidant activity was determined by the inhibition percentage ofconjugated diene formation in a linoleic acid emulsion system and bytrolox-equivalent antioxidant capacity (TEAC) assay. Their antioxidativecapacities for protecting human lymphocyte DNA fromH2O2-induced strand breaks was evaluated by cometassay.ResultsThe studied plants were found to be rich in flavonols, especially myricetinin BA, morin in MA, quercetin in HC, and kaemperol in CA. In addition,polyphenol abounded in BA and CA. The best conjugated diene formationinhibition percentage was found in the acidic methanolic extract of PA.Regarding TEAC, the best antioxidant activity was generated from the acidicmethanolic extract of HC. Water and acidic methanolic extracts of MA and HCboth had better inhibition percentages of tail DNA% and tail moment ascompared to the rest of the tested extracts, and significantly suppressedoxidative damage to lymphocyte DNA.ConclusionQuercetin and morin are important for preventing peroxidation and oxidativedamage to DNA, and the leaves of MA and HC extracts may have excellentpotential as functional ingredients representing potential sources ofnatural antioxidants.

Chilling stress and chilling tolerance of sweet potato as sensed by chlorophyll fluorescence

We studied changes in the chlorophyll (Chl) fluorescence components in chilling-stressed sweet potato (Ipomoea batatas L. Lam) cv. Tainung 57 (TN57, chilling-tolerant) and cv. Tainung 66 (TN66, chilling-susceptible). Plants under 12-h photoperiod and 400 µmol m−2 s−1 irradiance at 24/20 °C (day/night) were treated by a 5-d chilling period at 7/7 °C. Compared to TN66, TN57 exhibited a significantly greater basic Chl fluorescence (F0), maximum fluorescence (Fm), maximum fluorescence yield during actinic irradiation (Fm′ ), and the quantum efficiency of electron transport through photosystem 2, PS2 (ΦPS2). Chilling stress resulted in decrease in the potential efficiency of PS2 (Fv/Fm), ΦPS2, non-photochemical fluorescence quenching (NPQ), non-photochemical quenching (qN), and the occurrence of chilling injury in TN66. Chilling increased the likelihood of photoinhibition, characterized by a decline in the Chl fluorescence of both cultivars, and photoinhibition during low temperature stress generally occurred more rapidly in TN66.

Proteomics and transcriptomics of broccoli subjected to exogenously supplied and transgenic senescence-induced cytokinin for amelioration of postharvest yellowing ☆

UNLABELLED Previously, we investigated transgenic broccoli harboring senescence-associated-gene (SAG) promoter-triggered isopentenyltransferase (ipt), which encodes the key enzyme for cytokinin (CK) synthesis and mimics the action of exogenous supplied CK in delaying postharvest senescence of broccoli. Here, we used proteomics and transcriptomics to compare the mechanisms of ipt-transgenic and N(6)-benzylaminopurine (BA) CK treatment of broccoli during postharvest storage. The 2 treatments conferred common and distinct mechanisms. BA treatment decreased the quantity of proteins involved in energy and carbohydrate metabolism and amino acid metabolism, and ipt-transgenic treatment increased that of stress-related proteins and molecular chaperones and slightly affected levels of carbohydrate metabolism proteins. Both treatments regulated genes involved in CK signaling, sugar transport, energy and carbohydrate metabolism, amino acid metabolism and lipid metabolism, although ipt-transgenic treatment to a lesser extent. BA treatment induced genes encoding molecular chaperones, whereas ipt-transgenic treatment induced stress-related genes for cellular protection during storage. Both BA and ipt-transgenic treatments acted antagonistically on ethylene functions. We propose a long-term acclimation of metabolism and protection systems with ipt-transgenic treatment of broccoli and short-term modulation of metabolism and establishment of a protection system with both BA and ipt-transgenic treatments in delaying senescence of broccoli florets. BIOLOGICAL SIGNIFICANCE Transgenic broccoli harboring senescence-associated-gene (SAG) promoter-triggered isopentenyltransferase (ipt), which encodes the key enzyme for cytokinin (CK) synthesis and N(6)-benzylaminopurine (BA) CK treated broccoli both showed retardation of postharvest senescence during storage. The mechanisms underlying the two treatments were compared. The combination of proteomic and transcriptomic evidences revealed that the 2 treatments conferred common and distinct mechanisms in delaying senescence of broccoli florets. We propose a long-term acclimation of metabolism and protection systems with ipt-transgenic treatment of broccoli and short-term modulation of metabolism and establishment of a protection system with both BA and ipt-transgenic treatments in delaying senescence of broccoli florets. This article is part of a Special Issue entitled: Translational Plant Proteomics.

Identification of cucurbitacins and assembly of a draft genome for Aquilaria agallocha.

BackgroundAgarwood is derived from Aquilaria trees, the trade of which has come under strict control with a listing in Appendix II of the Convention on International Trade in Endangered Species of Wild Fauna and Flora. Many secondary metabolites of agarwood are known to have medicinal value to humans, including compounds that have been shown to elicit sedative effects and exhibit anti-cancer properties. However, little is known about the genome, transcriptome, and the biosynthetic pathways responsible for producing such secondary metabolites in agarwood.ResultsIn this study, we present a draft genome and a putative pathway for cucurbitacin E and I, compounds with known medicinal value, from in vitro Aquilaria agallocha agarwood. DNA and RNA data are utilized to annotate many genes and protein functions in the draft genome. The expression changes for cucurbitacin E and I are shown to be consistent with known responses of A. agallocha to biotic stress and a set of homologous genes in Arabidopsis thaliana related to cucurbitacin bio-synthesis is presented and validated through qRT-PCR.ConclusionsThis study is the first attempt to identify cucurbitacin E and I from in vitro agarwood and the first draft genome for any species of Aquilaria. The results of this study will aid in future investigations of secondary metabolite pathways in Aquilaria and other non-model medicinal plants.

The effect of red light and far-red light conditions on secondary metabolism in Agarwood

BackgroundAgarwood, a heartwood derived from Aquilaria trees, is a valuable commodity that has seen prevalent use among many cultures. In particular, it is widely used in herbal medicine and many compounds in agarwood are known to exhibit medicinal properties. Although there exists much research into medicinal herbs and extraction of high value compounds, few have focused on increasing the quantity of target compounds through stimulation of its related pathways in this species.ResultsIn this study, we observed that cucurbitacin yield can be increased through the use of different light conditions to stimulate related pathways and conducted three types of high-throughput sequencing experiments in order to study the effect of light conditions on secondary metabolism in agarwood. We constructed genome-wide profiles of RNA expression, small RNA, and DNA methylation under red light and far-red light conditions. With these profiles, we identified a set of small RNA which potentially regulates gene expression via the RNA-directed DNA methylation pathway.ConclusionsWe demonstrate that light conditions can be used to stimulate pathways related to secondary metabolism, increasing the yield of cucurbitacins. The genome-wide expression and methylation profiles from our study provide insight into the effect of light on gene expression for secondary metabolism in agarwood and provide compelling new candidates towards the study of functional secondary metabolic components.

Development of an In Planta system to monitor phosphorus status by agroinfiltration and agroinjection

Background and AimsAn optimal supply of phosphate (Pi) fertilizer is important to ensure crop yield and quality and to maintain agricultural sustainability. In this study, an in planta monitoring system was developed to conveniently monitor the P status of crops.MethodsA phosphate starvation-induced gene (TPSI1) from tomato (Solanum lycopersicum L.) was used to reflect P status in plants. Using agroinfiltration and agroinjection techniques, the GUS reporter gene driven by the TPSI1 promoter was transiently expressed in tobacco (Nicotiana benthamiana) leaves and tomato fruits to reflect their P status.ResultsWe showed that the vegetative growth of tobacco and the yield and quality of tomato were affected by the supply of Pi fertilizer. We further demonstrated that the expression of a GUS reporter gene driven by the TPSI1 promoter could accurately report on P status in tobacco leaves and tomato fruits in real time. Expression of the GUS reporter gene was independent of deficiencies in mineral nutrients other than P, demonstrating the specificity of this system.ConclusionsThe above results indicate that the employed agroinfiltration/agroinjection-based diagnostic system is useful for monitoring P status in plants.

Comparative proteomic analysis of cauliflower under high temperature and flooding stresses

Abstract High-temperature and waterlogging are major abiotic stresses that affect the yield and quality of cauliflower. Cauliflower cultivars ‘H41’ and ‘H69’ are tolerant to high temperature and flooding, respectively; however, ‘H71’ is sensitive to both stresses. The objectives of this study were to identify the proteins that were differentially regulated and the physiological changes that occurred during different time periods in ‘H41’, ‘H69’, and ‘H71’ when responding to treatments of flooding, 40°C, and both stresses combined. Changes in the leaf proteome were analyzed by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS) and identified by Mascot peptide mass fingerprint (PMF) and database searching. Stress treatments caused significant reductions in electrolyte leakage, chlorophyll fluorescence Fv/Fm, chlorophyll content, and water potential as stress times were prolonged. By the comparative proteomic analysis, 85 protein peaks that were differentially expressed in response to combination treatments at 0, 6, and 24h, 69 (33 in ‘H41’, 29 in ‘H69’, and 9 in ‘H71’) were identified, of which were cultivar specific. Differentially regulated proteins predominantly functioned in photosynthesis and to a lesser extent in energy metabolism, cellular homeostasis, transcription and translation, signal transduction, and protein biosynthesis. This is the first report that utilizes proteomics to discover changes in the protein expression profile of cauliflower in response to heat and flooding.

Transcriptomic and Proteomic Research To Explore Bruchid-Resistant Genes in Mungbean Isogenic Lines.

Mungbean (Vigna radiata (L.) Wilczek) is an important rotation legume crop for human nutrition in Asia. Bruchids (Callosobruchus spp.) currently cause heavy damage as pests of grain legumes during storage. We used omics-related technologies to study the mechanisms of bruchid resistance in seeds of the nearly isogenic lines VC1973A (bruchid-susceptible) and VC6089A (bruchid-resistant). A total of 399 differentially expressed genes (DEGs) were identified between the two lines by transcriptome sequencing. Among these DEGs, 251 exhibited high expression levels and 148 expressed low expression levels in seeds of VC6089A. Forty-five differential proteins (DPs) were identified by isobaric tags for relative and absolute quantification (iTRAQ); 21 DPs had higher abundances in VC6089A, and 24 DPs had higher abundances in VC1973A. According to transcriptome and proteome data, only three DEGs/DPs, including resistant-specific protein (g39185), gag/pol polyprotein (g34458), and aspartic proteinase (g5551), were identified and located on chromosomes 5, 1, and 7, respectively. Both g39185 and g34458 genes encode a protein containing a BURP domain. In previous research on bruchid molecular markers, the g39185 gene located close to the molecular markers of major bruchid-resistant locus may be a bruchid-resistant gene.

AFLP mapping of quantitative trait loci influencing seven head-related traits in broccoli (Brassica oleracea var. italica)

Summary This study identified amplified fragment length polymorphism (AFLP) markers linked to head-related traits in broccoli (Brassica oleracea var. italica) grown in both Winter (W) and Summer (S) in 2008 and in 2009. In total, 227 F2 plants derived from a cross between the heat-tolerant cultivar ‘Shin-Fa 1208’ (SF), and the heat-sensitive cultivar ‘Green Magic’ (GM), were grown in the field. Seven traits including floret height (FH), floret width (FW), head height (HH), head width (HW), head weight (HWT), top weight (TWT), and stem width (SW) were scored. Polymorphic bands were generated by AFLP, and the segregation ratios and distribution of the identified markers over the Brassica genome were compared. Nine linkage groups (LGs), ranging from 75.8 cM to146.5 cM in size, were constructed using 202 informative markers spanning a total of 1,034.5 cM. For the seven traits investigated, 73 quantitative trait loci (QTLs) were dispersed non-randomly in the broccoli genome. Thirty-one unique regions of the genome were detected across the population in Summer trial, and three QTL markers, located on LGs 1, 3, and 5, showed pleiotropic effects on these seven traits. Among these, the QTL linked to the d-64 (S) marker on LG 3 had effects on HH, HWT, and FW, and significant positive correlations existed among these three traits. Thus, the AFLP marker d-64 (S) may have potential for the simultaneous selection of large FW and HH, and increased HWT under high-temperature conditions.