Superoxide production by mitochondria isolated from green bell pepper fruit

Evidence is increasing to suggest that a wide range of environmentally induced plant disorders, including chilling injury, is mediated by reactive oxygen species produced during stress or upon relief from stress. Mitochondria were isolated from pericarp tissue of chilling-sensitive bell pepper fruit and their respiratory activity and ability to produce superoxide when supplied with NADH, succinate or malate-pyruvate were determined. Oxygen uptake rates were greater and less sensitive to cyanide with succinate than with NADH; rates increased and sensitivity to cyanide and respiratory control ratios (RCRs) decreased in fruit stored at 2°C. Disrupting mitochondrial membranes led to increased oxygen consumption with NADH and decreased consumption with succinate. resulting in RCRs of approximately 1 with both substrates. Superoxide production was greater with NADH than with either succinate or malatepyruvate. Superoxide dismutase and cyanide inhibited superoxide production almost completely. Antimycin A did not inhibit superoxide production with NADH, but did partially with succinate, especially in mitochondria sensitive to cyanide. Disrupting mitochondrial membranes enhanced superoxide production with NADH. Superoxide production by mitochondria isolated from fruit stored at 2°C increased with NADH and decreased with succinate. Results provide evidence that mitochondria may be a major source of superoxide in chilling-sensitive plant tissues exposed to low temperatures.     

辣椒细胞质雄性不育与活性氧代谢的关系

通过测定辣椒保护酶活性和膜脂过氧化产物,研究了辣椒胞质雄性不育三系与膜脂过氧化的关系.结果表明,从花粉母细胞减数分裂前至花粉成熟期,三系之间SOD活性无明显差异,不育系花药中的CAT和POD活性均显著或极显著低于保持系和恢复系,而O2-·产生效率、MDA含量则显著或极显著高于保持系和恢复系.同时,在花药发育过程中,由于质、核差异,保持系的CAT活性明显高于恢复系,SOD和POD活性保持系和恢复系之间无明显差异.     

Concurrent evolution of resistance and tolerance to potato virus Y in Capsicum annuum revealed by genome-wide association

We performed a genome-wide association study of pepper (Capsicum annuum) tolerance to potato virus Y (PVY). For 254 pepper accessions, we estimated the tolerance to PVY as the coefficient of regression of the fresh weight (or height) of PVY-infected and mock-inoculated plants against within-plant virus load. Small (strongly negative) coefficients of regression indicate low tolerance because plant biomass or growth decreases sharply as virus load increases. The tolerance level varied largely, with some pepper accessions showing no symptoms or fairly mild mosaics, whereas about half (48%) of the accessions showed necrotic symptoms. We found two adjacent single-nucleotide polymorphisms (SNPs) at one extremity of chromosome 9 that were significantly associated with tolerance to PVY. Similarly, in three biparental pepper progenies, we showed that the induction of necrosis on PVY systemic infection segregated as a monogenic trait determined by a locus on chromosome 9. Our results also demonstrate the existence of a negative correlation between resistance and tolerance among the cultivated pepper accessions at both the phenotypic and genetic levels. By comparing the distributions of the tolerance-associated SNP alleles and previously identified PVY resistance-associated SNP alleles, we showed that cultivated pepper accessions possess favourable alleles for both resistance and tolerance less frequently than expected under random associations, while the minority of wild pepper accessions frequently combined resistance and tolerance alleles. This divergent evolution of PVY resistance and tolerance could be related to pepper domestication or farmer's selection.     

Application of Vascular Puncture for Evaluation of Curtovirus Resistance in Chile Pepper and Tomato

Curtoviruses cause severe damage to tomatoes and peppers. Functional field resistance to curtoviruses in these plants is desirable but difficult to produce and difficult to screen for because it is time‐consuming and resistance could be achieved by developing resistance either to the virus or to insect feeding. To improve and speed curtovirus resistance testing in tomato (Solanum lycopersicum) and pepper (Capsicum annuum) plants, two puncture methods were developed and compared to leafhopper inoculation and feeding preference assays. The two puncture methods were adapted to introduce a modified Agrobacterium tumefaciens plasmid carrying a recombinant curtovirus into the meristem tissue of tomato plants and into newly germinated chile pepper seedlings. The puncture techniques were used to screen for resistance to curtoviruses in chile pepper and tomato breeding lines and varieties. Similarly, the peppers and tomatoes were assayed for curtovirus resistance using leafhopper inoculation and feeding preference, which was assessed by stylet sheath staining. Virus infection by puncture and leafhopper feeding was monitored using PCR and ELISA. ELISA was performed using an antibody to bacterially expressed coat protein. While pepper cvs Tabasco, NuMex Las Cruces cayenne and New Mexico 6‐4 were infected using both puncture and leafhopper inoculation methods, New Mexico 6‐4 had higher infection rates than the other two cultivars. Stylet sheath staining results suggest that leafhoppers prefer to feed on New Mexico 6‐4 rather than Tabasco and NuMex Las Cruces cayenne. Eight tomato cultivars were infected using meristem removal injection inoculation. Three tomatoes cultivars (CVF‐11, Saladmaster and Supersteak) were infected using leafhopper inoculation, although stylet sheath staining results suggested that the first two cultivars were not preferred by the insect vector. Our results suggest that puncture methods and leafhopper inoculation are successful in resistance screening, and both methods should be used as part of screening, because they assess different types of resistance.     

Thidiazuron induced somatic embryogenesis and plant regeneration in Capsicum annuum

An efficient protocol of direct somatic embryogenesis (without involving intermediate callus) has been developed from stem segments and shoot tips of Capsicum annuum L. Explants were cultured on Murashige and Skoog (MS) medium supplemented with thidiazuron (TDZ). Among the various concentration of TDZ tested, 0.5 μM was proved to be best for induction of somatic embryos. Induction, maturation and germination were achieved on the same medium. The shoots developed from somatic embryos were transferred for rooting to MS medium supplemented with indole-3-butyric acid (IBA). All the regenerated plants with 85 % survival rate were normal with respect to morphology and growth characteristics.     

Induction of plant defence compounds by Pseudomonas chlororaphis PA23 and Bacillus subtilis BSCBE4 in controlling damping-off of hot pepper caused by Pythium aphanidermatum

Bacillus subtilis strain BSCBE4 and Pseudomonas chlororaphis strain PA23 (=P. aureofaciens) were effective biocontrol agents against Pythium aphanidermatum, the causal agent of damping-off of hot pepper (Capsicum annum L.) in greenhouse vegetable production systems. Application of strains BSCBE4 and PA23 at the rate of 20 g kg^-1 of seed significantly increased the growth of hot pepper seedlings. The efficacies of various carriers in sustaining the population of these strains in storage were assessed. Both the antagonists survived up to 180 days of storage in peat and talc-based formulations. The two bacterial strains induced development of plant defence-related enzymes including phenylalanine ammonia lyase, peroxidase, polyphenol oxidase, phenol content, suppressed incidence of damping-off and increased growth of hot pepper seedlings.     

Cloning and functional annotation of rare mRNA species from drought-stressed hot pepper (<Emphasis Type="Italic">Capsicum annuum</Emphasis>)

To better understand gene expression at very low levels, we have designed a method to eliminate cDNA clones representing abundant mRNAs. A cDNA library for drought-stressed hot pepper (Capsicum annuum) (Choi et al., 2002) underwent double-negative screening, once with probes made from a drought-stressed plant, the second time, with probes from a non-stressed plant. The cDNA clones that showed very weak or negative signals were isolated for further analysis, which resulted in 1399 cDNA clones from about 20,000 screened clones. When nucleotide sequences were determined, we obtained 1142 tentative unique genes, with a redundancy rate of 20.41%. An homology database search for the deduced amino acid sequences revealed that about 79% of the cDNA clones could not be matched for functioning with previously characterized sequences. However, when these uncategorized clones were subjected to classification based on functional domains, most could be cited. Notably, clones with possible functions in RNA transport, protein synthesis, and regulation of protein activity showed a dramatic increase in appearance while those coding for transposable elements, viral proteins, and plasmid proteins occupied a much smaller portion compared with those in theArabidopsis thaliana genome. In addition, those coding for proteins targeted to the endoplasmic reticulum were dramatically more abundant in our clones compared with theArabidopsis database.     

Touch inhibits touch: sanshool-induced paradoxical tingling reveals perceptual interaction between somatosensory submodalities

Human perception of touch is mediated by inputs from multiple channels. Classical theories postulate independent contributions of each channel to each tactile feature, with little or no interaction between channels. In contrast to this view, we show that inputs from two sub-modalities of mechanical input channels interact to determine tactile perception. The flutter-range vibration channel was activated anomalously using hydroxy-α-sanshool, a bioactive compound of Szechuan pepper, which chemically induces vibration-like tingling sensations. We tested whether this tingling sensation on the lips was modulated by sustained mechanical pressure. Across four experiments, we show that sustained touch inhibits sanshool tingling sensations in a location-specific, pressure-level and time-dependent manner. Additional experiments ruled out the mediation of this interaction by nociceptive or affective (C-tactile) channels. These results reveal novel inhibitory influence from steady pressure onto flutter-range tactile perceptual channels, consistent with early-stage interactions between mechanoreceptor inputs within the somatosensory pathway.     

辣椒果实性状主基因+多基因遗传分析

以果实性状差异较大的一年生辣椒材料C.annuum B9431(P1)和灌木辣椒材料C.frutescens H108(P2)为亲本,构建4世代群体(P1、P2、F1、F2),应用数量性状主基因+多基因混合遗传模型方法对辣椒6个果实性状进行遗传分析,为辣椒果实性状QTL定位及分子标记辅助育种研究奠定理论基础。结果表明:6个果实性状均符合2对主基因+多基因遗传模型。其中,单果重量、果实纵径、果实横径、果形指数和果肉厚度5个性状均符合E-0模型,即2对加性-显性-上位性主基因+加性-显性-上位性多基因混合遗传模型;果柄长度最佳遗传模型为E-5,即2对完全显性主基因+加性-显性多基因模型。单果重量、果实纵径、果实横径、果形指数、果肉厚度和果柄长度主基因遗传率分别为87.64%、37.67%、82.46%、94.82%、83.33%和40.00%,多基因遗传率分别为7.50%、54.56%、10.53%、0.27%、12.96%和37.78%。