Chromoplast-Specific Proteins in Capsicum annuum

Chromoplasts are a common differentiation state of plastids in which the photosynthetic apparatus is absent and carotenoids accumulate to high levels. As a first step toward the isolation of chromoplast-specific genes, we have examined plastids of the bell pepper, Capsicum annuum L., for the presence of chromoplast-specific proteins. Intact chromoplasts were isolated from mature fruits of C. annuum var Emerald Giant, Golden Cal Wonder, and DNAP VS-12 by differential centrifugation followed by isopycnic sedimentation in gradients of silica sols. The plastids were then fractionated into soluble and membrane components and the proteins analyzed by one- and two-dimensional gel electrophoresis using isoelectric focusing, sodium dodecyl sulfate, and sodium dodecyl sulfate-urea gels. Two polypeptides with M_r of 35,000 and 58,000 accumulate to high levels in membrane fractions of chromoplasts of var Emerald Giant. These polypeptides are either not detectable or barely detectable in chloroplasts from immature fruits. Both polypeptides have been purified to near homogeneity. Yellow chromoplasts from var Golden Cal Wonder and red chromoplasts from var DNAP VS-12 contained the 35-kilodalton polypeptide, but not the 58-kilodalton species.     

Synthesis of Two Chromoplast-Specific Proteins During Fruit Development in Capsicum annuum

The time-course of accumuiation of two membrane proteins during fruit ripening was examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and western blots in tissue extracts of Capsicum annuum L., vars Emerald Giant, Albino, and DNAP VS-12. The proteins, named ChrA and ChrB, were previously shown to occur specifically in chromoplasts. Fruit development was divided into five stages based on changes in color. ChrA was not detectable in the first three stages, but accumulated to a high level in the fully mature, red fruit. ChrB was not detectable in the first, mature-green stage of fruit maturation, but was found in the second stage, when carotenoid accumulation first appeared, and in all later stages. The patterns of accumulation in chromoplasts that develop from proplastids or leucoplasts are similar to those in chromoplasts that develop from chloroplasts. We conclude that ChrA and ChrB are probably synthesized de novo during chromoplast development.     

Rhythmic Nitrate Reductase Activity in Leaves of Capsicum annuum L. and the Influence of Kinetin

In the expanding leaves of Capsicum annuum L. cv. California Wonder, two of the three peaks of nitrate reductase activity associated with the light period exhibit a circadian rhythm that persists in continuous light.     

在条锈菌侵染早期小麦初生叶内多聚核糖体水平与蛋白质合成能力的变化

小麦初生叶接种条锈菌毒性生理小种（ＣＹ２９）及其弱毒突变菌系（ＣＹ２９－ｍｕｔ３）后,呈不亲和反应的寄主叶片可溶性蛋白质合成能力在接种后２４ｈ显著高于未接种对照,但其后逐渐降低,直至接近对照;而呈亲和性反应的寄主叶片可溶性蛋白质合成在侵染早期与对照相近,但与膜结合蛋白质在９６ｈ时大大高于对照。对接种叶中核糖体的密度梯度分析证实：呈不亲和反应寄主叶片游离多聚核糖体及亲和反应的寄主内与膜结合多聚核糖体均有特异性增加。上述结果表明寄主的抗病和感病反应均与蛋白质合成能力的变化有关。     

Polyribosome Content in Young and Aged Wheat Leaves Subjected to Drought

The effect of a droughting treatment on the polyribosome contentof wheat leaves was markedly different on leaves of differentages. Growing leaves showed a large decrease in their polyribosomepopulation, while fully expanded leaves showed no loss of polyribosomescompared with controls. These results suggest that the largereductions in polyribosome population observed in growing tissuesare due to reductions in growth, rather than a direct effectof water stress.     

生物技术在辣椒育种研究上的应用

综述了国内外组织及器官培养、分子标记和基因工程三大生物技术在辣椒育种的研究与利用，并进行了展望。     

Peroxidases and the metabolism of capsaicin in Capsicum annuum L.

Capsaicinoids are acid amides of C₉ - C₁₁ branched-chain fatty acids and vanillylamine. These compounds are responsible for the pungency of the Capsicum species and of cultivars regarded as hot peppers. Moreover, it has been suggested that these compounds play an ecological role in seed dispersal. Because they are used in the pharmacological, food and pesticide industries, much attention has been paid on knowing how their accumulation is controlled, both in the fruit and in cell cultures. Such control involves the processes of biosynthesis, conjugation and catabolism. Recent progress has been made on the biosynthetic pathway, and several of the genes coding for biosynthetic enzymes have been cloned and expression studies performed. With regard to catabolism, cumulative evidence supports that capsaicinoids are oxidized in the pepper by peroxidases. Peroxidases are efficient in catalyzing in vitro oxidation of both capsaicin and dihydrocapsaicin. These enzymes are mainly located in placental and the outermost epidermal cell layers of pepper fruits, as occurs with capsaicinoids, and some peroxidases are present in the organelle of capsaicinoid accumulation, that is, the vacuole. Hence, peroxidases are in the right place for this function. The products of capsaicin oxidation by peroxidases have been characterized in vitro, and some of them have been found to appear in vivo in the Capsicum fruit. Details on the kinetics and catalytic cycle for capsaicin oxidation by peroxidases are also discussed.     

辣椒叶多糖抗氧化作用研究

目的:从辣椒叶中分离纯化活性多糖并考察其抗氧化活性。方法:采用水提醇沉法提取多糖,Sevag法脱蛋白,得辣椒叶粗多糖;分别使用超纯水、0.06 mol/L NaCl溶液、0.18 mol/L NaCl溶液作为洗脱液,通过DEAE-52离子交换柱色谱纯化得到三种辣椒叶多糖LD-0、LD-0.06、LD-0.18,测定多糖含量。DPPH、ABTS法检测多糖体外抗氧化作用。以小鼠血清、肝组织中总超氧化物歧化酶活性、过氧化氢酶活性及丙二醛含量为指标,考察小鼠灌胃LD-0.06多糖对脂质过氧化模型的影响。结果:辣椒叶粗多糖、LD-0、LD-0.06、LD-0.18多糖含量分别为9.92%、43.14%、82.97%、37.63%,其中LD-0.06多糖含量最高。体外抗氧化实验结果显示,三种结果均具备较好的清除能力,其中LD-0.06对ABTS+、DPPH·的清除效果最好,IC50值分别为0.58 mg/m L和0.60 mg/m L,结果与对照组在0.05水平具有显著性差异,说明辣椒叶多糖提取物是一种有效的天然抗氧化剂。在脂质过氧化模型小鼠体内,与模型组比较,LD-0.06多糖能显著增强小鼠血清和肝组织中的T-SOD与CAT活性,降低MDA的含量,且剂量越高,体内抗氧化能力越强。结论:辣椒叶多糖提取物具有一定的抗氧化作用,为进一步开发利用辣椒资源提供了理论依据。     

Carbon Partitioning and Export in Mature Leaves of Pepper (Capsicum annuum)

The partitioning of recently fixed carbon by mature pepper leaveshas been examined over a 10 h photoperiod using a constant specificradioactivity ¹⁴CO₂ labelling technique. Changes in the ratesof carbon partitioning into export, starch, sucrose and hexoseswere examined following changes in irradiance during the photoperiod.Leaves grown under 80 W m^–2 PAR were exposed to this irradiancefor the first 4 h of the photoperiod then the iiradiance wasdecreased. Leaves accumulated sufficient reserves in the first4 h to maintain export at the initial rate (approximately 20µg carbon cm^–2 leaf h^–1) over the following6 h of the photoperiod when the net photosynthesis rate (Pn)was decreased to 10% of the initial rate by the decreased irradiance.Export was initially maintained by the depletion of sucroseand hexose and then by carbon from the degradation of starchin the light. If leaves were exposed to low irradiance at the beginning ofthe photoperiod, then the export rate was linearly related tothe Pn during that period. When Pn exceeded that required tomaintain an export rate of approximately 20 µg carboncm^–2 h^–1, then more carbon was partitioned intostarch. At low initial irradiance, a greater proportion of photosynthatewas partitioned into export rather than starch and at high initialirradiancc the reverse occurred. There was a linear relationship between starch accumulationrate and Pn for all leaves but the relationship between Pn andexport rate was only significant for leaves with low levelsof reserve carbon. The results show that mature pepper leaves subjected to differentirradiances maintain constant export rates through alterationsof carbon partitioning. Export at low Pn is maintained at theexpense of sugar and starch reserves, with partitioning in highirradiance being predominantly to starch. Key words: Carbon partitioning, Starch, Export, Pepper (Capsicum annuum L.)     

Behaviour of the Phenolic Compounds on Capsicum annuum Leaves Infected with Phytophthora capsici

In Capsicum annuum leaves, some phenolic compounds, such as chlorogenic acid and various apigenin or luteolin glycosides, can be separated by HPLC technique. During infection with Phytophthora capsici (the agent of pepper mildew), the concentrations of these compounds decreased. A fungal glucosidasic activity may be implicated, producing apigenin aglycon in the necrotic areas. Some of the phenolic compounds were only present in intercellular spaces, where their concentration increasedduring fungal infection, mostly in the resistant pepper cultivar. The involvement of phenolics in C. annuum resistance is discussed in relation to oxidative metabolism.     

Properties of chlorophyllase from Capsicum annuum L. fruits

The in vitro properties of semi-purified chlorophyllase (chlorophyll-chlorophyllido hydrolase, EC 3.1.1.14) from Capsicum annuum fruits have been studied. The enzyme showed an optimum of activity at pH 8.5 and 50 degrees C. Substrate specificity was studied for chlorophyll (Chl) a, Chl b, pheophytin (Phe) a and Phe b, with Km values of 10.70, 4.04, 2.67 and 6.37 microM respectively. Substrate inhibition was found for Phe b at concentrations higher than 5 microM. Chlorophyllase action on Chl a' and Chl b' was also studied but no hydrolysis was observed, suggesting that the mechanism of action depends on the configuration at C-13(2) in the chlorophyll molecule, with the enzyme acting only on compounds with R132 stereochemistry. The effect of various metals (Mg2+, Hg2+, Cu2+, Zn2+ Co2+, Fe2+ and Fe3+) was also investigated, and a general inhibitory effect was found, this being more marked for Hg2+ and Fe2+. Functional groups such as -SH and -S-S- seemed to participate in the formation of the enzyme-substrate complex. Chelating ion and the carbonyl group at C3 appeared to be important in substrate recognition by the enzyme. The method for measuring Chlase activity, including HPLC separation of substrate and product, has been optimized.     

观赏辣椒一步成苗诱导条件的筛选

采用正交试验设计方法探讨6-BA、NAA和2,4-D对观赏辣椒一步成苗影响的结果表明:6-BA与NAA的浓度比对丛生芽诱导的影响最大。筛选出的观赏辣椒一步成苗的最适培养基为1/2MS 1.0mg·L-16-BA 0.5mg·L-1NAA 0.2mg·L-12,4-D,并获得移栽后的成活植株。     

Cytogenetic markers for the characterization of Capsicum annuum L. cultivars

Karyotypic studies with conventional staining have been unsuccessful due to the uniformity of Capsicum chromosomes. In this study, we found diagnostic chromosome characters that permit to characterize cultivars; this is the first cytological characterization of both rDNAs (18S and 5S) in a species of Capsicum using a genus-specific probe and the most exhaustive in C. annuum to date. The heterochromatic banding patterns enabled us to identify cultivars, and fluorescent in situ hybridization (FISH) showed one 5S rDNA locus largely conserved within the cultivars, whereas high variation in the number of 18S rDNA loci was observed. One of the most obvious differences is the presence of an additional active nucleolar organizer region in pair #12 and the dispersal of inactive 18S rDNA signals. These results indicate that fluorochrome banding together with silver impregnation and FISH procedures are very useful for the identification of chromosomes and the interpretation of chromosomal variation between cultivars. The functional role of this variation is still uncertain, but our results show that copy number variation of repetitive DNA during the course of evolution might provide an excellent experimental system for studying genome rearrangements accompanying functional divergence in domesticated C. annuum.     

Virus Diseases of Pepper (Capsicum annuum L.) in Ethiopia

Viruses occurring in hot pepper (Capsicum annuum L.) in Ethiopia during 1984–1986 were identified and their symptomatology, host range, serology, morphology and transmission described. Potyviruses were found to cause severe infections of pepper wherever the crop was observed. Disease incidence and yield loss in most surveyed areas were estimated at 40–100 % and 15–50 %, respectively. Besides potato Y and pepper veinal mottle viruses, a new potyvirus with distinctive properties was found widely distributed in Shewa and Welega Provinces. The virus, tentatively designated Ethiopian pepper mottle virus (EPMV), was shown to have an unusually narrow experimental host range and filamentous particles about 700–750 nm in length, and to be nonpersis-tently transmitted by aphids but not transmissible through pepper seeds. Cucumber mosaic virus was detected in some locations, but appeared to be of minor significance.     

EMS-induced lincomycin resistance in red pepper (Capsicum annuum L.)

Summary Ethyl methane sulphonate (EMS) is a potential mutagen to induce lincomycin resistance in Capsicum annuum. Mutagenized cotyledons were cultured on shoot regenerating medium containing lincomycin (100 mgl⁻¹). Approximately 14% of regenerated shoots were chlorophyll deficient and about 4% of regenerated shoots were green from mutaganized cotyledons. The regenerated green plants were resistant to lincomycin but sensitive to chloramphenicol, kanamycin, spectinomycin, and streptomycin. Reciprocal crosses were made between resistant and sensitive plants. Inheritance of lincomycin resistance was transmitted as a non-Mendelian trait. Lincomycin resistance is a first selectable and maternally inherited organelle encoded genetic marker described in chili pepper. Such mutants should be useful in designing biochemical selection schemes to recover somatic hybrids and cybrids.     

Marker development for erect versus pendant-orientated fruit in Capsicum annuum L

The erect habit of fruit setting is a unique characteristic of ornamental peppers and wild pepper species. The erect habit is known to be controlled by the up locus on pepper (Capsicum annuum L.) chromosome 12. The result of a genetic analysis using Saengryeog 211 (pendant), Saengryeog 213 (erect), and their F1 and BC1 progeny demonstrated that up is a recessive gene. To develop an up-linked marker, bulked segregant analysis (BSA) and amplified fragment length polymorphism (AFLP) were employed using 108 F2:3 individuals. The closest AFLP marker, A2C79, was located at a genetic distance of 1.7 cM from the up locus and was converted into a cleaved amplified polymorphic sequence (CAPS) marker. This marker was mapped at a genetic distance of 4.3 cM from the up locus. When the CAPS was applied to seven ornamental lines and 27 breeding lines with erect fruit, these genotypes of 28 lines were correctly predicted. Thus, the CAPS marker will be useful for marker-assisted selection (MAS) of pepper breeding lines with the up allele at the early seedling stage.