基于不同策略构建辣椒核心种质的研究

核心种质的构建为作物种质资源的高效利用提供了便利条件。以420份辣椒(Capsicum annuum L.)种质为试材,采用混合线性模型分析方法无偏地预测11个数量性状的基因型值,利用马氏距离计算种质间的遗传距离,分别采用3种聚类方法(中间距离法、类平均法和离差平方和法)、3种抽样方法(随机抽样法、优先抽样法和偏离度抽样法),按照25%的抽样比率构建辣椒核心种质库,采用均值、方差、极差和变异系数4个指标评价不同方法构建核心种质库的优劣。结果表明,类平均法优于中间距离法和离差平方和法,偏离度抽样法优于随机抽样法和优先抽样法。基于马氏距离、类平均法、偏离度抽样法获取的105份辣椒核心种质资源能够代表原群体的遗传多样性,为辣椒种质资源的高效利用提供了重要的理论依据。     

Genetic diversity and mixed infections of begomoviruses infecting tomato, pepper and cucurbit crops in Nicaragua

Begomoviruses were detected in Nicaraguan fields of tomato ( Lycopersicon esculentum ) and adjacently growing plants of pepper ( Capsicum annuum ), chilli pepper ( C . baccatum ), cushaw ( Cucurbita argyrosperma ) and Mexican fireplant ( Euphorbia heterophylla ) using polymerase chain reaction (PCR) and universal begomovirus primers. All tomato and Mexican fireplant plants showing symptoms were infected with begomoviruses, while only 30–46% of the pepper, chilli pepper and cushaw plants showing symptoms tested virus-positive. No begomoviruses were found in potato. The virus species were provisionally identified by sequencing 533 bp of the viral coat protein gene ( AV1 ). Tomato severe leaf curl virus (ToSLCV), Tomato leaf curl Sinaloa virus (ToLCSinV) and Pepper golden mosaic virus (PepGMV) were found to infect both tomato and pepper. A new provisional species designated Tomato leaf curl Las Playitas virus (ToLCLPV) was detected in a tomato plant. Squash yellow mottle virus (SYMoV) and PepGMV were found in cucurbits, the latter for the first time in this host. Euphorbia mosaic virus (EuMV) was detected in Mexican fireplant. Sequencing of a larger number of PCR-amplified clones from selected plants revealed intraspecific viral sequence variability, and also multiple begomovirus infections which could represent up to three species in a single tomato or cushaw plant. Phylogenetic grouping of virus sequences did not correlate with the host of origin.     

Selection forMeloidogyne incognita virulence against resistance genes from tomato and pepper and specificity of the virulence/resistance determinants

Experiments were designed to analyze the relationships between the root-knot nematodeMeloidogyne incognita and resistant tomato and pepper genotypes. From a natural avirulent isolate, near-isogenic nematode lineages were selected with virulence either against the tomatoMi resistance gene or the pepperMe3 resistance gene. Despite the drastic selection pressure used, nematodes appeared unable to overcome the pepperMe1 gene, therefore suggesting some differences in the resistance conferred byMe1 andMe3 in this species. Nematodes virulent onMi-resistant tomatoes were not able to reproduce onMe1-resistant nor onMe3-resistant peppers, and nematodes virulent onMe3-resistant peppers were not able to reproduce onMi-resistant tomatoes nor onMe1-resistant peppers. These results clearly demonstrate the specificity ofM. incognita virulence against resistance genes from both tomato and pepper, and indirectly suggest that gene-for-gene relationships could occur between these two solanaceous crops and the nematode.     

Effects of Temperature on Infection in Capsicum sp. and Nicotiana benthamiana by Impatiens Necrotic Spot Tospovirus

Infection by two isolates of impatiens necrotic spot tospovirus (INSV) under temperature regimes of 25/18°C (day/night) or 33°C (continuous) was studied in Capsicum annuum (systemically susceptible to tomato spotted wilt tospovirus, TSWV), C. chinense PI 152225 and PI 159236 (reacting hypersensitively to TSWV) and Nicotiana benthamiana (systemically susceptible to both tospoviruses). At 25/18°C infection was systemic in all hosts tested. At 33°C infection in N. benthamiana was systemic whereas in C. annuum and C. chinense it was restricted to the inoculated leaves. The result differed from that reported for TSWV, where high temperature made plants more susceptible, or caused no difference. Exchanging temperature regimes 6h to 4 days after inoculation did not affect the final results one month later, with plants being only locally infected at 33°C continuous regime, or systemically infected at 25/18°C alternate regime. The two INSV isolates were biologically and serologically stable for 5 passages in N. benthamiana held continuously at 33°C.     

Seedborne fungi detected on stored solanaceous berry seeds and their biological activities

We isolated 629 fungi from 1296 berry seeds of solanaceous plants, including tomato (Lycopersicon esculentum), eggplant (Solanum melongena), bell pepper (Capsicum annuum), and red pepper (Capsicum annuum var. annuum) preserved for long and short terms. The isolates were classified into 22 genera excluding unidentified fungi, and the fungal floras were divided into two types: the tomato–eggplant and pepper groups. The results of cluster analysis with unweighted pair-group method with arithmetic average also supported these groups. Most tomato seeds infested with Geotrichum candidum germinated and grew the same as uninfested seeds. Cladosporium sphaerospermum and Arthrinium sp. isolated from eggplant seeds strongly suppressed germination, and Penicillium variabile suppressed seminal root elongation on eggplant. Alternaria alternata, Botrytis cinerea, and Myrothecium verrucaria detected from red pepper or bell pepper seeds were pathogenic to the fruits and the seedlings after artificial inoculation.     

Assessment of real-time PCR as a method for determining the presence of <Emphasis Type="Italic">Verticillium dahliae</Emphasis> in different Solanaceae cultivars

Real-time PCR was used to detect and quantify Verticillium dahliae and to assess the susceptibility of four Capsicum annuum cultivars (Luesia, Padrón, SCM331 and PI201234) and the Capsicum chinense cv. C118 to this pathogen. The symptoms which developed after infection included stunting and yellowing, and were more acute in the cv. SCM331, which also suffered defoliation in later stages of the disease and in C118, which suffered severe stunting. Quantification of the pathogen DNA in roots 23 and 34 days post-inoculation (dpi) revealed that there were significantly higher amounts of Verticillium dahliae DNA in C118 than in the other cultivars, followed by SCM331, Padrón and PI201234. The lowest amounts of fungal DNA in roots were found in Luesia. In hypocotyls, the highest amounts of fungal DNA were found in SCM331, while Luesia, Padrón and PI201234 had much lower amounts, and C118 had intermediate levels. When a compatible versus an incompatible system was studied, using the near-isogenic tomato lines LA3030 (susceptible) and LA3038 (resistant to V. dahliae), we were able to detect fungal DNA in both lines. As expected, the fungus/plant DNA ratio was lower in LA3038 than in LA3030 and it decreased with time in LA3038. The amount of Verticillium dahliae DNA in the roots of LA3030 remained constant between days 23 and 34 post-inoculation, but increased 10-fold in collars. Finally, when real-time PCR was applied as a diagnostic method to samples from pepper plants, soil and water collected from farms in northwest Spain, we were able to detect V. dahliae DNA in these samples even when symptoms of the disease were not evident.     

Sulphur accumulation after Verticillium dahliae infection of two pepper cultivars differing in degree of resistance

Elemental sulphur levels, sulphur localization in stems, and levels of sulphate, glutathione and cysteine were studied in pepper ( Capsicum annuum ) cvs Yolo Wonder (higher resistance) and Luesia (lower resistance) after inoculation with Verticillium dahliae , the cause of vascular wilt. Accumulation of elemental sulphur (S⁰) was first detected 10 days after inoculation in Yolo Wonder (mean S⁰ level 7·3  µ g g⁻¹ DW), and 15 days after inoculation in Luesia (mean S⁰ level 3·3  µ g g⁻¹ DW). The maximum level was reached 21 days after inoculation in Yolo Wonder (14·1  µ g g⁻¹ DW). In control plants, elemental sulphur was not detected. SEM-EDX (scanning electron microscopy-energy dispersive X-ray microanalysis) indicated that the sulphur was not restricted to a specific location, but was dispersed throughout the vascular tissue. Sulphate levels showed a decline at the end of the experiment in inoculated plants, possibly related to the increase in sulphur levels seen in the two cultivars. The differences in sulphate levels between the two cultivars may be due to faster sulphate breakdown in cv. Yolo Wonder.     

Innovative liquid formulation of digestates for producing a biofertilizer based on Bacillus siamensis: Field testing on sweet pepper

A biofertilizer (BF) based on the plant growth promoting rhizobacterium (PGPR) Bacillus siamensis was produced using anaerobic digestate (AD) as the main ingredient of the growth medium, alongside a carbon source from residual origin. The use of residues for the growth of PGPR reduces the production costs of biofertilizers, but makes an assessment of the possible toxicity of residues for the bacteria or plants necessary. Therefore, the growth medium of PGPR was first optimized using the response surface methodology (RSM), followed by phytotoxicity tests and a field trial of the BF in a sweet pepper (Capsicum annuum L.) crop at two different locations. AD at 50% dilution, supplemented with 2.3% sugar beet molasses, was the optimum growth medium for producing the BF, with a bacterial concentration of 10⁹ cfu mL⁻¹. In the field trial, the treatments inoculated with BF and fertilized with decreased mineral N (80%) produced significantly better yields per ha than the controls with decreased N (80%) and full N (100%) without BF. This indicates improved efficiency of N use by the crop, as a consequence of the use of BF.     

辣椒种子中辣椒轻斑驳病毒的检测

采用三抗体夹心酶联（TAS-ELISA）法和反转录PCR（RT-RCR）扩增法分别对3份进口的包被种衣剂的辣椒种子进行了辣椒轻斑驳病毒（Pepper mild mottle virus,PMMoV）的检测。结果表明,两种方法均表明被检样品携带PMMoV,检测结果比较理想。在此基础上建立了一种快速、灵敏的从辣椒种子中直接检测PMMoV的检测体系,缩短了检测时间,提高了检测效率。     

小米辣辣椒素类物质含量测定

【目的】对小米辣Capsicum frutescens的辣椒素类物质合成规律进行研究.【方法】采用高效液相色谱法,对花后13个不同时期小米辣胎座、果肉的辣椒素和二氢辣椒素含量进行检测,结合SAS软件的多因素方差法进行分析.【结果和结论】辣椒素类物质在胎座和果肉中均能合成;其合成随花后果实发育逐渐增加到最高值,而后略微降低;其含量的高低与辣椒果实中组织部位和合成时期显著相关(P=0.00≤0.01),以胎座部位最多(22 d的质量分数达11.418 mg·g-1).