Begomovirus diversity in tomato crops in Costa Rica

Begomoviruses (Geminiviridae family) are characterized by their high recombination rate and a wide range of hosts, making their control difficult. In Costa Rica, various species of bipartite begomoviruses have been reported, which are Pepper golden mosaic virus (PepGMV), Tomato yellow mottle virus (ToYMoV), Tomato leaf curl Sinaloa virus (ToLCSiV) and the monopartite begomovirus Tomato yellow leaf curl virus (TYLCV). Since the TYLCV first report in Costa Rica, neither additional knowledge has been produced on how this begomovirus has spread in the country's territory nor on the distribution of the other bipartite species. A total of 429 tomato samples collected during the years 2015–2016 were used to study these aspects. Each sample was georeferenced and analysed with various techniques such as nucleic acid hybridization, polymerase chain reaction (PCR) and sequencing for the begomoviruses previously reported in Costa Rica. It was found that the presence/absence of the different species can vary, depending on the province. TYLCV is present in the six provinces analysed in this work, with a proportion from 3.7 to 86.6 per cent. Alajuela, Cartago, and Heredia are the provinces most affected by tomato-infecting begomoviruses. Fourteen different haplotypes of TYLCV were detected, but all were identified as TYLCV-IL. The distribution of TYLCV was related to the presence of the whitefly Bemisia tabaci MED, especially in the country's main tomato production areas. This information allows the phytosanitary surveillance services to develop strategies for the integrated management of the disease and to contribute data to the genetic improvement programmes of the crop.     

Characterisation and genetic diversity of pepper leafroll virus,a new bipartite begomovirus infecting pepper,bean and tomato in Peru

The complete genome of a novel bipartite begomovirus (genus Begomovirus, family Geminiviridae) was cloned from a severely diseased yellow Peruvian chili pepper (Capsicum baccatum cv. Pendulum) plant collected in the department of La Libertad, Northern Peru and full‐length sequenced. The two genomic components share a common region of 156 nucleotides with a 100% sequence identity. Analysis of the genome organisation and phylogenetic comparisons revealed that the virus is a typical New World begomovirus. The closest related begomovirus, an isolate of Tomato yellow vein streak virus (ToYVSV), shared only 76.8% nucleotide sequence identity for the DNA‐A component. Therefore, following species demarcation criteria of the International Committee on Taxonomy of Viruses, this virus isolate belongs to a new begomovirus species for which the name pepper leafroll virus (PepLRV) is proposed. Pepper plants infected with the cloned PepLRV isolate developed leaf roll symptoms similar to those observed in field‐infected plants suggesting this virus as the causal agent of the disease syndrome observed in the field. Widespread occurrence of PepLRV throughout Peru was demonstrated, infecting plants of diverse cultivated species such as tomato, pepper, common and pallar beans, and of the weed species Nicandra physaloides. Low genetic diversity was observed among PepLRV isolates present in this country with no evident geographical or temporal structure of the population, typical of a recent founder effect. This is the first report of a begomovirus infecting pepper and bean crops in Peru.     

Analysis of resistance in tomato and sweet pepper against the greenhouse whitefly using electrically monitored and visually observed probing and feeding behaviour

Different types of plant resistance against the greenhouse whitefly, Trialeurodes vaporariorum, were distinguished according to the tissue location of the resistance factors. The effects of resistance factors were compared by measuring the electrically monitored and visually observed probing and feeding behaviour of whiteflies on two resistant tomato genotypes (82207 and 82216), a susceptible tomato cultivar, and a non-host plant, sweet pepper. On sweet pepper, whiteflies displayed very short first probes, very long pathway probing and spent much time on non-feeding activities such as walking and standing still. Also, a high percentage of whiteflies rejected sweet pepper without ingesting substances from the phloem vessel. These data suggest a strong resistance that is based on the factors present in surface/epidermis and/or mesophyll layers of this plant. The behaviour of whiteflies on tomato 82207 was very different from that on sweet pepper. On tomato 82207 whiteflies apparently did not perceive resistance factors on the leaf surface and in the mesophyll. Resistance factors appeared to be present in the phloem tissue, because a higher number of phloem phases, longer phloem salivation periods and shorter phloem ingestion periods were observed when compared with the susceptible tomato cultivar. Tomato 82216 showed no clear resistance factor in the phloem. The importance of combined EPG and behavioural research for host-plant resistance studies are discussed.     

Growth, photosynthesis and chlorophyll fluorescence of sweet pepper plants as affected by the cultivation method

Different methods of sweet pepper cultivation (organic, integrated and conventional farming) were evaluated under greenhouse conditions. Organic treatment was defined as only 4‐kg manure application at the preplanting stage. Conventional and integrated treatments consisted of application of the manure dosage plus chemical fertiliser at 100% and 50% of the local farmers’ amount, respectively, to avoid nutrient depletion. Plant growth parameters such as shoot dry matter, total leaf area and leaf weight fraction were all reduced in the organic treatment compared with the conventional. Leaf expansion was dramatically reduced in the organic treatment 155 days after transplanting. Relative growth rates were significantly affected by the nitrogen concentration in each organ and were directly related to the cultivation method. Chlorophylls (a + b) contents in the leaves were reduced in the organic treatment and were directly correlated with the nondestructive quantification of chlorophylls using a portable chlorophyll meter. Net photosynthesis was also reduced in the organic treatment, but chlorophyll fluorescence was not affected. This study shows that biometric monitoring and fast and non‐invasive techniques of plant nutrient status analysis could help to improve growth as easy and useful tools to follow the nutrient status at different phenological stages, especially when no chemical fertilisers are applied.     

Carbohydrate metabolism during fruit development in sweet pepper (Capsicum annuum) plants

Growth, accumulation of sugars and starch, and the activity of enzymes involved in sucrose mobilization were determined throughout the development of sweet pepper fruits. Fruit development was roughly divided into three phases: (1) an initial phase with high relative growth rate and hexose accumulation, (2) a phase with declining growth rate and accumulation of sucrose and starch, and (3) a ripening phase with no further fresh weight increase and with accumulation of hexoses, while sucrose and starch were degraded. Acid and neutral invertase (EC 3.2.1.26) were closely correlated to relative growth rate until ripening and inversly correlated to the accumulation of sucrose. Acid invertase specifically increased during ripening, concurrently with the accumulation of hexoses. Sucrose synthase (EC 2.4.1.13) showed little correlation to fruit development, and in periods of rapid growth the activity of sucrose synthase was low compared to the invertases. However, during late fruit growth sucose synthase was more active than the invertases. We conclude that invertase activities determine the accumulation of assimilates in the very young fruits, and a reactivation of acid invertase is responsible for the accumulation of hexoses during ripening. During late fruit growth, before ripening, sucrose synthase is transiently responsible for the sucrose breakdown in the fruit tissue. Results also indicate that pyrophosphate-dependent phosphofructokinase (EC 2.7.1.90) and its activator fructose-2,6-bisphosphate (Fru2,6bisP) are involved in the regulation of the sink metabolism of the fruit tissue.     

Response of sweet pepper to phenols accumulated in greenhouse substrate

Prolonged cucumber cultivation in the same substrates leads to accumulation of phytotoxic phenolic compounds. Introduction of sweet pepper as an aftercrop eliminates substrate phytotoxicity. The aim of the study was to examine whether in sweet pepper detoxication of substrate phenols occurs by means of the glucosylation process. The examined materials were substrates differing in phytotoxicity level, and sweet pepper plants grown on these substrates. Substrate phytotoxicity was obtained by means of either repeated cucumber cultivation or by phenolic acid addition. During the vegetative growth phase of sweet pepper, the phytotoxicity and phenolic compound levels in the substrate, and the glucosylated phenol contents in above-ground plant parts were determined. Results showed that sweet pepper responds to an increased presence of phenols in the substrate by an intensified glucosylation.     

甜蛋白基因MBLII对番茄的遗传转化

以5～7d龄的 丽春 番茄无菌苗子叶作为外植体,研究了子叶外植体对抗生素卡那霉素的敏感性,抗生素卡那霉素对番茄筛选的适宜浓度为70mg/L.通过根癌农杆菌 Agrobacteriumtumefaciens 介导,成功地进行了马槟榔甜蛋白基因MBLII对番茄的遗传转化,获得转化番茄抗性植株,组织化学法有阳性表现、PCR特异扩增及Southern杂交检测出现特异条带,表明MBLII基因已顺利整合到转基因番茄植株的基因组.     

PCR-based assay for the detection of Xanthomonas euvesicatoria causing pepper and tomato bacterial spot

Aims:  To develop a PCR-based assay for Xanthomonas euvesicatoria detection in culture and in planta .
Methods and Results:  A fragment of 1600 bp specific for X. euvesicatoria was found by repetitive extragenic palindromic sequence-PCR. Among the primers designed on the basis of the partially sequenced fragment, the primers Xeu2.4 and Xeu2.5 direct amplification of the expected product (208 bp) for all the X. euvesicatoria strains and not for other related and unrelated phytopathogenic bacteria or saprophytic bacteria isolated from pepper and tomato phyllosphere. The assay permits the detection of X. euvesicatoria in pure culture, with a limit of detection of two bacterial cells and 1 pg of DNA per PCR, and in extracts obtained from asymptomatic inoculated tomato and pepper plants.
Conclusions:  Primers Xeu2.4 and Xeu2.5 provide a specific, sensitive and rapid assay for the detection of X. euvesicatoria in culture and in pepper and tomato plants.
Significance and Impact of the Study:  Because X. euvesicatoria is a quarantine organism in the European Union, and it is subjected to stringent international phytosanitary measures, this highly sensitivity PCR-based assay is suitable for its detection in pepper and tomato plant materials to avoid the introduction and spread of the bacterium.     

光质对温室甜椒干物质生产和分配指数的影响

为了研究不同光质对甜椒植株干物质生产与分配指数的影响,以甜椒茎有限生长品种"苏椒13号"为试验材料,于2010年计不同种彩色塑料薄膜(红、黄、蓝、绿、紫,白色为对照)试验。结果表明:不同光质处理的甜椒单位面积干物质生产量(Wt)与冠层截获的太阳光合有效辐射日积分(daily photosynthetic active radiation integral,PARi,MJ·m-2)之间的模型为Wt=22.07×e0.0054λPARi;单位面积植株总干重、果实的干物质分配指数、果实采收指数和单位面积果实产量均以红膜最高,紫膜最低;蓝膜的茎干物质分配指数最高,红膜最低;蓝膜、绿膜和紫膜的植株叶片干物质分配指数明显高于红膜和黄膜;不同光质处理对甜椒植株地上部分干物质分配指数的影响差异不显著;研究认为红膜和黄膜能够促进甜椒植株的干物质积累和果实发育,紫膜和蓝膜则有明显的抑制作用,该研究结果可为温室甜椒栽培的光质选择和环境调控提供决策依据。     

Differential expression of root-knot nematode resistance genes in tomato and pepper: evidence with Meloidogyne incognita virulent and avirulent near-isogenic lineages

Reproduction of artificially selected near isogenic Meloidogyne incognita lineages virulent and avirulent against the Mi resistance gene of tomato was assessed on host and resistant lines and cultivars of pepper. Egg mass production following inoculation of individual potted seedlings with second-stage juveniles was studied in experiments conducted in controlled environment. Artificially selected Mi-virulent nematode populations were unable to develop on resistant pepper lines PM 217 and PM 687. This suggests that the genetic systems governing resistance to root-knot nematodes are differently expressed in tomato and pepper, in spite of the very close phylogenetic relationships and structural genomic homologies occurring between these two vegetable crops. Moreover, these artificially selected nematode populations were also found unable to develop on the susceptible pepper cultivars California Wonder and Doux Long des Landes, while their pathogenicity was not significantly affected on susceptible tomatoes. Due to the existence of naturally virulent Meloidogyne populations, these results enhance the need for a better understanding of the mechanisms involved, in order to develop new forms of management of plant resistance to root-knot nematodes.     

Genetic diversity of begomoviruses infecting tomato plant in Saudi Arabia

Tomato is known as a highly valuable crop and grown worldwide for various uses. The cultivation and tomato production severely affected globally by several diseases caused by various pathogens. Begomoviruses causes yellow mosaic and leaf curl disease of tomato in the tropical, subtropical, temperate, and semi-arid regions. In Saudi Arabia, the tomato production adversely affected by disease caused by begomoviruses known as TYLCV and ToLCSDV. In this study, the pathogen was identified by Polymerase Chain Reaction using virus-specific primers and transmitted by whiteflies to healthy tomato seedlings. In a field survey, the tomato plants were exhibiting symptoms like viral infection. The infected leaf was randomly collected from various fields of tomato growing areas like Jeddah, Makkah, Tabuk, and Hail. The full-length viral genome was amplified by Rolling Circle Amplification technology (RCA) while betasatellites were amplified by PCR using universal betasatellites primers. The full-length viral genome (∼2.7 kb) and betasatellites (∼1.4 kb) were cloned and sequenced bi-directionally. The generated sequences were assembled and analyzed to find out the genetic variability by using bioinformatics tools and the genetic variability and phylogenetic relationships with selected begomoviruses were analyzed. The sequences showed the highest identity with an isolate of ToLCSDV and TYLCV. The nucleotide similarity and phylogenetic relationship showed the closest cluster with ToLCSDV and TYLCV. The data generated in this study elucidate that the causal organism is a variant of either TYLCV or ToLCSDV. The provided information from this study will be highly valuable for researchers and vegetable growers not only in Saudi Arabia but also in Arabian Peninsula.     

Genetic diversity of Bemisia tabaci cryptic species in Nigeria and their relationships with endosymbionts and acquired begomoviruses

Bemisia tabaci is a species complex of at least 44 cryptic species with a worldwide distribution. It is a serious pest of many crop plants as well as a successful vector of at least 100 begomoviruses. Using B. tabaci collected from cassava and tomato fields in the southwestern and north central regions of Nigeria, we determined nucleotide sequences from the mitochondrial cytochrome c oxidase subunit I (COI) of 23 B. tabaci samples, the 16S and 23S ribosomal DNA of endosymbionts, and the coat protein gene of geminiviruses ingested by the whiteflies. The COI analysis identified three different genetic groups including the indigenous Sub-Saharan Africa 1 subgroup 1 (SSA1-SG1) and 5 (SSA1-SG5, which was most prevalent), and an invasive cryptic species (Mediterranean). SSA1 was infected by five known secondary endosymbionts, Arsenophonus, Cardinium, Hamiltonella, Rickettsia, and Wolbachia, and co-infections with two or three endosymbionts were common. Five begomoviruses, okra enation leaf curl virus, squash leaf curl China virus, tobacco curly shoot virus, tomato leaf curl New Delhi virus, and tomato yellow leaf curl virus, were detected from 43.5% of the B. tabaci samples. However, cassava mosaic disease that causes devastating cassava yield losses was not detected in this study. This study improves the current understanding of the genetic diversity of B. tabaci cryptic species, and it reveals their relationships with endosymbionts and geminiviruses in the cassava and tomato fields of Nigeria.     

不同播期辣椒和番茄上烟粉虱成虫与捕食性天敌之间的关系

为了科学施药,合理利用和保护自然天敌进行烟粉虱的综合防治,用灰色系统分析方法、生态位分析方法和空间格局聚集强度指标分析方法,对春、秋两季辣椒和番茄田烟粉虱成虫及其主要捕食性天敌在数量、时间、空间格局等方面进行分析,结果表明:辣椒和番茄田烟粉虱成虫的捕食性天敌依次是粽管巢蛛、八斑球腹蛛、草间小黑蛛和小花蝽,两季辣椒田烟粉虱成虫的种群数量差异极显著(t=7.9063,t>t0.01).两季辣椒田中,八斑球腹蛛、异色瓢虫、粽管巢蛛、草间小黑蛛和龟纹瓢虫差异极显著(t>t0.01),锥腹肖蛸差异显著(t>t0.05).棚内天敌种类和数量较少.秋-冬季番茄田烟粉虱成虫数量显著高于辣椒田(t=2 4564,t>t0.01),而春-夏季两种作物上烟粉虱成虫种群数量没有明显差异,t=1.4628,t＜t0.05).     

广西番茄内生细菌的多样性和数量动态

为了探明内生细菌在番茄中的分布和数量变化规律,有目的地筛选防治番茄青枯病的内生细菌,我们对广西可培养的番茄内生细菌的类群和数量动态进行了调查。从广西部分县市采集的303个番茄样本中分离到624株内生细菌菌株,初步确定有芽孢杆菌(Bacillus)、假单胞菌(Pseudomonas)、黄单胞菌(Xanthomonas)、棒杆菌(Corynebacterium)、土壤杆菌(Agrobacterium)、微杆菌(Microbacterium)、肠杆菌(Enterobacter)和欧文氏菌(Erwinia)8个属,其中以芽孢杆菌、假单胞菌和土壤杆菌为芽孢杆菌为优势类群。番茄内生细菌在植株器官中的分布以根部数量最多,其次是茎和叶。内生细菌的总量在番茄生育期的变化趋势是从苗期到花期数量上升,而从结果期到成熟期数量逐渐下降。多数内生细菌种群的数量变化动态符合细菌总量的变化趋势,只有微杆菌在番茄植株整个生育期中始终保持下降的趋势。春季种植的番茄植株的内生细菌类群数量比秋季种植的少。     

The content of photosynthetic pigments and the light conditions in the fruits and leaves of sweet pepper

Investigations were carried out on the fruits of sweet pepper at its two development stages: on green fruits, on mature red and yellow fruits and on leaves. The content of the photosynthetic pigments and the optical properties were examined. In the green fruits when compared with leaves the content of the photosynthetic pigments is smaller by 40 to 50 % and the value of the ratio: chlorophyll a/b is lower. Chlorophyll is absent in mature fruits, while the content of carotenoids is a few times higher. The optical properties of green fruits and of the leaves in the PAR (photosynthetically active radiation) range are the same. In the range 700–1100 nm the green fruits show smaller reflectance and transmittance and a few times greater absorptance of irradiation which contributes to the warming up of the seed bag, while small absorptance of leaves in this range protects them against overheating. In mature fruits, in the PAR range, the reflectance and transmittance are higher, while the absorptance of irradiation in comparison with that of green fruits is smaller. In the range 700 – 1100 nm the changes are rather small and refer to the increase of reflectance and reduction of absorptance, while the transmittance of irradiation remains unchanged.     

Different transmission efficiencies may drive displacement of tomato begomoviruses in the fields in Taiwan

A progressive displacement of Tomato leaf curl Taiwan virus (ToLCTWV) by Tomato yellow leaf curl Thailand virus (TYLCTHV) from 2005 to 2009 has been recorded in tomato fields in Taiwan. Begomoviruses are exclusively transmitted by Bemisia tabaci complex, so we hypothesised that the displacement of tomato begomoviruses in the fields may be due to the invasion of a new virus/vector and the different transmission efficiencies of the viruses by the vectors. The objective of this research was to compare the transmission efficiency of TYLCTHV and ToLCTWV by the B and Q biotypes of B. tabaci complex. When transmission efficiency, virus retention in vector, and latent period for vector transmission were compared, the B biotype transmitted TYLCTHV and ToLCTWV more efficiently than did the Q biotype, and transmitted TYLCTHV more efficiently than ToLCTWV. The B biotype retained both viruses and remained infective throughout adulthood, but the Q biotype did not keep its infectivity, although it did retain both viruses lifelong. The B biotype transmitted TYLCTHV and ToLCTWV with the shortest latent period. In summary, B. tabaci B biotype and TYLCTHV is the best alliance for disease transmission, so we conclude that this may be one of drivers responsible for the displacement of ToLCTWV by TYLCTHV in tomato fields in Taiwan.     

Adaptability of tomato and pepper leaves to changes in photoperiod: Effects on the composition and function of the thylakoid membrane

The adaptability of the thylakoid membrane to extended photoperiod (from natural to 24 h) was studied using a photoperiod-sensitive species ( Lycopersicon esculentum Mill. cv. Trend) and a non-photoperiod-sensitive species ( Capsicum annuum L. cv. Delphin). Our results have shown that thylakoid membranes of both species adapt to an extended photoperiod by increasing their photosystem II to photosystem I ratio (PSII/PSI) in order to provide a more balanced energy distribution between both photosystems to improve quantum yield. In tomato plants, these results correspond with a lower chlorophyll (Chl) a/b ratio, a decrease in Chl associated with PSI light-harvesting chlorophyll a/b protein complexes and with an increase in Chl associated with PSII light-harvesting chlorophyll a/b protein complexes. In spite of these changes, the electron transport capacity through PSII and PSI per unit of Chl and the light saturation point of PSII remained unchanged. The inability of tomato plants to use supplemental light for an extended photoperiod is not the result of photoinhibitory conditions. In pepper plants a significant increase in electron transport capacity and in the light saturation point of PSII was found. There was a significant increase in CO₂ assimilation when the light period was increased from 12 to 24 h. In contrast to tomato, pepper plants adapt to a 24-h photoperiod by increasing their carboxylation capacity which is accompanied by an increase in electron transport capacity and the light saturation point.