Monitoring of co-infection virus and virus-like naturally in sweet pepper plant

Eight sweet pepper plant samples showing viral and viral like symptoms were collected from open field and used for detecting viral infections through biological, serological and biochemical methods. DAS-ELISA, DBIA and TPIA have relative effectiveness for detecting parenchymal viruses (CMV, TMV and PVY) and vascular virus (TYLCV), and the DAS-ELISA and TPIA are found more efficient (87.5%) than DBIA (78.1%). The examined leaf samples were found co-infected with different mixed types of viruses including (CMV, TMV, PVY and TYLCV), (CMV, PVY and TYLCV), (TMV, PVY and TYLCV) and (TMV and TYLCV) that enhanced different degrees of severe external symptoms. There are 2 out of 8 samples infected with Phytoplasma sp. by Diene’s stain and PCR using generated 16S rDNA gene primer with expected amplicon size of 680?bp. The co-infections with various viruses and phytoplasma has 12.5% frequency that reduced the levels of protein content, peroxidase and polyphenol oxidase activity quantitatively and qualitatively in 2 samples in comparison with other mixed categories. The sweet pepper plant can be considered as a reservoir for parenchymal and vascular viruses and Phytoplasma sp. due to the synergistic and antagonistic effects causing unusual and unpredictable biological and epidemiological, viral and viral-like via host biochemical effects.     

Low frequency of horizontal and vertical transmission of two begomoviruses through whiteflies exhibits little relevance to the vector infectivity

Transmissions of plant viruses between individuals of their vector insects through mating are rare events. Recently, three begomoviruses were found to be transmitted between males and females of the whitefly Bemisia tabaci through mating, and two viruses were shown to be transmitted transovarially to progeny. However, results between reports were not consistent. Here we examined the horizontal and vertical transmission of Tomato yellow leaf curl virus (TYLCV) and Tomato yellow leaf curl China virus (TYLCCNV) by the B and Q biotypes of B. tabaci, using virus isolates and whitefly colonies established recently in China. Both TYLCV DNA and TYLCCNV DNA were shown to be transmitted horizontally and vertically by each of the two biotypes of the whitefly, but frequency of transmission was usually low. In transovarial transmission, virus DNA was detected in eggs and nymphs but not in the adults of the first generation progeny, except in the combination of TYLCV and Q biotype whitefly where 2–3% of the offspring adults contained the virus DNA. We also showed that the first generation adults, which developed from eggs of viruliferous whiteflies, were not infective to plants. These results demonstrated that for the viruses and whiteflies tested here low frequency of horizontal and vertical transmission can be expected but these two modes of transmission are unlikely to have much epidemiological relevance in the field.     

Colored shading nets impede insect invasion and decrease the incidences of insect‐transmitted viral diseases in vegetable crops

Black shading nets are commonly used to protect agricultural crops from excessive solar radiation and wind, and for water saving. Recent studies have demonstrated that when black nets were replaced by either red, yellow, or pearl nets (ChromatiNets^?) of equivalent shading capacity, it increased the fruit yield and improved the quality of bell peppers (Capsicum annuum L.) and tomatoes (Solanum lycopersicum L.) (both Solanaceae). We studied the effects of these colored shading nets on the infestation by aphids [Myzus persicae (Sulzer) and Aphis gossypii Glover (both Hemiptera: Aphididae)] and whiteflies [Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae)], and the incidence of the viral diseases transmitted by these insects, for five consecutive years (2006–2010). These studies were conducted in the semi‐arid Besor region in southern Israel. The plants were grown in ‘walk‐in’ tunnels that were covered by several nets of 35% shading capacity in the range of photosynthetically active radiation. Although the shading nets permit free passage of these pests, the infestation levels of aphids and whiteflies in tunnels covered by either the yellow or pearl nets were consistently 2–3× lower than in tunnels covered by the black or red nets. In accordance with the pest results, when the incidences of Cucumber mosaic virus in pepper grown under the black or red nets ranged between 35 and 89%, they were 2–10× lower under the yellow or pearl nets. Similarly, when the incidences of necrotic Potato virus Y in tomato grown under black or red nets ranged between 42 and 50%, they were 2–3× lower under the yellow or pearl nets. Also, when the incidences of Tomato yellow leaf curl virus in tomato grown under the black or red nets ranged between 15 and 50%, but they were 2–4× lower under the yellow or pearl nets. Putative mechanisms of crop protection achieved by the yellow and pearl nets are discussed.     

Mapping in the era of sequencing: high density genotyping and its application for mapping TYLCV resistance in Solanum pimpinellifolium

Background

A RIL population between Solanum lycopersicum cv. Moneymaker and S. pimpinellifolium G1.1554 was genotyped with a custom made SNP array. Additionally, a subset of the lines was genotyped by sequencing (GBS).

Results

A total of 1974 polymorphic SNPs were selected to develop a linkage map of 715 unique genetic loci. We generated plots for visualizing the recombination patterns of the population relating physical and genetic positions along the genome.This linkage map was used to identify two QTLs for TYLCV resistance which contained favourable alleles derived from S. pimpinellifolium. Further GBS was used to saturate regions of interest, and the mapping resolution of the two QTLs was improved. The analysis showed highest significance on Chromosome 11 close to the region of 51.3 Mb (qTy-p11) and another on Chromosome 3 near 46.5 Mb (qTy-p3). Furthermore, we explored the population using untargeted metabolic profiling, and the most significant differences between susceptible and resistant plants were mainly associated with sucrose and flavonoid glycosides.

Conclusions

The SNP information obtained from an array allowed a first QTL screening of our RIL population. With additional SNP data of a RILs subset, obtained through GBS, we were able to perform an in silico mapping improvement to further confirm regions associated with our trait of interest. With the combination of different ~ omics platforms we provide valuable insight into the genetics of S. pimpinellifolium-derived TYLCV resistance.

Electronic supplementary material

The online version of this article (doi:10.1186/1471-2164-15-1152) contains supplementary material, which is available to authorized users.     

番茄黄化曲叶病毒的快速分子检测

番茄黄化曲叶病毒是当前世界范围内危害番茄生产的毁灭性病害。文章针对番茄黄化曲叶病毒全基因组序列的特异区段自主设计了1对特异性PCR引物(上游引物TYLCV-F：5′-ACGCATGCCTCTAATCCAGTGTA-3′, 下游引物TYLCV-R：5′-CCAATAAGGCGTAAGCGTGTAGAC-3′), 依据PCR扩增特异片段543 bp的有无可以快速、准确、高效、特异地检测出是否感染了TYLCV病毒, 这项技术可以方便地应用到工厂化育苗的带毒性检测、蔬菜大规模生产中植株发病情况的快速检测以及抗病毒育种, 从而为蔬菜安全可持续生产提供科技支撑。     

两个入侵烟粉虱隐种MEAM1与MED对番茄黄曲叶病毒水平传播能力的比较

【目的】烟粉虱 Bemisia tabaci 是番茄黄曲叶病毒（Tomato yellow leaf curl virus, TYLCV）在自然界的唯一传播媒介,除了可以直接取食获取TYLCV,烟粉虱还可以通过交配获取此植物病毒。虽然前人研究证明了烟粉虱Middle East-Asia Minor 1 （MEAM1）和Mediterranean （MED）隐种都可以通过交配在两性个体之间水平传播TYLCV,但有关MEAM1与MED隐种水平传播TYLCV的能力是否存在差异,不同研究的结果却并不一致。另外,目前尚无关于烟粉虱的水平传播行为是否有助于TYLCV在田间扩散的相关研究。【方法】从浙江、广东、云南和河南4个省份的田间采集MEAM1和MED隐种烟粉虱种群,在室内应用分子标记对各种群所属隐种鉴定后分别建立7个供试种群,然后观察每个种群内带毒成虫与不带毒成虫通过交配在不同性别个体之间水平传播TYLCV的能力,并选用采自广东的MEAM1隐种烟粉虱模拟群体交配实验探究通过交配获毒的烟粉虱个体是否具有传播TYLCV致健康番茄植株发病的能力。【结果】4个省内的MEAM1和MED隐种烟粉虱都可通过交配对TYLCV进行水平传播,但传播频率一般在10%以下。不同省份的MEAM1隐种种群之间在水平传播TYLCV的能力上无显著差异,不同省份的MED隐种种群之间也不存在显著差异。另外,同一省份的MEAM1隐种与MED隐种之间在水平传播TYLCV的能力上也不存在显著差异,而且在两个隐种中,带毒雄虫将病毒水平传播给不带毒雌虫与带毒雌虫将病毒水平传播给不带毒雄虫的频率没有显著差异。研究还表明,采自广东的MEAM1隐种烟粉虱个体通过交配水平获毒后不能致健康番茄植株发病。【结论】综合以上结果,我们推测TYLCV在中国境内两个入侵烟粉虱隐种MEAM1和MED各自种群内个体之间的水平传播概率较低,对该病毒在田间的扩散可能没有作用或作用不大。     

核黄素和接种番茄黄化曲叶病毒对番茄叶片防御酶活性的影响

以番茄品种‘1479’为材料,研究了喷施核黄素(Riboflavin)和接种番茄黄化曲叶病毒(TYLCV)对幼苗叶片中过氧化物酶(POD)、多酚氧化酶(PPO)、苯丙氨酸解氨酶(PAL)3种防御酶活性的影响。结果显示:(1)核黄素能显著降低番茄植株的番茄黄化曲叶病毒病的病情指数,并以2 mmol/L时诱导效果最佳,诱抗效果最高可达41.91%。(2)2mmol/L核黄素处理后96h内,番茄叶片的POD、PPO和PAL活性显著高于对照。(3)接种TYLCV后,核黄素处理和接种TYLCV处理均可诱导番茄叶片中防御酶活性显著增强。研究表明,核黄素处理可诱导POD、PPO和PAL活性的系统增强与番茄对TYLCV的诱导抗性密切相关。     

Replication of tomato yellow leaf curl virus (TYLCV) DNA in agroinoculated leaf discs from selected tomato genotypes

The leaf disc agroinoculation system was applied to study tomato yellow leaf curl virus (TYLCV) replication in explants from susceptible and resistant tomato genotypes. This system was also evaluated as a potential selection tool in breeding programmes for TYLCV resistance. Leaf discs were incubated with a head-to-tail dimer of the TYLCV genome cloned into the Ti plasmid ofAgrobacterium tumefaciens. In leaf discs from susceptible cultivars (Lycopersicon esculentum) TYLCV single-stranded genomic DNA and its double-stranded DNA forms appeared within 2–5 days after inoculation. Whiteflies (Bemisia tabaci) efficiently transmitted the TYLCV disease to tomato test plants following acquisition feeding on agroinoculated tomato leaf discs. This indicates that infective viral particles have been produced and have reached the phloem cells of the explant where they can be acquired by the insects. Plants regenerated from agroinfected leaf discs of sensitive tomato cultivars exhibited disease symptoms and contained TYLCV DNA concentrations similar to those present in field-infected tomato plants, indicating that TYLCV can move out from the leaf disc into the regenerating plant. Leaf discs from accessions of the wild tomato species immune to whitefly-mediated inoculation,L. chilense LA1969 andL. hirsutum LA1777, did not support TYLCV DNA replication. Leaf discs from plants tolerant to TYLCV issued from breeding programmes behaved like leaf discs from susceptible cultivars.The Hebrew University of Jerusalem, Faculty of Agriculture, Department of Field and Vegetable Crops     

Chromosomal rearrangements between tomato and Solanum chilense hamper mapping and breeding of the TYLCV resistance gene Ty-1

Tomato yellow leaf curl disease, a devastating disease of Solanum lycopersicum (tomato), is caused by a complex of begomoviruses generally referred to as Tomato yellow leaf curl virus (TYLCV). Almost all breeding for TYLCV resistance has been based on the introgression of the Ty-1 resistance locus derived from Solanum chilense LA1969. Knowledge about the exact location of Ty-1 on tomato chromosome 6 will help in understanding the genomic organization of the Ty-1 locus. In this study, we analyze the chromosomal rearrangement and recombination behavior of the chromosomal region where Ty-1 is introgressed. Nineteen markers on tomato chromosome 6 were used in F(2) populations obtained from two commercial hybrids, and showed the presence of a large introgression in both. Fluorescence in situ hybridization (FISH) analysis revealed two chromosomal rearrangements between S. lycopersicum and S. chilense LA1969 in the Ty-1 introgression. Furthermore, a large-scale recombinant screening in the two F(2) populations was performed, and 30 recombinants in the Ty-1 introgression were identified. All recombination events were located on the long arm beyond the inversions, showing that recombination in the inverted region was absent. Disease tests on progenies of informative recombinants with TYLCV mapped Ty-1 to the long arm between markers MSc05732-4 and MSc05732-14, an interval overlapping with the reported Ty-3 region, which led to the indication that Ty-1 and Ty-3 may be allelic. With this study we prove that FISH can be used as a diagnostic tool to aid in the accurate mapping of genes that were introgressed from wild species into cultivated tomato.