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High-Throughput Sequencing Identified Multiple Fig Viruses and Viroids Associated with Fig Mosaic Disease in Iraq. 高通量测序鉴定出与伊拉克无花果马赛克病有关的多种无花果病毒和病毒株。
IF 1.8 3区 农林科学 Q2 PLANT SCIENCES Pub Date : 2024-10-01 DOI: 10.5423/PPJ.OA.04.2024.0068
Nabeel Al-Kaeath, Shrooq Zagier, Osamah Alisawi, Fadhal Al Fadhal, Naima Mahfoudhi

Mosaic is the most common viral disease affecting fig plants. Although the Fig mosaic virus is the leading cause of mosaic disease, other viruses are also involved. High-throughput sequencing was used to assess viral infections in fig plants with mosaic. The genomic DNA and total RNAseq of mosaic-symptomatic fig leaves were sequenced using the Illumina platform. The analysis revealed the presence of fig badnavirus 1 (FBV-1), grapevine badnavirus 1 (GBV-1), citrus exocortis viroid (CEVd), and apple dimple fruit viroid (ADFVd). The FBV-1 and GBV-1 sequences were 7,140 bp and 7,239 bp long, respectively. The two genomes encode one open reading frame containing five major protein domains. The viroids, CEVd and ADFVd, were 397 bp and 305 bp long. Phylogenetic analyses revealed a close relationship between FBV-1 and Iranian isolates of the same species, while GBV-1 was closely related to Russian grapevine badnavirus isolates (Tem64, Blu17, KDH48, and Pal9). CEVd was closely related to other Iraqi isolates, while ADFVd was strongly related to a Spanish isolate. A registered endogenous pararetrovirus, caulimovirus-Fca1, with a size of 7,556 bp, was found in the RNA transcripts with a low expression level. This integrant was also detected in the genomes of the two lines 'Horaishi' (a female line) and 'Caprifig 6085' (a male line). Phylogenetic analyses revealed that caulimovirus-Fca1 was distinct from two other clades of different endogenous virus genera.

马赛克是影响无花果植物的最常见病毒病。虽然无花果马赛克病毒是导致马赛克病的主要原因,但其他病毒也参与其中。我们利用高通量测序技术评估了无花果植株马赛克病的病毒感染情况。利用Illumina平台对有花叶病症状的无花果叶片的基因组DNA和总RNAseq进行了测序。分析发现了无花果坏疽病毒1(FBV-1)、葡萄坏疽病毒1(GBV-1)、柑橘外皮层病毒(CEVd)和苹果酒窝果病毒(ADFVd)。FBV-1 和 GBV-1 的序列长度分别为 7,140 bp 和 7,239 bp。这两个基因组编码一个开放阅读框,包含五个主要蛋白质结构域。病毒体 CEVd 和 ADFVd 的长度分别为 397 bp 和 305 bp。系统进化分析表明,FBV-1与伊朗的同种分离物关系密切,而GBV-1与俄罗斯的葡萄坏疽病毒分离物(Tem64、Blu17、KDH48和Pal9)关系密切。CEVd 与伊拉克的其他分离株关系密切,而 ADFVd 与西班牙的一个分离株关系密切。在 RNA 转录本中发现了一个注册的内源性副逆转录病毒 caulimovirus-Fca1,大小为 7,556 bp,表达水平较低。在两个品系 "Horaishi"(雌性品系)和 "Caprifig 6085"(雄性品系)的基因组中也检测到了这种整合体。系统进化分析表明,花椰菜病毒-Fca1 与其他两个不同内源病毒属的支系截然不同。
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引用次数: 0
Investigation of Antimicrobial Compounds Produced by Endolichenic Fungi in Different Culture Media. 研究内生真菌在不同培养基中产生的抗菌化合物
IF 1.8 3区 农林科学 Q2 PLANT SCIENCES Pub Date : 2024-10-01 DOI: 10.5423/PPJ.NT.06.2024.0087
Jaycee Augusto G Paguirigan, Eunah Jeong, Kyo Bin Kang, Jae-Seoun Hur, Wonyong Kim

Continuous use of synthetic fungicides has led to explosive emergence of fungicide-resistant microbes. Therefore, there are urgent needs for environmentally friendly antimicrobial agents with novel modes of action. This study investigated endolichenic fungi (ELF) as a source of antimicrobial compounds against various plant pathogens. We utilized an One Strain MAny Compounds (OSMAC) approach to enhance the chemical diversity of fourteen ELF. This involved cultivation of ELF in four growth media and subsequently assessing antimicrobial activities of culture extracts. Nearly half of the culture extracts exhibited antimicrobial activity against a Gram-positive bacterium, but showed minimal activity against Gram-negative bacteria tested. Notably, culture extracts from two ELF, Chaetomium globosum and Nodulisporium sp., demonstrated significant inhibitory effects against plant pathogenic fungi. LC-MS/MS-based metabolome profiling confirmed the presence of known bioactive compounds like cyclic dipeptides and chaetoglobosins. These findings highlight the effectiveness of combining OSMAC and metabolomics for identifying antimicrobial agents for agricultural use.

合成杀真菌剂的持续使用导致对杀真菌剂产生抗药性的微生物大量出现。因此,迫切需要具有新型作用模式的环境友好型抗菌剂。本研究将内生真菌(ELF)作为抗各种植物病原体的抗菌化合物来源进行了研究。我们采用了一种 "一个菌株多种化合物"(OSMAC)的方法来提高十四种内生真菌的化学多样性。这包括在四种生长培养基中培养 ELF,然后评估培养提取物的抗菌活性。近一半的培养提取物对革兰氏阳性菌具有抗菌活性,但对革兰氏阴性菌的活性极低。值得注意的是,两种 ELF(Chaetomium globosum 和 Nodulisporium sp.)的培养物提取物对植物病原真菌有显著的抑制作用。基于 LC-MS/MS 的代谢组分析证实了已知生物活性化合物的存在,如环二肽和茶黄素。这些发现凸显了结合 OSMAC 和代谢组学鉴定农用抗菌剂的有效性。
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引用次数: 0
Assessing Cold Plasma's Impact on Banana Growth and Fusarium Wilt Control. 评估冷等离子体对香蕉生长和镰刀菌枯萎病控制的影响
IF 1.8 3区 农林科学 Q2 PLANT SCIENCES Pub Date : 2024-10-01 DOI: 10.5423/PPJ.OA.05.2024.0083
Priya Rajakumar, Nadiya Akmal Baharum, Afiqah Insyirah Lutfi, Najiah Mohd Sadali, Muhamad Shakirin Mispan, Lim Lian Kuang, Yap Seong Ling, Norzulaani Khalid, Nur Ardiyana Rejab

Bananas (Musa spp.), which serve millions of people worldwide, face a serious threat from Fusarium wilt (FW) disease caused by Fusarium oxysporum f. sp. cubense (Foc). Developing disease-resistant varieties particularly through breeding is challenging due to banana's seedless nature (parthenocarpic). As an alternative, cold plasma (CP) technology, has the potential to be used for crop improvement. Our study demonstrates a favourable impact of CP on the growth performance of banana (Berangan cultivar, AAA) in terms of height, leaf number and stem diameter. CP-treated plants also displayed delayed disease progression as well as lower disease severity indicated by slightly lower value of leaf symptoms index and rhizome discoloration index compared to the control plants. Additionally, quantitative real-time polymerase chain reaction analysis revealed differential expression of several defence (PR1, WRKY22, PAL, and CEBiP) and growth (Cytochrome P450, NAC68, and CAT) related genes in CP-treated plants, particularly in conjunction with Foc infection. These findings shed light on the potential use of CP in managing FW in banana and offer insights into possible mechanism behind improved traits.

香蕉(Musa spp.)为全世界数百万人提供服务,但却面临着由 Fusarium oxysporum f. sp. cubense(Foc)引起的镰刀菌枯萎病(FW)的严重威胁。由于香蕉无籽(孤雌生殖)的特性,特别是通过育种开发抗病品种具有挑战性。作为一种替代方法,冷等离子体(CP)技术有可能用于作物改良。我们的研究表明,冷等离子体对香蕉(Berangan 栽培品种,AAA)在高度、叶片数和茎直径方面的生长表现有有利影响。与对照植株相比,氯化石蜡处理植株的叶片症状指数和根茎褪色指数值略低,这表明氯化石蜡处理植株还能延缓病害发展,降低病害严重程度。此外,定量实时聚合酶链式反应分析表明,在 CP 处理的植株中,特别是在 Foc 感染的情况下,一些防御基因(PR1、WRKY22、PAL 和 CEBiP)和生长相关基因(细胞色素 P450、NAC68 和 CAT)的表达存在差异。这些发现揭示了氯化石蜡在管理香蕉凋萎病虫害方面的潜在用途,并深入揭示了性状改善背后的可能机制。
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引用次数: 0
Investigation of Tissue-Specific Distribution and Genetic Variation of Alfalfa Mosaic Virus and Chinese Artichoke Mosaic Virus in Chinese Artichoke (Stachys affinis miq.). 紫花苜蓿花叶病毒和中国朝鲜蓟花叶病毒在中国朝鲜蓟(Stachys affinis miq.)
IF 1.8 3区 农林科学 Q2 PLANT SCIENCES Pub Date : 2024-08-01 DOI: 10.5423/PPJ.OA.06.2024.0094
Ji-Soo Park, Dong-Joo Min, Tae-Seon Park, You-Seop Shin, Jin-Sung Hong

The Chinese artichoke (Stachys affinis syn. S. sieboldii) is a widely cultivated crop, and its rhizome is used as a medicinal vegetable. To investigate the causes of viral diseases in Chinese artichokes, the infection rates of four virus species infecting Chinese artichoke were investigated. Since the Chinese artichoke propagates through its tuber, this study aimed to determine whether viral transmission to the progeny is possible through the tuber, by identifying the virus present in the tuber and investigating its accumulation. First, reverse transcription polymerase chain reaction analysis was performed to detect viruses using total RNA extracted from the flowers, leaves, and tubers of Chinese artichoke plants. Alfalfa mosaic virus (AMV) and Chinese artichoke mosaic virus (ChAMV) had high infectivity in Chinese artichoke and most plants were simultaneously infected with AMV and ChAMV. These viruses were present in all tissues, but their detection frequency and accumulation rates varied across different tissues of the Chinese artichoke. Also, we sequenced the coat protein (CP) genes of AMV and ChAMV to investigate genetic variations of virus between the leaf and tuber. It provides information on CP gene sequences and genetic diversity of isolates identified from new hosts of AMV and ChAMV. This study offers valuable insights into the distribution and spread of the ChAMV and AMV within Chinese artichoke plants, which have implications for the management and control of viral infections in crops.

中国朝鲜蓟(Stachys affinis syn. S. sieboldii)是一种广泛种植的作物,其根茎可作为药用蔬菜。为了研究中国朝鲜蓟病毒病的成因,我们调查了感染中国朝鲜蓟的四种病毒的感染率。由于中国朝鲜蓟通过块茎繁殖,本研究旨在通过鉴定块茎中的病毒并调查其积累情况,确定病毒是否可能通过块茎传播给后代。首先,利用从中国朝鲜蓟植株的花、叶和块茎中提取的总 RNA 进行反转录聚合酶链反应分析,以检测病毒。紫花苜蓿花叶病毒(AMV)和中国朝鲜蓟花叶病毒(ChAMV)对中国朝鲜蓟的感染率很高,大多数植株同时感染了AMV和ChAMV。这些病毒存在于所有组织中,但在中国朝鲜蓟不同组织中的检测频率和累积率有所不同。此外,我们还对 AMV 和 ChAMV 的衣壳蛋白(CP)基因进行了测序,以研究病毒在叶片和块茎之间的遗传变异。该研究提供了有关 CP 基因序列以及从 AMV 和 ChAMV 的新宿主中鉴定出的分离株的遗传多样性的信息。这项研究为了解 ChAMV 和 AMV 在中国朝鲜蓟植株中的分布和传播提供了宝贵的信息,对管理和控制农作物病毒感染具有重要意义。
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引用次数: 0
Comprehensive Metatranscriptomic Analysis of Plant Viruses in Imported Frozen Cherries and Blueberries. 进口冷冻樱桃和蓝莓中植物病毒的元转录组学综合分析。
IF 1.8 3区 农林科学 Q2 PLANT SCIENCES Pub Date : 2024-08-01 DOI: 10.5423/PPJ.OA.06.2024.0088
Ga-Eun Lee, Hyo-Jeong Lee, Rae-Dong Jeong

The possibility of new viruses emerging in various regions worldwide has increased due to a combination of factors, including climate change and the expansion of international trading. Plant viruses spread through various transmission routes, encompassing well-known avenues such as pollen, seeds, and insects. However, research on potential transmission routes beyond these known mechanisms has remained limited. To address this gap, this study employed metatranscriptomic analysis to ascertain the presence of plant viruses in imported frozen fruits, specifically cherries and blueberries. This analysis aimed to identify pathways through which plant viruses may be introduced into countries. Virome analysis revealed the presence of six species of plant viruses in frozen cherries and blueberries: cherry virus A (CVA), prunus necrotic ringspot virus (PNRSV), prune dwarf virus (PDV), prunus virus F (PrVF), blueberry shock virus (BlShV), and blueberry latent virus (BlLV). Identifying these potential transmission routes is crucial for effectively managing and preventing the spread of plant viruses and crop protection. This study highlights the importance of robust quality control measures and monitoring systems for frozen fruits, emphasizing the need for proactive measures to mitigate the risk associated with the potential spread of plant viruses.

由于气候变化和国际贸易扩大等多种因素,世界各地区出现新病毒的可能性增加了。植物病毒的传播途径多种多样,包括花粉、种子和昆虫等众所周知的途径。然而,对这些已知机制之外的潜在传播途径的研究仍然有限。为了填补这一空白,本研究采用了元转录组学分析来确定进口冷冻水果(特别是樱桃和蓝莓)中是否存在植物病毒。这项分析旨在确定植物病毒可能进入各国的途径。病毒组分析表明,冷冻樱桃和蓝莓中存在六种植物病毒:樱桃病毒 A (CVA)、梅花坏死环斑病毒 (PNRSV)、梅花矮小病毒 (PDV)、梅花病毒 F (PrVF)、蓝莓休克病毒 (BlShV) 和蓝莓潜伏病毒 (BlLV)。确定这些潜在的传播途径对于有效管理和预防植物病毒传播及作物保护至关重要。这项研究强调了对冷冻水果采取强有力的质量控制措施和监控系统的重要性,强调了采取积极措施降低植物病毒潜在传播风险的必要性。
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引用次数: 0
Potentiality of Beneficial Microbe Bacillus siamensis GP-P8 for the Suppression of Anthracnose Pathogens and Pepper Plant Growth Promotion. 有益微生物暹罗芽孢杆菌 GP-P8 抑制炭疽病病原体和促进辣椒植株生长的潜力。
IF 1.8 3区 农林科学 Q2 PLANT SCIENCES Pub Date : 2024-08-01 DOI: 10.5423/PPJ.OA.01.2024.0022
Ji Min Woo, Hyun Seung Kim, In Kyu Lee, Eun Jeong Byeon, Won Jun Chang, Youn Su Lee

This study was carried out to screen the antifungal activity against Colletotrichum acutatum, Colletotrichum dematium, and Colletotrichum coccodes. Bacterial isolate GP-P8 from pepper soil was found to be effective against the tested pathogens with an average inhibition rate of 70.7% in in vitro dual culture assays. 16S rRNA gene sequencing analysis result showed that the effective bacterial isolate as Bacillus siamensis. Biochemical characterization of GP-P8 was also performed. According to the results, protease and cellulose, siderophore production, phosphate solubilization, starch hydrolysis, and indole-3-acetic acid production were shown by the GP-P8. Using specific primers, genes involved in the production of antibiotics, such as iturin, fengycin, difficidin, bacilysin, bacillibactin, surfactin, macrolactin, and bacillaene were also detected in B. siamensis GP-P8. Identification and analysis of volatile organic compounds through solid phase microextraction/gas chromatography-mass spectrometry (SPME/GC-MS) revealed that acetoin and 2,3-butanediol were produced by isolate GP-P8. In vivo tests showed that GP-P8 significantly reduced the anthracnose disease caused by C. acutatum, and enhanced the growth of pepper plant. Reverse transcription polymerase chain reaction analysis of pepper fruits revealed that GP-P8 treated pepper plants showed increased expression of immune genes such as CaPR1, CaPR4, CaNPR1, CaMAPK4, CaJA2, and CaERF53. These results strongly suggest that GP-P8 could be a promising biocontrol agent against pepper anthracnose disease and possibly a pepper plant growth-promoting agent.

本研究旨在筛选出对尖孢褐斑病菌(Colletotrichum acutatum)、褐斑病菌(Colletotrichum dematium)和球孢褐斑病菌(Colletotrichum coccodes)的抗真菌活性。在体外双重培养试验中发现,从辣椒土壤中分离出的细菌 GP-P8 对测试病原体有效,平均抑制率为 70.7%。16S rRNA 基因测序分析结果显示,有效的细菌分离物为暹罗芽孢杆菌。此外,还对 GP-P8 进行了生化鉴定。结果显示,GP-P8 能产生蛋白酶和纤维素、嗜苷酸、磷酸盐溶解、淀粉水解和吲哚-3-乙酸。使用特定引物,还在暹罗芽孢杆菌 GP-P8 中检测到了参与生产抗生素的基因,如 iturin、fengycin、difficidin、bacilysin、bacillibactin、surfactin、macrolactin 和 bacillaene。通过固相微萃取/气相色谱-质谱(SPME/GC-MS)对挥发性有机化合物的鉴定和分析表明,GP-P8 分离物产生了乙酰丙酮和 2,3-丁二醇。体内试验表明,GP-P8 能显著减轻由 C. acutatum 引起的炭疽病,并促进辣椒植株的生长。对辣椒果实进行的逆转录聚合酶链反应分析表明,GP-P8 处理过的辣椒植株的免疫基因(如 CaPR1、CaPR4、CaNPR1、CaMAPK4、CaJA2 和 CaERF53)的表达量有所增加。这些结果有力地表明,GP-P8 可能是一种很有前景的防治辣椒炭疽病的生物防治剂,也可能是一种促进辣椒植株生长的药剂。
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引用次数: 0
RT-RPA Assay Combined with a Lateral Flow Strip to Detect Soybean Mosaic Virus. RT-RPA 分析法与侧流带相结合检测大豆花叶病毒。
IF 1.8 3区 农林科学 Q2 PLANT SCIENCES Pub Date : 2024-08-01 DOI: 10.5423/PPJ.OA.02.2024.0027
Bong Geun Oh, Ju-Yeon Yoon, Ho-Jong Ju

Soybean (Glycine max L.) is one of the most widely planted and used legumes in the world, being used for food, animal feed products, and industrial production. The soybean mosaic virus (SMV) is the most prevalent virus infecting soybean plants. This study developed a diagnostic method for the rapid and sensitive detection of SMV using a reverse transcription-recombinase polymerase amplification (RT-RPA) technique combined with a lateral flow strip (LFS). The RT-RPA and RT-RPA-LFS conditions to detect the SMV were optimized using the selected primer set that amplified part of the VPg protein gene. The optimized reaction temperature for the RT-RPA primer and RT-RPA-LFS primer used in this study was 38℃ for both, and the minimum reaction time was 10 min and 5 min, respectively. The RT-RPA-LFS was as sensitive as RT-PCR to detect SMV with 10 pg/μl of total RNA. The reliability of the developed RT-RPA-LFS assay was evaluated using leaves collected from soybean fields. The RT-RPA-LFS diagnostic method developed in this study will be useful as a diagnostic method that can quickly and precisely detect SMV in the epidemiological investigation of SMV, in the selection process of SMV-resistant varieties, on local farms with limited resources.

大豆(Glycine max L.)是世界上种植和使用最广泛的豆科植物之一,可用于食品、动物饲料产品和工业生产。大豆花叶病毒(SMV)是感染大豆植物最普遍的病毒。本研究利用反转录-重组聚合酶扩增(RT-RPA)技术结合侧流条带(LFS)开发了一种快速灵敏检测 SMV 的诊断方法。利用扩增部分 VPg 蛋白基因的引物组对检测 SMV 的 RT-RPA 和 RT-RPA-LFS 条件进行了优化。本研究中使用的 RT-RPA 引物和 RT-RPA-LFS 引物的优化反应温度均为 38℃,最短反应时间分别为 10 分钟和 5 分钟。在检测 10 pg/μl 总 RNA 的 SMV 时,RT-RPA-LFS 的灵敏度与 RT-PCR 不相上下。利用从大豆田中采集的叶片对所开发的 RT-RPA-LFS 检测方法的可靠性进行了评估。本研究开发的 RT-RPA-LFS 诊断方法可作为一种诊断方法,在资源有限的地方农场进行 SMV 流行病学调查和抗 SMV 品种选育过程中快速、准确地检测 SMV。
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引用次数: 0
Light- and Relative Humidity-Regulated Hypersensitive Cell Death and Plant Immunity in Chinese Cabbage Leaves by a Non-adapted Bacteria Xanthomonas campestris pv. vesicatoria. 非适应细菌野油菜黄单胞菌 pv. vesicatoria 对光照和相对湿度调控的大白菜叶片超敏细胞死亡和植物免疫作用
IF 1.8 3区 农林科学 Q2 PLANT SCIENCES Pub Date : 2024-08-01 DOI: 10.5423/PPJ.OA.03.2024.0057
Young Hee Lee, Yun-Hee Kim, Jeum Kyu Hong

Inoculation of Chinese cabbage leaves with high titer (107 cfu/ml) of the non-adapted bacteria Xanthomonas campestris pv. vesicatoria (Xcv) strain Bv5-4a.1 triggered rapid leaf tissue collapses and hypersensitive cell death (HCD) at 24 h. Electrolyte leakage and lipid peroxidation markedly increased in the Xcv-inoculated leaves. Defence-related gene expressions (BrPR1, BrPR4, BrChi1, BrGST1 and BrAPX1) were preferentially activated in the Xcv-inoculated leaves. The Xcv-triggered HCD was attenuated by continuous light but accelerated by a dark environment, and the prolonged high relative humidity also alleviated the HCD. Constant dark and increased relative humidity provided favorable conditions for the Xcv bacterial growth in the leaves. Pretreated fluridone (biosynthetic inhibitor of endogenous abscisic acid [ABA]) increased the HCD in the Xcv-inoculated leaves, but exogenous ABA attenuated the HCD. The pretreated ABA also reduced the Xcv bacterial growth in the leaves. These results highlight that the onset of HCD in Chinese cabbage leaves initiated by non-adapted pathogen Xcv Bv5-4a.1 and in planta bacterial growth was differently modulated by internal and external conditional changes.

给大白菜叶片接种高滴度(107 cfu/ml)的野油菜黄单胞菌(Xanthomonas campestris pv. vesicatoria,Xcv)菌株 Bv5-4a.1,24 小时后会引发叶片组织迅速塌陷和超敏细胞死亡(HCD)。与防御相关的基因表达(BrPR1、BrPR4、BrChi1、BrGST1 和 BrAPX1)在 Xcv 接种的叶片中优先被激活。Xcv 触发的 HCD 在持续光照下会减弱,但在黑暗环境中会加速,长时间的高相对湿度也会减轻 HCD。持续的黑暗和相对湿度的增加为 Xcv 细菌在叶片中的生长提供了有利条件。预处理氟啶酮(内源脱落酸 [ABA] 的生物合成抑制剂)增加了 Xcv 接种叶片的 HCD,但外源 ABA 减轻了 HCD。预处理的 ABA 还能减少叶片中 Xcv 细菌的生长。这些结果表明,非适应性病原菌 Xcv Bv5-4a.1 在大白菜叶片中引发的 HCD 以及植物体中细菌的生长受内部和外部条件变化的调节是不同的。
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引用次数: 0
Development and Application of Reverse Transcription Nanoplate-Based Digital PCR Assay for Sensitive and Accurate Detection of Rice Black-Streaked Dwarf Virus in Cereal Crops. 基于反转录纳米片的数字 PCR 检测方法的开发与应用,用于灵敏、准确地检测谷类作物中的水稻黑条矮缩病病毒。
IF 1.8 3区 农林科学 Q2 PLANT SCIENCES Pub Date : 2024-08-01 DOI: 10.5423/PPJ.NT.03.2024.0048
Hyo-Jeong Lee, Hae-Jun Kim, Sang-Min Kim, Rae-Dong Jeong

The emergence of rice black-streaked dwarf virus (RBSDV) poses a significant threat to global cereal crop cultivation, necessitating the urgent development of reliable detection and quantification techniques. This study introduces a reliable approach for the precise and sensitive quantification of the RBSDV in cereal crop samples, employing a reverse transcription digital polymerase chain reaction (RT-dPCR) assay. We assessed the specificity and sensitivity of the RT-dPCR assay proposed for precise RBSDV detection and quantification. Our findings demonstrate that RT-dPCR was specific for detection of RBSDV, with no cross-reactivity observed with other viruses infecting cereal crops. The RT-dPCR sensitivity was over 10 times that of RT-quantitative PCR (RT-qPCR). The detection limit of RT-dPCR was 0.096 copies/μl. In addition, evaluation of RT-dPCR assay with field samples was conducted on 60 different cereal crop samples revealed that RT-dPCR (45/60) exhibited superior accuracy compared with RT-qPCR (23/60). In this study, we present a specific and accurate RT-dPCR assay for the detection and quantification of RBSDV.

水稻黑条矮缩病病毒(RBSDV)的出现对全球谷类作物种植构成了重大威胁,因此迫切需要开发可靠的检测和定量技术。本研究采用一种反转录数字聚合酶链反应(RT-dPCR)检测方法,为精确、灵敏地定量检测谷类作物样本中的 RBSDV 引入了一种可靠的方法。我们评估了为精确检测和量化 RBSDV 而提出的 RT-dPCR 检测方法的特异性和灵敏度。我们的研究结果表明,RT-dPCR 对检测 RBSDV 具有特异性,与感染谷类作物的其他病毒没有交叉反应。RT-dPCR 的灵敏度是 RT 定量 PCR(RT-qPCR)的 10 倍以上。RT-dPCR 的检测限为 0.096 个拷贝/微升。此外,通过对 60 份不同的谷类作物样本进行田间样本 RT-dPCR 分析评估发现,与 RT-qPCR(23/60)相比,RT-dPCR(45/60)表现出更高的准确性。在本研究中,我们提出了一种用于检测和定量 RBSDV 的特异而准确的 RT-dPCR 分析方法。
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引用次数: 0
Identification and Characterization Colletotrichum spp. Causing Mango Dieback in Indonesia. 导致印度尼西亚芒果枯萎病的 Colletotrichum spp.
IF 1.8 3区 农林科学 Q2 PLANT SCIENCES Pub Date : 2024-08-01 DOI: 10.5423/PPJ.NT.04.2024.0061
Khaerani Nurlaelita, Arif Wibowo, Ani Widiastuti

Dieback disease in mango trees has been observed in Indonesia, particularly in Java Island, with the causal agent remaining unidentified. One of the important pathogens that are responsible for causing mango dieback is Colletotrichum. Field surveys were conducted in various mango cultivating areas in Java Island, Indonesia to assess prevalence of Colletotrichum as dieback disease pathogen. Eleven Colletotrichum isolates were recovered from symptomatic dieback twigs and morphologically characterized. Genetic diversity fingerprint analysis was carried out using rep-PCR. Phylogenetic analysis identified isolates as belonging to Colletotrichum asianum and Colletotrichum cairnsense using partial sequences of four gene regions, including ITS, ACT, GAPDH, and TUB2. Pathogenicity tests on mango seedlings cv. Arumanis showed that all fungal isolates were responsible for causing dieback symptoms. Subsequently, symptomatic tissue was reisolated to fulfill Koch's Postulate. This study represented new funding for two species of Colletotrichum causing mango dieback in Indonesia.

在印度尼西亚,特别是爪哇岛,发现了芒果树的枯萎病,但病原菌仍未确定。Colletotrichum 是导致芒果枯萎病的重要病原体之一。在印度尼西亚爪哇岛的多个芒果种植区进行了实地调查,以评估枯萎病病原体 Colletotrichum 的流行情况。从有症状的枯萎病树枝上分离出 11 株 Colletotrichum,并对其进行了形态鉴定。使用 rep-PCR 进行了遗传多样性指纹分析。利用四个基因区的部分序列,包括 ITS、ACT、GAPDH 和 TUB2,通过系统发育分析确定分离株属于 Colletotrichum asianum 和 Colletotrichum cairnsense。对 Arumanis 品种的芒果幼苗进行的致病性测试表明,所有真菌分离物都能引起枯萎症状。随后,对有症状的组织进行了重新分离,以验证科赫推定。这项研究为印尼两种引起芒果枯萎病的 Colletotrichum 提供了新的资金来源。
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Plant Pathology Journal
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