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Pathotype Classification of Plasmodiophora brassicae Isolates Using Clubroot-Resistant Cultivars of Chinese Cabbage 白菜抗棍棒病品种分离的芸苔菌病型分类
Pub Date : 2016-10-01 DOI: 10.5423/PPJ.OA.04.2016.0081
Hun Kim, Eun Ju Jo, Y. Choi, K. Jang, G. Choi
Clubroot disease caused by Plasmodiophora brassicae is one of the most serious diseases in Brassica crops worldwide. In this study, the pathotypes of 12 Korean P. brassicae field isolates were determined using various Chinese cabbage including 22 commercial cultivars from Korea, China, and Japan, and 15 inbred lines. All P. brassicae isolates exhibited the typical clubroot disease on non-clubroot resistant cultivar, indicating that the isolates were highly pathogenic. According to the reactions on the Williams’ hosts, the 12 field isolates were initially classified into five races. However, when these isolates were inoculated onto clubroot-resistant (CR) cultivars of Chinese cabbage, several isolates led to different disease responses even though the isolates have been assigned to the same race by the Williams’ host responses. Based on the pathogenicity results, the 12 field isolates were reclassified into four different groups: pathotype 1 (GN1, GN2, GS, JS, and HS), 2 (DJ and KS), 3 (HN1, PC, and YC), and 4 (HN2 and SS). In addition, the CR cultivars from Korea, China, and Japan exhibited distinguishable disease responses to the P. brassicae isolates, suggesting that the 22 cultivars used in this study, including the non-CR cultivars, are classified into four different host groups based on their disease resistance. Combining these findings, the four differential hosts of Chinese cabbage and four pathotype groups of P. brassicae might provide an efficient screening system for resistant cultivars and a new foundation of breeding strategies for CR Chinese cabbage.
由芸苔Plasmodiophora brassicae引起的棒根病是世界上危害芸苔作物最严重的病害之一。本研究以韩国、中国和日本的22个商品白菜品种和15个自交系为材料,对12株韩国芸苔菌大田分离株进行了病型鉴定。所有菌株在非根茎病抗性品种上均表现出典型的根茎病,表明菌株具有高致病性。根据威廉姆斯夫妇宿主的反应,这12个野外分离株最初被分为5个种族。然而,当这些分离株接种到白菜的抗大白菜(CR)品种上时,几个分离株导致了不同的疾病反应,即使这些分离株被威廉姆斯的宿主反应分配到同一种。根据致病性结果,将12株田间分离株重新划分为病原型1 (GN1、GN2、GS、JS和HS)、2 (DJ和KS)、3 (HN1、PC和YC)和4 (HN2和SS) 4组。此外,来自韩国、中国和日本的CR品种对芸苔菌分离株表现出不同的疾病反应,这表明本研究中使用的22个品种(包括非CR品种)根据其抗病性可分为4个不同的宿主群体。综上所述,白菜的4种不同寄主和甘蓝的4个病型群可为抗性品种的筛选提供一个有效的筛选体系,并为CR大白菜的育种策略提供新的基础。
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引用次数: 25
Stemphylium Leaf Blight of Garlic (Allium sativum ) in Spain: Taxonomy and In Vitro Fungicide Response 西班牙大蒜茎叶枯病:分类及体外杀菌剂反应
Pub Date : 2016-10-01 DOI: 10.5423/PPJ.OA.03.2016.0063
Laura Gálvez, J. Gil-Serna, M. García, C. Iglesias, D. Palmero
The most serious aerial disease of garlic is leaf blight caused by Stemphylium spp. Geographical variation in the causal agent of this disease is indicated. Stemphylium vesicarium has been reported in Spain, whereas S. solani is the most prevalent species recorded in China. In this study, Stemphylium isolates were obtained from symptomatic garlic plants sampled from the main Spanish production areas. Sequence data for the ITS1–5.8S–ITS2 region enabled assignation of the isolates to the Pleospora herbarum complex and clearly distinguished the isolates from S. solani. Conidial morphology of the isolates corresponded to that of S. vesicarium and clearly discriminated them from S. alfalfae and S. herbarum on the basis of the size and septation pattern of mature conidia. Conidial morphology as well as conidial length, width and length:width ratio also allowed the Spanish isolates to be distinguished from S. botryosum and S. herbarum. Control of leaf blight of garlic is not well established. Few studies are available regarding the effectiveness of chemical treatments to reduce Stemphylium spp. incidence on garlic. The effectiveness of nine fungicides of different chemical groups to reduce Stemphylium mycelial growth in vitro was tested. Boscalid + pyraclostrobin (group name, succinate dehydrogenase inhibitors + quinone outside inhibitors), iprodione (dicar-boximide), and prochloraz (demethylation inhibitors) were highly effective at reducing mycelial growth in S. vesicarium with EC50 values less than 5 ppm. In general, the effectiveness of the fungicide was enhanced with increasing dosage.
大蒜最严重的空气病害是由茎叶枯病引起的蒜叶枯病,并指出了该病病原的地理差异。在西班牙已有报道,而在中国最常见的是水仙。在本研究中,从西班牙主要产区的有症状大蒜植株中分离得到了茎门菌。ITS1-5.8S-ITS2区域的序列数据表明,分离菌株属于Pleospora herbarum复合体,与S. solani有明显区别。分离菌株的分生孢子形态与水仙花的分生孢子形态一致,并根据成熟分生孢子的大小和分隔模式将其与紫花苜蓿和草本植物区分开来。分生孢子的形态以及分生孢子的长度、宽度和长宽比也使西班牙菌株与S. botryosum和S. herbarum区分开来。大蒜叶枯病的防治尚不完善。关于化学处理降低大蒜茎蚜发病率的有效性的研究很少。研究了不同化学基团的9种杀菌剂体外抑制茎草菌丝生长的效果。Boscalid + pyraclostrobin(组名,琥珀酸脱氢酶抑制剂+醌外抑制剂),iprodione(二氯肟胺)和prochloraz(去甲基化抑制剂)在EC50值小于5 ppm的情况下对S. vesicarium的菌丝生长非常有效。总的来说,杀菌剂的效果随着用量的增加而增强。
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引用次数: 20
Postharvest Disease Control of Colletotrichum gloeosporioides and Penicillium expansum on Stored Apples by Gamma Irradiation Combined with Fumigation 伽玛辐照联合熏蒸对贮藏苹果炭疽菌和膨胀青霉采后病害的防治
Pub Date : 2016-10-01 DOI: 10.5423/PPJ.OA.03.2016.0062
W. Cheon, Young Soo Kim, K. Balaraju, Bong-Su Kim, Byeong-Ho Lee, Y. Jeon
To study the control of postharvest decay caused by Colletotrichum gloeosporioides and Penicillium expansum, gamma irradiation alone or in combination with fumigation was evaluated to extend the shelf life of apples in South Korea. An irradiation dose of 2.0 kGy resulted in the maximum inhibition of C. gloeosporioides and P. expansum spore germination. The gamma irradiation dose required to reduce the spore germination by 90% was 0.22 and 0.35 kGy for C. gloeosporioides and P. expansum, respectively. Microscopic observations revealed that when the fungal spores were treated with gamma irradiation (4.0 kGy), conidial germination was stopped completely resulting in no germ tube formation in C. gloeosporioides. Treatment with the eco-friendly fumigant ethanedinitrile had a greater antifungal activity against C. gloeosporioides and P. expansum in comparison with the non-treated control under in vitro conditions. The in vitro antifungal effects of the gamma irradiation and fumigation treatments allowed us to further study the effects of the combined treatments to control postharvest decay on stored apples. Interestingly, when apples were treated with gamma irradiation in combined with fumigation, disease inhibition increased more at lower (< 0.4 kGy) than at higher doses of irradiation, suggesting that combined treatments reduced the necessary irradiation dose in phytosanitary irradiation processing under storage conditions.
为了研究对绿孢炭疽菌和膨胀青霉的采后腐烂的控制,在韩国研究了单独辐照或联合熏蒸对苹果保质期的影响。2.0 kGy的辐照剂量对黄芽孢杆菌和大芽孢杆菌的萌发抑制作用最大。γ辐照剂量对黄芽孢粉和大芽孢粉的萌发率降低90%,分别为0.22和0.35 kGy。显微镜下观察发现,伽马辐射(4.0 kGy)对真菌孢子的影响使孢子萌发完全停止,没有芽管形成。在体外条件下,环保型熏蒸剂乙二腈处理对黄孢霉和芽孢霉具有较强的抑菌活性。辐照和熏蒸处理的体外抑菌效果为进一步研究联合处理对贮藏苹果采后腐烂的防治效果提供了条件。有趣的是,当苹果用伽马辐射和熏蒸联合处理时,较低剂量(< 0.4 kGy)的疾病抑制作用比较高剂量的辐照更强,这表明联合处理减少了储存条件下植物检疫辐照加工所需的辐照剂量。
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引用次数: 12
Feasible Management of Southern Corn Leaf Blight via Induction of Systemic Resistance by Bacillus cereus C1L in Combination with Reduced Use of Dithiocarbamate Fungicides 蜡样芽孢杆菌C1L诱导系统抗性与减少使用二硫代氨基甲酸酯杀菌剂联合治理南方玉米叶枯病的可行性
Pub Date : 2016-10-01 DOI: 10.5423/PPJ.OA.02.2016.0044
Yingqi Lai, Peili Lin, Chao-Ying Chen, Chien-Jui Huang
Dithiocarbamate fungicides such as maneb and mancozeb are widely used nonsystemic protectant fungicides to control various plant fungal diseases. Dithiocarbamate fungicides should be frequently applied to achieve optimal efficacy of disease control and avoid either decline in effectiveness or wash-off from leaf surface. Dithiocarbamates are of low resistance risk but have the potential to cause human neurological diseases. The objective of this study was to develop a strategy to effectively control plant disease with reduced use of dithiocarbamtes. Southern corn leaf blight was the model pathosystem for the investigation. When corn plants were drench-treated with Bacillus cereus C1L, a rhizobacterium able to induce systemic resistance in corn plants against southern leaf blight, frequency of spraying dithiocarbamate fungicides could be decreased. The treatment of B. cereus C1L was able to protect maize from southern leaf blight while residues of dithiocarbamates on leaf surface were too low to provide sufficient protection. On the other hand, frequent sprays of mancozeb slightly but significantly reduced growth of corn plants under natural conditions. In contrast, application of B. cereus C1L can significantly promote growth of corn plants whether sprayed with mancozeb or not. Our results provide the information that plant disease can be well controlled by rhizobacteria-mediated induced systemic resistance in combination with reduced but appropriate application of dithiocarbamate fungicides just before a heavy infection period. An appropriate use of rhizobacteria can enhance plant growth and help plants overcome negative effects caused by dithiocarbamates.
二硫代氨基甲酸酯类杀菌剂是广泛应用于防治各种植物真菌病害的非全身性杀菌剂。二硫代氨基甲酸酯杀菌剂应经常使用,以达到最佳的防病效果,避免药效下降或从叶片表面脱落。二硫代氨基甲酸酯耐药风险低,但有可能引起人类神经系统疾病。本研究的目的是制定一种策略,以有效地控制植物病害,减少使用二硫代氨基甲酸酯。以南方玉米叶枯病为研究对象。用蜡样芽孢杆菌(Bacillus cereus C1L)对玉米植株进行淋雨处理,可以减少二硫代氨基甲酸酯杀菌剂的喷洒频率。蜡样芽孢杆菌C1L处理对玉米南部叶枯病有一定的保护作用,但二硫代氨基甲酸酯在叶片表面的残留量过低,不能提供足够的保护。另一方面,在自然条件下,频繁喷施代森锰锌对玉米植株生长有轻微但显著的抑制作用。与此相反,无论是否喷施代森锰锌,蜡状芽孢杆菌C1L均能显著促进玉米植株的生长。我们的研究结果提供了根瘤菌介导的诱导系统抗性与在严重侵染期前减少但适当使用二硫代氨基甲酸酯杀菌剂相结合可以很好地控制植物病害的信息。适当使用根瘤菌可以促进植物生长,帮助植物克服二硫代氨基甲酸酯引起的负面影响。
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引用次数: 7
Differential Control Efficacies of Vitamin Treatments against Bacterial Wilt and Grey Mould Diseases in Tomato Plants 维生素处理对番茄青枯病和灰霉病的不同防治效果
Pub Date : 2016-10-01 DOI: 10.5423/PPJ.OA.03.2016.0076
J. Hong, Hyeon Ji Kim, Heesoo Jung, H. Yang, Do Hoon Kim, C. Sung, Chang-Jin Park, Seog-Won Chang
Bacterial wilt and grey mould in tomato plants are economically destructive bacterial and fungal diseases caused by Ralstonia solanacearum and Botrytis cinerea, respectively. Various approaches including chemical and biological controls have been attempted to arrest the tomato diseases so far. In this study, in vitro growths of bacterial R. solanacearum and fungal B. cinerea were evaluated using four different vitamins including thiamine (vitamin B1), niacin (vitamin B3), pyridoxine (vitamin B6), and menadione (vitamin K3). In planta efficacies of the four vitamin treatments on tomato protection against both diseases were also demonstrated. All four vitamins showed different in vitro antibacterial activities against R. solanacearum in dose-dependent manners. However, treatment with 2 mM thiamine was only effective in reducing bacterial wilt of detached tomato leaves without phytotoxicity under lower disease pressure (106 colony-forming unit [cfu]/ml). Treatment with the vitamins also differentially reduced in vitro conidial germination and mycelial growth of B. cinerea. The four vitamins slightly reduced the conidial germination, and thiamine, pyridoxine and menadione inhibited the mycelial growth of B. cinerea. Menadione began to drastically suppress the conidial germination and mycelial growth by 5 and 0.5 mM, respectively. Grey mould symptoms on the inoculated tomato leaves were significantly reduced by pyridoxine and menadione pretreatments one day prior to the fungal challenge inoculation. These findings suggest that disease-specific vitamin treatment will be integrated for eco-friendly management of tomato bacterial wilt and grey mould.
番茄青枯病和灰霉病是由番茄青枯病(Ralstonia solanacearum)和番茄灰霉病(Botrytis cinerea)分别引起的具有经济破坏性的细菌和真菌病害。迄今为止,人们已经尝试了包括化学和生物控制在内的各种方法来控制番茄病害。本研究采用硫胺素(维生素B1)、烟酸(维生素B3)、吡哆醇(维生素B6)、甲萘酮(维生素K3)四种不同的维生素,对细菌茄白葡萄球菌(R. solanacearum)和真菌B. cinerea的体外生长进行了研究。在植物试验中,还证实了四种维生素处理对番茄防治这两种病害的效果。4种维生素对茄青霉的体外抑菌活性均呈剂量依赖性。然而,在较低的病害压力(106菌落形成单位[cfu]/ml)下,2 mM硫胺素处理仅能有效减少离体番茄叶片的青枯病,且无植物毒性。维生素处理也有不同程度地降低了灰葡萄球菌的离体孢子萌发和菌丝生长。4种维生素均能轻微降低菌丝萌发,硫胺素、吡哆醇和甲萘醌均能抑制灰葡萄球菌菌丝生长。美那酮对孢子萌发和菌丝生长的抑制作用分别为5 mM和0.5 mM。在真菌攻毒接种前1天,吡哆醇和美萘酮预处理显著降低了接种番茄叶片上的灰霉病症状。这些研究结果表明,针对番茄青枯病和灰霉病的生态管理应结合特定疾病的维生素处理。
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引用次数: 12
Control of Basal Stem Rot Disease in Oil Palm by Supplementation of Calcium, Copper, and Salicylic Acid 补充钙、铜和水杨酸防治油棕基底腐病
Pub Date : 2016-10-01 DOI: 10.5423/PPJ.OA.03.2016.0052
M. Shahul, Hamid Rahamah Bivi, Adamu Saidu Paiko, A. Khairulmazmi, M. Akhtar, A. S. Idris
Continuous supplementation of mineral nutrients and salicylic acid (SA) as foliar application could improve efficacy in controlling basal stem rot (BSR) disease in oil palm seedling. It is revealed from the results that the highest disease severity index (58.3%) was recorded in T8 treatments at 9 months after inoculation. The best disease control was achieved by T7 treatments (calcium/copper/SA [Ca/Cu/SA]) (5.0%) followed by T1 (5.5%), T5 (5.8%), T3 (8.3%), T6 (8.3%), T4 (13.3%), and T2 (15.8%) treatments. Continuous supplementation of Ca/Cu/SA was found to be the most effective in controlling the disease and the high performance liquid chromatography results showed the detection of ergosterol at very low concentration in the treated samples. Moreover, the transmission electron microscopy analysis results clearly indicated that T7 treatment was also enhancing lignification, which was responsible for the thickness of the secondary cell walls and middle lamella compared to untreated samples. It was therefore, concluded that continuous supplementation of minerals nutrients and SA could effectively suppress disease severity by reducing ergosterol activity and also improve the process of lignification in the treated plants. Furthermore, this treatment also managed to delay the onset of BSR symptoms and promote the growth of the seedlings and eventually suppress the BSR disease.
叶面连续添加矿质营养素和水杨酸(SA)可以提高油棕幼苗根腐病的防治效果。结果显示,接种后9个月T8组疾病严重程度指数最高,为58.3%。T7处理(钙/铜/SA [Ca/Cu/SA])防治效果最好(5.0%),其次是T1(5.5%)、T5(5.8%)、T3(8.3%)、T6(8.3%)、T4(13.3%)和T2(15.8%)处理。连续添加Ca/Cu/SA对防治效果最好,高效液相色谱分析结果表明,处理后的样品中麦角甾醇含量极低。此外,透射电镜分析结果清楚地表明,与未处理的样品相比,T7处理也增强了木质素化,这是导致次级细胞壁和中间薄片厚度增加的原因。由此可见,持续补充矿物质营养素和SA可通过降低麦角甾醇活性有效抑制病害严重程度,并改善处理植株的木质素化过程。此外,该处理还成功地延缓了BSR症状的出现,促进了幼苗的生长,并最终抑制了BSR疾病。
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引用次数: 37
Occurrence of Toxigenic Fusarium vorosii among Small Grain Cereals in Korea 产毒镰刀菌在韩国小粒谷物中的发生
Pub Date : 2016-10-01 DOI: 10.5423/PPJ.OA.05.2016.0123
Theresa Lee, Ji-Seon Paek, Kyung Ah Lee, Soohyung Lee, Jung-Hye Choi, Hyeonheui Ham, S. Hong, J. Ryu
Fusarium graminearum species complex (FGSC) causes Fusarium head blight in small grain cereals. To date, four species (F. graminearum, F. asiaticum, F. boothii, and F. meridionale ) belonging to FGSC frequently occur in Korean cereals. In addition, we first reported the occurrence of additional species (F. vorosii ) within FGSC, which was isolated from barley, corn, and rice in Korea. Phylogenetic analysis of the Fusarium isolates of this group using combined multi-gene sequences confirmed species identification. Moreover, the macroconidia produced by these isolates were morphologically similar to those of the F. vorosii holotype. Chemical analysis indicated that the F. vorosii isolates produced various trichothecenes such as nivalenol and deoxynivalenol with their acetyl derivatives along with zearalenone. Pathogenicity tests demonstrated that all of the F. vorosii isolates examined were pathogenic on barley, corn, and rice with variation in aggressiveness. This study is the first report of F. vorosii in Korean cereals, their pathogenicity towards barley and corn, and their ability to produce trichothecenes and zearalenone.
小麦赤霉病菌种复合体(FGSC)在小粒谷物中引起赤霉病。迄今为止,属于FGSC的四种(F. graminearum, F. asiatium, F. boothii和F. meridionale)经常出现在朝鲜谷物中。此外,我们首次报道了在韩国从大麦、玉米和水稻中分离到的FGSC中出现的其他物种(F. vorosii)。对该组镰刀菌分离株进行多基因序列联合系统发育分析,证实了该菌株的物种鉴定。此外,这些菌株产生的大分生孢子在形态上与vorosii全型菌株相似。化学分析表明,vorosii分离株可产生多种毛霉烯类化合物,如雪腐镰刀菌醇和脱氧雪腐镰刀菌醇及其乙酰衍生物以及玉米赤霉烯酮。致病性试验表明,所有分离的vorosii菌株都对大麦、玉米和水稻具有致病性,但侵袭性有所不同。本研究首次报道了韩国谷物中vorosii菌对大麦和玉米的致病性,以及它们产生毛霉烯和玉米赤霉烯酮的能力。
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引用次数: 13
Development of Nested PCR, Multiplex PCR, and Loop-Mediated Isothermal Amplification Assays for Rapid Detection of Cylindrocladium scoparium on Eucalyptus 巢式PCR、多重PCR和环介导等温扩增技术在桉树上的快速检测
Pub Date : 2016-10-01 DOI: 10.5423/PPJ.OA.03.2016.0065
T. Qiao, Jing Zhang, Shujiang Li, Shan Han, T. Zhu
Eucalyptus dieback disease, caused by Cylindrocladium scoparium, has occurred in last few years in large Eucalyptus planting areas in China and other countries. Rapid, simple, and reliable diagnostic techniques are desired for the early detection of Eucalyptus dieback of C. scoparium prior to formulation of efficient control plan. For this purpose, three PCR-based methods of nested PCR, multiplex PCR, loop-mediated isothermal amplification (LAMP) were developed for detection of C. scoparium based on factor 1-alpha (tef1) and beta-tubulin gene in this study. All of the three methods showed highly specific to C. scoparium. The sensitivities of the nested PCR and LAMP were much higher than the multiplex PCR. The sensitivity of multiplex PCR was also higher than regular PCR. C. scoparium could be detected within 60 min from infected Eucalyptus plants by LAMP, while at least 2 h was needed by the rest two methods. Using different Eucalyptus tissues as samples for C. scoparium detection, all of the three PCR-based methods showed much better detection results than regular PCR. Base on the results from this study, we concluded that any of the three PCR-based methods could be used as diagnostic technology for the development of efficient strategies of Eucalyptus dieback disease control. Particularly, LAMP was the most practical method in field application because of its one-step and rapid reaction, simple operation, single-tube utilization, and simple visualization of amplification products.
近年来,桉树枯梢病在中国和其他国家的大片桉树种植区发生。在制定有效的防治计划之前,需要快速、简单、可靠的诊断技术来早期检测桉树枯梢病。为此,本研究建立了基于1- α因子(tef1)和β -微管蛋白基因的三种PCR检测方法:巢式PCR、多重PCR、环介导等温扩增(LAMP)。3种方法均对鸢尾草具有较高的特异性。巢式PCR和LAMP的灵敏度远高于多重PCR。多重PCR的敏感性也高于常规PCR。LAMP法可在60 min内检测到桉叶枯病菌,其余两种方法至少需要2 h。以不同桉树组织作为检测样本,三种PCR方法的检测结果均明显优于常规PCR。基于本研究结果,我们认为三种基于pcr的方法均可作为诊断技术,用于开发有效的桉树枯梢病防治策略。其中LAMP法具有一步反应快、操作简单、单管利用、扩增产物可视化简单等优点,是现场应用中最实用的方法。
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引用次数: 6
Deep Sequencing Analysis of Apple Infecting Viruses in Korea 韩国苹果感染病毒的深度测序分析
Pub Date : 2016-10-01 DOI: 10.5423/PPJ.OA.04.2016.0104
I. Cho, D. Igori, Seungmo Lim, G. Choi, J. Hammond, Hyoun-Sub Lim, J. Moon
Deep sequencing has generated 52 contigs derived from five viruses; Apple chlorotic leaf spot virus (ACLSV), Apple stem grooving virus (ASGV), Apple stem pitting virus (ASPV), Apple green crinkle associated virus (AGCaV), and Apricot latent virus (ApLV) were identified from eight apple samples showing small leaves and/or growth retardation. Nucleotide (nt) sequence identity of the assembled contigs was from 68% to 99% compared to the reference sequences of the five respective viral genomes. Sequences of ASPV and ASGV were the most abundantly represented by the 52 contigs assembled. The presence of the five viruses in the samples was confirmed by RT-PCR using specific primers based on the sequences of each assembled contig. All five viruses were detected in three of the samples, whereas all samples had mixed infections with at least two viruses. The most frequently detected virus was ASPV, followed by ASGV, ApLV, ACLSV, and AGCaV which were withal found in mixed infections in the tested samples. AGCaV was identified in assembled contigs ID 1012480 and 93549, which showed 82% and 78% nt sequence identity with ORF1 of AGCaV isolate Aurora-1. ApLV was identified in three assembled contigs, ID 65587, 1802365, and 116777, which showed 77%, 78%, and 76% nt sequence identity respectively with ORF1 of ApLV isolate LA2. Deep sequencing assay was shown to be a valuable and powerful tool for detection and identification of known and unknown virome in infected apple trees, here identifying ApLV and AGCaV in commercial orchards in Korea for the first time.
深度测序已经产生了来自5种病毒的52个contigs;从8个小叶和/或生长迟缓的苹果样品中鉴定出苹果萎绿叶斑病病毒(ACLSV)、苹果茎沟病毒(ASGV)、苹果茎点蚀病毒(ASPV)、苹果绿皱伴随病毒(AGCaV)和杏潜伏病毒(ApLV)。与5个病毒基因组的参考序列相比,组装的contigs核苷酸(nt)序列的一致性为68% ~ 99%。其中,asv和ASGV序列以52个contigs最为丰富。采用RT-PCR方法,根据各序列的特异性引物,确定了5种病毒的存在。在三个样本中检测到所有五种病毒,而所有样本都混合感染了至少两种病毒。检测到最多的病毒是ASPV,其次是ASGV、ApLV、ACLSV和AGCaV,这些病毒在混合感染中也有发现。AGCaV在编号为1012480和93549的组装序列中被鉴定出来,与AGCaV分离株Aurora-1的ORF1序列同源性分别为82%和78%。ApLV在ID 65587、1802365和116777三个组装片段中被鉴定出来,分别与ApLV分离物LA2的ORF1序列同源性分别为77%、78%和76%。深度测序技术被证明是检测和鉴定受感染苹果树中已知和未知病毒的一种有价值和强大的工具,本研究首次鉴定了韩国商业果园中的ApLV和AGCaV。
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引用次数: 13
Establishment of a Simple and Rapid Gene Delivery System for Cucurbits by Using Engineered of Zucchini yellow mosaic virus 利用西葫芦黄花叶病毒工程技术建立一种简便快速的西葫芦基因传递系统
Pub Date : 2016-02-01 DOI: 10.5423/PPJ.NT.08.2015.0173
Mi-Hyung Kang, Jang-Kyun Seo, Hoseong Choi, Hongsoo Choi, Kook Hyung Kim
The infectious full-length cDNA clone of zucchini yellow mosaic virus (ZYMV) isolate PA (pZYMV-PA), which was isolated from pumpkin, was constructed by utilizing viral transcription and processing signals to produce infectious in vivo transcripts. Simple rub-inoculation of plasmid DNAs of pZYMV-PA was successful to cause infection of zucchini plants (Cucurbita pepo L.). We further engineered this infectious cDNA clone of ZYMV as a viral vector for systemic expression of heterologous proteins in cucurbits. We successfully expressed two reporter genes including gfp and bar in zucchini plants by simple rub-inoculation of plasmid DNAs of the ZYMV-based expression constructs. Our method of the ZYMV-based viral vector in association with the simple rub-inoculation provides an easy and rapid approach for introduction and evaluation of heterologous genes in cucurbits.
从南瓜中分离得到西葫芦黄花叶病毒(ZYMV)分离物PA (pZYMV-PA),利用病毒转录和加工信号,构建了具有传染性的西葫芦黄花叶病毒(ZYMV)体内转录物全长cDNA克隆。pZYMV-PA质粒dna的简单摩擦接种成功地引起西葫芦(Cucurbita pepo L.)植株的侵染。我们进一步设计了这个传染性的ZYMV cDNA克隆作为异源蛋白在葫芦中的系统表达的病毒载体。我们通过简单地摩擦接种基于zymv表达构建的质粒dna,成功地在西葫芦植物中表达了gfp和bar两个报告基因。我们建立的基于zymv的病毒载体与简单的摩擦接种相结合的方法,为葫芦中外源基因的导入和评价提供了一种简便、快速的方法。
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引用次数: 10
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The Plant Pathology Journal
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