大豆褐斑粒与大豆花叶病毒若干株系的关系

 近年来发生的大豆褐斑粒是由大豆感染大豆花叶病毒的黄斑株系和顶枯株系等所致。人工接种上述病毒证明了这一点。种子斑驳的严重度与植株发病的严重度有某种相关性。
病株所结种子并不一定全部为褐斑粒。褐斑粒或病株所结无病状种子都可以传病,带毒种子产生疫苗的多少视环境条件、大豆品种及病毒的株系而异。     

Detection of plum pox potyvirus using monoclonal antibodies to structural and non-structural proteins1

Eleven monoclonal antibodies specific to plum pox potyvirus (PPV) coat protein were obtained by hybridoma technology from Spanish PPV isolates. In addition, two monoclonal antibodies specific for PPV cylindrical inclusions (CIP non-structural proteins) were obtained. The monoclonal antibodies specific for PPV coat protein were assayed by DASI ELISA against 81 PPV isolates. At least nine different epitopes were found and 21 distinct serological patterns of reaction (serogroups) were established using nine selected monoclonal antibodies against the collection of PPV isolates, indicating the high variability of coat protein among PPV isolates. Changes in epitope composition were observed after aphid and mechanical transmission, indicating the occurrence of mixtures of isolates in field trees. Monoclonal antibody 5B reacted with all PPV isolates assayed, with very high affinity, using DASI ELISA. This method was compared with immunocapture-PCR on field samples in spring, and showed very good coincidence of results. The efficiency of PPV detection can be slightly increased using monoclonal antibodies specific to cylindrical inclusions mixed with monoclonal antibodies against structural proteins, and using mixtures of monoclonal antibodies against different epitopes of coat protein. ELISA-I and immunoprinting-ELISA were able to detect CIP and PPV in extracts and tissue section, respectively, of woody plants. Two monoclonal antibodies offer the possibility of distinguishing between Marcus and Dideron PPV types (M or D). These D-specific monoclonal antibodies can be used in routine tests with high affinity.     

Indian peanut clump virus (IPCV) infection on wheat and barley: symptoms, yield loss and transmission through seed

Wheat and barley crops were shown to be susceptible to Indian peanut clump virus (IPCV) under field conditions. In wheat, the Hyderabad isolate of IPCV (IPCV-H) induced symptoms resembling the rosette caused by soil-borne wheat mosaic virus, and these were apparent only three weeks after emergence. Early-infected plants were severely stunted and dark green, with chlorotic streaks on the youngest leaves, which turned necrotic as the plants aged; most of these plants died. Late-infected plants were also stunted and were conspicuous in the field because of their dark green appearance as a result of delayed maturity. The virus was detected by ELISA and nucleic acid hybridization in all plants with symptoms. These plants usually produced fewer tillers than healthy ones. Spikes were malformed, often did not emerge from the flag leaf, and they contained few, shrivelled seeds. Grain yield was decreased, on average, by 58%. In barley, IPCV-H caused severe stunting and general leaf chlorosis. As the plants aged, the leaves became necrotic and the few infected plants that reached maturity produced small spikes. IPCV-H antigens were detected by ELISA in every wheat seed from infected plants and the virus was transmitted through wheat seed at a frequency of 0.5–1.3%. Storage at 4°C for more than a year did not affect seed transmission frequency. The virus was detected in leaves and roots of seed-transmitted plants. Seed transmission was not detected in barley. The Durgapura isolate (IPCV-D) was detected in wheat crops (cv. RR-21) at 3 different locations in Rajasthan State, India. Infected plants showed reduced growth without any overt symptoms.     

Significance of the heterologous encapsidation of zucchini yellow mosaic potyvirus in transgenic plants expressing plum pox potyvirus capsid protein1

Transgenic Nicotiana benthamiana plants expressing the coat protein of an aphid-transmissible strain of plum pox potyvirus (PPV-D) were infected with an aphid non-transmissible strain of another potyvirus, zucchini yellow mosaic potyvirus (ZYMV-NAT). Non-viruliferous Myzus persicae could acquire and transmit ZYMV-NAT from these plants but not from infected N. benthamiana control plants (not transformed, or transformed by the vector alone). Immunosorbent electron microscopy experiments using the decoration technique revealed that ZYMV-NAT virus particles in the infected transgenic plants expressing the PPV coat protein could be coated not only with ZYMV antibodies but also, on segments of the particles, with PPV antibodies. This suggests that aphid transmission of ZYMV-NAT occurred through heterologous encapsidation, and reveals a potential risk of releasing genetically engineered plants expressing viral coat proteins into the environment.     

黄瓜花叶病毒CA株系在花生上流行的研究

 据1985~1990年在密云县试验观察,黄瓜花叶病毒(CMV)在花生上流行受病毒种传率、苗期蚜虫发生数量及苗期温度和雨量的影响。花生苗期和花针初期为易感病时期,随后抗性有所增强,但不明显。苗期和花针期发病植株,CMV种传率最高,之后带毒率有所下降。CMV种传率高低与病害流行关系,在同一年份呈极显著正相关关系,而在年度间未达显著水平。花生田间蚜虫发生盛期与病害发生高峰期是吻合的,蚜虫发生数量与病害流行关系显著。根据花生种传率和苗期蚜虫发生数量两因素6年次资料配合的病害流行预测式,其回报可靠度的置信范围在93%左右。     

A model system for plant-virus interaction—infectivity and seed transmission of <Emphasis Type="Italic">Cherry leaf roll virus</Emphasis> (CLRV) in <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>

The wide natural incidence of Cherry leaf roll virus (CLRV) in deciduous forest trees and nurseries in northern Europe is believed to have occurred, apart from occasional mechanical spread and transmission through grafting, mainly by seed transmission. The mode of the vertical transmission and its role in the epidemiology of the virus has not been investigated, basically due to the inconvenient host-pathogen combinations studied to date. With the aim of obtaining an appropriate system for identification of viral genes and products participating in infection processes and seed transmission of CLRV, we performed infection and seed transmissibility tests with CLRV in Arabidopsis thaliana plants. Two phylogenetically and serologically different CLRV isolates were tested. Both of them were found able to infect A. thaliana plants, exhibited clear symptoms of the infection and spread systemically in the plants. Infection of the seeds and of a remarkable number of seedlings generated from infected seeds was possible for two consecutive generations. These results, for first time, report seed transmission of CLRV in the model plant A. thaliana and allow the assumption to be made of embryo invasion during seed transmission. Furthermore, first indications are given that genetically diverse CLRV isolates exhibit different abilities for vertical transmission in A. thaliana. The CLRV-A. thaliana model system is suitable for investigating viral invasion of developing plant organs and meristematic tissue, a prerequisite for successful virus dissemination via vertical transmission through seed.     

Seed transmission of Melon necrotic spot virus and efficacy of seed-disinfection treatments

Rates of seed transmission of Melon necrotic spot virus (MNSV) were estimated in seedlings grown from commercial melon ( Cucumis melo ) cv. Galia F₁ seeds. Seedlings at the cotyledon stage and adult plants were assayed for MNSV by DAS-ELISA and RT-PCR. None of the seedling groups tested positive for MNSV by ELISA. The proportion of seedlings infected with MNSV was at least 7 and 8% in seed lots 05 and 06, respectively, as estimated from RT-PCR analysis of grouped seedlings. Fourteen and eight grouped samples (10 seedlings per group), of a total of 200 and 100 seedlings, respectively, grown from infected seeds were MNSV-positive in seed lots 05 and 06, respectively, corresponding to seed-to-seedling transmission rates of 11·3 and 14·8%, respectively. Several seed-disinfection treatments were evaluated for their ability to prevent seed transmission of MNSV. The results suggest that a treatment of 144 h at 70°C can be used to eradicate MNSV in melon seeds without hindering germination.     

<Emphasis Type="Italic">Indian citrus ringspot virus:</Emphasis> localization of virus in seed tissues and evidence for lack of seed transmission

Indian citrus ringspot disease is an important viral disease in kinnow mandarin orchards where disease incidence up to 100% has been recorded. The disease is caused by Indian citrus ringspot virus (ICRSV), a positive sense flexuous RNA virus. The transmission of ICRSV is generally through budwood. Association of ICRSV with pollens of naturally infected flowers from cv. ‘Kinnow’ mandarins has been shown previously and this study demonstrates the presence of ICRSV in seed tissues. DAC-ELISA revealed the presence of virus in seed coats but not in embryo and endosperm of seeds collected from the fruits of ICRSV-infected Kinnow plants. Of the infected seed coats, 18% were found to harbor the virus. The seedlings in the grow-out test did not show any symptom for 2 years and the virus could not be detected in seedlings by DAC-ELISA and RT-PCR. The present study indicated that ICRSV could be localized in the testa of seeds but its transmission to progeny was not observed.     

Evaluation of Seed Transmission of Turnip yellow mosaic virus and Tobacco mosaic virus in Arabidopsis thaliana

ABSTRACT The mechanism of virus transmission through seed was studied in Arabidopsis thaliana infected with Turnip yellow mosaic virus (TYMV) and Tobacco mosaic virus (TMV). Serological and biological tests were conducted to identify the route by which the viruses reach the seed and subsequently are located in the seed. Both TYMV and TMV were detected in seed from infected plants, however only TYMV was seed-transmitted. This is the first report of transmission of TYMV in seed of A. thaliana. Estimating virus seed transmission by grow-out tests was more accurate than enzyme-linked immunosorbent assay due to the higher frequency of antigen in the seed coat than in the embryo. Virus in the seed coat did not lead to seedling infection. Thus, embryo invasion is necessary for seed transmission of TYMV in A. thaliana. Crosses between healthy and virus-infected plants indicated that TYMV from either the female or the male parent could invade the seed. Conversely, invasion from maternal tissue was the only route for TMV to invade the seed. Pollination of flowers on healthy A. thaliana with pollen from TYMV-infected plants did not result in systemic infection of healthy plants, despite TYMV being carried by pollen to the seed.     

莴苣花叶病毒病的研究Ⅱ病害的分布、损失、寄主和传播

 莴苣花叶病在山东普遍发生,其中以泰安等地比较严重。苗期和发棵前期发病的损失最大;人工接种证明,3叶期和7叶期感染的病株全无食用价值。在测定的15科63种植物中,有4科14钟植物表现感病。种子带毒率为1-2%,不同品种和不同生育期感染的病株种子带毒率有很大差异,开花后发病的植株种子不带毒。室内测定,萝卜蚜(Rhopalosiphum pseudobrassicae),棉蚜(Aphis gassypii)和大戟长管蚜(Macrosiphum euphorbiae)均可传毒。桃蚜取食15秒钟即可获毒或传毒,单头蚜传毒率为40%左右,获毒1次可连续传毒2-3株。     

Seed transmission of Sweet potato leaf curl virus in sweet potato (Ipomoea batatas)

Sweet potato leaf curl virus (SPLCV) infects sweet potato and is a member of the family Geminiviridae (genus Begomovirus). SPLCV transmission occurs from plant to plant mostly via vegetative propagation as well as by the insect vector Bemisia tabaci. When sweet potato seeds were planted and cultivated in a whitefly‐free greenhouse, some sweet potato plants started to show SPLCV‐specific symptoms. SPLCV was detected by PCR from all leaves and floral tissues that showed leaf curl disease symptoms. More than 70% of the seeds harvested from SPLCV‐infected sweet potato plants tested positive for SPLCV. SPLCV was also identified from dissected endosperm and embryos. The transmission level of SPLCV from seeds to seedlings was up to 15%. Southern blot hybridization showed SPLCV‐specific single‐ and double‐stranded DNAs in seedlings germinated from SPLCV‐infected seeds. Taken altogether, the results show that SPLCV in plants of the tested sweet potato cultivars can be transmitted via seeds and SPLCV DNA can replicate in developing seedlings. This is the first seed transmission report of SPLCV in sweet potato plants and also, to the authors' knowledge, the first report of seed transmission for any geminivirus.     

大豆花叶病的流行因素及综合防治

1985年设置流行观察圃,以4个品种、4种播期、两种种子传毒率的种子,系统观察各因素对病害流行的作用。另设综合防治田,品种与观察圃相同;采取隔离种植、种传率低、适期播种及早期拔除病苗等措施。1986年用相同的4个品种,设5个隔离试区,各播种不同种传率(0—33％)的种子,测定播种种子传毒率对流行的影响。结果证明:传毒种子是当地最主要的侵染来源,田间杂草不传病,蚜虫传播距离多在100米以内,花期以后被侵染的植株,其种传率显著降低。播种种子传毒率与田间流行程度及收获种子传毒率相关显著。所用防病措施效果明显,可控制田间病情指数为4％、种传率为0％、种子斑驳率低于4％。     

Disinfection of pepper seed infected with different strains of capsicum mosaic virus by trisodium phosphate and dry heat treatment

Disinfection of pepper seed infected with capsicum mosaic virus (CaMV) by immersion in 100 g/1 Na₃ P0₄ solution was compared with dry heat treatment at 76°C. The virus content of the seed varied with the CaMV strains used to infect the pepper cultivars and the time of harvest of seeds from infected plants. Immersion times in Na₃PO₄ had to be increased from 15 min to 2 h to obtain near-complete virus inactivation; these treatments had no effects on germination.
Heating seed in an oven at 76°C for 3 days following a waiting period of 3 months after harvest always eliminated all the virus present, but adversely affected germination. This resulted in delayed emergence and a reduction in the number of seedlings suitable for further raising. The viability of heat-treated seeds also decreased with continued storage after treatment.
There were inconsistent differences in germination of seed from healthy plants and plants infected with the CaMV strains P8 or P11. The possibility of internal seed infection and practical consequences are discussed.     

Variation in the pathogenicity of two Turnip mosaic virus isolates in wild UK Brassica rapa provenances

Brassica rapa can be infected with Turnip mosaic virus (TuMV) as a result of manual inoculation or aphid transmission, but infected plants have not been found in the field. In this study, B. rapa plants grown from seed collected from two field sites in southern England were mechanically inoculated with one of two distinct isolates (pathotypes) of TuMV under glasshouse conditions. These had either been isolated from Brassica oleracea growing wild in Wales, (GBR 83, pathotype 3) or Dorset (GBR 98, pathotype 1). Use of ELISA as an index of infection in manually inoculated B. rapa showed that although seed provenance had a small effect on the proportion of plants infected, the biggest factor was the virus isolate. Both virus isolates infected both lines of B. rapa , but invaded at different rates, although both resulted in easily discernible symptoms. The severity of symptoms was not related to amounts of virus in the infected plants. A significantly greater proportion of plants were infected with GBR 83 at 45 days post-inoculation (d.p.i.) than GBR 98. but GBR 98 caused significantly more severe and obvious symptoms as well as greater mortality at 119 d.p.i., in plants from both sites than GBR 83.     

蚕豆染色病毒(BBSV)在中国的发生与鉴定

 在1985年4月蚕豆病毒病的调查中,先后在四川、湖北、江苏和浙江4省的秋播蚕豆试验区内都发现一种由种子带毒的蚕豆染色病毒(BBSV)。所有病株都发生在由叙利亚国际旱地农业研究中心供种的试验区内,约10-40%的小区或品系发病,株发病率在0-18%之间。病害是随种子传入我国的。
病株为系统感染,病株叶片上的症状可以是轻花叶、斑驳、褪色斑或畸形,有的小叶正常而无明显病变。病毒粒体为等径球状体,直径28毫微米。病毒样本能与英g国染色病毒的抗血清发生沉淀反应,但与真花叶病毒的抗血清不发生反应。在病株上未发现能传毒的象甲,也无象甲的为害状,病毒病尚未扩散。在试验区以外的农田里未发现有蚕豆染色病毒存在。     

Presence of Xylella fastidiosa in Sweet Orange Fruit and Seeds and Its Transmission to Seedlings

ABSTRACT Xylella fastidiosa, a xylem-limited bacterium, causes several economically important diseases in North, Central, and South America. These diseases are transmitted by sharpshooter insects, contaminated budwood, and natural root-grafts. X. fastidiosa extensively colonizes the xylem vessels of susceptible plants. Citrus fruit have a well-developed vascular system, which is continuous with the vascular system of the plant. Citrus seeds develop very prominent vascular bundles, which are attached through ovular and seed bundles to the xylem system of the fruit. Sweet orange (Citrus sinensis) fruit of cvs. Pera, Natal, and Valencia with characteristic symptoms of citrus variegated chlorosis disease were collected for analysis. X. fastidiosa was detected by polymerase chain reaction (PCR) in all main fruit vascular bundles, as well as in the seed and in dissected seed parts. No visual abnormalities were observed in seeds infected with the bacterium. However, the embryos of the infected seeds weighed 25% less than those of healthy seeds, and their germination rate was lower than uninfected seeds. There were about 2,500 cells of X. fastidiosa per infected seed of sweet orange, as quantified using real-time PCR techniques. The identification of X. fastidiosa in the infected seeds was confirmed by cloning and sequencing the specific amplification product, obtained by standard PCR with specific primers. X. fastidiosa was also detected in and recovered from seedlings by isolation in vitro. Our results show that X. fastidiosa can infect and colonize fruit tissues including the seed. We also have shown that X. fastidiosa can be transmitted from seeds to seedlings of sweet orange. To our knowledge, this is the first report of the presence of X. fastidiosa in seeds and its transmission to seedlings.     

Incidence of Chalara elegans in groundnut seed samples and seed transmission of blackhull

The incidence of Chalara elegans ( =Thielaviopsis basicola ) was determined in seven commercial groundnut seed samples in the Vaalharts area of South Africa. It was demonstrated that 1.19% of the seeds were infected by the pathogen. Surface contamination of visually healthy seeds from the same samples was confirmed by testing seed washings for viable propagules of C. elegans on raw carrot dises. Naturally infected seed planted in autoclaved soil in the greenhouse resulted in 34.4% severely diseased plants. These results indicate that seed-borne inoculum may be an important factor in the transmission of this disease.     

Comparison of extraction procedures and determination of the detection threshold for Clavibacter michiganensis ssp. michiganensis in tomato seeds

Several seed extraction procedures, used for detection of Clavibacter michiganensis ssp. michiganensis ( Cmm ) in naturally infected and artificially infested tomato seed lots were evaluated. Extraction methods that included grinding the seeds were significantly better at detecting the pathogen in three different seed lots than methods that used only soaking. The detection threshold of Cmm in relation to seed sample size was determined by adding naturally infected seeds into samples of three different sizes. Cmm was detected by agar plating assay, on three media (CNS, mSCM, D₂ANX), and by direct PCR from seeds and Bio-PCR (bacteria cultured on agar media prior to PCR). In samples of 10 000 seeds containing one infected seed, Cmm could be detected only by Bio-PCR and in only one replicate out of five. In samples containing five or 10 infected seeds per 10 000 seeds, three of five and five of five replicates, respectively, were detected by the three detection methods. In samples of 5000 seeds, one infected seed could be detected in all five replicates only after adding a concentration step. A high correlation ( R ² = 0·9448) between artificially infested seeds and the disease incidence was found. Seed lots infested with less than 58 colony-forming units (CFU) per g did not cause disease under glasshouse conditions, whereas lots with about 1000 CFU g⁻¹ caused disease in 78 plants out of 2000.