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1.
Characterization of Rhizoctonia solani from potato in Great Britain   总被引:1,自引:1,他引:1  
One hundred and thirty five isolates of Rhizoctonia solani were obtained from British potato crops between 2001 and 2003. Isolates were assigned to anastomosis group (AG) using conventional PCR assays for AG2-1 or AG3 or through the observation of hyphal interactions, where appropriate. A previously published primer set was modified in this study to enhance specificity for AG3PT. Most of the isolates (92·6%) belonged to AG3PT whilst some (6·7%) belonged to AG2-1. Only one isolate recovered (0·7%) belonged to AG5. Isolates of AG2-1 were diverse, with variation in both the length of the rDNA intergenic spacer 1 (IGS1) region and the categories of hyphal interaction observed between pairings of AG2-1 isolates. No variation in the length of the rDNA IGS1 region was observed amongst the AG3 isolates collected. Tests carried out on potato stems with a sub-set of the isolates revealed a wide range of aggressiveness amongst AG2-1 isolates. Sequencing of the rDNA internal transcribed spacer (ITS) region of the AG2-1 isolates and construction of a neighbour joining tree with other AG2-1 sequences available indicated that AG2-1 isolates with the short IGS1 region were closely related. This is the first investigation which provides evidence of the relative AG composition of R. solani populations causing disease in potato crops in Great Britain.  相似文献   

2.
Stem canker and black scurf are diseases of potato caused by the fungus Rhizoctonia solani . Spatiotemporal experimentation and empirical modelling were applied for the first time to investigate the effect of antagonistic Trichoderma harzianum on the dynamics of soilborne R. solani on individual potato plants. Trichoderma harzianum reduced the severity of symptoms, expressed as 'rhizoctonia stem lesion index' (RSI), during the first 7 days post-inoculation when the inoculum of R. solani was placed at certain distances (30–60 mm) from the host. For example, with inoculum at 40 mm from the host, RSI was 6 and 40 with and without T. harzianum , respectively. At later observation times, the antagonistic effect was overcome. Trichoderma harzianum reduced the severity of black scurf on progeny tubers. Furthermore, the mean number of progeny tubers per potato plant was reduced by the biocontrol treatment (means of 6·5 ± 1·1 and 9·9 ± 2·7 tubers per plant with and without T. harzianum , respectively), as was the proportion of small (0·1–20·0 g) tubers (48% and 66% with and without T. harzianum , respectively). Additionally, there were fewer malformed and green-coloured tubers in pots treated with T. harzianum than in those without T. harzianum .  相似文献   

3.
A total of 119 isolates of Rhizoctonia were collected from stem canker lesions, stolon and root lesions, hymenia on stems, or from black scurf on tubers of potato plants ( Solanum tuberosum ) in Finland (latitudes 60–67°N). All isolates except three belonged to anastomosis group 3 (AG-3) of R. solani , as determined by phylogenetic analysis of the internal transcribed spacer sequences (ITS1 and ITS2) of ribosomal RNA (rRNA) genes. Sensitivity of the 119 isolates to the fungicide flutolanil was tested in vitro (EC50 values 0·14–0·75  µ g active ingredient mL−1). The isolates also varied considerably in growth rate (5·1–14·8 mm day−1). The severity of disease caused by 99 isolates was determined based on the proportion of potato sprouts affected by lesions, discoloration or death, which was c . 1–60%. Only two isolates that were able to cause severe symptoms showed particularly low sensitivity to the fungicide and rapid growth rate. One isolate each of anastomosis groups AG-2-1 and AG-5 and an unknown, binucleate Rhizoctonia sp. were detected. The AG-5 isolate and the binucleate isolate caused mild symptoms on potato sprouts, whereas the AG-2-1 isolate was not pathogenic. Taken together, AG-3 of R. solani was the predominant causal agent of the stem canker and black scurf diseases of potato in Finland and showed considerable variability in disease severity, fungicide sensitivity and growth rate in vitro .  相似文献   

4.
为明确宁夏马铃薯主栽品种对黑痣病的抗性水平,利用田间和室内接种相结合的方法对供试马铃薯品种的抗病性进行评价。室内接种试验表明,供试的20份马铃薯材料中没有免疫及高抗品种,接种量与抗性水平呈负相关,其中‘陇薯7号’,‘黑美人’,‘青薯9号’在接种量为5~15g水平上表现为中抗。通过聚类分析,接种量为5g和10g时,以欧式距离13作为最佳聚类分割点,供试品种可分为3个类群。田间接种综合评价分析表明,不同品种间抗性存在显著差异,通过聚类分析,以欧式距离13作为最佳聚类分割点,试验品种可分为4个高感品种、11个中感品种和5个中抗品种。  相似文献   

5.
由立枯丝核菌引起的马铃薯黑痣病是一种常见的土壤和种薯传播病害,在世界各地均有发生。本试验对分离纯化的67个丝核菌菌株进行融合群测定,结果表明:67个菌株都属于AG3融合群;对其中来自6个不同地域的11个菌株进行ITS测定,11个菌株的相似度为99%。8种药剂对马铃薯黑痣病菌的毒力测定结果表明,20%氟胺·嘧菌酯水分散粒剂50mg/L和25g/L咯菌腈悬浮种衣剂50mg/L的抑菌效果较好,室内抑菌效果可达100%。  相似文献   

6.
为明确碳酸氢盐对马铃薯黑痣病菌的毒力及田间防效,采用菌丝生长速率法,测定了碳酸氢盐对马铃薯黑痣病菌的毒力;并通过盆栽防治试验及田间防治试验,研究了碳酸氢盐对马铃薯黑痣病的防治效果。结果表明:碳酸氢铵抑制马铃薯黑痣病菌菌丝生长的有效中浓度EC50仅为0.71g/L,碳酸氢钾EC50为3.8g/L;在PSA培养基中同时加入0.79g/L碳酸氢铵、1g/L碳酸氢钾与在PSA培养基中含1.58g/L碳酸氢铵的抑菌效果一样;尿素对马铃薯黑痣病没有防治效果。碳酸氢铵与碳酸氢钾混配对该病有良好的防治作用,相对尿素最高防治效果可达到63.4%。覆膜栽培能够提高碳酸氢盐的防效和马铃薯产量。碳酸氢盐可用于田间马铃薯黑痣病的防治。  相似文献   

7.
8.
An extensive study was conducted to determine where in the production chain Rhizoctonia solani became associated with UK module-raised Brassica oleracea plants. In total, 2600 plants from 52 crops were sampled directly from propagators and repeat sampled from the field. Additional soil, compost and water samples were collected from propagation nurseries and screened using conventional agar isolation methods. No isolates of R. solani were recovered from any samples collected from propagation nurseries. Furthermore, nucleic acid preparations from samples of soil and compost from propagation nurseries gave negative results when tested for R. solani using real-time PCR. Conversely, R. solani was recovered from 116 of 1300 stem bases collected from field crops. All the data collected suggested R. solani became associated with B. oleracea in the field rather than during propagation. Parsimony and Bayesian phylogenetic studies of ribosomal DNA suggested the majority of further classified isolates belonged to anastomosis groups 2-1 (48/57) and AG-4HGII (8/57), groups known to be pathogenic on Brassica spp. in other countries. Many R. solani isolates were recovered from symptomless plant material and the possibilities for such an association are discussed.  相似文献   

9.
The infection process ofRhizoctonia solani AG-3 was studied on potato sprouts, cv. Bintje, in growth chamber trials at 15 °C. Initially hyphae ofR. solani grew predominantly in the longitudinal direction of the sprouts (runner hyphae). They tended to follow the junctions between epidermis cells as was observed by SEM. The hyphae formed side-branches mainly half-way of the subterranean parts of the sprouts. They branched several times with short swollen cells to form infection cushions. Lesions developed only underneath the infection cushions and were first observed five days after inoculation. The necrotic area was proportional to the area covered with infection cushions on the sprouts. Depth of the lesions could extend up to the vascular bundle. Sprouts were colonized only in healthy tissue in the epidermal layer underneath the infection cushion and in necrotic tissue. A few days after appearance of the lesions,R. solani formed brown, uninfective mycelium on and in the circumference of these lesions.Aldicarb did not influence any part of the infection process. Ethoprophos delayed the emergence of sprouts, but increased the number of sprouts per tuber. As soon as sprouts had emerged, growth was considerably promoted by ethoprophos. Ethoprophos delayed the appearance of lesions and reduced their size. Oxamyl showed the same effects to a smaller extent.As the size of lesions appears to be proportional to the size of the infection cushions, any agents that change the size of the infection cushions, such as pesticides or antagonists, may alter the severity of the disease.Samenvatting Het infectieproces vanRhizoctonia solani AG-3 werd bestudeerd op aardappelspruiten, cv. Bintje, in een klimaatcel bij 15C. Aanvankelijk groeide de schimmel met runnerhyfen voornamelijk in de lengterichting van de spruit. Via SEM kon waargenomen worden, dat de hyfen hierbij vooral over de begrenzingen van de epidermiscellen groeiden. Het mycelium vormde veel zijvertakkingen, bestaande uit iets gezwollen korte cellen, welke voornamelijk halverwege op het ondergrondse deel van de spruit gevormd werden. Een dichte massa van deze cellen vormde een infectiekussentje. Lesies, welke vanaf vijf dagen na inoculatie werden waargenomen, bevonden zich slechts onder spruitoppervlak bezet met infectiekussentjes. De lesiegrootte was recht evenredig met het spruitoppervlak dat bezet was met infectiekussentjes. De diepte van de lesies reikte tot aan de vaatbundels. De spruit werd alleen door de schimmel gekoloniseerd in gezond epidermisweefsel onder het infectiekussentje en in necrotisch weefsel. Enkele dagen na verschijning van lesies vormde R.solani bruin, niet infectieus, mycelium op en rondom de lesies.Aldicarb had geen effect op het infectieproces. Ethoprophos vertraagde de opkomst en verhoogde het aantal tot ontwikkeling gekomen spruiten per knol in gestoomd zand. Direct na opkomst had ethoprophos echter een sterk groeistimulerend effect. Ethoprophos vertraagde de lesievorming en reduceerde de lesiegrootte, vergeleken met onbehandelde planten. Oxamyl vertoonde deze effecten in geringere mate.Daar de lesiegrootte direct gecorreleerd blijkt met de grootte van het infectiekussentje, mag verwacht worden dat elke beïnvloeding van de ontwikkeling van het mycelium van R.solani, bijvoorbeeld door pesticiden of antagonisten, een verandering van de lesiegrootte ten gevolge heeft.  相似文献   

10.
由立枯丝核菌(Rhizoctonia solani)引起的马铃薯黑痣病由于常年连作和马铃薯种植规模扩大,病害发生越来越严重[1,2].目前马铃薯黑痣病的防治主要依靠农药的施用[3,4],但化学药剂的频繁使用会导致抗药性的产生、同时带来农药残留和环境污染等问题[5].从棉花根际土壤分离得到的多粘类芽胞杆菌Sx3可产生杀镰...  相似文献   

11.
马铃薯立枯丝核菌生物学特性研究   总被引:3,自引:0,他引:3  
采用生长速率法研究了黑龙江省马铃薯立枯丝核菌(Rhizoctonia solani Kühn)菌丝生长的条件。结果表明病菌菌丝在10~35℃均能生长,适宜温度是25~30℃,30℃菌丝生长最快。在pH为3~12的范围内病菌菌丝均能生长,以pH 5~7菌丝生长速度较快。黑暗对菌丝生长有促进作用,光照可促进菌核的形成。不同碳源、氮源对立枯丝核菌菌丝生长的影响均较明显,可溶性淀粉和酵母粉对菌丝生长有显著的促进作用。立枯丝核菌菌丝体及菌核的致死温度分别为49℃和51℃。明确了黑龙江省马铃薯立枯丝核菌生长的环境条件,可为病害防治提供理论依据。  相似文献   

12.
During a spring survey in 1991, 130 isolates of R. solani were collected in 25 commercial flower bulb fields from diseased plants occurring in bare patches. On the basis of hyphal fusion frequency and pathogenicity to flower bulbs, tulip isolates were provisionally assigned to AG 2-t to distinguish these isolates from AG 2-1 isolates which were non-pathogenic to bulbs. Hyphal fusion frequency of a subgroup of 7 AG 2-t isolates was highly variable when paired with 7 AG 2-1 isolates (2-75%), thus making assignment of AG 2-t isolates to AG 2-1 inconclusive. The mean hyphal fusion frequency among AG 2-t isolates was 65% (±6%) indicating AG 2-t to be a relatively homogeneous group. Hyphal fusion frequency among AG 2-1 isolates was highly variable with a mean 51% (±25%) indicating AG 2-1 to be a heterogeneous group. The optimum growth temperature for AG 2-t and AG 2-1 isolates on malt peptone agar was 20-25 °C. The host range of AG 2-t and two AG 2-1 isolates comprised tulip, iris, hyacinth and lily at both 9 and 18 °C, and cruciferous, sugarbeet and lettuce seedlings at 18 °C. Six other AG 2-1 isolates were pathogenic to cruciferous seedlings, but not to any of the bulbous crops. The tested narcissus, Tagetes patula, tomato, potato, wheat, leek and maize cultivars were not susceptible to AG 2-t and AG 2-1 isolates. Statistical analysis using a proportional-odds model revealed significant differences in aggressiveness between R. solani AG 2-t isolates and differences in susceptibility between tulip and iris cultivars. At 18 °C, but not at 9 °C, isolates representing AG 2-2, AG 4, AG 5 and AG BI were pathogenic to bulbous crops. In addition to bare patch causing AG 2-t isolates, other anastomosis groups may cause disease in field grown tulips. For the development of optimal crop rotation schedules, the impact of bulb rot causing isolates under field conditions needs further study.  相似文献   

13.
The acceleration of black scurf development after haulm destruction was mainly due to changes in the exudation of volatiles from tubers. Volatile products from decomposing potato roots and stolons and, probably, unstable substances in the tuber exudate as well, further promoted sclerotium formation.Sclerotium production byR. solani AG-3 was investigated on agar media, periderm strips and harvested tubers, which were exposed to the volatile exudates from growing subterranean potato plant parts. The volatile exudate from growing tubers contained both inhibitory and stimulatory substances which were not identified definitely. Inhibition dominated during tuber growth, decreased when plants were yellowing and disappeared after the shoots were excised. When the inhibitory components were trapped by KOH, the non-trapped volatile tuber exudates from young growing plants were as stimulatory as those from plants after haulm killing. CO2 might be an inhibitor as tuber respiration was negatively correlated to black scurf formation. Tests in vitro suggested that inhibition of sclerotium formation by CO2 can be overcome by stimulatory nutrients. Sclerotium production on agar media was not stimulated by ethylene, although volatiles from harvested ripe apples were very stimulatory.The results imply that after haulm killing, the increase in black scurf development may be prevented by loosening the soil and quick separation of tubers from plant residues thus preventing accumulation of the stimuli.Samenvatting De vluchtige exsudaten van aardappelknollen, die nog aan de plant bevestigd zaten, beinvloedden de produktie van sclerotiën doorRhizoctonia solani AG-3 op agarplaten en op geoogste knollen, die geincubeerd waren in een plant-aarde systeem. Onder dezelfde proef- omstandigheden was de sclerotiënvorming op geincubeerde losse knollen veel hoger dan op de agarplaten, maar op peridermstrips juist lager. Wellicht dragen dus naast stabiele ook instabiele knolexsudaten bij tot de vorming van lakschurft.Het vluchtige knolexsudaat van jonge planten bleek zowel stimulerende als remmende componenten te bevatten. Als de remmende fractie met KOH werd weggevangen, stimuleerden de resterende uitademingsprodukten van jonge groeiende knollen de vorming van sclerotiën even sterk als de uitademingsprodukten van oude afrijpende knollen na loofvernietiging. Tijdens de knolgroei overheerste de invloed van de remmende exsudaten, maar dat nam af als de plant vergeelde en verdween na loofvernietigen. Toename van lakschurft na loofdoding berust dus vooral op het wegvallen van de remmende componenten. In de praktijk zou na loofdoding de effectiviteit van de stimulerende exudaten verminderd kunnen worden door de grond van de teeltrug los te maken waardoor ze niet kunnen ophopen aan het knoloppervlak.De remmende fractie kon worden weggevangen met KOH, wat betekent dat het gaat om koolzuur of een andere zure component. Inderdaad bleek de produktie van koolzuur door knollen geleidelijk af te nemen tijdens de veroudering en zeer snel na loofdoding. Daarnaast is bekend dat lakschurft geleidelijk toeneemt bij veroudering van de plant, en zeer snel na loofdoding. In vitro leek koolzuur de sclerotiënvorming alleen op wateragar te remmen, maar niet op een voedzamer medium. De sclerotiënvorming op agarmedia werd sterk gestimuleerd door gasvormige produkten van appels. Echter, er werd geen bevestiging gevonden voor het idee dat het stress-produkt ethyleen sclerotiënvorming stimuleert. Na het loofafknippen lekte er een veel grotere hoeveelheid water uit de knollen dan uit knollen van intacte planten. Water kan het benutten van voedzame stoffen door de schimmel bevorderen. Daardoor kan een manier van loofdoden die de stolon breekt de kans op lakschurft verkleinen.Gasvormige produkten van afstervende wortels en stolonen hadden geen invloed op de sclerotiënvorming op agarplaten. Maar wel versterkten zij de lichte stimulering die uitging van afrijpende knollen. In de praktijk zouden jonge potaardappelen dus milieuvriendelijker beschermd kunnen worden tegen zowel virusinfectie als zware lakschurft-vorming met een nieuw-te-ontwikkelen methode van groen-rooien die de stolonen breekt en de knollen op het veld laat afharden in losse grond en gescheiden van de overige planteresten.  相似文献   

14.
To investigate the ability of black dot symptoms to develop on infected potato tubers during storage, the growth of Colletotrichum coccodes was followed in vitro on malt agar at temperatures ranging from 5–27°C, and in vivo on artificially infected potato tubers kept at 5, 10 and 15°C. In vitro , 13 isolates from different geographical origins grew at all temperatures tested; growth started with a delay of 10 days at 5°C and of 4 days at 10°C, and was fastest at 27°C. All isolates had similar growth patterns and produced conidia and sclerotia at all temperatures. Minitubers were successfully infected at 5, 10 and 15°C by depositing either a mycelial plug or a drop of conidial suspension on the tuber surface. Sclerotia were observed after 7 days at the point of inoculation. Symptoms extended in all cases, although more slowly at 5 and 10 than at 15°C. Latent infections were detected in up to 21% of tubers without black dot symptoms at harvest. These results show that latent infections by C. coccodes are probably quite frequent, and that the pathogen is able to develop at low temperatures in controlled conditions. This suggests that black dot symptoms can increase during storage if stores are not adequately managed.  相似文献   

15.
During the 1990s, silver scurf (causal agent Helminthosporium solani ) emerged as an economically important disease of table stock and processing potatoes ( Solanum tuberosum ). The pathogen attacks the periderm of the potato tuber causing blemishes. The disease cycle of silver scurf has both field and storage phases. Primary infection occurs in the field and high relative humidity favours the spread and increase of silver scurf in potato stores. Control of the disease by chemical and cultural practices remains difficult. Increase in disease has been attributed to H. solani isolates resistant to the postharvest fungicide thiabendazole (TBZ). Polymerase chain reaction (PCR)-based detection methods for H. solani and TBZ-resistant isolates are rapid and more specific than traditional identification. This review discusses the biology of the pathogen, epidemiology of the disease, detection of the pathogen and integrated control measures for the management of silver scurf in both field and potato tuber stores.  相似文献   

16.
马铃薯黑痣病立枯丝核菌(Rhizoctonia solani)菌丝存在遗传分化现象。从定西市马铃薯产区采集茎部和薯块病样80份进行融合群测定。结果表明,80株菌株中与标准菌株AG3融合的占总数的70.0%、与AG4-HG-Ⅱ融合的占15.0%、与AG2-1融合的占7.5%。为了有效控制马铃薯黑痣病,采用生长速率法在室内选取9种杀菌剂进行药效评价。结果表明:30%噻呋酰胺悬浮剂EC50最低,为0.015 2μg/mL;其次是40%氟胺·异菌脲悬浮剂、3%多抗霉素可湿性粉剂和250g/L嘧菌酯悬浮剂,其EC50分别为0.064 1、0.129 3和0.176 7μg/mL;EC50最大的是25.75%多抗·福美双可湿性粉剂,为2.930 4μg/mL。田间拌种,30%噻呋酰胺悬浮剂和250g/L嘧菌酯悬浮剂防效较好,达70%以上,值得在生产中推广。  相似文献   

17.
The sensitivity of a bioassay in detecting soil inoculum of Colletotrichum coccodes and Helminthosporium solani was examined using potato minitubers and microplants. Tests were conducted on soils which were collected from fields in which the interval after a previous potato crop differed, and which were also artificially infested with conidia or microsclerotia. For C. coccodes , determining plant infection based on the occurrence of infected roots after 9–12 weeks was a sensitive method for detecting and quantifying the amount of inoculum in soil. Infestations of less than 0·4 microsclerotia per g soil were detected in artificially infested soils. A semiselective medium, developed for isolating C. gloeosporioides from pepper, detected soil infestations by C. coccodes as low as nine conidia or one microsclerotium per g soil in artificially infested soil. For H. solani , infection on minitubers was a sensitive measure, with soil inoculum of fewer than 10 conidia per g soil being detected. Soil infestation could be quantified by assessing the percentage surface area of minitubers covered by sporulating lesions, which was strongly related to the amount of soil infestation. The results of these bioassay tests were compared with published results for real-time quantitative PCR assays on the same soils. The two methods were in good agreement in artificially infested soils, but the bioassay appeared to be more sensitive with naturally infested soils.  相似文献   

18.
为了探索马铃薯黑痣病高效防控技术,本研究选择240 g/L噻呋酰胺SC、20%嘧菌酯WG、10%苯醚甲环唑ME、0.3%四霉素AS、100万孢子/g寡雄腐霉WP、1%申嗪霉素SC等6种杀菌剂进行室内毒力测定和田间拌种防效试验.结果显示,6种杀菌剂对马铃薯黑痣病菌均有抑菌作用,其中1%申嗪霉素SC、20%嘧菌酯WG、0....  相似文献   

19.
A somatic cell selection approach, developed using thaxtomin A as a positive selection agent, recovered 113 potato cell colonies of cv. Iwa from which 39 regenerated plants within a single subculture period. In glasshouse pathogenicity trials, 13 of these regenerated variants exhibited significantly greater resistance to common scab disease than the unselected parent cultivar, both in mean tuber surface coverage with lesions and the proportion of disease free tubers. The best variant had on average an 85–86% lower disease score (or 91–92% less estimated tuber surface cover) than the unselected parent cultivar. The disease-resistant variants varied in their response to thaxtomin A in detached leaflet and tuber slice bioassays. This suggested that the cell selection procedure and thaxtomin A screening had limited success in selecting for toxin tolerance, and that thaxtomin A tolerance was not necessarily the driving factor for induction of disease resistance observed. Variation in relative necrosis scores between leaflet and tuber assays for individual variants possibly suggests differential expression of thaxtomin A tolerance in the different tissues. Yield estimates from the glasshouse grown potatoes showed the majority of disease-resistant variants had tuber yields equivalent to the unselected parent cultivar. This suggests that selection for disease resistance was not associated with negative tuber yield attributes and that these variants may have commercial merit and are worthy of further agronomic testing.  相似文献   

20.
Interactions between Verticillium dahliae and Colletotrichum coccodes , major causal agents of potato early dying (PED) syndrome, were studied in four potato cultivars that differ in their susceptibility to these pathogens . Aseptic plantlets of Nicola, Desiree, Alpha and Cara were inoculated with identical concentrations of each pathogen or with a mixture of the pathogens, and grown for 4 weeks in a monitored growth chamber. Coinoculation of Nicola with both pathogens caused more severe foliar disease symptoms and crown rot and greater C. coccodes colonization, than inoculation with each pathogen separately. Significant reductions in weight and height were also observed in plants coinoculated with both pathogens, as compared with plants inoculated with each pathogen separately or noninoculated plants. In Desiree, more roots were covered with C. coccodes sclerotia and disease symptoms were significantly more severe in plants inoculated with both pathogens together. However, plant weight and height were similar to those of plants inoculated with C. coccodes only. In Alpha, disease symptoms and levels of sclerotia in the roots were not affected by simultaneous inoculation with both pathogens. Weight and height of all plants were similar, whether inoculated with each pathogen separately or with both pathogens together. In Cara, plants inoculated with the mixture or either pathogen alone were smaller than the noninoculated control. Disease symptoms and occurrence of sclerotia were similar in plants inoculated with the combination and with a single pathogen. Compared with the effects of inoculation with either pathogen, simultaneous inoculation with both pathogens can, in some cultivars, increase the incidence of PED syndrome and thus severely decrease yields.  相似文献   

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