Cacao resistance to Phytophthora: Effect of pathogen species, inoculation depths and pod maturity

Two species of Phytophthora (P. palmivora and P. capsici) and inoculations at two depths (3 mm and 9 mm) were tested each on 10 clones of Theobroma cacao to determine their effects on pod resistance. Ripe and unripe pods were also assessed to determine the influence of physiological status of the pod on the expression of resistance. The two pathogens tested (P. palmivora and P. capsici) differed significantly in their reactions on pods, with P. palmivora being more aggressive than P. capsici. However, the lack of interaction between clones and pathogen species and the similarity in the ranking of clones based on lesion size suggested that selection for resistant clones can be based on one of the two pathogens, preferably the more aggressive one. Pod reactions differed between inoculation depths (3 mm and 9 mm), and between pod maturity stages (ripe and unripe pods) with relatively larger lesions being recorded at 9 mm depth and on unripe pods as compared to those observed at 3 mm depth and on unripe pods, respectively. The magnitude of increase in lesion sizes, however, varied with genotypes, indicating that inoculation depth and pod maturity stage should be standardized in screening cacao germplasm for resistance to Phytophthora.     

Rapid Screening of Cacao Genotypes for Field Resistance to Phytophthora palmivora Using Leaves, Twigs and Roots

Black pod, caused by Phytophthora spp. is one of the most important diseases of cacao occurring worldwide. Losses due to black pod caused by P. palmivora are still moderate in Côte d'Ivoire but P. megakarya causes high losses in Ghana and other Central African countries. Variation in field attack has been observed between cacao genotypes, but evaluation of pod losses is unsuitable for obtaining rapid progress in breeding. Results of inoculation tests using young detached leaves, twigs and roots, obtained from field and nursery plants, are presented here and compared to field resistance of similar genotypes observed over a 10-year period. Nine different Upper Amazon Forastero genotypes were tested together with progenies obtained by crossing these with the susceptible check IFC5 (Amelonado genotype). Rank correlations between the early screening tests and the level of field attack were positive and mostly significant (r=0.58–0.95). The coefficient of correlation was slightly higher for leaves (r=0.88) and roots (r=0.89) than for twigs (r=0.76). Also, resistance of the different plant organs was correlated (r=0.6–0.9). Resistance of the Upper Amazon parents was well correlated with the resistance of their cross progenies (r=0.7–0.9), suggesting that resistance is highly heritable. Resistance of leaves and twigs from the nursery was better correlated with field resistance than resistance of leaves and twigs from the field, which might result from more uniform growing conditions in the nursery. Inoculation of leaves appears the most suitable early screening method for black pod resistance. Application of this test in breeding more resistant cacao cultivars is discussed.     

Foliar resistance of cacao (Theobroma cacao) to Phytophthora palmivora as an indicator of pod resistance in the field: the effect of light intensity and time of day of leaf collection

Resistance of cacao leaves to Phytophthora palmivora was studied with regard to the time of leaf collection (morning, afternoon) and the degree of exposure of the leaves to light in the field (low, medium and high). The efficiency of leaf disc inoculations in predicting field resistance of nine clones was compared with that of detached and attached pod inoculations. Significant effects were observed, with leaves exposed to high light intensity and collected early in the afternoon showing highest susceptibility. The effect of time of leaf collection was reduced when leaves were stored overnight and leaf discs prepared and inoculated the following day, as compared to inoculations on the day of collection. Interactions between the main factors were significant, though less substantial than the clone effects. The most significant correlations with pod resistance ( r  = 0·70 to 0·97) were obtained for leaves collected early in the morning and exposed to intermediate shade conditions in the canopy. For other treatments, the correlations with pod resistance were still positive ( r  = 0·23 to 0·83) but often not significant. Pod inoculations in the laboratory were better correlated with field resistance ( r  = 0·92) than pod inoculations in the field ( r  = 0·72). Detached pod inoculations were also better correlated with leaf disc inoculations than those of attached pods. The results confirm the validity of laboratory inoculations of leaves and pods to assess field resistance to Phytophthora . Standardization of the leaf disc test is essential to obtain reliable results.     

A Laboratory Assay for Phytophthora infestans Resistance in Various Solanum Species Reflects the Field Situation

Physiological and molecular research on resistance responses of Solanum tuberosum cultivars and partially resistant Solanum species to Phytophthora infestans requires a reliable resistance test that can be used in the laboratory. Laboratory tests performed on detached leaves and intact plants were compared with field tests for similarity of late blight reactions. Detached leaves from field-grown plants were as resistant as detached leaves from climate chamber-grown plants when challenged with P. infestans. However, detached leaves incubated in covered trays at high relative humidity were more susceptible than detached leaves kept in open trays or leaves on intact plants. The incubation conditions of detached leaves in covered trays rather than detachment itself appeared to affect the resistance expression. Detached leaves of some wild Solanum genotypes became partially infected, whereas intact plants were completely resistant when inoculated. Inoculation of leaves on intact plants, however, resulted in lower infection efficiencies. These limitations should be taken into account when choosing the appropriate inoculation method for specific purposes. For resistance screening, laboratory tests proved to be a good alternative for field tests. The ranking of resistance levels for twenty plant genotypes was similar under laboratory and field conditions.     

Foliar resistance of cacao (Theobroma cacao) to Phytophthora palmivora as an indicator of pod resistance in the field: interaction of cacao genotype, leaf age and duration of incubation

The effects were studied of four leaf development stages (LDS) and three durations of incubation (DI) on the accuracy of leaf-disc tests on eight cacao (cocoa) clones (C) for predicting field resistance to phytophthora pod rot caused by Phytophthora palmivora . The clones were known to possess different general combining abilities (GCA) for pod resistance in the field, evaluated monthly at harvest over a 9-year period. Disease severity (DS) was affected strongly by C, DI and LDS, with increasing levels of significance. Two- and three-way interaction effects were smaller than the clone effect, but still significant. Clone effects were most significant for LDS3 (i.e. leaves 50–60 days old) and for DI5 and DI7 (observations made 5 and 7 days after inoculation, respectively). Coefficients of rank correlation between DS and field results were significant for seven of the 12 treatments, with highest values obtained again for treatments LDS3/DI5 ( r = 0·87) and LSD3/DI7 ( r = 0·93). Pooling of data for different LDS and DI treatments did not further improve the correlation with field results. However, these correlations were improved (from an average of 0·74 to 0·88) when the GCA values for field resistance were based on weekly observations, carried out in one year, including losses of pods and cherelles. It was concluded that, when carried out in a standardized manner and under optimal conditions, the leaf-disc test may explain 75–90% of the genetic variation for field resistance of cacao genotypes to P. palmivora .     

Limited morphological,physiological and genetic diversity of Phytophthora palmivora from cocoa in Papua New Guinea

In Papua New Guinea (PNG) cocoa (Theobroma cacao) is one of the most important cash crops grown in the tropical lowland and island regions. As in most cocoa‐growing areas, phytophthora black pod and canker cause significant yield losses. Cocoa breeding activities in PNG are focused in East New Britain province where disease control recommendations are also developed. This study tested the hypothesis that there was no diversity in the Phytophthora palmivora population causing black pod on cocoa by characterizing the variation in pathogen populations within and between the five major cocoa‐growing areas. Diseased pods were sampled hierarchically from the five locations and additional isolates were collected from soil, stem and leaf lesions, or retrieved from culture collections. Morphological characters showed continuous variation within the range described for P. palmivora. Genetic analysis revealed that the isolates belonged to one dominant clonal lineage, with restricted distributions of several other subpopulations. Lowest diversities were found in the geographically isolated Karkar Island and East Sepik province. Soil isolates showed greater genetic diversity than isolates from cocoa lesions. Intra‐farm variation was as much as inter‐farm or inter‐province variation. Both mating types were detected, although no strong evidence of sexual recombination was observed. The analysis revealed limited geographic, temporal or host specialization, suggesting continuous selection for pathogenicity from a genetic pool of P. palmivora. These findings have significant implications on the deployment of cocoa genotypes, enforcement of inter‐province quarantine and sustainable disease management strategies.     

云南白背飞虱不同种群对杀虫剂的抗药性及变化趋势

为了掌握云南不同稻区白背飞虱Sogatella furcifera(Horváth)种群对常用杀虫剂的抗药性及其发展趋势,2012-2015年,连续4年采用稻茎浸渍法在室内测定了滇西沧源、滇西南勐海和滇东师宗3个稻区田间白背飞虱对5种常用杀虫剂的抗药性。云南不同稻区田间白背飞虱对5种药剂的抗性水平总体在0.9~23.4倍,其中对毒死蜱的抗药性最高,除少数年份外,LC502 mg/L,抗性倍数10倍,抗药性达到了中等水平,年度间变化幅度较大;对噻嗪酮的抗药性呈下降趋势,滇西沧源和滇西南勐海种群对噻嗪酮的抗药性由2012年的中抗水平(11.0倍和23.4倍)下降到2015年的4.6倍和7.5倍;3个稻区种群4年对吡虫啉、吡蚜酮和噻虫嗪的抗性倍数最高达7.2、6.5和5.5倍,总体表现为无抗或低水平抗性。由于噻嗪酮和毒死蜱在云南稻区的频繁和广泛使用,田间白背飞虱已对其产生抗药性,建议限制这两种杀虫剂的使用频率或与不同作用方式的药剂轮换使用,其余药剂可以有计划地选用。     

枯草芽胞杆菌VD18R19在胡椒上的定殖动态及促生作用和对胡椒瘟病的防治效果

生防芽胞杆菌Bacillus subtilis定殖能力是决定其应用效果的关键因素。目前,芽胞杆菌在胡椒上的定殖情况鲜有报道。本研究以生防芽胞杆菌VD18R19为对象,研究其在胡椒根系和叶片表面的定殖动态及其促生长作用和对胡椒瘟病的防治效果。首先利用荧光质粒p AD43-25标记菌株VD18R19;然后通过荧光观测和平板计数方法检测标记菌株在胡椒根系和叶片表面的定殖形态和数量动态;最后通过盆栽试验分别评估了该菌株的促生长作用和对胡椒瘟病的防治效果。结果表明,菌株VD18R19荧光标记成功,且标记具有良好的遗传稳定性(120世代后高达85.33%);该菌株在胡椒根际偏好于根毛区定殖,定殖数量呈先升后降趋势,在处理后第9 d的定殖数量最多(9.88×106 CFU/g),整个30 d的定殖密度均维持在106 CFU/g水平;叶片定殖呈迅速下降趋势,处理后第9 d的定殖密度仅5.13×104 CFU/g,在21 d后基本检测不到该菌株;在不接种病原菌的情况下,菌株VD18R19处理的胡椒苗的株高、叶片数和地上部分鲜重等均显著好于对照,其促生率分别为61.71%、36.18%和106.43%;胡椒瘟病病原菌Phytophthora capsici叶片接种后,菌株VD18R19处理的胡椒苗在发病率和病斑大小上均显著低于对照,对胡椒瘟病的防治效果达76.86%。以上研究结果揭示了芽胞杆菌VD18R19在胡椒上的定殖动态及其防治效果,对该菌的开发应用有重要指导意义。     

利用诱导因子抗新疆甜瓜疫霉病的田间效果

用化学诱导因子和灭活的生物诱导因子在田间自然条件下诱导了新疆甜瓜产生对疫霉病的整体抗性。1996年的试验结果表明，用化学诱导因子诱导处理，最高诱抗效果可达79．40％，平均诱抗效果在40％以上；生物诱导因子诱导处理，最高诱抗效果为68．2％，平均诱抗效果为30％以上。1997年的试验基本上验证了1996年的结果，化学诱导因子最高诱抗效果可达60％，平均诱抗效果为35．86％，经t测验分析，诱抗效果显著。     

Irvingia, a forest host of the cocoa black-pod pathogen, Phytophthora megakarya, in Cameroon

Phytophthora megakarya is an increasingly important invasive pathogen of cocoa in West Africa, where it forms part of the black-pod disease complex together with the more widespread, but less aggressive, P. palmivora . Its purported centre of diversity, based on evidence from molecular studies and mating types, lies on the Cameroon–Nigeria border, from where it has spread throughout both countries and more recently into Ghana and the Ivory Coast. A survey undertaken in an ancient primary forest (Korup National Park) in western Cameroon, adjacent to the Nigerian border, identified Irvingia sp., close to I. gabonensis (Irvingiaceae), as a wild host of P. megakarya , using both morphological and molecular characterization. ITS fingerprinting of two additional Phytophthora isolates, obtained during a supplementary survey in central Cameroon, showed identical banding patterns for the isolate from a cocoa pod, whilst the isolate from an unknown forest fruit proved to belong to P. heveae . This is the first authenticated record of P. heveae from the African continent. Subsequent AFLP analysis grouped the two P. megakarya isolates with other Cameroonian isolates held in the CABI Genetic Resource Collection, but separated them from the Ghanaian isolates.     

小豆疫霉茎腐病病原菌鉴定及抗病资源筛选

在黑龙江省佳木斯市合江地区农科所发现一种小豆疫霉茎腐病,典型症状是在茎部产生红棕色病斑,有时可在病茎和病荚产生白色霉层,严重时发病植株萎蔫、死亡.病原菌游动孢子囊卵圆形至倒梨形,无乳突,在孢囊柄上不脱落,平均大小为47μm×30μm,长宽比为1.6∶1.有性生殖为同宗配合,藏卵器圆形,平均直径为36.8μm,卵孢子平均直径为29.7μm,平均壁厚为2.9μm;雄器球形至卵圆形,围生,平均大小为16.4μm×15.7μm.生长温度为9～37℃,最适生长温度为24～27℃,生长抑制温度为39℃.完全抑制菌丝生长的孔雀绿浓度为5μg/ml,恶霉灵浓度为100μg/ml时对菌丝的生长相对抑制率为20%.病原菌无伤接种时只侵染小豆,伤口接种对绿豆、豇豆和菜豆具有不同程度的致病性.根据形态、生理、寄主范围和病害症状,病原菌被鉴定为豇豆疫霉菌小豆专化型(Phytophthora vignae f. sp. adzukcola).对70份小豆资源的抗性进行了室内接种评价,有7份资源表现抗病.     

贵州省烟草黑胫病菌对甲霜灵的抗药性

为了明确贵州地区烟草黑胫病菌对甲霜灵的抗药性水平, 从贵州省12个市、县采集黑胫病病株, 经分离纯化鉴定, 获得98株黑胫病菌株, 采用生长速率法测定其对甲霜灵的敏感性。结果表明, 采自不同地区的菌株对甲霜灵的敏感性差异较大, 各供试菌株的EC50 值分布范围为0.419 2～20.486 1 μg/mL; 各地区平均EC50 在0.551 9～10.986 0 μg/mL之间; 最低抗药性水平为1.00, 最高抗药性水平为48.87, 相差48.87倍, 各地区平均抗药性水平在1.32～26.21之间。遵义、兴仁、贵定、赫章和兴义有抗甲霜灵菌株出现, 除遵义部分菌株的抗性水平表现为高抗外, 其余菌株对甲霜灵产生的抗性均处于中抗水平。     

On-farm selection for quality and resistance to pest/diseases of cocoa in Sulawesi: (ii) quality and performance of selections against Phytophthora pod rot and vascular-streak dieback

The cocoa industry in Sulawesi, the main region of cocoa production in Indonesia, is threatened by destructive diseases, including vascular-streak dieback (VSD) caused by the basidiomycete Oncobasidium theobromae and stem canker and Phytophthora pod rot (PPR) or black pod, caused by Phytophthora palmivora. Using the considerable genetic diversity of cocoa on farms, host resistance was identified and tested with the participation of farmers. Forty-nine local and international cocoa selections with promising resistance characteristics (as well as susceptible controls) were side-grafted onto mature cocoa in a replicated trial with single-tree plots. Developing grafts were assessed in the dry season for severity of VSD infection, scored from 0 (no infection) to 4 (graft death). All of the 49 clones in the trial became infected with VSD in at least some replicates. Average severity varied from 0.2 to 1.6. Potential VSD-resistance was found in eight clones, including DRC 15, KA2 106 and a local Sulawesi selection, VSD2Ldg. Some of the most susceptible clones were local Sulawesi selections from areas with a history of little or no VSD. Thirty-four pod-bearing clones were evaluated over a 2-year period for yield, quality and resistance to natural infections of PPR. Cumulative PPR incidence for all clones was 22% but varied from 8.6 to 43% among clones. Clones with less than 15% PPR incidence were designated as resistant, including DRC 16 and local Sulawesi selections, Aryadi 1, Aryadi 3 and VSD1Ldg. Scavina 12 was moderately resistant in the trial with a PPR incidence of 23%. Cumulative incidences of the mirid, Helopeltis spp., determined in the same evaluation period, indicated that DRC16 was the most susceptible clone with an incidence of 52% in ripe pods and 23% in immature pods. In comparison, KKM4 showed evidence of resistance to Helopeltis spp., with incidences of 34 and 0.8% in ripe and immature pods, respectively. The impact of diseases and pests (including cocoa pod borer) on bean losses and bean quality varied between clones but generally the bean size (or bean count) was affected more than the fat content or shell content.     

知母提取物诱导马铃薯植株抗晚疫病作用机制初探

为明确知母提取物诱导马铃薯抗晚疫病的作用,本研究测试了经知母提取物处理并接种晚疫病菌后马铃薯植株的超氧化物歧化酶（SOD）、苯丙氨酸解氨酶(PAL)、β 1,3 葡聚糖酶和几丁质酶活性的变化。结果表明,经过知母提取物处理后的马铃薯植株,在受到晚疫病菌侵染后24～96 h,SOD和PAL活性显著升高并持续处于高活性水平;在接种48 h后β 1,3 葡聚糖酶活性开始显著升高,直到接种后96 h一直维持在较高活性水平;而无论是接种晚疫病菌还是经过知母提取物处理,几丁质酶活性都没有发生显著的变化。     

Foliar aggressiveness of Northern Ireland isolates of Phytophthora infestans on detached leaflets of three potato cultivars

The aggressiveness of 20 Northern Ireland single-lesion isolates of Phytophthora infestans was compared following their inoculation onto detached leaflets of three potato cultivars chosen on the basis of their differing levels of race-nonspecific resistance to late blight: Bintje (highly susceptible); Cara (moderately resistant); and Stirling (more resistant). Five isolates from outside Northern Ireland were included for comparative purposes: two from the Republic of Ireland; two from the USA (representing the US-1 and US-8 clonal lineages); and one from Mexico. To control the variation between tests, a balanced incomplete block design was used, as opposed to either a complete block design or the method of inclusion of standard isolates, where such variation would have increased the error. Highly significant variation for disease parameters, including latent period, infection frequency, area under the lesion expansion curve (AULEC) and sporulation capacity, was found between isolates. These differences were much more marked on the cultivars exhibiting higher levels of race-nonspecific resistance. There was a significant interaction between isolate and cultivar for all parameters assessed and, overall, no one isolate was the most aggressive across all three potato cultivars. However, a group comprising seven of the 20 Northern Ireland isolates was consistently found to exhibit the highest levels of aggression towards all three cultivars for each of the disease parameters. These results demonstrate that significant variation for foliar aggressiveness exists within the Northern Ireland population of P. infestans , and indicate the importance of selecting appropriately aggressive isolates for evaluation of host resistance to late blight within breeding programmes.     

疫霉菌对霜脲氰抗性遗传及对霜脲氰和甲霜灵的交互抗性

就疫霉菌对霜脲氰抗性的产生和遗传，以及疫霉菌对霜脲氰和甲霜灵的交互抗性进行了研究。结果表明，苎麻疫霉容易对霜脲氰产生抗药性突变，但抗性在连续３代单游动孢子分离过程中逐渐丧失。大雄疫霉对霜脲氰不易产生抗性突变，恶疫霉、大雄疫霉和苎麻疫霉对甲霜灵和霜脲氰不存在交互抗性     

甘薯根腐病抗病性室内鉴定方法的研究

将PDA 平面培养的甘薯根腐病菌研磨成浆,按一定比例加入蛭石中,接菌最适浓度为每200 g蛭石加1/ 2 培养皿菌浆,随后栽入薯苗,在25 ℃下培养25 ～30 d 后检查。以薯苗叶片枯黄程度及根部黑根死根多少划分病级,以病级平均数确定品种抗性,同时对抗感品种接种根腐病菌后过氧化物酶活性进行测定。试验证明,上述方法可以简便、快速、准确地测定品种对根腐病的抗性,同时也证明过氧化物酶活性作为量化分级标准的可行性。     

甘薯根腐病抗病性室内鉴定方法的研究

将PDA 平面培养的甘薯根腐病菌研磨成浆,按一定比例加入蛭石中,接菌最适浓度为每200 g蛭石加1/ 2 培养皿菌浆,随后栽入薯苗,在25 ℃下培养25 ～30 d 后检查。以薯苗叶片枯黄程度及根部黑根死根多少划分病级,以病级平均数确定品种抗性,同时对抗感品种接种根腐病菌后过氧化物酶活性进行测定。试验证明,上述方法可以简便、快速、准确地测定品种对根腐病的抗性,同时也证明过氧化物酶活性作为量化分级标准的可行性。     

Assessment of inoculation methods for screening black alder resistance to Phytophthora ×alni

Identification of resistance to Phytophthora ×alni could provide the basis for a management strategy against alder decline in riparian ecosystems in Europe. This study aimed to test methods to evaluate the resistance of riparian alders to the disease, and to screen alder genotypes for resistance. Phytophthora ×alni isolates were compared for their aggressiveness (lesion length on stem) and sporulation capacity (sporangia). While no difference in lesion lengths was found between isolates, sporangia production was dependent on isolate, highlighting the need for careful selection of isolates used for zoospore inoculation methods. Inoculation tests carried out at different periods of the year revealed a seasonal change in susceptibility to the disease, with the period from June to September being the most efficient for inoculation tests. Stem‐wounded inoculations, carried out on excised shoots, were unreliable for evaluating the level of resistance of alder genotypes to P. ×alni infection, with divergent results between two successive years or between two inoculation periods during the same year. In contrast, a method that mimics the natural conditions of infection, based on flooding of rooted cuttings in artificially infected river water, was found promising. Another method, based on the inoculation of foliated terminal shoots with zoospore suspensions, was found to be repeatable and could be used for high‐throughput analyses. Altogether, the results show a continuous resistance response from highly susceptible to moderately resistant genotypes. This suggests that breeding might be a useful strategy to manage alder decline caused by P. ×alni.     

辣椒疫病抗性的浸根接种鉴定方法

为建立规模化的辣椒疫病苗期抗性鉴定技术, 将辣椒疫霉人工接种于抗疫病辣椒品种‘ICPN21-03’和感病辣椒品种‘B16144’, 研究了接种菌液浓度?接种时期?病情调查时期和接种方法等对人工接种鉴定效果的影响?结果表明, 接种菌液游动孢子浓度为10个/mL时, 感病品种病情指数为73.37, 可以满足寄主发病的需要; 1 ~ 10真叶期接种均能使植株发病, 但5 ~ 6真叶期接种能反映抗感病品种真实抗性; 接种5 d调查抗感病品种病情指数分别为9.32和73.37, 可明显区别抗感病品种?使用浸根法进行鉴定, 需菌量小, 对接种环境要求小?用37个辣椒品种进行验证, 鉴定结果表明该苗期抗性鉴定技术可很好地对辣椒品种进行抗性水平分级鉴定?