In vitro screening for resistance to apple proliferation in Malus species

A screening system for apple proliferation resistance was developed, based on in vitro graft‐inoculation with the causal agent ‘Candidatus Phytoplasma mali’. For this, in vitro cultures of the field‐resistant apomictic genotypes Malus sieboldii, H0909, D2212 and the susceptible Malus × domestica genotypes Golden Delicious and rootstock M9 were established, as well as in vitro cultures of Rubinette and Golden Delicious infected with ‘Ca. P. mali’ strains PM4 and PM6, respectively. Healthy in vitro shoots were inoculated by micrografting with infected shoots used as graft tip. After 6 weeks graft contact no significant differences for graft quality were observed between healthy and infected grafts. Mortality of grafts and transmission rates were not significantly different among the different genotypes. The phytoplasma concentration in inoculated shoots was determined at different times post‐inoculation (p.i.) by quantitative real‐time PCR. Infected M. sieboldii and D2212 had lower phytoplasma concentration than the susceptible controls and showed no symptoms. H0909 showed an intermediate behaviour exhibiting lower phytoplasma concentrations with strain PM4 but growth was affected. The dynamics of phytoplasma concentration reached a maximum at 6–8 months p.i. for all genotypes but the values for 3–5 and 10–12 months p.i. were similar. The resistance of M. sieboldii and D2212 was confirmed in vitro. A significant difference in phytoplasma concentration was observed between strains PM4 and PM6.     

Screening Luffa germplasm and advanced breeding lines for resistance to Tomato leaf curl New Delhi virus

The two cultivated Luffa species can be severely infected by Tomato leaf curl New Delhi virus (ToLCNDV) with up to 100% yield loss. Here, 52 Luffa genotypes were screened for ToLCNDV resistance after natural field infection. Mean vulnerability index (VI) ranged from 0.00 to 75.33; genotypes IIHR-137 and IIHR-138 had no symptoms (VI 0), 16 genotypes were resistant (VI 0–25), 15 were moderately resistant (VI 26–50), and 19 were moderate to susceptible (VI?>?50). Ten of the most resistant genotypes and five susceptible checks were then challenge-inoculated using whiteflies or sap in an insect-proof net house; only IIHR-137 [L. cylindrica (L.) Roem.] was symptomless (VI 0.00), and 3–5% of plants of IIHR-138 [L. cylindrica (L.) Roem.] and IIHR-Sel-1 [L. acutangula (L.) Roxb.] had only mild symptoms; genotype Arka Prasan was most susceptible (VI 80.96). Asymptomatic plants were confirmed as infected using polymerase chain reaction. Susceptible genotypes rapidly developed leaf curling, then a severe mosaic 10 days post-inoculation. The resistant inbred lines identified are good candidates for a breeding program for ToLCNDV-resistant cultivars.

     

Screening for blast resistance in rice using AMMI models to understand G x E interaction in Guyana

In rice, totally one hundred and three genotypes were evaluated for resistance against blast disease in three hot spot locations of Guyana. The experiments were conducted in spring and autumn seasons of 2015, 2016 and 2017. The modified upland blast nursery (UBN) method was adopted for screening blast resistance. The Additive Main effects and Multiplicative Interaction (AMMI) analysis was applied to understand the interaction between genotype (G) and environment (E). The results of all seasons at three locations showed the consistent expression of high resistance by genotype FL 127 followed by FG12–08 and FG12–273. The AMMI analysis of variance (ANOVA) revealed that, resistance was dominated by the genotype main effect. The magnitude of genotypes by environment (G x E) sum of square (SS) was 3.64 times smaller than genotype SS, and 6.74 times larger than environment SS. The analysis has indicated that the differences of the genotypes across the environment were not substantial. All of the seven environment vectors appeared to be positively correlated. The screening at Onverwagt Back, autumn 2016 (E7) and Black Bush Polder, spring 2015 (E5) were observed to be the most representative environment. This was clearly revealed by smaller average - environment axis (AEA) angle. Likewise, E4 (Canje, autumn 2015) was observed closer to the “ideal test environment”, thus suggested as an ideal environment for blast resistant screening. Also, genotypes clustered near the origin have showed stable blast disease resistant status and contributed little to both G and G x E. These genotypes proposed as useful ones for breeding disease resistant varieties and hybrids in rice.     

Evaluation of Brassica species for resistance to Rhizoctonia solani and binucleate Rhizoctonia (Ceratobasidum spp.) under controlled environment conditions

Isolates of R. solani AG 2–1, AG 8, AG 10 and binucleate Rhizoctonia (Ceratobasidium spp.) were tested for virulence on Brassica crops in growth chamber experiments. Isolate virulence and genotype resistance were determined based on percent of seedling survival, shoot length reduction, and shoot fresh weight. Isolates had significant effects on all tested measurements, compared to the non-inoculated controls. Rhizoctonia solani AG 2–1 appears to be the most aggressive pathogen on all tested genotypes followed by R. solani AG 8, binucleate Rhizoctonia and R. solani AG 10, respectively. Genotype by isolate interaction effects were found to be significant for percent of seedling survival and shoot length reduction. None of the tested genotypes exhibited any level of resistance to R. solani AG 2–1, but three promising genotypes with moderate levels of resistance to R. solani AG 10, R. solani AG 8 and binucleate Rhizoctonia were identified. Moderate heritability (0.57) was observed for the percent of seedling survival in the resistant genotype KS4022.     

The reliability of leaf bioassays for predicting disease resistance on fruit: a case study on grapevine resistance to downy and powdery mildew

This study was designed to assess the reliability of grapevine leaf bioassays for predicting disease resistance on fruit in the field. The efficacy of various grapevine quantitative trait loci (QTLs) for conferring resistance to downy and powdery mildew was evaluated in bioassays and in a 2‐year field experiment for downy mildew. The resistance genes studied were inherited from Muscadinia rotundifolia (Rpv1 and Run1) and from American Vitis species through cv. Regent (QTLRgP and QTLRgD). In bioassays, genotypes carrying Run1 blocked powdery mildew development at early stages. Genotypes combining Run1 with QTLRgP displayed no greater level of resistance. For downy mildew, genotypes carrying Rpv1 and/or QTLRgD were more resistant than the susceptible cv. Merlot, and showed a high level of leaf resistance in the field (<10% severity). Disease levels on bunches were much higher than those on leaves, with a high variability between Rpv1 genotypes (1–48%). A Bayesian decision theory framework predicted that an OIV‐452 threshold of 5 in leaf bioassays allowed accurate selection of grapevine genotypes (P = 0·83) with satisfactory disease severity on bunches. Therefore, this study validates that the use of early bioassays on leaves, as currently performed by grapevine breeders, ensures a satisfactory level of resistance to downy mildew of bunches in the field.     

Genotype-isolate interaction for resistance to black stem in sunflower (Helianthus annuus)

Two experiments were undertaken to determine the partial resistance of sunflower genotypes to seven isolates of Phoma macdonaldii . In the first experiment, 28 genotypes, including recombinant inbred lines and their parents, M6 mutant lines developed by gamma irradiation, and some genotypes from different geographical origins, were used. The experiment consisted of a split-plot design with three replications, each with 12 seedlings per genotype per isolate, in controlled conditions. Seven days after inoculation, plantlets were scored on a 1–9 scale for percentage necrotic area. Highly significant differences were observed among genotypes, isolates and their interactions. The presence of a differential interaction between genotypes and P. macdonaldii isolates was confirmed in a second experiment using 12 genotypes representing large variability for partial resistance to P. macdonaldii isolates, as identified in the first experiment. Inbred lines B454/03, ENSAT-B5 and LC1064C were the most susceptible sunflower genotypes, whereas two American lines SDR19 and SDR18 presented high partial resistance to all P. macdonaldii isolates studied. The least and most aggressive isolates were MA6 and MP3, respectively. Isolates interacted differentially with sunflower genotypes. This study identified two genotypes (AS613 and PAC2) presenting specific resistance to isolate MP8. The results also showed a wide range of isolate-nonspecific partial resistances among the lines tested. The information presented here could assist sunflower breeders to choose parents of crosses for breeding of durable resistance to phoma black stem disease.     

Screening Lactuca for resistance to Myzus persicae

A macrotest was developed to screen rapidly many genotypes ofLactuca in the greenhouse for resistance toMyzus persicae. In this test the estimated number of aphids on certain leaves of plants artificially exposed to aphid infestation was representative for the total number of aphids per plant. Significant differences in resistance between genotypes were observed and several partially resistant PIVT numbers were found among 645 accessions tested in 1973 and 1974. Also a microtest with leaf cages was developed in which biomass after seven to eight days of larval development and net larvae production during four to seven days functioned as criteria of resistance. Besides differences between genotypes, also within plants and between plants of the same genotype significant differences were found to occur. After testing nineteen genotypes with different levels of resistance it appeared that there was a clear relationship between results from both types of tests.     

Stability of resistance to Phytophthora infestans in potato: an international evaluation

Ten institutions in nine countries joined together to test the stability of resistance of 14 potato genotypes to the oomycete pathogen Phytophthora infestans in three separate trials. Seven of the genotypes were tested in one trial involving seven locations, and all 14 were tested in two subsequent trials, each involving eight locations. Stability of resistance was tested with nonparametric tests and with an additive main effects and multiplicative interaction (AMMI) model. Overall, resistance to P. infestans was robust; resistant genotypes were consistently resistant in all locations and trials. The nonparametric analysis indicated that specific genotypes were basically stable across sites for resistance. In trial 3, the Z statistic for overall stability was significant at 0·05%, indicating a significant level of interaction across the trial, but there were no significant interactions for specific genotypes in this trial. The genotype by environment (G × E) effect of the AMMI model was highly significant in both trials, but the mean square of G × E was less than 10% of the genotype effect in each trial. The first two principal components (PCA1 and PCA2) of the AMMI analyses together explained 75 and 80% of the interaction effects in trials 2 and 3, respectively. Based on both nonparametric and AMMI analyses, Ecuador and Argentina were locations of relatively high interaction effects for both trials 2 and 3, although in Ecuador this interaction was not associated with any particular potato genotype. Other locations also had high interaction effects, but these occurred in only one trial. The genotypes Chata Blanca and, to a lesser extent, Torridon were relatively unstable in trials 2 and 3, but in the case of Torridon, resistant, this did not represent a significant loss of resistance.     

Insecticide resistance traits differ among and within host races in Aphis gossypii

BACKGROUND: The polyphagous cotton‐melon aphid Aphis gossypii Glover is structured into geographically widespread host races comprising a few clones specialised on Cucurbitaceae, cotton, eggplant or pepper. To assess insecticide resistance among and within host races, leaf disc bioassays were conducted on aphid clones collected from Cucurbitaceae (genotypes C4 and C9), cotton (genotypes Burk and Ivo), eggplant (genotype Auber) and pepper (genotype PsP4). Molecular diagnostic (PCR‐RFLP) and enzyme assays were also performed to detect the basic mechanisms underlying insecticide resistance. RESULTS: All six clones were susceptible to acetamiprid (neonicotinoid) or carbosulfan (carbamate). Conversely, all clones were resistant to dimethoate (organophosphate) (RF = 4.1–38.1) and carried mutation S431F in the acetylcholinesterase gene. Auber, PsP4 and Burk also carried mutation A302S in this gene, which possibly conferred moderate resistance (RF = 3.7–6.8) to profenofos and monocrotophos (organophosphates). Auber and Burk were highly resistant (RF = 41.2 and 473 respectively) to cypermethrin (pyrethroid). This resistance was likely associated with point mutation super‐kdr (M918L) in the voltage‐gated sodium channel gene (para gene) or metabolic detoxification mediated by esterase and oxidase enzymes. CONCLUSION: Multiple resistance to a broad range of insecticides and multiple mechanisms of resistance in some clones could explain to some extent the low genetic diversity observed within A. gossypii host races. Copyright © 2009 Society of Chemical Industry     

Phenotyping Castanea hybrids for Phytophthora cinnamomi resistance

Castanea sativa is susceptible to Phytophthora spp., a serious root pathogen causing ink disease, while C. crenata and C. mollissima show resistance to infection. Interspecific controlled crosses were established for introgression of resistance genes from the resistant species into the susceptible C. sativa, and two mapping populations were created. Phytophthora cinnamomi resistance of each progeny was evaluated by root and excised shoot inoculation tests. The number of days of survival after root inoculation was the best discriminator of resistance to P. cinnamomi while the percentage of shoots with internal lesions was the symptom most associated with survival. The lesion progression rate in the excised shoot inoculation test was strongly and negatively correlated with survival in the root inoculation test. The excised shoot inoculation test appears to be a reliable approach for screening the resistance of chestnut genotypes to P. cinnamomi. Strong genetic correlations were obtained between survival and ink disease symptoms and among symptoms, indicating that common or linked genes might influence resistance to P. cinnamomi. The most resistant genotypes selected from this study will be tested for other commercial variables, such as ease of vegetative propagation and stock–scion compatibility.     

Genetic resistance to bacterial blight disease in Persian walnut

Bacterial blight disease of Persian walnut (Juglans regia, L.), caused by Xanthomonas arboricola pv. juglandis (Xaj), leads to significant nut losses in northern, central and western areas of Iran. To identify the natural sources of resistance to disease in the endemic walnut genotypes of Iran, sixteen walnut genotypes, collected from different areas of Hamedan province, were inoculated with Xaj in a randomized complete block design with five replicates for each genotype. Two-year old genotypes were gently sprayed with a suspension of bacteria adjusted to approximately 2 × 10⁹ cfu ml⁻¹ of distilled water in May. Infected leaves were rated for disease 28 and 42 days after inoculation, using a 0 to 5 severity scale, based on the number, size and distribution of lesions on the leaves. Data analyses showed that there were variations among genotypes in response to pathogen. Upon inoculation by bacterial suspension genotype 94 showed the highest resistance to both disease incidence and its progress after 4–6 weeks of infection. Genotype 65 showed high susceptibility to disease and genotype 69 showed high susceptibilities both to disease incidence and its progress after 4–6 weeks of infection.     

玉米抗南方锈病种质标记基因型鉴定与遗传多样性分析

为阐明玉米抗南方锈病种质的标记基因型和遗传背景,利用7个与玉米抗南方锈病3个基因连锁的SSR标记鉴定了38份抗病玉米种质的标记基因型,并采用40个多态性SSR标记对39份抗南方锈病的玉米自交系和6个标准测验种进行了遗传多样性研究。结果表明,7个与抗病基因连锁的SSR标记将38份抗病种质鉴定为17种标记基因型,表明可能存在多样的抗性基因组合方式;辽2204等9份种质仅扩增出齐319的标记基因型,沈136和W456仅扩增出W2D的标记基因型;种质LO932未扩增出与齐319、P25和W2D相同的标记,可能携带新的抗南方锈病基因;相近遗传背景的抗性种质分属不同的标记基因型,表明抗病种质可能携带的抗南方锈病基因在育种选择中发生了分离。40对多态性SSR引物在45份自交系中共检测出115个等位基因变异,平均每对引物检测到2.88个等位基因,变异范围为2~4;平均多态性信息含量为0.4649,变化范围为0.1258~0.6951;通过UPGMA聚类分析,39份抗病材料被划分到以标准测验种为代表的6个杂种优势亚群中,与系谱分析基本一致,这为在育种中合理利用抗源提供了信息。     

Sources of genetic resistance in cowpea (Vigna unguiculata (L.) Walp.) to cowpea aphid-borne mosaic potyvirus

Fifty-one cowpea (Vigna unguiculata (L.) Walp.) genotypes were tested by mechanical inoculation with seven geographically diverse isolates of cowpea aphid-borne mosaic potyvirus to identify resistant sources. Of 51 genotypes three, TVu-401, TVu-1582 and TVu-1593 were found immune to all the seven isolates. Forty-five genotypes gave different reactions to individual isolates. Several immune, resistant and tolerant genotypes against each isolate were identified. A considerable evidence of pathogenic variability among the virus isolates was also observed.     

四川58个小麦品种苗期抗条锈基因推导及成株期抗性表现

为了解四川小麦品种所含抗条锈基因及其在田间的抗性表现,将29个毒性谱各异的小麦条锈菌鉴别菌株于苗期接种35个已知抗条锈基因载体品系和58份四川小麦品种（系）,通过抗病谱对比分析和各品种（系）系谱分析,推导了四川小麦品种（系）的抗条锈基因型。在田间,选用中国小麦条锈菌优势小种CY32、CY33、Su-4和对Yr10、Yr24和Yr26致病的贵农22致病类型田间突变株进行混合接种,在乳熟期对各品种（系）的抗条锈性进行了鉴定。结果显示,Yr2、Yr5、Yr7、Yr9、Yr10、Yr24、Yr27、YrSpp、YrAlba等9个基因以单基因或基因组合的形式存在于44个四川小麦品种（系）中。有6个品种（系）含有Yr9基因,10个品种（系）含有Yr10基因,11个品种（系）含有Yr24基因,其中1个同时含有Yr10和Yr24基因。23个品种（系）含有未知基因及其组合。共有19个小麦品种（系）成株期对包含上述混合菌种在内的田间流行菌系表现抗病,其中5个品种抗性受成株抗性基因控制。     

Investigation of in-vitro methods for measuring eggplant resistance to Verticillium dahliae

An assay based on electrolyte leakage (LEA) from callus tissue of Solanum spp. treated with culture filtrates of Verticillium dahliae was useful for screening eggplant genotypes for resistance to Verticillium wilt. Loss of electrolytes from the most resistant genotypes was significantly lower than that from the susceptible genotypes when callus tissue was incubated with a 10% filtrate solution of the virulent isolate 122. No difference was detected with filtrate solutions of the avirulent isolate 809 of the fungus. Among the tested genotypes, the S. melongena lines CCR3 and SM19/14 and the wild species S. torvum were the most resistant in the glasshouse tests and released the smallest amounts of electrolytes. The most susceptible cultivars, Lunga Violetta and Giant of China, produced the greatest leakage of electrolytes. LEA performed on stem segments failed to distinguish between susceptible and resistant genotypes of eggplant.     

Epidemiological and histological components of crown rust resistance in oat genotypes

Crown rust, caused by Puccinia coronata f. sp. avenae, can cause significant damage in all regions where oats (Avena sativa L.) are cultivated. The primary means of controlling crown rust has been through genetic resistance, although in most cases resistance has been quickly overcome by the pathogen. More durable partial or non-specific resistance may possess different mechanisms from those underlying genes with specific effects. We studied the epidemiological and histological components of crown rust resistance with potential use in plant protection. Among the components evaluated, pustule density showed the clearest effect on resistance, while the latent period was not an important component. Cell death associated with the accumulation of autofluorescent and phenolic compounds was common in the resistant genotypes, but temporally distinct for the genotypes studied. Genotype Pc68/5*Starter, which has race-specific resistance, showed rapid cell death that prevented the development of pathogen colonies. Conversely, with cultivar URS 21 and genotypes 04B7113-1 and 04B7119-2, cell death and associated accumulation of autofluorescent and phenolic compounds was delayed until pathogen colonies were already established. Pathogen colonies developed normally in susceptible plants genotypes, and had usually produced sporogenic tissue by 5 days after inoculation. The data suggest that the resistance mechanisms, especially hypersensitivity and phenolic compound production, active in resistant plants are similar but may be differently expressed over time. The temporal variation in the expression of hypersensitivity and phenolic compound production reflects the level of field resistance in these genotypes.     

Variation for resistance to kernel infection and toxin accumulation in winter wheat infected with Fusarium graminearum

Host resistance is the main way to control Fusarium head blight (FHB) in wheat. Despite improved levels of resistance to infection and spread in vegetative tissue, the toxin deoxynivalenol (DON) can still accumulate to unacceptable concentration levels. In this study, our objectives were to assess the genetic variation for resistance to kernel infection (RKI) and resistance to toxin accumulation (RTA) and their role in controlling DON. We collected spikes with different levels of visual symptoms from each of 32 wheat genotypes and at four environments and determined DON and fungal biomass (FB) from each sample. We assessed RKI by regressing FB on the level of visual symptoms and RTA by regressing DON on FB for each genotype. Significant genetic effects were found for RKI and RTA. Some genotypes consistently had low FB in their grain despite increasing visual symptoms suggesting RKI. Additionally, some genotypes consistently had low DON in their grain despite increasing FB levels suggesting a higher RTA in these genotypes. The variation for RKI and RTA explained a significant fraction of the variation for DON among genotypes with moderate visual symptoms using independent grain samples. Although RKI and RTA were significantly correlated (r = 0.58, P = 0.05), RTA was more predictive of DON accumulation because it modeled 32 to 44% of the genotype sum of squares for DON, while only 9 to 10% were predicted using RKI. Thus, variation for RTA was important in explaining variation for DON among genotypes with acceptable levels of resistance to fungal infection and spread. This work indicates that there is a need to develop a better understanding of RTA and rapid screening methods for this trait.     

A strategy to identify sources of quantitative resistance in pathosystems involving disease escape and physiological resistance: the case study of rice sheath blight

An epidemiology‐based strategy using rice sheath blight (ShB) as a biological model was developed that enables identification of sources of resistance. A set of 163 cultivated rice genotypes, including genotypes which had been reported to express partial resistance to ShB, and a few genotypes reported as very susceptible, were assessed using two complementary methods. First, microfield experiments allowed measurement of disease intensification at, and spread from, inoculated sources, along with morphological traits of each genotype. Secondly, detached tiller tests allowed measurement of the physiological resistance to the disease under conditions where morphology does not come into play. Multivariate analysis involving hierarchical cluster analysis, followed by multiple correspondence analysis, indicated that levels of physiological resistance, groups of plant morphology and disease levels in microfields were associated. Results from logistic regressions further indicated that a decreased number of lesions measured on detached tillers increased the odds of a rice genotype belonging to the group with low disease intensity in microfields. The combined results from microfield and detached tiller tests allowed identification of 23 genotypes with low level of disease intensity, which may be used as sources of resistance to ShB in breeding programmes. The results suggest that this strategy, which combines the crop stand and the individual tiller scales, may be applied to the identification of sources of resistance to a range of diseases with similar life cycle traits.     

Investigations on the role of cuticular wax in resistance to powdery mildew in grapevine

Cuticular wax on the plant epidermis inhibits or enhances prepenetration events of powdery mildew (Erysiphe necator Schwein). We examined the role of cuticular leaf and berry waxes as a resistance mechanism in four grapevine genotypes (Italia?×?Mercan-174, Gürcü, Isabella, Özer Karas?) resistant to powdery mildew after natural infection and inoculation. To understand cuticular wax properties, we determined the amount of wax and antifungal effects of thin layer chromatography (TLC) fractions from cuticular leaf and berry waxes, then assessed the chemical composition of fractions with different antifungal activities using gas chromatography/mass spectrometry (GC/MS). Susceptible genotypes Cabernet Sauvignon and Italia were used for comparison. Resistant and sensitive genotypes did not differ significantly in the total amount of wax on leaves and berries; however, 27 fatty acids, 26 alkanes, 6 terpenes, 4 indole derivatives and 4 ketones, and 3 amides, 3 phenols and 3 steroids were detected in fractions with high antifungal activity (≥75% inhibition of germination) in leaf and/or berry cuticular waxes of resistant genotypes only. These compounds may be evaluated as markers for powdery mildew resistance during genotype selection in a grapevine breeding program.     

Evaluation of soybean genotypes for resistance against the rust-causing fungus Phakopsora pachyrhizi in East Africa

Soybean rust, caused by the biotrophic fungus Phakopsora pachyrhizi, is the most important foliar disease of soybean (Glycine max) worldwide. Deployment of resistant soybean cultivars is the best option for managing this disease. Genes conferring resistance to P. pachyrhizi have been identified, but pathotypes of the rust fungus overcoming these resistance genes have also been found. To identify novel resistance genes, soybean genotypes from both local and international sources were screened at multiple locations in Tanzania and Uganda in 2016 and 2017. The results from this screening revealed that infection types, disease severities, and sporulation levels varied among the genotypes and locations. The majority of the genotypes had tan-coloured (TAN) lesions and developed moderate sporulation, implying susceptibility, while only seven of the 71 lines had reddish-brown (RB) lesions and showed low disease severities in all of the screening environments. We identified seven genotypes that were the most resistant to rust in the most locations over the two years. These genotypes will be useful for further studies and, ultimately, for rust management, as they show broad resistance to various pathotypes of the rust fungus.