DNA marker-assisted evaluation of potato genotypes for potential resistance to potato cyst nematode pathotypes not yet invading into Japan

One of major objectives of crop breeding is conferring resistance to diseases and pests. However, large-scale phenotypic evaluation for many diseases and pests is difficult because strict controls are required to prevent their spread. Detection of disease resistance genes by using DNA markers may be an alternative approach to select potentially resistant accessions. Potato (Solanum tuberosum L.) breeders in Japan extensively use resistance gene H1, which confers nearly absolute resistance to potato cyst nematode (Globodera rostochiensis) pathotype Ro1, the only pathotype found in Japan. However, considering the possibility of accidental introduction of the other pathotypes, breeding of resistant varieties is an important strategy to prevent infestation by non-invading pathotypes in Japan. In this study, to evaluate the prevalence of resistance genes in Japanese genetic resources, we developed a multiplex PCR method that simultaneously detects 3 resistance genes, H1, Gpa2 and Gro1-4. We revealed that many Japanese varieties possess not only H1 but Gpa2, which are potentially resistant to other pathotypes of potato cyst nematode. On the other hand, no genotype was found to have the Gro1-4, indicating importance of introduction of varieties having Gro1-4. Our results demonstrate the applicability of DNA-marker assisted evaluation of resistant potato genotypes without phenotypic evaluation.     

Pathotype-specificity in potato cyst nematodes,a reconsideration

Summary Within the two potato cyst nematode (PCN) species, Globodera rostochiensis and G. pallida, eight pathotypes are presently discerned by seven differential Solanum clones, whereby resistance is defined as a Pf/Pi ratio <1.0, and susceptibility as a ratio above 1.0. This pathotyping is felt to be unsatisfactory. Reconsidering the original data and data published by others resulted in the conclusion that there are only three reliably recognizable Ro-pathotypes Ro1/Ro4, Ro2/Ro3 and Ro5. The differential clones 60.21.19 and 65.346.19 differentiate only between the two species. Within G. pallida it does not seem possible to identify pathotypes reliably. The resistance-virulence pattern is of a quantitative nature fitting a polygenic system. There appear to be pathotype-specific effects, which are not large enough to identify and classify pathotypes reliably. It is therefore suggested to rename the Ro-pathotypes into Ro1 (old Ro1 and Ro4), Ro3 (old Ro2 and Ro3) and Ro5 (old Ro5) and to speak of more and less virulent Pa-populations.The rigorous requirements for resistance in various countries present great problems to the breeders and result in the removal of much valuable partial resistance. Especially with G. pallida these rigorous requirements are probably an obstacle rather than an encouragement to solve the pallida problem.     

Characterization of resistance to Globodera rostochiensis pathotype Ro1 in cultivated and wild potato species accessions from the Vavilov Institute of Plant Industry

Potato cyst nematodes (PCN) collected in six localities in the Leningrad region of North West Russia were identified as Globodera rostochiensis pathotype Ro1 and were used for subsequent resistance tests. Seventy‐nine accessions of cultivated and closely related wild potato species from the VIR collection in Russia were screened on resistance to G. rostochiensis pathotype Ro1 and on the presence of molecular markers for H1 and Gro1‐4 resistance genes. No associations were detected between the resistance level of diploid and tetraploid Andean and tetraploid Chilean potato landraces (indigenous cultivars) and their related wild species and their geographical distribution or presence of PCR‐based markers that are associated with the H1 and Gro1‐4 genes. At the same time, all susceptible genotypes lacked such markers. New sources of resistance were found and could be used in breeding.     

New sources of resistance to potato cyst-nematodes in the Commonwealth Potato Collection

Summary Accessions obtained from the Commenwealth Potato Collection were evaluated for resistance to European populations of potato cyst-nematodes (PCN). With over 36% of the current collection assessed, resistance to pathotypes of both Globodera rostochiensis and G. pallida were identified in 16 species from Argentina, Bolivia or Peru. Resistance to all PCN pathotypes was particularly prevalent in species from the Andean regions of Bolivia and North Argentina, supporting the hypothesis of co-evolution of the potato and PCN in these regions.     

New sources of resistance to the potato cyst nematodes Globodera pallida and G. rostochiensis in the Commonwealth Potato Collection

The Commonwealth Potato Collection contains 77 species following the incorporation of the personal collection of Professor JG Hawkes. The material originates from several expeditions to the Americas and represents a potentially valuable source of resistance to pests and pathogens, including potato cyst nematodes (PCN). The germplasm from the Hawkes collection screened in this study included taxa mainly from Mexico, Bolivia, Argentina and Peru. Seedlings of 198 accessions, from 63 different species were screened for reaction to the PCN species G. pallida and G. rostochiensis. Of the 198 accessions tested, 56% contained G. pallida-resistant clones and 53% contained clones resistant to G. rostochiensis. Twelve species in this survey are believed to contain novel, previously unreported sources of resistance. The geographical distribution of resistance indicated a contrast between the two PCN species. Although, as in previous studies, resistance to G. rostochiensis was centred on Bolivia and Argentina, G. pallida resistance was found across the geographical range of tuber-bearing Solanum. The possibility of cross-resistance arising from selective pressure from the more northern species G. mexicana is discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

The Importance of the Bolivian Wild Potato Species in Breeding for Globodera pallida Resistance

Screening for resistance to the potato cyst nematode, Globodera pallida, in potatoes from. Bolivia, was carried out in 1983 and 1984, using a mixture of four nematode populations representing pathotypes Pa₁, Pa₂ and Pa₃ From the 66 accessions of 17 species and subspecies evaluated, highly resistant genotypes were identified in 21 accessions from seven species. All had Pf/Pi values of 2 or less, whereas the susceptible control, Solanum tuberosum cv. ‘Disiree’ had Pf/Pi values of more than 2G in both tests. Two diploid wild species, S. brevicaule and S. leptophyes, showed the best resistant. The geographical distributional of resistant populations and the evolution of resistance in wild potato populations are discussed.     

Variation in resistance level of potato genotypes and virulence level of potato cyst nematode populations

Summary In two experiments, using different testing methods, the number of newly formed cysts was determined on nine potato genotypes with resistance from various sources. Ten potato cyst nematode (PCN) populations were used in these experiments. Rank correlation between numbers of cysts over potato genotype-PCN population combinations for both experiments was high (r_s = 0.90). Dendrograms for PCN populations and potato genotypes were constructed, based on a simultaneous hierarchical clustering procedure for potato genotype-PCN population interaction terms. Several virulence groups could be identified within Globodera rostochiensis as well as within G. pallida. Host genotypes, derived from the same sources of resistance, were clustered in different resistance groups.     

The importance of wild potato species resistant to the potato cyst nematode,Globodera pallida,pathotypes P4A and P5A,in potato breeding. I. Resistance studies

Summary Seven wild diploid potato species, Series Tuberosa, representing 1023 clones were screened for resistance to the potato cyst nematode, Globodera pallida. Over 25% of the clones were resistant to pathotype P₄A and almost 30% were resistant to pathotype P₅A. The resistance in hybrid progenies of these and other resistant species with cultivated potatoes was evaluated, and over 2200 seedlings were screened. High frequencies of resistance (>50%) to P₄A were found in progenies with Solanum leptophyes, S. vernei, S. gourlayi and S. capsicibaccatum, whereas resistance to P₅A was found in these species as well as S. sparsipilum. The importance of nematode resistant wild species for potato breeding is discussed.     

The importance of wild potato species resistant to the potato cyst nematode,Globodera pallida,pathotypes P4A and P5A,in potato breeding. II. The crossability of resistant species

Summary Over 2400 pollinations were made to investigate the crossability relationships between cultivated potatoes, and wild diploid species from Series Tuberosa and Circaeifolia, as well as wild polyploid species in Series Tuberosa and Longipedicellata resistant to potato cyst nematode, Globodera pallida pathotypes P₄A and P₅A. Wild diploids in Series Tuberosa crossed easily with cultivated diploid species, except with Solanum lignicaule where most pollinations failed, and seed set was extremely low (0.2 seeds per pollination or less). It is suggested that this species is 1EBN. S. capsicibaccatum is clearly isolated from Series Tuberosa, but can form hybrids with S. lignicaule, which can act as a bridging species to S. tuberosum haploids. S. gourlayi and S. oplocense can be crossed with both subspecies of S. tuberosum, but S. papita, Series Longipedicellata is reproductively isolated from the tetraploid cultigens. The crossability data are discussed in the light of germplasm utilisation for breeding potato varieties resistant to potato cyst nematode.     

Improvement of diagnostic markers for resistance to Globodera pallida and application for selection of resistant germplasms in potato breeding

Occurrence of pale potato cyst nematode, Globodera pallida (Stone) Behrens, was first recorded in Japan in 2015. Among several control measures, cultivation of resistant potato (Solanum tuberosum L.) varieties is the most effective in cost and environmental impact. As no G. pallida-resistant varieties have yet been developed in Japan, great emphasis is being placed on screening of germplasm possessing the resistance and development of the resistant varieties. In this study, we first improved previously reported DNA markers linked to the G. pallida resistance loci (GpaIV^s_adg and Gpa5) and then used these to screen more than 1,000 germplasms to select several candidate germplasms with resistance. We performed inoculation testing on the selected candidates and identified several resistant germplasms to the Japanese G. pallida population. Furthermore, we developed a simultaneous detection method combining three DNA markers linked to G. pallida and Globodera rostochiensis (Wollenweber) Behrens resistance loci. We validated the ability of C237-I marker to select resistant allele of GpaIV^s_adg and predict the presence of resistance in a Japanese breeding population. Resistant germplasms identified in this study could potentially be used to develop G. pallida-resistant varieties. The marker evaluation methods developed in this study will contribute to the efficient development of resistant varieties.     

Durability of resistance against potato cyst nematodes

A requirement for evaluating the effectiveness of major resistance genes (R-genes) is detailed knowledge of the genetic variability of the nematode populations in an area. Obtaining insight in the genetic variation in an area is a tour de force, because qualitative data are not sufficient allele frequency data at well studied loci are required. Another important step in predicting the durability of major R-genes is to study the parasite at the molecular level. Also for nematodes it seems that R-genes are part of a molecular surveillance system recognising foreign molecules. For potato cyst nematodes it is becoming clear that numerous proteins are secreted to manipulate the host plant and it may be assumed that some of these secretory proteins are also recognised by the host. Knowledge of the molecular nature and function of these avirulence gene products may reveal clues to predict the durability of an R-gene. R-genes that recognise avirulence gene products that have a crucial function in the nematode maybe very durable. When the current molecular models for gene-for-gene interactions are correct, the durability of an R-gene may be predicted by studying the dispensability factor and functional constraints of the avirulence gene products. This revised version was published online in August 2006 with corrections to the Cover Date.     

马铃薯不同品种(系)和稻、薯轮作模式对根结线虫病的防治效果

根结线虫病是马铃薯连作减产的主要原因之一。为研究根结线虫病的农艺防治措施,本文开展了品种抗性评价、稻、薯轮作和土壤淹水下的线虫消减动态研究。抗性评价试验表明, 18个品种(系)中没有筛选到同时抗南方根结线虫和爪哇根结线虫的材料,品系1002-1和品种丽薯6号相对抗性较好,青薯9号和合作88相对敏感。马铃薯根结线虫发病土壤中可同时检测到卵、幼虫和成虫等不同形态。水旱轮作试验表明,经一茬水稻栽培可使土壤中根结线虫虫态完全消失,再种植马铃薯也不再发生根结线虫病,且马铃薯有效产量显著提高,青薯9号、合作88和丽薯6号的单株有效产量较对照分别提高50.5%、43.7%和26.0%,差异均极显著, 1002-1单株有效产量与对照差异不显著。由此可见,稻、薯轮作根结线虫防治效果明显。为进一步探索水旱轮作的防治机制,观察了土壤淹水进程中各种虫态消减动态。结果表明,在淹水过程中,根结线虫的卵、幼虫和成虫不同虫态此消彼长,要淹水60 d以上,土壤中才检测不到所有形态虫体。水早轮作中水稻生长期淹水时间足够长,满足防治线虫的淹水条件。因此结合抗病品种选用进行稻、薯轮作是防治马铃薯根结线虫病既绿色环保又可持续有...     

Multiple resistance to diseases and pests in potatoes

Summary The potato has more characters of economic importance that need to be considered by the breeder than any other temperate crop. In Europe these include resistance to at least twelve major diseases and pests. Highest priority has been given to resistance to late blight (Phytophthora infestans), virus diseases (particularly those caused by potato leafroll virus and potato virus Y) and potato cyst nematode (Globodera rostochiensis andG. pallida). Useful sources of resistance are available and early generation screening techniques have been developed to allow positive selection for multiple resistance and the breeding value of clones used as parents to be determined. Progress in restriction fragment length polymorphism technology should result in more efficient selection in the future.     

Effectiveness of quantitative resistance conferred by the genetic background of pepper in the control of root‐knot nematodes and influence onto durability of Me1‐ and Me3‐resistant genes in greenhouse conditions

In pepper (Capsicum annuum), the major genes (R‐genes) Me1 and Me3 confer resistance against root‐knot nematodes (Meloidogyne spp.). The combination of R‐genes and quantitative resistance factors in the same genotype is considered a good breeding strategy for increasing the durability of R‐genes. To ascertain this hypothesis, five pepper inbred lines, differing in their quantitative resistance level, were combined with Me1 or Me3 genes in F₁ hybrids. The resistance of inbred lines and F₁ hybrids was evaluated in a greenhouse with soil naturally infected by M. incognita in two successive growing years. In both years, lines carrying Me3 were less infected by the nematode when combined with quantitative resistance. An increase in nematode infection was observed in the second growing year in lines carrying Me1 or Me3, independently of quantitative resistance. The infection level recorded in inbred lines without R‐genes was similar in both years. The effectiveness of quantitative resistance controlling M. incognita is confirmed in greenhouse conditions, although the durability of Me1 and Me3 when combined with quantitative resistance factors was not seen to increase.     

Genetic analysis of resistance to leaf-rust (Puccinia recondita f. sp. tritici) pathotypes in the durum wheats 'PBW 34' and 'DWL 5023'

Genetic analysis of leaf-rust resistance was conducted on two durum wheats. Triticum durum cvs. ‘PBW 34’ and ‘DWL 5023’ were crossed with the leaf-rust-susceptible durum wheat ‘Malvi Local’. The F₁, F₂ and F₃ generations were tested against leaf-rust pathotypes 1, 77A and 108. In ‘PBW 34’, a single dominant gene was effective against each of the pathotypes 1 and 108, whereas two independently inherited dominant genes were effective against pathotype 77A. In ‘DWL 5023’, two independently inherited dominant genes were operative against pathotypes 1 and 77A, whereas a single dominant gene was identified as being operative against pathotype 108. Allelic tests on F₂ generation and joint segregation analysis on F₃ generation seedlings, suggested that two different genes in each cultivar are effective against these three leaf-rust pathotypes. Cultivar ‘PBW 34’ has Lrd1 and Lrd2 genes whereas Lrd1 and Lrd3 genes are present in ‘DWL 5023’.     

Genetic Analysis of Resistance to Xanthomonas campestris pv. oryzae (Ishiyama) Dye in a Rice Breeding Line ARC 10464

The genetics of resistance to three Indian pathotypes of Xanthomonas campestris pv. oryzae, namely, IX01, IX08 and IX09, was studied in a landrace of Indica rice ‘ARC 10464’. Resistance to each of the two pathotypes IX01 and IX09 was governed by two independently-inherited dominant genes while a single dominant gene was operative against patho-type 1X08. The joint segregation tests conducted on F₂ plant progenies (F₃ families) using pathotypes IX01, IX08 and IX09 suggested that the gene/(s) effective against each of the pathotypes are different.     

Genotype × environment interaction for resistance to the white potato cyst nematode (Globodera pallida,pathotype E) in Solanum vernei x S. Tuberosum hybrids

Summary The standard root-ball test for assessing quantitative resistance of Globodera pallida in host material derived from Solanum vernei has produced variable results. This study of two sets of clones shows that genotype x environment interactions are responsible and that linear regressions (b) of phenotypic means on environments may enable good predictions to be made of the effects of the interactions. A relationship between the regression value (b) and phenotypic mean is shown for this material. The implication of this for a screening procedure in a potato breeding programme aimed at increasing potato cyst nematode resistance in adapted cultivars of S. tuberosum is discussed.     

Differential resistances to anthracnose in Capsicum baccatum as responding to two Colletotrichum pathotypes and inoculation methods

Chili anthracnose, caused by Colletotrichum spp., is one of the major diseases to chili production in the tropics and subtropics worldwide. Breeding for durable anthracnose resistance requires a good understanding of the resistance mechanisms to different pathotypes and inoculation methods. This study aimed to investigate the inheritances of differential resistances as responding to two different Colletotrichum pathotypes, PCa2 and PCa3 and as by two different inoculation methods, microinjection (MI) and high pressure spray (HP). Detached ripe fruit of Capsicum baccatum ‘PBC80’ derived F₂ and BC₁s populations was assessed for anthracnose resistance. Two dominant genes were identified responsible for the differential resistance to anthracnose. One was responsible for the resistance to PCa2 and PCa3 by MI and the other was responsible for the resistance to PCa3 by HP. The two genes were linked with 16.7 cM distance.     

Mapping and candidate gene identification of loci determining tolerance to greenbug (Schizaphis graminum, Rondani) in barley

Greenbug is one of the most aggressive pests of barley and wheat. In Argentina, yield losses of wheat, barley, oat and sorghum crops caused by greenbug are chronic and at times severe. Since Marker Assisted selection for greenbug resistance genes in barley is very limited, the purpose of the current study was to map greenbug resistance genes in doubled haploid (DH) lines and to identify candidate genes. A set of DH lines of the Oregon-Wolfe Barley (OWB) mapping population derived from the cross between OWB_DOM and OWB_REC and both parental lines were screened for tolerance to greenbug. There was significant variation among the DH lines in foliar area (FA), dry weight (DW) and chlorophyll contents (Ch) between infested and control DH lines. Three main QTLs were identified. These QTLs explained 82 % of the FA, 80 % of DW and 58 % of Ch variability of infested plants. The initial and final FA and DW of controls and final DW of infested plants were associated with the same molecular markers on chromosome 2H (Vrs1, BmAc0144f, GBR259, GBS705). The final FA of infested plants was significantly linked to molecular markers on chromosome 5H (GBRO986, GBR518, GBM1483, GBR1082). The positive alleles were provided by OWB_DOM. The content of chlorophyll of infested plants was associated with the marker loci Ris44, GBR1608, GBR1637N and GBS0785 on chromosome 7H, with the positive alleles provided by OWB_REC. Both parents contributed to different tolerance traits. The QTLs found in this population are new greenbug resistance loci. A sequence homology search was performed to derive the putative function of the genes linked to the QTLs.     

Development of elite rice restorer lines in the genetic background of R022 possessing tolerance to brown planthopper, stem borer, leaf folder and herbicide through marker-assisted breeding

Rice leaf folder, stem borer and brown planthopper (BPH) are the most devastating rice insect pests. Developing and planting insect-resistant rice varieties is the most economical and effective measure for controlling these pests. BPH can be controlled with native BPH-resistance genes in rice, while at present rice leaf folder and stem borer can only be controlled through planting transgenic Bt rice. In this study, the breeding of a new restorer line KR022 possessing stacked BPH-resistance genes Bph14 and Bph15, Bt gene cry1C and glufosinate-resistance gene bar, is reported for the first time. A rice restorer line R022 with BPH-resistance genes Bph14 and Bph15 was used as a recurrent parent to cross with the transgenic rice T1C-19 of cry1C and bar genes during the breeding process. The restorer line KR022 was developed from the backcross populations of R022 and T1C-19 through molecular marker-assisted selection and glufosinate-resistance selection. The cry1C and bar genes were found to integrate on chromosome 11 of KR022, and the genome recovery of KR022 was up to 95.8 % of the R022 genome. The quantification of Cry1C protein expression showed that it was expressed at different levels in the leaf, stem, panicle, endosperm, and root of KR022 and its hybrid rice. The insect-resistance evaluation indicated that KR022 and its hybrid rice had good resistance to rice leaf folder and stem borer, both in laboratory settings and in the field. Furthermore, they exhibited increased resistance to BPH at both the seedling and mature stage. The field trial showed there was no significant difference in key agronomic traits between KR022 and its recurrent parent R022, and four hybrids from KR022 yield much higher than the control II-You 838. Moreover, KR022 and its hybrid rice were found to have resistance to the herbicide glufosinate. These results demonstrate that KR022 is effective as a rice restorer line for the breeding of “green super rice”, possessing multiple tolerances to rice BPH, stem borer, leaf folder and glufosinate.